Differentiation of Fanconi anemia and aplastic anemia using mitomycin C test in Tunisia

Fanconi anemia (FA) is a recessive chromosomal instability syndrome that is clinically characterized by multiple symptoms. Chromosome breakage hypersensitivity to alkylating agents is the gold standard test for FA diagnosis. In this study, we provide a detailed laboratory protocol for accurate assessment of FA diagnosis based on mitomycin C (MMC) test. Induced chromosomal breakage study was successful in 171 out of 205 aplastic anemia (AA) patients. According to the sensitivity of MMC at 50 ng/ml, 38 patients (22.22%) were diagnosed as affected and 132 patients (77.17%) as unaffected. Somatic mosaicism was suspected in an 11-year-old patient with a FA phenotype. Twenty-six siblings of FA patients were also evaluated and five of them (19.23%) were diagnosed as FA. From this study, a standard protocol for diagnosis of FA was developed. It is routinely used as a diagnostic test of FA in Tunisia.     

Nucleotide variation at the CHALCONE ISOMERASE locus in Arabidopsis thaliana

An approximately 1.9-kb region encompassing the CHI gene, which encodes chalcone isomerase, was sequenced in 24 worldwide ecotypes of Arabidopsis thaliana (L.) Heynh. and in 1 ecotype of A. lyrata ssp. petraea. There was no evidence for dimorphism at the CHI region. A minimum of three recombination events was inferred in the history of the sampled ecotypes of the highly selfing A. thaliana. The estimated nucleotide diversity theta(TOTAL) = 0.004, theta(SIL) = 0. 005 was on the lower part of the range of the corresponding estimates for other gene regions. The skewness of the frequency spectrum toward an excess of low-frequency polymorphisms, together with the bell-shaped distribution of pairwise nucleotide differences at CHI, suggests that A. thaliana has recently experienced a rapid population growth. Although this pattern could also be explained by a recent selective sweep at the studied region, results from the other studied loci and from an AFLP survey seem to support the expansion hypothesis. Comparison of silent polymorphism and divergence at the CHI region and at the Adh1 and ChiA revealed in some cases a significant deviation of the direct relationship predicted by the neutral theory, which would be compatible with balancing selection acting at the latter regions.     

Natural variation in Arabidopsis lyrata vernalization requirement conferred by a FRIGIDA indel polymorphism

Species share homologous genes to a large extent, but it isnot yet known to what degree the same loci have been targetsfor natural selection in different species. Natural variationin flowering time is determined to a large degree by 2 genes,FLOWERING LOCUS C and FRIGIDA, in Arabidopsis thaliana. Here,we examine whether FRIGIDA has a role in differences in floweringtime between and within natural populations of Arabidopsis lyrata,a close outcrossing perennial relative of A. thaliana. We found2 FRIGIDA sequence variants producing potentially functionalproteins but with a length difference of 14 amino acids. Thesevariants conferred a 15-day difference in flowering time inan association experiment in 2 Scandinavian populations. Thedifference in flowering time between alleles was confirmed withtransformation to A. thaliana. Because the north European late-floweringpopulations harbor both late- and early sequence variants atintermediate frequencies and the late-flowering variant is mostfrequent in the southern early flowering European population,other genetic factors must be responsible for the floweringtime differences between the populations. The length polymorphismoccurs at high frequencies also in several North American populations.The occurrence of functional variants at intermediate frequenciesin several populations suggests that the variation may be maintainedby balancing selection. This is in contrast to A. thaliana,where independent loss-of-function mutations at the FRIGIDAgene are responsible for differences between populations andlocal adaptation.     

Biomineralization Consolidation of Fresco Wall Paintings Samples by Bacillus sphaericus

Bacterial-induced mineralization has been explored for protection and consolidation of degraded limestone, concrete and plaster by precipitation of calcium carbonate. It is the first time that Bacillus sphaericus was used for consolidating the nonsterilized decayed wall paintings samples by immersing them in sterile nutritional media. The B. sphaericus used in this study produced urease, which catalyzes the hydrolysis of urea (CO(NH₂)₂) into ammonium (NH₄) and carbonate (CO₃^?2) leading to the precipitation of calcium carbonate. The effect of B. sphaericus on wall paintings was determined by recording the evolution of culture media chemistry and examining the treated wall paintings under a scanning electron microscope to show the structural and morphological evolution of calcium carbonate that was investigated in wall paintings models.     

Azaindoles as potent CRTH2 receptor antagonists

A new class of 7-azaindole analogs of MK-7246 as potent and selective CRTH2 antagonists is reported. The SAR leading to the identification of the optimal azaindole regioisomer as well as the pharmacokinetics and off-target activities of the most potent antagonists are disclosed.     

