AMP Affects Intracellular Ca2+ Signaling, Migration, Cytokine Secretion and T Cell Priming Capacity of Dendritic Cells

The nucleotide adenosine-5′-monophosphate (AMP) can be released by various cell types and has been shown to elicit different cellular responses. In the extracellular space AMP is dephosphorylated to the nucleoside adenosine which can then bind to adenosine receptors. However, it has been shown that AMP can also activate A1 and A2a receptors directly. Here we show that AMP is a potent modulator of mouse and human dendritic cell (DC) function. AMP increased intracellular Ca2+ concentration in a time and dose dependent manner. Furthermore, AMP stimulated actin-polymerization in human DCs and induced migration of immature human and bone marrow derived mouse DCs, both via direct activation of A1 receptors. AMP strongly inhibited secretion of TNF-α and IL-12p70, while it enhanced production of IL-10 both via activation of A2a receptors. Consequently, DCs matured in the presence of AMP and co-cultivated with naive CD4+CD45RA+ T cells inhibited IFN-γ production whereas secretion of IL-5 and IL-13 was up-regulated. An enhancement of Th2-driven immune response could also be observed when OVA-pulsed murine DCs were pretreated with AMP prior to co-culture with OVA-transgenic naïve OTII T cells. An effect due to the enzymatic degradation of AMP to adenosine could be ruled out, as AMP still elicited migration and changes in cytokine secretion in bone-marrow derived DCs generated from CD73-deficient animals and in human DCs pretreated with the ecto-nucleotidase inhibitor 5′-(alpha,beta-methylene) diphosphate (APCP). Finally, the influence of contaminating adenosine could be excluded, as AMP admixed with adenosine desaminase (ADA) was still able to influence DC function. In summary our data show that AMP when present during maturation is a potent regulator of dendritic cell function and point out the role for AMP in the pathogenesis of inflammatory disorders

Where It’s at Really Matters: In Situ In Vivo Vascular Endothelial Growth Factor Spatially Correlates with Electron Paramagnetic Resonance pO2 Images in Tumors of Living Mice

Purpose: Tumor microenvironments show remarkable tumor pO_{2} heterogeneity, as seen in prior EPR pO_{2} images (EPROI). pO_{2} correlation with hypoxia response proteins is frustrated by large rapid pO2 changes with position. Procedures: To overcome this limitation, biopsies stereotactically located in the EPROI were used to explore the relationship between vascular endothelial growth factor A (VEGF) concentrations in living mouse tumors and the local EPROI pO_{2}. Results: Quantitative ELISA VEGF concentrations correlated (p = 0.0068 to 0.019) with mean pO_{2}, median pO_{2}, and the fraction of voxels in the biopsy volume with pO_{2} less than 3, 6, and 10 Torr. Conclusions: This validates EPROI hypoxic fractions at the molecular level and provides a new paradigm for the assessment of the relationship, in vivo, between hypoxia and hypoxia response proteins. When translated to human subjects, this will enhance understanding of human tumor pathophysiology and cancer response to therapy

