Application of Genetic Bit Analysis (GBATM) for allelic selection in plant breeding

Genetic Bit Analysis (GBA) is a relatively new technique developed to score single-nucleotide polymorphisms among alleles. It is currently being used as a genetic diagnostic tool in human paternity tests as well as in pedigree analysis of farm animals. GBA relies on the polymerase chain reaction and enzyme-linked colorimetry to distinguish differences among genotypes. The identification of a single-nucleotide polymorphism which distinguished the plastome of cytoplasmic male-sterile onion varieties from the plastome of fertile lines provided a model system for testing the utility of GBA in plants. Our results demonstrate that GBA permits rapid and accurate allele determination in onion breeding lines, resulting in accurate prediction of sterility at the seedling stage. The use of 96-well microtiter plates and automated liquid-handlers allowed semi-automation of Genetic Bit Analysis.     

Effect of AMP and its analogues on the environment of phosphorylase b coenzyme

Using fluorescence and SH-groups titration techniques we have proved existence of an inverse relationship betweenthe rate constants of bimolecular deactivation of phosphorylase b coenzyme by iodide ion in the presence of AMP and its analogues and the shielding rate constants of slow cysteine, C-108, by the diverse nucleotides, The greater the shielding rate constants, the smaller the rate constant of deactivation by iodied ion. The analogues we have used were AMP, IMP, 2′dAMP, 2′AMP and 3′AMP.     

LINE-1 and inflammatory gene methylation levels are early biomarkers of metabolic changes: association with adiposity

We analyzed whether global and inflammatory genes methylation can be early predictors of metabolic changes and their associations with the diet, in a cross-sectional study (n?=?40). Higher global methylation was associated to adiposity, insulin resistance, and lower quality of the diet. Methylation of IL-6, SERPINE1 and CRP genes was related to adiposity traits and macronutrients intake. SERPINE1 hypermethylation was also related to some metabolic alterations. CRP methylation was a better predictor of insulin resistance than CRP plasma concentrations. Global and inflammatory gene promoter hypermethylation can be good early biomarkers of adiposity and metabolic changes and are associated to the quality of the diet.     

Protective Effect of <Emphasis Type="Italic">Lactobacillus casei</Emphasis> on DMH-Induced Colon Carcinogenesis in Mice

The administration of probiotics is a promising approach to reduce the prevalence of colon cancer, a multifactorial disease, with hereditary factors, as well as environmental lifestyle-related risk factors. Biogenic polyamines, putrescine, spermidine, and spermine are small cationic molecules with great roles in cell proliferation and differentiation as well as regulation of gene expression. Ornithine decarboxylase is the first rate-limiting enzyme for polyamine synthesis, and upregulation of ornithine decarboxylase activity and polyamine metabolism has been associated with abnormal cell proliferation. This paper is focused on studying the protective role of Lactobacillus casei ATCC 393 in a chemically induced mouse model of colon carcinogenesis, directing our attention on aberrant crypt foci as preneoplastic markers, and on polyamine metabolism as a possible key player in carcinogenesis. BALB/c mice were administered 1,2-dimethylhydrazine dihydrochloride (DMH) to induce colon cancer (20 mg/kg body weight, subcutaneous, twice a week for 24 weeks). L. casei ATCC 393 was given orally (10⁶ CFU, twice a week), 2 weeks before DMH administration. Hematoxylin and eosin staining, high-performance liquid chromatography, and Western blotting were used to evaluate aberrant crypt foci, urinary polyamines, and ornithine decarboxylase expression in the colon. The experimental data showed that the preventive administration of L. casei ATCC 393 may delay the onset of cancer as it significantly reduced the number of DMH-induced aberrant crypt foci, the levels of putrescine, and the expression of ornithine decarboxylase. Hence, this probiotic strain has a prospective role in protection against colon carcinogenesis, and its antimutagenic activity may be associated with the maintenance of polyamine metabolism.     

