HLA-A and -B alleles and haplotypes in hemochromatosis probands with HFE C282Y homozygosity in central Alabama

BACKGROUND: We wanted to quantify HLA-A and -B allele and haplotype frequencies in Alabama hemochromatosis probands with HFE C282Y homozygosity and controls, and to compare results to those in other populations. METHODS: Alleles were detected using DNA-based typing (probands) and microlymphocytotoxicity (controls). RESULTS: Alleles were determined in 139 probands (1,321 controls) and haplotypes in 118 probands (605 controls). In probands, A*03 positivity was 0.7482 (0.2739 controls; p =< 0.0001; odds ratio (OR) 7.9); positivity for B*07, B*14, and B*56 was also increased. In probands, haplotypes A*03-B*07 and A*03-B*14 were more frequent (p < 0.0001, respectively; OR = 12.3 and 11.1, respectively). The haplotypes A*01-B*60, A*02-B*39, A*02-B*62, A*03-B*13, A*03-B*15, A*03-B*27, A*03-B*35, A*03-B*44, A*03-B*47, and A*03-B*57 were also significantly more frequent in probands. 37.3% of probands were HLA-haploidentical with other proband(s). CONCLUSIONS: A*03 and A*03-B*07 frequencies are increased in Alabama probands, as in other hemochromatosis cohorts. Increased absolute frequencies of A*03-B*35 have been reported only in the present Alabama probands and in hemochromatosis patients in Italy. Increased absolute frequencies of A*01-B*60, A*02-B*39, A*02-B*62, A*03-B*13, A*03-B*15, A*03-B*27, A*03-B*44, A*03-B*47, and A*03-B*57 in hemochromatosis cohorts have not been reported previously

Senescent changes in parafoveal color appearance: saturation as a function of stimulus area.

The chromatic content (saturation) of monochromatic stimuli (480, 505, 577, and 650 nm) was scaled as a function of field size at three different retinal locations by 58 observers ranging from 18 to 83 yr of age. The different retinal locations (6 deg nasal, 2.5 deg inferior and 6 deg temporal eccentricity) were chosen according to anatomical studies demonstrating different degrees of senescent losses of cones or ganglion cells. Nine field sizes were tested, ranging from 0.0096 to 0.96 deg in diameter. The subjects used a percentage scale to judge the saturation of the flashed stimulus presentations (2 s on, 5 s off). The data analysis demonstrated that older observers require larger field sizes than younger observers to perceive hue as well as larger field sizes to reach the same level of scaled saturation. The spatial dependency of color appearance for younger and older observers was not correlated with senescent losses in retinal cells reported for the different retinal locations. The data were modeled by using an impulse-response function (i.e., Naka-Rushton equation) so that perceptive fields could be compared to electrophysiological measures of receptive fields or dendritic fields of retinal and cortical cells

Senescent changes in parafoveal color appearance: saturation as a function of stimulus area.

The chromatic content (saturation) of monochromatic stimuli (480, 505, 577, and 650 nm) was scaled as a function of field size at three different retinal locations by 58 observers ranging from 18 to 83 yr of age. The different retinal locations (6 deg nasal, 2.5 deg inferior and 6 deg temporal eccentricity) were chosen according to anatomical studies demonstrating different degrees of senescent losses of cones or ganglion cells. Nine field sizes were tested, ranging from 0.0096 to 0.96 deg in diameter. The subjects used a percentage scale to judge the saturation of the flashed stimulus presentations (2 s on, 5 s off). The data analysis demonstrated that older observers require larger field sizes than younger observers to perceive hue as well as larger field sizes to reach the same level of scaled saturation. The spatial dependency of color appearance for younger and older observers was not correlated with senescent losses in retinal cells reported for the different retinal locations. The data were modeled by using an impulse-response function (i.e., Naka-Rushton equation) so that perceptive fields could be compared to electrophysiological measures of receptive fields or dendritic fields of retinal and cortical cells

Photopigment optical density of the human foveola and a paradoxical senescent increase outside the fovea.

Photopigment optical density (OD) of middle-(M) and long-(L) wavelength-sensitive cones was determined to evaluate the hypothesis that reductions in the amount of photopigment are responsible for age-dependent sensitivity losses of the human cone pathways. Flicker thresholds were measured at the peak and tail of the photoreceptor's absorption spectrum as a function of the intensity of a bleaching background. Photopigment OD was measured at 0 (fovea), 2, 4, and 8 deg in the temporal retina by use of a 0.3-deg-diameter test spot. Seventy-two genetically characterized dichromats were studied so that the L- and M-cones could be analyzed separately. Subjects included 28 protanopes with M- but no L-cones and 44 deuteranopes with L- but no M-cones (all male, age range 12-29 and 55-83 years). Previous methods have not provided estimates of photopigment OD for separate cone classes in the foveola. In this study, it was found that foveolar cones are remarkably efficient, absorbing 78% of the available photons (OD = 0.65). Photopigment OD decreased exponentially with retinal eccentricity independently of age and cone type. Paradoxically, the OD of perifoveal cones increased significantly with age. Over the 70-year age range of our participants, the perifoveal M- and L-cones showed a 14% increase in capacity to absorb photons despite a 30% decrease in visual sensitivity over the same period

Variety of genotypes in males diagnosed as dichromatic on a conventional clinical anomaloscope.