Påverkan på EKG-registrering vid ändrat kroppsläge och elektrodplacering

Elektrokardiografi, EKG är den mest använda kardiovaskulära undersökningen för studering av hjärtats funktion, patologi i hjärtats retledningssystem och rytm. För att registrera och undersöka hjärtats elektriska aktivitet, används de standardiserade 12-avledningssystemet (12-SL) med distal placering av extremitetsavledningarna på underarmar och underben. På grund av muskelstörningar som kan förekomma under bland annat ett arbetsprov, är 12-SL inte alltid optimalt. Därför har andra modifierade system för elektrodplacering av extremitetselektroderna som Mason- Likar och Lund-avledningssystemet, ersatt med en omplacering proximalt på överarmar och höftkam. Tidigare studier har visat en förändring i EKG-registreringen som kan leda till falska patologier eller missvisande av det befintliga patologierna. Studier har även visat en förändring i EKG-registreringar som kan uppkomma vid olika kroppspositioner. Syftet med denna studie var därför att studera den potentiella skillnaden som uppstår mellan 12-SL och en proximal omplacering enligt Lund-systemet med en kroppspositionsändring. Studien inkluderade 31 deltagare remitterade till ett ordinärt arbetsprov, där en registrering skedde vid distal- och proximal koppling samt med patienten i sittande läge. Utifrån erhållna EKG-registreringar jämfördes R-vågsamplituden, QRS-durationen samt PQ tiden. Studien visade en statistisk signifikant skillnad i alla extremitetsavledningar vid en förflyttning av elektroderna för R-vågamplituden samt för avledning V5 och –aVL vid ändring av kroppsposition. Ingen signifikant förändring kunde observeras för PQ-tiden och QRS-durationen.Electrocardiography, ECG is the most used cardiovascular method to study cardiac function, pathology of the heart's retinal system and rhythm. To record and investigate the electrical activity of the heart, the standard 12-lead system (12-SL) with distal placement of the limb leads on the forearms and lower legs are commonly used. Due to muscular artifact that can occur in connection with exercise ECGs, 12-SL is not always optimal for use. Therefore, other systems such as Mason-Likar and the Lund-system have replaced with a proximal repositioning of the limb electrodes. Previous studies have shown a change in ECG registration that can lead to false pathology and alarms when modifying the standardized 12-lead system. Previous studies have also shown a possible change in ECG-registrations that can occur with a different body positions. The purpose of this research was to study the potential change that arises between 12-SL and a proximal relocation according to the Lund system with a difference in a body position. The study included 31 participants referred to the regular work test, where a registration was made at distal and proximal connection as well as with the patient in a sitting position. Based on the obtained ECG, the R- wave amplitudes, QRS- durations and PQ- intervals was measured and compared. Results showed a statistically significant difference in all limb leads at and displacements of the electrodes for the R-wave amplitude as well as for the V5 and -aVL with a different body position. No significant change was observers for the PQ- time and QRS- duration

Påverkan på EKG-registrering vid ändrat kroppsläge och elektrodplacering

Elektrokardiografi, EKG är den mest använda kardiovaskulära undersökningen för studering av hjärtats funktion, patologi i hjärtats retledningssystem och rytm. För att registrera och undersöka hjärtats elektriska aktivitet, används de standardiserade 12-avledningssystemet (12-SL) med distal placering av extremitetsavledningarna på underarmar och underben. På grund av muskelstörningar som kan förekomma under bland annat ett arbetsprov, är 12-SL inte alltid optimalt. Därför har andra modifierade system för elektrodplacering av extremitetselektroderna som Mason- Likar och Lund-avledningssystemet, ersatt med en omplacering proximalt på överarmar och höftkam. Tidigare studier har visat en förändring i EKG-registreringen som kan leda till falska patologier eller missvisande av det befintliga patologierna. Studier har även visat en förändring i EKG-registreringar som kan uppkomma vid olika kroppspositioner. Syftet med denna studie var därför att studera den potentiella skillnaden som uppstår mellan 12-SL och en proximal omplacering enligt Lund-systemet med en kroppspositionsändring. Studien inkluderade 31 deltagare remitterade till ett ordinärt arbetsprov, där en registrering skedde vid distal- och proximal koppling samt med patienten i sittande läge. Utifrån erhållna EKG-registreringar jämfördes R-vågsamplituden, QRS-durationen samt PQ tiden. Studien visade en statistisk signifikant skillnad i alla extremitetsavledningar vid en förflyttning av elektroderna för R-vågamplituden samt för avledning V5 och –aVL vid ändring av kroppsposition. Ingen signifikant förändring kunde observeras för PQ-tiden och QRS-durationen.Electrocardiography, ECG is the most used cardiovascular method to study cardiac function, pathology of the heart's retinal system and rhythm. To record and investigate the electrical activity of the heart, the standard 12-lead system (12-SL) with distal placement of the limb leads on the forearms and lower legs are commonly used. Due to muscular artifact that can occur in connection with exercise ECGs, 12-SL is not always optimal for use. Therefore, other systems such as Mason-Likar and the Lund-system have replaced with a proximal repositioning of the limb electrodes. Previous studies have shown a change in ECG registration that can lead to false pathology and alarms when modifying the standardized 12-lead system. Previous studies have also shown a possible change in ECG-registrations that can occur with a different body positions. The purpose of this research was to study the potential change that arises between 12-SL and a proximal relocation according to the Lund system with a difference in a body position. The study included 31 participants referred to the regular work test, where a registration was made at distal and proximal connection as well as with the patient in a sitting position. Based on the obtained ECG, the R- wave amplitudes, QRS- durations and PQ- intervals was measured and compared. Results showed a statistically significant difference in all limb leads at and displacements of the electrodes for the R-wave amplitude as well as for the V5 and -aVL with a different body position. No significant change was observers for the PQ- time and QRS- duration