Neonatal immunization with a single dose of recombinant BCG expressing subunit S1 from pertussis toxin induces complete protection against Bordetella pertussis intracerebral challenge

The currently used pertussis vaccines are highly efficacious; however, neonates are susceptible to whooping cough up to the sixth month. In agreement, DTP-immunized neonate mice were not protected against intracerebral challenge with Bordetella pertussis. Neonate mice immunized with either DTP or a recombinant-BCG strain expressing the genetically detoxified S1 subunit of pertussis toxin do not show a humoral immune response against PT. On the other hand, rBCG-Pertussis induces higher PT-specific IFN-gamma production and an increase in both IFN-gamma(+) and TNF-alpha(+)-CD4(+)-T cells than the whole cell pertussis vaccine and confers protection against a lethal intracerebral challenge with B. pertussis.     

Synthesis of new palladium(II) complexes containing tetradentate-nitrogen donor ligands: Combined structural studies by NMR spectroscopy and X-ray crystallography

Treatment of the tetradentate (NN′N′N) N-alkylaminopyrazole ligands 3,6-dimethyl-1,8-(3,5-dimethyl-1-pyrazolyl)-3,6-diazaoctane (ddad) and 1,4-bis[2-(3,5-dimethyl-1-pyrazolyl)ethyl]piperazine (bedp) with [PdCl₂(CH₃CN)₂] in a 1:1 M/L ratio in acetonitrile produces [Pd₂Cl₄(L)] and [PdCl₂(L)] (L = ddad and bedp). Treatment of the corresponding complex [PdCl₂(L)] (L = ddad, bedp) in the presence of AgBF₄ in CH₂Cl₂/methanol (2:1) or NaBF₄ in acetonitrile gives [Pd(L)](BF₄)₂. The Pd(II) complexes have been characterised by elemental analyses, conductivity measurements, IR and ¹H and ¹³C{¹H} NMR spectroscopies when possible. The X-ray structure of the complex [Pd(ddad)]Cl₂ · 3H₂O has been determined. The Pd(II) is coordinated to the ddad ligand by two nitrogen atoms of pyrazolyl groups and two nitrogen atoms of the amine groups, in a slightly distorted square-planar geometry.     

Adjustments of water use efficiency by stomatal regulation during drought and recovery in the drought-adapted Vitis hybrid Richter-110 (V. berlandieri x V. rupestris)

The hybrid Richter-110 (Vitis berlandieri x Vitis rupestris) (R-110) has the reputation of being a genotype strongly adapted to drought. A study was performed with plants of R-110 subjected to water withholding followed by re-watering. The goal was to analyze how stomatal conductance (g(s)) is regulated with respect to different physiological variables under water stress and recovery, as well as how water stress affects adjustments of water use efficiency (WUE) at the leaf level. Water stress induced a substantial stomatal closure and an increase in WUE, which persisted many days after re-watering. The g(s) during water stress was mainly related to the content of ABA in the xylem and partly related to plant hydraulic conductivity but not to leaf water potential. By contrast, low g(s) during re-watering did not correlate with ABA contents and was only related to a sustained decreased hydraulic conductivity. In addition to a complex physiological regulation of stomatal closure, g(s) and rate of transpiration (E) were strongly affected by leaf-to-air vapor pressure deficit (VPD) in a way dependent of the treatment. Interestingly, E increased with increasing VPD in control plants, but decreased with increasing VPD in severely stressed plants. All together, the fine stomatal regulation in R-110 resulted in very high WUE at the leaf level. This genotype is revealed to be very interesting for further studies on the physiological mechanisms leading to regulation of stomatal responsiveness and WUE in response to drought.     

Biotransformation of the fungistatic compound (R)-(+)-1-(4′-chlorophenyl)propan-1-ol by Botrytis cinerea

The biotransformation of the fungistatic agent (R)-(+)-1-(4′-chlorophenyl)propan-1-ol (1) by the phytopathogen Botrytis cinerea has been studied. The main reaction pathways involved hydroxylations on several positions as well as condensations with secondary metabolites of the fungus. The antifungal activity of compound 1 against B. cinerea has also been determined.