Significance of the hydrophobic residues 225–230 of apoA-I for the biogenesis of HDL

We studied the significance of four hydrophobic residues within the 225–230 region of apoA-I on its structure and functions and their contribution to the biogenesis of HDL. Adenovirus-mediated gene transfer of an apoA-I[F225A/V227A/F229A/L230A] mutant in apoA-I^−/− mice decreased plasma cholesterol, HDL cholesterol, and apoA-I levels. When expressed in apoA-I^−/− × apoE^−/− mice, approximately 40% of the mutant apoA-I as well as mouse apoA-IV and apoB-48 appeared in the VLDL/IDL/LDL. In both mouse models, the apoA-I mutant generated small spherical particles of pre-β- and α4-HDL mobility. Coexpression of the apoA-I mutant and LCAT increased and shifted the-HDL cholesterol peak toward lower densities, created normal αHDL subpopulations, and generated spherical-HDL particles. Biophysical analyses suggested that the apoA-I[225–230] mutations led to a more compact folding that may limit the conformational flexibility of the protein. The mutations also reduced the ability of apoA-I to promote ABCA1-mediated cholesterol efflux and to activate LCAT to 31% and 66%, respectively, of the WT control. Overall, the apoA-I[225–230] mutations inhibited the biogenesis of-HDL and led to the accumulation of immature pre-β- and α4-HDL particles, a phenotype that could be corrected by administration of LCAT.     

Quinidine: an “Endangered Species” Drug Appropriate for Management of Electrical Storm in Brugada Syndrome

Brugada syndrome is an inherited channelopathy associated with an increased risk of syncope and sudden cardiac death. In rare cases it can be manifested with electrical storm. We report two cases of Brugada syndrome that presented with electrical storm and were treated successfully with oral quinidine, an "endangered species" drug.     

Novel lipophilic amidate oxorhenium and oxotechnetium complexes as potential brain agents: synthesis, characterization and biological evaluation

Novel oxorhenium and oxotechnetium complexes based on the tetradentate 1-(2-hydroxybenzamido)-2-(pyridinecarboxamido)benzene, H3L, ligand have been synthesized and characterized herein. Thus, by reacting equimolar quantities of the triply deprotonated ligand L3- with the suitable MO3+ precursor, the following neutral MOL complexes could be easily produced following similar synthetic routes: M = Re (1), M = 99gTc (2), and M = 99mTc (3). Complexes 1 and 2, prepared in macroscopic amounts, were chemically characterized and their structure determined by single-crystal X-ray analysis. They are isostructural metal chelates, adopting a distorted square pyramidal geometry around the metal. The N3O donor atom set of the tetradentate ligand defines the basal plane and the oxygen atom of the M = O core occupies the apex of the pyramid. Complex 3 forms quantitatively at tracer level by mixing the H3L ligand with Na99mTcO4 generator eluate in aqueous alkaline media and using tin chloride as reductant in the presence of citrate. Its structure was established by chromatographic comparison with prototypic complexes 1 and 2 using high-performance liquid chromatographic techniques. When challenged with excess glutathione in vitro, complex 3 is rapidly converted to hydrophilic unidentified metal species. Tissue distribution data after administration of complex 3 in vivo revealed a significant uptake and retention of this compound in brain tissue.     

Variability in susceptibility among cucumber and tomato strains of Tetranychus urticae Koch to 2-tridecanone from tomato trichomes: effects of host plant shift

The effects of secondary plant compounds on different host races/strains of a herbivorous arthropod are not easy to interpret based on dose–response tests alone. This difficulty arises because the responses in a population to a given dose are dictated not only by genetic factors, but also by factors such as feeding history, age and plant. To discriminate between these possibilities different strains of a herbivorous mite (Tetranychus urticae Koch) were exposed to one relevant toxin from glandular hairs on tomato (the methyl ketone, 2-tridecanone) and these strains had a known feeding history on either or both of two host plants (tomato and cucumber). It was hypothesized that tomato is a relatively hostile host plant to spider mites partly due to methyl ketones from glandular hairs and that consequently there will be stronger selection on the tomato strains than on the cucumber strains. However, the expected differences between the tomato and cucumber strains did not show up; three spider mite strains collected from tomato and two strains collected from cucumber appeared to be equally susceptible to 2-tridecanone. This unexpected result cannot result from selection for resistance to 2-tridecanone, but it may be the consequence of cross-resistance to other ketones in cucumber or the bio-accumulation (i.e. metabolic load) of 2-tridecanone prior to the toxicity test. To test this hypothesis, one of the tomato strains was released on cucumber for different time intervals and one of the cucumber strains on tomato. It was found that the resistance of the tomato strain to 2-tridecanone increased 6 months after transfer to cucumber. This increased resistance cannot be the result of selection because 2-tridecanone is absent from cucumber. Hence, it may be due either to selection for resistance to another ketone in cucumber, possibly leading to cross-resistance, or to the absence of 2-tridecanone bioaccumulation on cucumber. Transfer of the cucumber strain from cucumber to tomato also increased the resistance to 2-tridecanone. As this was accompanied by high mortality directly after the transfer, selection for resistance may have played a role. Alternatively, the increased resistance may be due to induction of resistance to secondary plant compounds of tomato, including 2-tridecanone. In conclusion, experiments on host plant transfer show that the tomato strain and the cucumber strain are not equal in their resistance to 2-tridecanone.     

