Salivary Cortisol Mediates Effects of Poverty and Parenting on Executive Functions in Early Childhood

In a predominantly low-income population-based longitudinal sample of 1,292 children followed from birth, higher level of salivary cortisol assessed at ages 7, 15, and 24 months was uniquely associated with lower executive function ability and to a lesser extent IQ at age 3 years. Measures of positive and negative aspects of parenting and household risk were also uniquely related to both executive functions and IQ. The effect of positive parenting on executive functions was partially mediated through cortisol. Typical or resting level of cortisol was increased in African American relative to White participants. In combination with positive and negative parenting and household risk, cortisol mediated effects of African American ethnicity, income-to-need, and maternal education on child cognitive ability.

Effect of Quercetin on Paraoxonase 2 Levels in RAW264.7 Macrophages and in Human Monocytes––Role of Quercetin Metabolism

There is increasing evidence that the intracellular antioxidant enzyme paraoxonase 2 (PON2) may have a protective function in the prevention of atherogenesis. An enhancement of PON2 activity by dietary factors including flavonoids is therefore of interest. In the present study we determined the effect of quercetin on paraoxonase 2 levels in cultured murine macrophages in vitro and in overweight subjects with a high cardiovascular risk phenotype supplemented with 150 mg quercetin/day for 42 days in vivo. Supplementation of murine RAW264.7 macrophages in culture with increasing concentrations of quercetin (1, 10, 20 μmol/L) resulted in a significant increase in PON2 mRNA and protein levels, as compared to untreated controls. Unlike quercetin, its glucuronidated metabolite quercetin-3-glucuronide did not affect PON2 gene expression in cultured macrophages. However the methylated quercetin derivative isorhamnetin enhanced PON2 gene expression in RAW264.7 cells to similar extent like quercetin. Although supplementing human volunteers with quercetin was accompanied by a significant increase in plasma quercetin concentration, dietary quercetin supplementation did not change PON2 mRNA levels in human monocytes in vivo. Current data indicate that quercetin supplementation increases PON2 levels in cultured monocytes in vitro but not in human volunteers in vivo

Assessing genetic polymorphisms using DNA extracted from cells present in saliva samples

<p>Abstract</p> <p>Background</p> <p>Technical advances following the Human Genome Project revealed that high-quality and -quantity DNA may be obtained from whole saliva samples. However, usability of previously collected samples and the effects of environmental conditions on the samples during collection have not been assessed in detail. In five studies we document the effects of sample volume, handling and storage conditions, type of collection device, and oral sampling location, on quantity, quality, and genetic assessment of DNA extracted from cells present in saliva.</p> <p>Methods</p> <p>Saliva samples were collected from ten adults in each study. Saliva volumes from .10-1.0 ml, different saliva collection devices, sampling locations in the mouth, room temperature storage, and multiple freeze-thaw cycles were tested. One representative single nucleotide polymorphism (SNP) in the catechol-<it>0</it>-methyltransferase gene (COMT rs4680) and one representative variable number of tandem repeats (VNTR) in the serotonin transporter gene (5-HTTLPR: serotonin transporter linked polymorphic region) were selected for genetic analyses.</p> <p>Results</p> <p>The smallest tested whole saliva volume of .10 ml yielded, on average, 1.43 ± .77 μg DNA and gave accurate genotype calls in both genetic analyses. The usage of collection devices reduced the amount of DNA extracted from the saliva filtrates compared to the whole saliva sample, as 54-92% of the DNA was retained on the device. An "adhered cell" extraction enabled recovery of this DNA and provided good quality and quantity DNA. The DNA from both the saliva filtrates and the adhered cell recovery provided accurate genotype calls. The effects of storage at room temperature (up to 5 days), repeated freeze-thaw cycles (up to 6 cycles), and oral sampling location on DNA extraction and on genetic analysis from saliva were negligible.</p> <p>Conclusions</p> <p>Whole saliva samples with volumes of at least .10 ml were sufficient to extract good quality and quantity DNA. Using 10 ng of DNA per genotyping reaction, the obtained samples can be used for more than one hundred candidate gene assays. When saliva is collected with an absorbent device, most of the nucleic acid content remains in the device, therefore it is advisable to collect the device separately for later genetic analyses.</p

Persister cell phenotypes contribute to poor patient outcomes after neoadjuvant chemotherapy in PDAC

