Cognate microRNA-Gene Regulatory Networks Mediating Glioblastoma Oncogenic Properties

Malignant gliomas, including the most common and fatal form glioblastoma (GBM, WHO grade IV astrocytoma), remain a challenge to treat. In the United States and Europe, more than 30,000 patients per year are newly diagnosed with GBM. Despite ongoing trials, the best currently available multimodal treatment approaches include surgical resection followed by concomitant and adjuvant radiation (RT) and temozolomide (TMZ) therapy, resulting in a low median overall survival (OS) rate ranging from 12.2 - 15.9 months. The important role of genetic and epigenetic changes in DNA, RNA, and protein alteration as well as epigenetic changes secondary to the tumor microenvironment and outside selection pressure (therapeutic interventions), are increasingly being recognized. In GBM treatment, the focus is shifting toward a more patient-centered (personalized) therapy. In this regard, in particular, microRNAs are being increasingly studied. MicroRNAs are non¬protein coding small RNAs that serve as negative gene regulators by binding to a specific sequence in the promoter region of a target gene, thus regulating gene expression. A single microRNA potentially targets hundreds of genes; thus, microRNAs and their cognate target genes have important roles as tumor suppressors and oncogenes as well as regulators of various cancer- specific cellular features, such as proliferation, apoptosis, invasion, and metastasis. The identification of distinct microRNA-gene regulatory networks in GBM patients can be expected to provide novel therapeutic insights by identifying candidate patients for targeted therapies. To this end, in this work we identified and validated clinically relevant and meaningful novel gene- microRNA regulatory networks that correlated with MR tumor phenotypes, histopathology, and patient survival and response rates to therapy. - Le traitement des gliomes malins, y compris sous leur forme la plus commune et meurtrière, le glioblastome (GBM, ou astrocytome de grade IV selon l'OMS), demeure à ce jour un défi. Aux États-Unis et en Europe, un nouveau diagnostic de GBM est prononcé dans plus de 30Ό00 cas par an. En dépit de tests en cours, les meilleures approches thérapeutiques combinées actuellement disponibles comprennent la résection chirurgicale de la tumeur, suivie d'une radiothérapie adjuvante ainsi que d'un traitement au temozolomide (RT/TMZ), thérapies dont résulte une médiane de survie globale basse (overall survival, OS), comprise entre 12.2 et 15.9 mois. On reconnaît de plus en plus le rôle majeur de l'ADN, de l'ARN et de l'altération des protéines ainsi que des modifications épigénétiques, secondaires par rapport au microenvironnement de la tumeur et à la pression de sélection extérieure (les interventions thérapeutiques). Dans le traitement du GBM, le centre d'intérêt se déplace vers une thérapie centrée sur le cas individuel du patient. Dans ce but, en particulier les microARN sont de plus en plus analysés. Les microARN sont de petits ARN non-codants (les protéines) qui servent de régulateurs négatifs de gènes en s'attachant à une séquence spécifique dans la région promotrice d'un gène-cible, régulant ainsi l'expression du gène. Un seul microARN cible potentiellement des centaines de gènes; on a ainsi découvert que les microARN et leurs gènes-cibles apparentés ont une fonction importante en tant que suppresseurs de tumeurs et d'oncogènes, ainsi que comme régulateurs de diverses caractéristiques cellulaires spécifiques du cancer, comme la prolifération, l'apoptose, l'invasion et la métastase. On peut s'attendre à ce que l'identification de réseaux microARN régulateurs de gènes, distincts selon les patients de GBM, fournisse une approche thérapeutique inédite par la détermination des patients susceptibles de réagir favorablement à des thérapies ciblées.

Wound-healing gene family expression differences between fetal and foreskin cells used for bioengineered skin substitutes.