Reliability and Validity of a 20-s Alternative to the Wingate Anaerobic Test in Team Sport Male Athletes

The intent of this study was to evaluate relative and absolute reliability of the 20-s anaerobic test (WAnT₂₀) versus the WAnT₃₀ and to verify how far the various indices of the 30-s Wingate anaerobic test (WAnT₃₀) could be predicted from the WAnT₂₀ data in male athletes. The participants were Exercise Science majors (age: 21.5±1.6 yrs, stature: 0.183±0.08 m, body mass: 81.2±10.9 kg) who participated regularly in team sports. In Phase I, 41 participants performed duplicate WAnT₂₀ and WAnT₃₀ tests to assess reliability. In Phase II, 31 participants performed one trial each of the WAnT₂₀ and WAnT₃₀ to determine the ability of the WAnT₂₀ to predict components of the WAnT₃₀. In Phase III, 31 participants were used to cross-validate the prediction equations developed in Phase II. Respective intra-class correlation coefficients (ICC) for peak power output (PPO) (ICC = 0.98 and 0.95) and mean power output (MPO) (ICC 0.98 and 0.90) did not differ significantly between WAnT₂₀ and WAnT₃₀. ICCs for minimal power output (PO_min) and fatigue index (FI) were poor for both tests (range 0.53 to 0.76). Standard errors of the means (SEM) for PPO and MPO were less than their smallest worthwhile changes (SWC) in both tests; however, PO_min and FI values were “marginal,” with SEM values greater than their respective SWCs for both tests values. Stepwise regression analysis showed that MPO had the highest coefficient of predictability (R = 0.97), with PO_min and FI considerable lower (R = 0.71 and 0.41 respectively). Cross-validation showed insignificant bias with limits of agreement of 0.99±1.04, 6.5±92.7 W, and 1.6±9.8% between measured and predicted MPO, PO_min, and FI, respectively. WAnT₂₀ offers a reliable and valid test of leg anaerobic power in male athletes and could replace the classic WAnT₃₀.     

Response of antioxidative enzymes and apoplastic bypass transport in Thlaspi caerulescens and Raphanus sativus to cadmium stress

A hydroponics experiment using hyperaccumulator Thlaspi caerulescens (alpine pennycress) and non-specific accumulator Raphanus sativus (common radish) was conducted to investigate the short-term effect of increasing Cd concentrations (0, 25, 50, 75, 100 microM) on metal uptake, chlorophyll content, antioxidative enzymes, and apoplastic bypass flow. As expected, T. caerulescens generally showed better resistance to metal stress, which was reflected by higher Cd accumulation within plant tissues with no signs of chlorosis, or wilt. Glutathione reductase (GR) and superoxide dismutase (SOD) activities in fresh leaves were monitored as the plant metal-detoxifying response. In general, both plant species exhibited an increase trend of GR activity before declining at 100 microM likely due to excessive levels of phytotoxic Cd. SOD activity exhibited almost a similar variation pattern to GR and decreased also at 100 microM Cd. For both plant species, fluorescent PTS uptake (8-hydroxy-1,3,6-pyrenetrisulphonic acid) increased significantly with metal level in exposure solutions indicating that Cd has a comparable effect to drought or salinity in terms of the gain of relative importance in apoplastic bypass transport under such stress conditions.     

New indole amide derivatives as potent CRTH2 receptor antagonists

A new series of indole amide acting as hCRTH2 receptor ligands had been explored and are described herein. Several amide derivatives displaying low nanomolar activity in hCRTH2 binding and whole blood assays were identified. They were found to behave as a full antagonists, exhibiting good selectivity over related prostaglandin receptors. Also, prototypical compounds in this novel series which displayed acceptable CYP profiles and were orally bioavailable in rats were identified.     

The comparative effects of metals on the hatching of earthworm cocoons

To establish the use of Metaphire posthuma as a sensitive model for ecotoxicological studies, the comparative effects of five metals on the hatching profiles of the cocoons of the earthworms, Metaphire posthuma, Eisenia foetida and Perionyx excavatus, were studied. The cocoons of the three species of earthworms were exposed to copper, chromium (III), chromium (VI), lead and zinc at 1.25, 2.5, 5.0 and 10.0 ppm. Viable cocoons were incubated at 20 +/- degrees C by using the immersion method. The results indicated that the inhibition of cocoon hatching was concentration dependent. The normal hatching, delayed hatching and non-viability of cocoons were recorded. At a concentration of 1.25 ppm, there was almost no effect on the hatching of the cocoons of all three species of earthworms, except when exposed to chromium (VI), but higher concentrations (2.5 and 5.0 ppm) caused severe effects. It was concluded that M. posthuma was more sensitive than the other two species, and that it is a suitable model for use in ecotoxicity testing.