NPM1c, une cible dans le traitement des leucémies aigues myéloïdes (LAM)

Acute myeloid leukemia (AML) is a genetically heterogeneous disease. Despite improvements in understanding the biology of AML, survival rates remain quite low. Prognosis of AML patients largely depends on acquired cytogenetic and molecular abnormalities. Nucleophosmin-1 (NPM1) is an essential gene encoding for a nucleocytoplasmic shuttling protein mainly localized to the nucleolus. Among several functions, NPM1 plays major roles in stabilization of the p14ARF tumor suppressor protein, regulation of ribosome biogenesis, control of centrosome duplication, response to stress stimuli and P53 activation. NPM1 is one of the most frequently mutated genes in AML accounting for around one third of patients (NPM1c AML). In mutant NPM1c proteins, critical tryptophan residues in the C-terminus are lost and a de novo nuclear export signal is created. This leads to ectopic and aberrant accumulation of NPM1c, along with normal NPM1, in the cytoplasm of AML blasts, thus playing a major role in leukemogenesis. The promyelocytic leukemia protein (PML) tumor suppressor organizes nuclear bodies (NB) domains that control proteolysis and P53-driven senescence. PML NBs are disorganized in NPM1c AML. Retinoic acid (RA), a hormone favoring differentiation of myeloid cells, increases survival of chemotherapy-treated NPM1c AMLs. We and others previously reported that RA and Arsenic trioxide (ATO) synergize to induce NPM1c degradation and to inhibit growth and induce apoptosis of NPM1c cells. Importantly, combined RA/ATO treatment significantly reduced bone marrow blasts in some NPM1c AML patients and restored the subnuclear localization of both NPM1 and PML. The main objective of the current work was to unravel the biochemical pathways driving NPM1c catabolism and to elucidate the molecular mechanisms of RA/ATO anti-leukemic activities. We demonstrate that PML is required for RA/ATO-induced NPM1c degradation. In primary NPM1c-AML blasts, RA rapidly upregulates the initial low basal PML expression through Pin-1 inhibition prior to NPM1c clearance. This RA-induced PML stabilizes P53 and primes blasts for ATO-driven nuclear body reformation, yielding hyper-activation of P53. RA/ATO combination elicits PML-dependent responses, associated with in vivo P53 activation in NPM1c AML xenografts. Importantly, RA/ATO-initiated a transient complete remission in a NPM1c-AML patient demonstrating clinical relevance of this combination. We also demonstrate that ATO-driven NPM1c degradation happens via the PML/SUMO/RNF4/Proteasome pathway, which cooperates with direct RA-induced ubiquitination. By establishing the mechanisms underlying RA and ATO sensitivity of NPM1c AMLs, our studies identify a striking parallelism with APL, paving the way to the crafting of curative targeted therapies in this category of AML patients.La leucémie aiguë myéloïde (LAM) est une maladie très hétérogène du point de vue génétique. Malgré les avancées dans la caractérisation moléculaire de la LAM, les taux de survie des patients restent assez modestes. Le pronostic de ces patients dépend en grande partie des anomalies cytogénétiques et moléculaires acquises. La nucléophosmine-1 (NPM1) est un gène essentiel codant pour une protéine nucléocytoplasmique, qui fait la navette entre le nucléole, le noyau et le cytoplasme