For tissue engineering, several cell types and tissues have been proposed as starting material. Allogenic skin products available for therapeutic usage are mostly developed with cell culture and with foreskin tissue of young individuals. Fetal skin cells offer a valuable solution for effective and safe tissue engineering for wounds due to their rapid growth and simple cell culture. By selecting families of genes that have been reported to be implicated in wound repair and particularly for scarless fetal wound healing including transforming growth factor-beta (TGF-beta) superfamily, extracellular matrix, and nerve/angiogenesis growth factors, we have analyzed differences in their expression between fetal skin and foreskin cells, and the same passages. Of the five TGF-beta superfamily genes analyzed by real-time reverse transcription-polymerase chain reaction, three were found to be significantly different with sixfold up-regulated for TGF-beta2, and 3.8-fold for BMP-6 in fetal cells, whereas GDF-10 was 11.8-fold down-regulated. For nerve growth factors, midkine was 36-fold down-regulated in fetal cells, and pleiotrophin was 4.76-fold up-regulated. We propose that fetal cells present technical and therapeutic advantages compared to foreskin cells for effective cell-based therapy for wound management, and overall differences in gene expression could contribute to the degree of efficiency seen in clinical use with these cells.

Widespread age-related differences in the human brain microstructure revealed by quantitative magnetic resonance imaging.

A pressing need exists to disentangle age-related changes from pathologic neurodegeneration. This study aims to characterize the spatial pattern and age-related differences of biologically relevant measures in vivo over the course of normal aging. Quantitative multiparameter maps that provide neuroimaging biomarkers for myelination and iron levels, parameters sensitive to aging, were acquired from 138 healthy volunteers (age range: 19-75 years). Whole-brain voxel-wise analysis revealed a global pattern of age-related degeneration. Significant demyelination occurred principally in the white matter. The observed age-related differences in myelination were anatomically specific. In line with invasive histologic reports, higher age-related differences were seen in the genu of the corpus callosum than the splenium. Iron levels were significantly increased in the basal ganglia, red nucleus, and extensive cortical regions but decreased along the superior occipitofrontal fascicle and optic radiation. This whole-brain pattern of age-associated microstructural differences in the asymptomatic population provides insight into the neurobiology of aging. The results help build a quantitative baseline from which to examine and draw a dividing line between healthy aging and pathologic neurodegeneration.

Flight energetics in relation to sexual differences in the mating behaviour of a mayfly, Siphlonurus aestivalis

Mayflies (Ephemeroptera) are known to have short adult life-spans. Adults are unable to feed, and they utilize reserves stored during their aquatic larval stage. Energy reserves (fat, glycogen, and free sugars) of mature larvae, subimagoes and imagoes of both sexes of Siphlonurus aestivalis Eaton were compared. All the stages of both sexes had low glycogen and free sugar contents, and the only significant change occurred during the transformation of the mature larva to subimago when almost all the reserves of free sugars were used up. Glycogen and free sugars may serve as energy sources permitting individuals to swim and fly out of the water during emergence. Fat made up most of the energy reserves of mature larvae and was the main source of energy used during the final development of both sexes. Young adult males had high fat reserves which they used as a source of energy for their swarming flights. In contrast, females did not seem to use a significant amount of fat for flight. This difference is probably related to the different mating strategies of the sexes in this species. Males perform long flights waiting for females, whereas females perform only brief flights to mate and reproduce.

Valproate treatment of human cord blood CD4-positive effector T cells confers on them the molecular profile (microRNA signature and FOXP3 expression) of natural regulatory CD4-positive cells through inhibition of histone deacetylase.

