Origins of Allium ampeloprasum horticultural groups and a molecular phylogeny of the section Allium (Allium: Alliaceae)

The subgenus Allium section Allium includes economically important species, such as garlic and leek, as well as other polyploid minor crops. Phylogenetic studies within this section, with a focus on horticultural groups within A. ampeloprasum, were performed on 31 accessions of 17 species using the nuclear ribosomal DNA internal transcribed spacer (ITS) region and the chloroplast trnL-F and trnD-T regions. The results confirmed the monophyly of section Allium. Four main clades were identified on all ITS analyses but the relationships among those and the remaining species studied within section Allium remained unresolved. Trees based on cpDNA recovered two major clades and a topology only partly congruent with that of the ITS tree. Intra-individual polymorphism of the ITS region proved useful in tracking putative parent species of polyploid taxa. The allopolyploid origin of great headed garlic (GHG), A. iranicum and A. polyanthum was confirmed. No signs of hybridization in leek or kurrat were detected but possible introgression events were identified in pearl onion and bulbous leek. Although GHG is often used as a garlic substitute, molecular analysis revealed only a distant relationship with garlic. We also clarified the previous incorrect classification of cultivated forms within A. ampeloprasum, by showing that leek, kurrat, pearl onion, and bulbous leek should be considered separately from GHG.     

Structural basis of trypsin inhibition and entomotoxicity of cospin, serine protease inhibitor involved in defense of Coprinopsis cinerea fruiting bodies

Cospin (PIC1) from Coprinopsis cinerea is a serine protease inhibitor with biochemical properties similar to those of the previously characterized fungal serine protease inhibitors, cnispin from Clitocybe nebularis and LeSPI from Lentinus edodes, classified in the family I66 of the MEROPS protease inhibitor classification. In particular, it exhibits a highly specific inhibitory profile as a very strong inhibitor of trypsin with K(i) in the picomolar range. Determination of the crystal structure revealed that the protein has a β-trefoil fold. Site-directed mutagenesis and mass spectrometry results have confirmed Arg-27 as the reactive binding site for trypsin inhibition. The loop containing Arg-27 is positioned between the β2 and β3 strands, distinguishing cospin from other β-trefoil-fold serine protease inhibitors in which β4-β5 or β5-β6 loops are involved in protease inhibition. Biotoxicity assays of cospin on various model organisms revealed a strong and specific entomotoxic activity against Drosophila melanogaster. The inhibitory inactive R27N mutant was not entomotoxic, associating toxicity with inhibitory activity. Along with the abundance of cospin in fruiting bodies of C. cinerea and the lack of trypsin-like proteases in the C. cinerea genome, these results suggest that cospin and its homologs are effectors of a fungal defense mechanism against fungivorous insects that function by specific inhibition of serine proteases in the insect gut.     

Contraction wave in axial direction in free wall of guinea pig left ventricle

Mechanical activation of the normal left ventricle (LV) is not simultaneous; however, the potential consequences of the ejection function of the ventricle are not entirely known. We studied contraction of the LV free wall to determine whether it reveals a contraction wave in the axial direction during ejection. Seven guinea pig hearts in situ were studied via thoracotomy. In each heart, the ventricular and aortic pressures were measured by two microtipped manometers (2-Fr, Millar). Contraction of the LV free wall was assessed with a video system (Dalsa D6-0256 camera and EPIX PIXCI D32 frame grabber; acquisition rate, 500 frames/s), and 15-18 epicardial markers were used to divide the region into 20-25 triangular areas. The area sizes were studied during contraction to locate the position of the contraction wave. For each triangular area, two variables were determined as follows: the time (t(c)) from the end of diastole until the size of the area reached 80% of maximum size reduction (normalized with the duration of systole) and the normalized latitude (L(ax)) of the area (determined at the end of diastole). A relationship between these two variables was determined by regression analysis. We found that the t(c) at which the contraction wave reached a triangular area was in positive correlation with the L(ax) value for that triangular area with a slope of 0.25 +/- 0.09 and a linear correlation coefficient of 0.41 +/- 0.08. Thus contraction in the guinea pig LV free wall occurs progressively from apex to base with successive areas reaching 80% contraction.     