Neoadjuvant chemotherapy can improve the survival of individuals with borderline and unresectable pancreatic ductal adenocarcinoma; however, heterogeneous responses to chemotherapy remain a significant clinical challenge. Here, we performed RNA sequencing (n = 97) and multiplexed immunofluorescence (n = 122) on chemo-naive and postchemotherapy (post-CTX) resected patient samples (chemoradiotherapy excluded) to define the impact of neoadjuvant chemotherapy. Transcriptome analysis combined with high-resolution mapping of whole-tissue sections identified GATA6 (classical), KRT17 (basal-like) and cytochrome P450 3A (CYP3A) coexpressing cells that were preferentially enriched in post-CTX resected samples. The persistence of GATA6hi and KRT17hi cells post-CTX was significantly associated with poor survival after mFOLFIRINOX (mFFX), but not gemcitabine (GEM), treatment. Analysis of organoid models derived from chemo-naive and post-CTX samples demonstrated that CYP3A expression is a predictor of chemotherapy response and that CYP3A-expressing drug detoxification pathways can metabolize the prodrug irinotecan, a constituent of mFFX. These findings identify CYP3A-expressing drug-tolerant cell phenotypes in residual disease that may ultimately inform adjuvant treatment selection

Modera, observa, escuta e foca-te na conversa de grupo: uma reflexão crítica

A técnica focus group consiste numa abordagem metodológica de natureza qualitativa, cuja recolha de dados se processa através de uma conversa com um grupo de participantes sobre um determinado tema. Esta obedece a um conjunto de procedimentos técnicos associados. Com este trabalho, temos o objetivo de caracterizar alguns desses aspetos aquando da implementação desta técnica em estudos da área da educação. Para atingir tal propósito, procedemos a uma revisão da literatura de 12 artigos empíricos. Da análise efetuada, constatamos um notório desfasamento entre o que o que é proferido na literatura e o veiculado nas investigações. Verificamos que subsiste a inexistência de aspetos relacionados com: i) processos de preparação e implementação da moderação; ii) local onde decorreu a sessão; iii) guião da moderação; iv) indicação e função do moderador e do observador. Esta subvalorização pode comprometer a qualidade dos dados recolhidos. Não obstante, cientes de que a ausência da descrição dos momentos específicos, bem como de aspetos inerentes a esses nos artigos, não implica a sua adoção na investigação. Do exposto resulta, evidentemente, a necessidade de incluir os aspetos supramencionados em artigos científicos.publishe

Aggression as an equifinal outcome of distinct neurocognitive and neuroaffective processes

AbstractEarly onset aggression precipitates a cascade of risk factors, increasing the probability of a range of externalizing and internalizing psychopathological outcomes. Unfortunately, decades of research on the etiological contributions to the manifestation of aggression have failed to yield identification of any risk factors determined to be either necessary or sufficient, likely attributable to etiological heterogeneity within the construct of aggression. Differential pathways of etiological risk are not easily discerned at the behavioral or self-report level, particularly in young children, requiring multilevel analysis of risk pathways. This study focuses on three domains of risk to examine the heterogeneity in 207 urban kindergarten children with high levels of aggression: cognitive processing, socioemotional competence and emotion processing, and family context. The results indicate that 90% of children in the high aggression group could be characterized as either low in verbal ability or high in physiological arousal (resting skin conductance). Children characterized as low verbal, high arousal, or both differed in social and emotional competence, physiological reactivity to emotion, and aspects of family-based contextual risk. The implications of this etiologic heterogeneity of aggression are discussed in terms of assessment and treatment.</jats:p

Term Decidual Cell Thrombomodulin and Endothelial Protein C Receptor Expression Is Upregulated by Thrombin Irrespective of Ovarian Hormonal Priming

Abstract It is well known that normal pregnancy is associated with changes in the hemostatic system that favors a prothrombotic milieu. Moreover, pregnancies complicated by adverse pregnancy outcomes (APO), including fetal loss, intrauterine growth restriction, preeclampsia and abruptio placentae frequently have evidence of uteroplacental thrombosis, vascular damage, and fibrin deposition, upon pathologic examination. The protein C (PC) system [includes; PC, protein S (PS), thrombomodulin (TM), endothelial protein C receptor (EPCR)] is pivotal in the maintenance of the hemostatic balance. TM and EPCR are both transmembrane glycoproteins and are central to the generation of APC, down-regulation of thrombin (FIIa) formation, and down-regulation of the pro-inflammatory process. In trophoblasts TM EPCR are involved in regulating fibrin formation, trophoblast proliferation and apoptosis. Decidual cells (DCs) are located at the maternal fetal interface, an area that must undergo substantial remodeling of the extracellular matrix. We hypothesized that the DCs would express components of the PC system to regulate thrombin since it is involved in extracellular matrix degradation. DCs were harvested and prepared from term placentas from Cesarean sections from 5 uncomplicated pregnancies. The amnion was removed, and the decidua was scraped from the chorion, and digested. The DCs were purified using a Percoll Gradient, seeded on gelatin coated dishes, and passaged until free of white cells. Confluent DCs were incubated with vehicle control or primed in serum-containing medium with 10–8 M estradiol(E2), or 10–7 M medroxy-progesterone acetate(MPA), or E2+MPA. DCs were switched to defined medium with corresponding control and steroids with or without FIIa. After 6 hours, real time PCR was performed on extracted RNA. In the hormonally unprimed condition, thrombin tended to increase the expression of TM an average of 5 fold increase (n=5). The combination of E2+MPA revealed an average increase in the expression of TM in response to FIIa of 5.24 fold. This effect was neutralized by the addition of hirudin. In the hormonally unprimed condition, FIIa increased EPCR expression in response to FIIa, average of 2.18 (n=5). There was a slight further increase with the addition of FIIa 2.7, 2.1, 2.5 fold in the DCs primed with E2, MPA, E2+MPA, respectively. The effect was neutralized by the addition of hirudin. A western blot analysis of TM protein expression, a pattern similar to real time PCR was observed. The lack of differential response of FIIa to hormonal priming may be protective, particularly in the peripartum period, where there are significant hormonal fluctuations, yet FIIas regulatory mechanisms must be preserved. Further studies are underway to assess the interaction of the PC system, FIIa and the effect of gestational age on DCs. Derangements of PC system and DCs may predispose to APO. Table 1 N=5 No stimulant T T + H No Stimulant T T+H Real time PCR normalized with beta-actin. T=thrombin (2.5U/ml), H=Hirudin (3U/ml). Column 2 to 4 are DCs without sterois priming. Column 5 to 7 are DCs primed with E2 + MPA. TM 1 +/− 0 5 +/− 5.24 1.08 +/− 0.43 3.08 +/− 1.49 5.24 +/− 2.28 1.85 +/− 2.2 EPCR 1 +/− 0 2.18 +/− 1.38 1.02 +/− 0.17 2.96 +/− 1.90 2.48 +/− 0.77 2.45 +/− 2.2