Regulatory T cells (Tregs) play a key role in immune system homeostasis and tolerance to antigens, thereby preventing autoimmunity, and may be partly responsible for the lack of an appropriate immune response against tumor cells. Although not sufficient, a high expression of forkhead box P3 (FOXP3) is necessary for their suppressive function. Recent reports have shown that histones deacetylase inhibitors increased FOXP3 expression in T cells. We therefore decided to investigate in non-Tregs CD4-positive cells, the mechanisms by which an aspecific opening of the chromatin could lead to an increased FOXP3 expression. We focused on binding of potentially activating transcription factors to the promoter region of FOXP3 and on modifications in the five miRs constituting the Tregs signature. Valproate treatment induced binding of Ets-1 and Ets-2 to the FOXP3 promoter and acted positively on its expression, by increasing the acetylation of histone H4 lysines. Valproate treatment also induced the acquisition of the miRs Tregs signature. To elucidate whether the changes in the miRs expression could be due to the increased FOXP3 expression, we transduced these non-Tregs with a FOXP3 lentiviral expression vector, and found no changes in miRs expression. Therefore, the modification in their miRs expression profile is not due to an increased expression of FOXP3 but directly results from histones deacetylase inhibition. Rather, the increased FOXP3 expression results from the additive effects of Ets factors binding and the change in expression level of miR-21 and miR-31. We conclude that valproate treatment of human non-Tregs confers on them a molecular profile similar to that of their regulatory counterpart.

Degree of previous cutting in explaining the differences in diameter distributions between mature managed and natural Norway spruce forests

Abstract

Pediatric and adult eosinophilic esophagitis: similarities and differences.

Early in the 1990s, several case series described adults suffering from dysphagia and children with refractory reflux symptoms, both accompanied by an eosinophil-predominant infiltration, thereby conclusively distinguishing it from gastroesophageal reflux disease. Eosinophilic esophagitis (EoE) was recognized as its own entity in the adult and in the pediatric literature. In the last decade, evidence has accumulated that EoE represents a T-helper (Th)2-type inflammatory disease. Remodeling of the esophagus is a hallmark of EoE, leading to esophageal dysfunction and bolus impaction. Familial occurrence and disease association with single-nucleotide polymorphisms underscore the influence of genetics in this disease. Eosinophilic esophagitis may affect individuals at any age, although the clinical presentation is highly age dependent. There is a significant allergic bias in the EoE population, with the majority of patients having concurrent allergic rhinitis, asthma, eczema, and/or a history of atopy. One noteworthy difference is that in children, EoE seems to be primarily a food antigen-driven disease, whereas in adults, mainly aeroallergen sensitization has been observed. Treatment modalities for EoE include the 3Ds: drugs, diet, and dilation. The crucial question of whether adult and pediatric EoE are different phenotypes of one single entity or whether we are confronted with two different diseases is still open. Here, we review similarities and differences between EoE in adults and children.

Intergenerational differences in cardiovascular risk factor levels in Switzerland

Background: There is little information regarding cardiovascular risk factor (CV RF) trends in Switzerland. We aimed at assessing generation differences in CV RFs by comparing CV RFs levels within selected age groups separated by a 20 year time lag. Design: Two population-based surveys. Methods: Data from the Monica (1984-1986) and Colaus (2004-2006) surveys were used. Analyses were stratified by sex and age groups (35-44, 45-54, 55-64 and 65-75 years). Results: No changes were found for BMI levels and status between surveys: in men, 26}3, 26}3, 27}4 and 27}4 kg/m2 for age groups 35-44, 45-54, 55-64 and 65-74, respectively, in MONICA, vs. 26}4, 26}4, 27}4 and 28}4 kg/m2 in COLAUS, p=NS, in women: 24}4, 26}4, 26}4 and 26}5 kg/m2 in MONICA, vs. 24}5, 25}5, 26}5 and 26}5 kg/m2 in COLAUS, p=NS. Similar results were found after adjusting for education. Smoking prevalence increased in men: 28, 30, 22 and 15% for age groups 35-44, 45-54, 55-64 and 65-74, respectively, in MONICA, vs. 35, 29, 28 and 21% in COLAUS. In women, changes differed according to age: 39, 26, 16 and 18%, in MONICA vs. 28, 30, 22 and 15% in COLAUS. Blood pressure decreased in the younger age groups and remained constant in the older ones: in men, systolic blood pressure was 129}15, 133}16, 138}18 and 143}21 mm Hg in MONICA, vs. 125}12, 129}15, 137}16 and 144}19 mm Hg in COLAUS, p<0.01. Similar findings were obtained after adjusting for education. Prevalence of hypertension increased, due to an increase in the prevalence of treated subjects, in men : 4, 8, 16 and 19% for age groups 35-44, 45-54, 55-64 and 65-74, respectively, in MONICA, vs. 5, 14, 31 and 46% in COLAUS, p<0.05; in women: 2, 10, 16, and 24% in MONICA, vs. 4, 12, 24, and 34% in COLAUS, p<0.05. This increase was stronger in men: 14, 17, 23 and 31% for age groups 35-44, 45-54, 55-64 and 65-74, respectively, in MONICA vs. 10, 21, 41 and 55% in COLAUS, p<0.01 and smaller in women: 6, 15, 24 and 44% in MONICA vs. 6, 16, 30 and 42% in COLAUS, p=NS. Similar findings were obtained after adjusting for education. Conclusion: With the exception of BMI, the newer Swiss generations appear to have a worse CV profile than the older generations. This is especially true regarding smoking and hypertension.