Factors affecting direct organogenesis from flower explants of Allium giganteum

A novel method is described for the propagation of Allium giganteum Regel using direct organogenesis resulting in multiple shoot structures formed on mature flower buds or ovaries. A two step induction and differentiation procedure, similar to that described earlier in onion, was tested. Flowers were inoculated on the induction medium for 6 days and extracted ovaries were placed on the differentiation medium. Optimal formation of multiple shoot structures was obtained using modified BDS medium containing 50 g l^–1 sucrose solidified by a mixture of agar/gellan-gum, with 8.88 M benzylaminopurine (BA) and 9.05 M 2,4 dichlorophenoxyacetic acid (2,4-D) in induction medium and 9.08 M tidiazuron (TDZ) in the differentiation medium. Five plant sources obtained from different European retailers of ornamental bulbs were tested separately. All tested genotypes produced multiple organogenic structures, although induction percentages clustered in two distinctive groups. Shoots formed tended to become dormant, and attempts to improve their growth and rooting included treatment with fluridone. Dormancy was partly broken when shoots were briefly dipped in 1 M fluridone. Genetic analysis of plant sources using random amplified polymorphic DNA method showed that 5 retailers actually distribute only two different clones, one of them more and the other less responsive to shoot organogensis.     

Rutin in buckwheat herbs grown at different UV-B radiation levels: comparison of two UV spectrophotometric and an HPLC method

Rutin is an antioxidant with many interesting pharmacological effects. It can also be found in buckwheat (Fagopyrum esculentum Moench). UV radiation stimulates the activity of enzymes of the phenylpropanoid pathway and there is some evidence that it influences the rutin content in plants. The aim of the present research was (1) to examine the influence of different levels of UV-B radiation on rutin content and (2) to compare the results obtained by three analytical methods. The plants were grown under three UV-B levels: reduced, ambient and enhanced, simulating 17% ozone depletion. Analyses were performed by HPLC and two spectrophotometric methods. In one, the absorbancies were measured at 420 nm with and without the addition of AlCl(3). In another method the concentration was calculated from absorbancies at 352.5 nm and 366.5 nm according to the Official Methods of Analysis of AOAC International. The highest amounts of rutin were found in flowers, followed by leaves and stems. A comparison of the different treatments revealed that the highest amounts of rutin were in plants grown under ambient radiation, followed by the plants cultivated under enhanced UV-B and then under reduced UV-B radiation. Treatments caused more effect on leaves than on flowers. Leaves developed under ambient light conditions contained 97% more rutin than leaves grown under reduced UV-B radiation. In flowers, the contents differed by 19% only. The results obtained using the three methods showed a good correlation, but the absolute differences were surprisingly high. The AOAC and the AlCl(3) methods gave, on average, 140% and 30% higher results than HPLC, respectively.     

Hip biomechanics in orthopaedic clinical practice

Radiographic and clinical studies, coupled with biomechanical assessment of the hip, are important tools for predicting the development of osteoarthitis of the hip. In order to better understand the treatment of hip dysplasia, it is necessary to determine the contact stress in the hip joint. In this study, a three-dimensional mathematical model was used to determine hip joint contact stress. Because of the discrepancy in the results of analyses of different radiographic indicators of hip dysplasia, the calculation of hip joint contact stress is proposed for a more accurate assessment of the severity of hip dysplasia.     

The small world in biophysical systems structural properties of glycolysis and the TCA cycle in Escherichia coli

It has been shown that the central metabolic network of Escherichia coli is of the small-world type. In this paper, we present that the metabolic network of glycolysis and TCA cycle as a part of the E. coli metabolism is also a small-world network. We found that the hubs of the studied network are consistent with those found in the complete metabolic network. The evolutionary meaning of this finding is discussed.     

Humanized mice as a model for rheumatoid arthritis

Genetic susceptibility to rheumatoid arthritis (RA), a common autoimmune disease, is associated with certain HLA-DR4 alleles. Treatments are rarely curative and are often tied to major side effects. We describe the development of a humanized mouse model wherein new, less toxic, vaccine-like treatments for RA might be pretested. This model includes four separate transgenes: HLA-DR*0401 and human CD4 molecules, a RA-related human autoantigenic protein (HCgp-39), and a T-cell receptor (TCRalphabeta) transgene specific for an important HCgp-39 epitope, eliciting strong Th1 responses in the context of HLA-DR*0401.