mais demeure localisée principalement dans le nucléole. NPM1 joue plusieurs rôles dont la stabilisation de la protéine suppresseur de tumeur p14ARF, la régulation de la biogenèse des ribosomes, le contrôle de la duplication des centrosomes, la réponse aux stimuli de stress et l’activation de P53. NPM1 est considéré comme un des gènes les plus fréquemment mutés dans la LAM, représentant environ un tiers des patients (LAM avec NPM1c). Dans les protéines mutées NPM1c, deux résidus de tryptophane à l'extrémité C-terminale sont perdus et un signal d'exportation nucléaire est créé. Cela conduit à une accumulation ectopique et aberrante de la protéine NPM1c, ainsi que la protéine normale NPM1, dans le cytoplasme des blastes. Ceci joue un rôle majeur dans la leucémogénèse et l’établissement de la LAM. La protéine de la leucémie promyélocytaire (PML) organise des corps nucléaires (CN) qui contrôlent la protéolyse et la sénescence induite par la voie P53. Les CN PML sont désorganisés dans la LAM avec NPM1c. L'acide rétinoïque (AR) est une hormone favorisant la différenciation des cellules myéloïdes. L’addition de l’AR à la chimiothérapie augmente la survie des patients LAM avec mutation NPM1c. Nous avons précédemment démontré que l’AR et le trioxyde d’Arsenic (ATO) fonctionnent en synergie pour induire la dégradation de la protéine NPM1c, inhiber la croissance et induire l’apoptose des cellules leucémiques exprimant NPM1c. D’une manière importante, l’association AR/ATO a entrainé la baisse significative du nombre de blastes dans la moelle osseuse chez certains patients atteints de LAM avec NPM1c, et a conduit à la restauration de la localisation nucléaire de NPM1 et de PML. L’objectif principal du travail en cours était de dévoiler les voies biochimiques conduisant au catabolisme de NPM1c et d’élucider les mécanismes moléculaires de l’activité anti-leucémique de l’association AR/ATO. Nous démontrons que PML est nécessaire pour la dégradation de NPM1c induite par l’association AR/ATO. Dans les blastes primaires de patients LAM avec NPM1c, l’AR augmente rapidement le taux d'expression basal de PML, via l'inhibition de Pin-1, et avant la dégradation de la NPM1c. l’induction de PML par l’AR conduit à la stabilisation de P53 et prépare pour la reformation des CN par l'ATO, pour induire par la suite une hyper-activation de P53. En outre, l’association AR/ATO induit des réponses dépendantes de PML, associées à l'activation de P53 in vivo dans les souris xénogreffées avec des cellules LAM NPM1c. D’une manière très importante, l’association AR/ATO a induit une rémission complète, quoique transitoire, chez un patient LAM avec NPM1c démontrant l’efficacité clinique de cette association. Nous démontrons également que la dégradation de NPM1c induite par l'ATO se produit par le biais de la voie PML/ SUMO/RNF4/Protéasome, qui coopère avec une ubiquitination directe induite par l’AR. Nos études identifient un parallélisme flagrant avec la leucémie aiguë promyélocytaire, et ouvrent des horizons pour les applications cliniques de thérapies ciblées contre les LAM avec NPM1c. De plus, l'activation de Pin-1 et la perte de PML dans plusieurs types de tumeurs suggèrent qu’au-delà des LAM avec NPM1c, l'axe AR/ATO/Pin-1/PML/P53 pourrait être plus largement exploité au niveau thérapeutique, surtout dans les tumeurs malignes sensibles à l’association AR/ATO