Differences in therapeutic alliance when working with an interpreter: a preliminary study

This preliminary study aims to analyze the therapeutic alliance in a crosscultural triadic setting, where there is a therapist and a client who speak a different language but are able to interact thanks to an interpreter/cultural mediator. The participants' (therapists, clients, interpreters) representations associated with the notion of therapeutic alliance, and the level of alliance between each group was obtained and compared. Clients (N = 9) were all from Albanese origin. The results show that the three groups of participants give particular meanings to the alliance and tend to evaluate their alliance level differently. The interpreter's mediating role in the construction of the therapeutic alliance is discussed.

Specialized home palliative care for adults and children: differences and similarities.

OBJECTIVE: To compare the provision of specialized home palliative care (SHPC) by the adult and pediatric SHPC teams at the Munich University Hospital. METHODS: All patients treated by one of the SHPC teams and their primary caregivers were eligible for the prospective nonrandomized survey. We analyzed the demographics, the underlying diseases, duration and impact of SHPC on symptom control and quality of life (QOL) as well as the caregivers' burden and QOL. RESULTS: Between April 2011 and June 2012, 100 adult and 43 pediatric patients were treated consecutively; 60 adults (median age, 67.5 years; 55% male) and 40 children (median age, 6 years, 57% male) were included in the study. Oncologic diseases were dominant only in the adult cohort (87 versus 25%, p<0.001). The median period of care was higher in the pediatric sample (11.8 versus 4.3 weeks; NS). Ninety-five percent of adult and 45% of pediatric patients died by the end of the study (p<0.001), 75% and 90% of them at home, respectively. The numbers of significant others directly affected by the patient's disease was higher in children (mean 3.4 versus 1.2; p<0.001). The QOL of adult patients and children (p<0.05 for both), as well as of their primary caregivers (p<0.001 for both) improved during SHPC, while the caregivers' burden was lowered (p<0.001 for both). CONCLUSIONS: Our results show important differences in several clinically relevant parameters between adults and children receiving SHPC. This should assist in the development of age-group specific SHPC concepts that effectively address the specific needs of each patient population.

Evaluation of Racial Differences in Seat Belt and Child Restraint Use: A Review of Current Literature, 2004

This report presents the results of a literature review conducted to evaluate differences in seat belt use by race. A literature review was conducted to evaluate overall seat belt use, racial differences in seat belt use, overall child restraint use, racial differences in child restraint use, and information about seat belt and child restraint use specific to Iowa. A number of national studies and regional studies were found and are presented. Mixed results were found as to whether racial differences exist in both seat belt use and child restraint use. However, in the course of the literature review, several items that are of interest to safety in Iowa have emerged, although little data specific to Iowa was encountered. First, national seat belt use appears to be lower among African-Americans than for Caucasians or Hispanics. Second, national crash rates among Hispanics appear to be higher than those for Caucasians, particularly when population and lower vehicle miles traveled (VMT) are considered. One issue that should be considered throughout this literature review is that the Hispanic population may be higher than reported due to large numbers of undocumented persons who do not appear in population estimates, driver’s license, or other databases.