Protein S Free Antigen Levels and the Occurrence of IgG and IgM Antibodies to Protein S in Normal Pregnancy

Abstract The evaluation of thrombophilia (TP) in pregnancy is central to the assessment for the risk for adverse pregnancy outcomes (APO) and thrombosis. Antibodies to protein S (PS) have been described in patients with autoimmune diseases and have also been related to thrombosis and APO in pregnancy. Therefore several major issues exist in the evaluation of PS in pregnancy for TP and the risk for APO and thrombosis. What are the ranges of free PS antigen (FPSAG) levels in normal pregnancy (NP)? Do antibodies to PS occur in so called NP and do they affect the levels of PS? We studied 82 1st trimester (TRI) samples, 64 2nd TRI samples, and 78 3rd TRI samples from normal pregnancies to evaluate the FPSAG levels and antibodies against PS both IgG and IgM. We also tested 50 normal non-pregnant donors for antibodies against PS to establish the non-pregnant reference range. The data are show in table 1. It reveals that even in NP the FPSAG levels can decrease to as low as 28% in 1st and 2nd TRI, and 20% in 3rd TRI. These values are significantly below the non-pregnant values. The differences of decreased of PS between 1st vs 3rd, and 2nd vs 3rd TRI are statistically significant (p<0.05). We have previously reported that PS lower than 30% in the 2nd to 3rd TRI was associated with a higher risk for APO. If additional TP factors are present, PS levels in lower NP reference range may increase the risk for APO and thrombosis. The data also shows that there is an increase in PS IgG and IgM antibodies during gestation compared to non-pregnant controls. The number of patients with IgG and IgM antibodies above the 3SD of control value increase with gestation (3% vs 8% vs 9% for IgG, and 13% vs 26% vs 31% for IgM, [1st TRI, 2nd TRI, 3rd TRI respectively]). The lack of overall correlation of antibodies with the protein level is similar to the finding of Gris et al with protein Z and APO. Do the antibodies affect the function but not the level of the factors responsible for the natural inhibition of the prothrombotic process? Do these antibodies become part of the pathogenesis of APO and thrombosis in pregnancy associated with the anticardiolipin syndrome? These questions need to be addressed by larger studies of normal and adverse pregnancy patients. The increase in the number of NP subjects with high levels of PS-IgG and IgM antibodies with gestation might indicate an enhanced immune response to components of the hemostatic system in some patients who might become more vulnerable to APO. The low values of FPSAG in NP add to the difficulty of assessing for a pre-existing deficiency state. However, if an inherited deficiency is present the FPSAG should be in the lower ranges of those we have established for the three TRI. This need to be defined by the assessment of a larger group of PS deficiency patients during pregnancy. Table 1 shows the mean and standrad deviation of FPSAG levels and the titer of PS IgG and IgM antibodies 1st trimester 2nd trimester 3dr trimester CTR CTR= non-pregnant control FPSAG 38.8+/−10.5% 35.9+/−8.3% 27.9+/−7.0% 88+/−19% Anti-PS IgG 0.29+/−0.35OD 0.45+/−0.25OD 0.44+/−0.31OD 0.27+/−0.20OD Anti-PS IgM 0.48+/−0.38OD 0.66+/−0.42OD 0.61+/−0.36OD 0.26+/−0.15O