Platelet and Red Blood Cell Counts, as well as the Concentrations of Uric Acid, but Not Homocysteinaemia or Oxidative Stress, Contribute Mostly to Platelet Reactivity in Older Adults

Purpose. The goal of this study was to estimate the hierarchical contribution of the most commonly recognized cardiovascular risk factors associated with atherogenesis to activation and reactivity of blood platelets in a group of men and women at ages 60-65. Methods. Socioeconomic and anthropometric data were taken from questionnaires. Blood morphology and biochemistry were measured with standard diagnostic methods. Plasma serum homocysteine was measured by immunochemical method. Plasma concentrations of VCAM, ICAM, total antioxidant status, and total oxidant status were estimated with commercial ELISA kits. Markers of oxidative stress of plasma and platelet proteins (concentrations of protein free thiol and amino groups) and lipids (concentrations of lipid peroxides) and generation of superoxide anion by platelets were measured with colorimetric methods. Platelet reactivity was estimated by impedance aggregometry with arachidonate, collagen, and ADP as agonists. Expression of selectin-P and GPIIb/IIIa on blood platelets was tested by flow cytometry. Results. Platelet aggregation associated significantly negatively with HGB and age and significantly positively with PLT, MPV, PCT, PDW, and P-LCR. When platelet reactivity (“cumulative platelet reactivity_aggregation”) was analyzed in a cumulated manner, the negative association with serum concentration of uric acid (Rs=−0.169, p=0.003) was confirmed. Multivariate analysis revealed that amongst blood morphological parameters, platelet count, plateletcrit, and number of large platelets and uric acid are the most predictive variables for platelet reactivity. Conclusions. The most significant contributors to platelet reactivity in older subjects are platelet morphology, plasma uricaemia, and erythrocyte morphology

Hunting for the LCT-13910*T Allele between the Middle Neolithic and the Middle Ages Suggests Its Absence in Dairying LBK People Entering the Kuyavia Region in the 8th Millennium BP

<div><p>Populations from two medieval sites in Central Poland, Stary Brześć Kujawski-4 (SBK-4) and Gruczno, represented high level of lactase persistence (LP) as followed by the LCT-13910*T allele’s presence (0.86 and 0.82, respectively). It was twice as high as in contemporaneous Cedynia (0.4) and Śródka (0.43), both located outside the region, higher than in modern inhabitants of Poland (0.51) and almost as high as in modern Swedish population (0.9). In an attempt to explain the observed differences its frequency changes in time were followed between the Middle Neolithic and the Late Middle Ages in successive dairying populations on a relatively small area (radius ∼60km) containing the two sites. The introduction of the T allele to Kuyavia 7.4 Ka BP by dairying LBK people is not likely, as suggested by the obtained data. It has not been found in any of Neolithic samples dated between 6.3 and 4.5 Ka BP. The identified frequency profile indicates that both the introduction and the beginning of selection could have taken place approx. 4 millennia after first LBK people arrived in the region, shifting the value of LP frequency from 0 to more than 0.8 during less than 130 generations. We hypothesize that the selection process of the T allele was rather rapid, starting just after its introduction into already milking populations and operated <i>via</i> high rates of fertility and mortality on children after weaning through life-threatening conditions, favoring lactose-tolerant individuals. Facing the lack of the T allele in people living on two great European Neolithization routes, the Danubian and Mediterranean ones, and based on its high frequency in northern Iberia, its presence in Scandinavia and estimated occurrence in Central Poland, we propose an alternative Northern Route of its spreading as very likely. None of the successfully identified nuclear alleles turned out to be deltaF508 CFTR.</p></div

Suggested Northern Route of LCT-13910*T spreading.

<p>Contrasted is time-dependent occurrence of the T allele along west-east gradient from Iberia (0.27; >5 Ka BP [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122384#pone.0122384.ref066" target="_blank">66</a>]), through Scandinavia (0.05; >4 Ka BP [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122384#pone.0122384.ref067" target="_blank">67</a>]), up to Kuyavia and the Chełmno land (<4 Ka BP as predicted by us), with its simultaneous absence along the Danubian and Mediterranean Routes.</p

Location of the explored Polish archaeological sites.

<p>Location of the explored Polish archaeological sites.</p

The value of the Fisher exact test.

<p>Comparison of the T allele frequency found at studied archaeological sites. Statistically significant differences after Bonferroni are typed in boldface.</p><p>The value of the Fisher exact test.</p

Mean allele frequency calculated for two chosen scenarios from Fig. 2A (only curves falling into confidence intervals were taken into account) along with their probabilities marked on the interpolated version of 2D part of Fig. 2A (curve 1—probability 0.027, curve 2—probability 0.297).

<p>Mean allele frequency calculated for two chosen scenarios from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122384#pone.0122384.g002" target="_blank">Fig. 2A</a> (only curves falling into confidence intervals were taken into account) along with their probabilities marked on the interpolated version of 2D part of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122384#pone.0122384.g002" target="_blank">Fig. 2A</a> (curve 1—probability 0.027, curve 2—probability 0.297).</p

Confidence intervals for the T allele and LP frequency found at studied archaeological sites and included in numerical calculations.

<p>Confidence intervals for the T allele and LP frequency found at studied archaeological sites and included in numerical calculations.</p

Canonical, Non-Canonical and Atypical Pathways of Nuclear Factor кb Activation in Preeclampsia

Although higher nuclear factor &kappa;B (NF&kappa;B) expression and activity is observed in preeclamptic placentas, its mechanism of activation is unknown. This is the first study to investigate whether the canonical, non-canonical, or atypical NF&kappa;B activation pathways may be responsible for the higher activation of NF&kappa;B observed in preeclamptic placentas. The study included 268 cases (130 preeclamptic women and 138 controls). We studied the expression of the genes coding for NF&kappa;B activators (NIK, IKK&alpha;, IKK&beta;, and CK2&alpha;) and inhibitors (I&kappa;B&alpha; and I&kappa;B&beta;) using RT-PCR in real time. The RT-PCR results were verified on the protein level using ELISA and Western blot. To determine the efficiency of the pathways, the ratios of activator(s) to one of the inhibitors (I&kappa;B&alpha; or I&kappa;B&beta;) were calculated for each studied pathway. The preeclamptic placentas demonstrated significantly lower IKK&alpha; and CK2&alpha; but higher I&kappa;B&alpha; and I&kappa;B&beta; protein levels. In addition, the calculated activator(s) to inhibitor (I&kappa;B&alpha; or I&kappa;B&beta;) ratios suggested that all studied pathways might be downregulated in preeclamptic placentas. Our results indicate that preeclamptic placentas may demonstrate mechanisms of NF&kappa;B activation other than the canonical, non-canonical, and atypical forms. In these mechanisms, inhibitors of NF&kappa;B may play a key role. These observations broaden the existing knowledge regarding the molecular background of preeclampsia development