The hypothesis that dichromatic behavior on a clinical anomaloscope can be explained by the complement and arrangement of the long- (L) and middle-wavelength (M) pigment genes was tested. It was predicted that dichromacy is associated with an X-chromosome pigment gene array capable of producing only a single functional pigment type. The simplest case of this is when deletion has left only a single X-chromosome pigment gene. The production of a single L or M pigment type can also result from rearrangements in which multiple genes remain. Often, only the two genes at the 5' end of the array are expressed; thus, dichromacy is also predicted to occur if one of these is defective or encodes a defective pigment, or if both of them encode pigments with identical spectral sensitivities. Subjects were 128 males who accepted the full range of admixtures of the two primary lights as matching the comparison light on a Neitz or Nagel anomaloscope. Strikingly, examination of the L and M pigment genes revealed a potential cause for a color-vision defect in all 128 dichromats. This indicates that the major component of color-vision deficiency could be attributed to alterations of the pigment genes or their regulatory regions in all cases, and the variety of gene arrangements associated with dichromacy is cataloged here. However, a fraction of the dichromats (17 out of 128; 13%) had genes predicted to encode pigments that would result in two populations of cones with different spectral sensitivities. Nine of the 17 were predicted to have two pigments with slightly different spectral peaks (usually &lt; or = 2.5 nm) and eight had genes which specified pigments identical in peak absorption, but different in amino acid positions previously associated with optical density differences. In other subjects, reported previously, the same small spectral differences were associated with anomalous trichromacy rather than dichromacy. It appears that when the spectral difference specified by the genes is very small, the amount of residual red-green color vision measured varies; some individuals test as dichromats, others test as anomalous trichromats. The discrepancy is probably partly attributable to testing method differences and partly to a difference in performance not perception, but it seems there must also be cases in which other factors, for example, cone ratio, contribute to a person's ability to extract a color signal from a small spectral difference

FOXO-dependent expression of the proapoptotic protein Bim: pivotal role for apoptosis signaling in endothelial progenitor cells

Endothelial progenitor cells (EPCs) contribute to postnatal neovascularization. Risk factors for coronary artery disease reduce the number of EPCs in humans. Since EPC apoptosis might be a potential mechanism to regulate the number of EPCs, we investigated the effects of oxidative stress and HMG-CoA-reductase inhibitors (statins) on EPC apoptosis. Atorvastatin, mevastatin, or VEGF prevented EPC apoptosis induced by H2O2. The antiapoptotic effect was reversed by inhibition of the PI3K/Akt pathway. Forkhead transcription factors (FOXO1, FOXO3a, FOXO4) exert proapoptotic effects and are phosphorylated and, thereby, inactivated by Akt. Therefore, we elucidated the involvement of forkhead transcription factors. Atorvastatin induced the phosphorylation of the predominant forkhead factor FOXO4 in EPCs. In addition, atorvastatin reduced the expression of the proapoptotic forkhead-regulated protein Bim in a PI3K-dependent manner. Consistently, overexpression of FOXO4 activated the Bim promoter as determined by reporter gene expression and stimulated the expression of Bim, resulting in an increased EPC apoptosis. Statins failed to prevent EPC apoptosis induced by overexpression of Bim or nonphosphorylatable FOXO4, suggesting that the protective effects of statins depend on this pathway. In summary, our results show that FOXO-dependent expression of Bim plays a pivotal role for EPC apoptosis. Statins reduce oxidative stress-induced EPC apoptosis, inactivate FOXO4, and down-regulate Bim

Synergistic effects of chemotherapeutic drugs in lymphoma cells are associated with down-regulation of inhibitor of apoptosis proteins (IAPs), prostate-apoptosis-response-gene 4 (Par-4), death-associated protein (Daxx) and with enforced caspase activation.

Cytotoxic drugs mediate apoptotic tumor cell death by influencing key regulator proteins of programmed cell death. In clinical practice cytotoxic drug combinations are desired to potentiate tumor cell kill and to minimize side effects. Nevertheless, the molecular mechanisms underlying synergistic and antagonistic effects on tumor cells are still poorly understood. In order to elucidate these molecular mechanisms we established models of synergistic and antagonistic drug combinations within the same lymphoma cell lines. By combination index method we demonstrated that bendamustine in combination with either doxorubicin or mitoxantrone caused antagonistic effects on disruption of mitochondrial membrane potential as well as on the rate of apoptosis. In contrast the combination of bendamustine with cladribine acted synergistically on these parameters. By using the IC(50) (dosages causing 50% rate of apoptosis) the synergistic effect of the combination of bendamustine and cladribine was associated with an enhanced mitochondrial release of cytochrome c and Smac/DIABLO, by down-regulation of x-linked inhibitor of apoptosis (XIAP), cIAP1, Par-4 and Daxx as well as by a significantly increased activation of caspases-3, -6, -7, -8 and -9. At the same rate of apoptosis (IC(50)), the antagonistic combinations did not increase the release of cytochrome c or Smac/DIABLO, nor down-regulate the expression of XIAP, cIAP1, Par-4 and Daxx, nor increase the activation of caspases. The role of down-regulation of IAPs and of enforced caspase activation for synergism in this model was supported by the observation, that broad spectrum inhibition of caspases re-established expression of XIAP. Our study is the first to outline the molecular alterations caused by synergistic and antagonistic drug combinations within the same lymphoma cell model. The above described mechanisms were already assessable at a point where the effects of synergistic or antagonistic combinations could not yet be discriminated quantitatively by the level of apoptosis rate of the lymphoma cells