Genetic skeletal disorders of the fetus and infant: Pathologic and molecular findings in a series of 41 cases

BACKGROUND: Genetic skeletal disorders of the fetus and infant are a large group of genetic disorders, comprising the groups formerly assigned as skeletal dysplasias (osteochondrodysplasias), dysostoses, and malformation syndromes with a skeletal component. Genetic skeletal disorders may be prenatally detected by ultrasonography or result in intrauterine or early postnatal death, constituting one difficult diagnostic field met by the pathologist who performs the perinatal autopsy. METHODS: In this retrospective study, we have gathered radiologic, physical, histopathologic, and molecular data regarding 41 cases of genetic skeletal disorders diagnosed among 1980 fetal and perinatal autopsies over a 10‐year period. RESULTS: Our series of cases were classified according to the 2006 Nosology and Classification of Genetic Skeletal Disorders. The overall frequency of genetic skeletal disorders was 1:48 autopsies. The FGFR3 group and osteogenesis imperfecta type 2 were the more frequently encountered disorders. The mean gestational age at autopsy was 21.9 weeks (range, 12–37 weeks). A final diagnosis was obtained in 95% of cases. Genetic skeletal disorders were detected by prenatal ultrasound in 90% of cases, with a correct typing of the disorder achieved in only 34%. Molecular analysis was confirmative in 5 cases. CONCLUSIONS: The central role of the perinatal pathologist in collaboration with specialized services is essential for the correct interpretation of the radiologic, physical, and histopathologic findings, to accurately classify specific types of genetic skeletal disorders and enable genetic counseling. Birth Defects Research (Part A), 2009. © 2009 Wiley‐Liss, Inc.     

Expression and promoter methylation status of hMLH1, MGMT,APC, and CDH1 genes in patients with colon adenocarcinoma

Colorectal cancer (CRC) is the third most common cancer in men and the second in women worldwide. CRC development is the result of genetic and epigenetic alterations accumulation in the epithelial cells of colon mucosa. In the present study, DNA methylation, an epigenetic event, was evaluated in tumoral and matching normal epithelium in a cohort of 61 CRC patients. The results confirmed and expanded knowledge for the tumor suppressor genes hMLH1, MGMT, APC, and CDH1. Promoter methylation was observed for all the examined genes in different percentage. A total of 71% and 10% of the examined cases were found to be methylated in two or more and in all genes, respectively. mRNA and protein levels were also evaluated. Promoter methylation of hMLH1, MGMT, APC, and CDH1 genes was present at the early stages of tumor’s formation and it could also be detected in the normal mucosa. Correlations of the methylated genes with patient’s age and tumor’s clinicopathological characteristics were also observed. Our findings suggest that DNA methylation is a useful marker for tumor progression monitoring and that promoter methylation in certain genes is associated with more advanced tumor stage, poor differentiation, and metastasis.     

Combining immunological and molecular data to assess phylogenetic relations of some Greek Podarcis species

Most recent molecular studies revealed the phylogeny of Greek Podarcis species, which for years remained elusive, due to discordant data produced from various chromosomal, complement fixation and protein studies. In this report, we analyzed cellular immune responses of spleen-derived lymphocytes from six allopatric Podarcis species encountered in Greece, by assessing two-way mixed lymphocyte reaction (MLR)-induced proliferation. On the basis of stimulation indices (S.I.) as determined from cultures set up from xenogeneic splenocytes coincubated in pairs, we generated a phylogenetic tree, fully consistent with the phylogenetic relationships of Podarcis as determined by parallel analyses based on partial mitochondrial (mt) DNA sequences. Although the exact mechanisms triggering lymphocyte responses in lizard two-way xenogeneic MLR are not fully understood, our results show the potential use of cell-mediated immune responses as an additional approach to mtDNA analysis, for species delimitation within specific lizard taxa.     

Effects of basketball training on maximal oxygen uptake, muscle strength, and joint mobility in young basketball players

The purpose of this study was to examine the effects of prolonged basketball skills training on maximal aerobic power, isokinetic strength, joint mobility, and body fat percentage, in young basketball players, and controls of the same age. Twenty basketball players and 18 control boys participated in the study. Basketball players participated both in their school's physical education program and in a children's basketball team training program. Controls participated only in their school's physical education program. All subjects were tested every 6 months (18 months total, 11(1/2), 12, 12(1/2), 13 years old) for VO(2)max, peak torque values of the quadriceps and hamstrings at 180 and 300 degrees x s(-1) and range of motion of the knee and hip joints. Body fat percentage was assessed at the beginning and the end of the experimental period. Results showed that the basketball group had lower heart rate values in all ages and higher VO(2) values in the initial test compared with the control in submaximal intensity. The VO(2)max was altered in both groups on the final test, when compared to the initial test. However, the basketball group had a higher VO(2)max on each of the 6-month follow-up measurements, compared to the control group (p < 0.001). At the end of the 18-month follow-up period no significant differences were observed in isokinetic strength and joint mobility of the lower limbs between the 2 groups. On the contrary, the boys of the trained group had significantly lower percentage body fat values, compared to controls. In conclusion, regular basketball training increased aerobic power and decreased body fat percentage of prepubescent boys, while it did not affect muscle strength and joint mobility of the lower limbs. The major implication suggested by the findings of the present study is that, in order to improve the basic physical components, specific training procedures should be incorporated during the basketball training sessions. It is recommended that all children should be involved in some type of cardiovascular and resistance training program.