Påverkan på EKG-registrering vid ändrat kroppsläge och elektrodplacering

Elektrokardiografi, EKG är den mest använda kardiovaskulära undersökningen för studering av hjärtats funktion, patologi i hjärtats retledningssystem och rytm. För att registrera och undersöka hjärtats elektriska aktivitet, används de standardiserade 12-avledningssystemet (12-SL) med distal placering av extremitetsavledningarna på underarmar och underben. På grund av muskelstörningar som kan förekomma under bland annat ett arbetsprov, är 12-SL inte alltid optimalt. Därför har andra modifierade system för elektrodplacering av extremitetselektroderna som Mason- Likar och Lund-avledningssystemet, ersatt med en omplacering proximalt på överarmar och höftkam. Tidigare studier har visat en förändring i EKG-registreringen som kan leda till falska patologier eller missvisande av det befintliga patologierna. Studier har även visat en förändring i EKG-registreringar som kan uppkomma vid olika kroppspositioner. Syftet med denna studie var därför att studera den potentiella skillnaden som uppstår mellan 12-SL och en proximal omplacering enligt Lund-systemet med en kroppspositionsändring. Studien inkluderade 31 deltagare remitterade till ett ordinärt arbetsprov, där en registrering skedde vid distal- och proximal koppling samt med patienten i sittande läge. Utifrån erhållna EKG-registreringar jämfördes R-vågsamplituden, QRS-durationen samt PQ tiden. Studien visade en statistisk signifikant skillnad i alla extremitetsavledningar vid en förflyttning av elektroderna för R-vågamplituden samt för avledning V5 och –aVL vid ändring av kroppsposition. Ingen signifikant förändring kunde observeras för PQ-tiden och QRS-durationen.Electrocardiography, ECG is the most used cardiovascular method to study cardiac function, pathology of the heart's retinal system and rhythm. To record and investigate the electrical activity of the heart, the standard 12-lead system (12-SL) with distal placement of the limb leads on the forearms and lower legs are commonly used. Due to muscular artifact that can occur in connection with exercise ECGs, 12-SL is not always optimal for use. Therefore, other systems such as Mason-Likar and the Lund-system have replaced with a proximal repositioning of the limb electrodes. Previous studies have shown a change in ECG registration that can lead to false pathology and alarms when modifying the standardized 12-lead system. Previous studies have also shown a possible change in ECG-registrations that can occur with a different body positions. The purpose of this research was to study the potential change that arises between 12-SL and a proximal relocation according to the Lund system with a difference in a body position. The study included 31 participants referred to the regular work test, where a registration was made at distal and proximal connection as well as with the patient in a sitting position. Based on the obtained ECG, the R- wave amplitudes, QRS- durations and PQ- intervals was measured and compared. Results showed a statistically significant difference in all limb leads at and displacements of the electrodes for the R-wave amplitude as well as for the V5 and -aVL with a different body position. No significant change was observers for the PQ- time and QRS- duration

Differences in Population Estimates Between an Administrative System and Census: The Case of Israel

The component method of population estimation relies on census counts, vital statistics, and internal and international migration. In Israel, all individuals' events are registered. To apply the component method, individuals' records from the census and the administrative data are aggregated to give population estimates. However, this method disconnects individual from aggregated characteristics, and differences appear in individual characteristics between census and administrative data. The Israel Central Bureau of Statistics conducted individual follow-ups. The pros and cons of each method are presented. On average, individual estimates on statistical areas are larger than aggregate estimates, and the differences are stable after several years. The individual estimates are preferable under at least four situations: in conducting a registry of emigrants; in conducting a census of institutions; where administrative files are available; and where census method is suitable to this method.statistical areas, population estimates, census, component method, population registry, administrative files,

The decline in consanguineous marriage among Muslims in Israel: The role of education

Background: There are two explanations for the inverse relationship between consanguinity and women's education. The female empowerment hypothesis posits that better-educated women will demand more freedom in choosing a marriage partner, whereas the role incompatibility hypothesis posits that school enrollment may prevent women from fulfilling spousal obligations. Objective: This article presents estimates of the relative contributions of school enrolment and educational attainment to the decline in consanguineous marriage. Methods: Our data comes from three rounds of the Palestinians in Israel Socio-Economic Survey. Using multinomial logistic regression analysis, we estimated discrete-time event history models to assess the effects of education on the probability of a consanguineous marriage. To test the two hypotheses we constructed two distinct education vectors for each woman from information on the number of years of schooling. The first charts yearly enrolment in education, whereas the second vector reflects actual attainment in each year. Results: Between 1975‒1979 and 2005‒2010, consanguineous marriage declined by almost 60Š. The rise in the age of leaving school explains about a third of the decline. Educational attainment did not contribute to the decline. Contribution: To the best of our knowledge, this is the first study to present estimates of the relative contributions of school enrolment and educational attainment to the decline in consanguineous marriage