Cross-country differences in disclosure quality: a study of fair value disclosures by european real estate companies

Companies are under IAS 40 required to report fair values of investment properties on the balance sheet or to disclose them in the notes. The standard requires also that companies have to disclose the methods and significant assumptions applied in determining fair values of investment properties. However, IAS 40 does not include any illustrative examples or other guidance on how to apply the disclosure requirements. We use a sample with publicly traded companies from the real estate sector in the EU. We find that a majority of the companies use income based methods for the measurement of fair values but there are considerable cross-country variations in the level of disclosures about the assumptions used in determining fair values. More specifically, we find that Scandinavian and German origin companies disclose more than French and English origin companies. We also test whether disclosure quality is associated with enforcement quality measured with the “Rule of Law” index according to Kaufmann et al. (2010), and associated with a secrecy- versus transparency-measure based on Gray (1988). We find a positive association between disclosure and earnings quality and a negative association with secrecy.

Streptozotocin induces a shift towards T regulatory cell responses in vivo

Objectives: Streptozotocin (STZ) induced diabetes is currently the most commonly used animalmodel for islet transplantation.However, STZtreatment and the ensuing hyperglycemia were both shown to affect the immune response, including an apparent induction of lymphopenia. The aim of this study was to evaluate the respective effect of STZ and hyperglycemia on the immune system in STZ induced diabetic C57BL/6 mice. Methods: Phenotypes and levels of T and B cells were analyzed by flow cytometry in blood and spleen over time. The effect of hyperglycemia was further characterized by insulin replacement, islet transplantation and by using Rip (rat insulin promoter) DTR (dipheteria tocin receptor) transgenic mice. Results: STZ but not hyperglycemia was toxic for splenocytes in vitro, whereas hyperglycemia correlated with diabetes associated blood and spleen lymphopenia in vivo. Moreover, independently of hyperglycemia, STZ lead to a relative increase of T regulatory cells which retained their suppressive capacity in vitro. Conclusion: These data suggest thatSTZand the ensuing acute hyperglycemia have major direct and indirect effects on immune homeostasis. Thus, high caution needs to be exercised in the interpretation of the results of tolerance induction and/or immunosuppressive protocols in STZ-induced diabetes and islet transplantation models.

Oligogalacturonide defense signals in plants: large fragments interact with the plasma membrane in vitro.

Oligogalacturonides are plant cell wall-derived regulatory molecules which stimulate defense gene expression during pathogenesis. In vitro, these compounds enhance the phosphorylation of an approximately 34-kDa protein (pp34) in purified plasma membranes from potato and tomato leaves. We now show that polygalacturonate-enhanced phosphorylation of pp34 occurs in plasma membranes purified from tomato roots, hypocotyls, and stems and from undifferentiated potato cells. Furthermore, a similar phosphorylation is detected in leaf plasma membranes from soybean, a plant distantly related to tomato. Purified oligogalacturonides 13 to at least 26 residues long stimulate pp34 thiophosphorylation in vitro. This stimulation pattern differs from the induction of many known defense responses in vivo, where a narrower range of smaller fragments, between approximately 10 and 15 residues long, are active. On the basis of these differences we suggest that observed effects of applied exogenous oligogalacturonides on defense responses may not necessarily reflect the situation during pathogenesis. The cell wall could act as a barrier to many exogenous oligo- and polygalacturonides as well as other large regulatory ligands.