bluuebunny/crossref_metadata_2025 huggingface dataset:
Transient Enhanced Expression of Cdk5 Activator p25 after Acute and Chronicd‐Amphetamine Administration
Authors: Mlewski Estela Cecilia , Krapacher Favio Ariel , Ferreras Soledad , Paglini Gabriela
Published: August 2009

The cellular and molecular mechanisms of sensitization in the addictive process are still unclear. Recently, chronic treatment with cocaine has been shown to upregulate the expression of cyclin‐dependent kinase 5 (cdk5) and its specific activator, p35, in the striatum, as a downstream target gene of ΔFosB, and has been implicated in compensatory adaptive changes associated with psychostimulants. Cdk5 is a serine/threonine kinase and its activation is achieved through association with a regulatory subunit, either p35 or p39. P35 is cleaved by the protease calpain, which results in the generation of a truncated product termed p25, which contains all elements necessary for cdk5 activation. The cdk5/p35 complex plays an essential role in neuronal development and survival. It has also been involved in neuronal trafficking and transport and in dopaminergic transmission, indicating its role either in presynaptic and postsynaptic signaling. In this study we report that the cdk5/p35 complex participates in acute and chronicd‐amphetamine (AMPH)‐evoked behavioral events, and we show a surprisingly transient enhanced expression of p25 and a lasting increased expression of p35 in dorsal striatal synaptosomes after acute and chronic AMPH administration. Pak1, a substrate for cdk5, is also enriched in the synaptosomal fraction of acute AMPH‐treated rats. Our data suggest that the transient upregulation of p25 may regulate the activity of cdk5 in phosphorylating particular substrates, such as Pak1, implicated in the compensatory adaptive morphophysiologic changes associated with the process of behavioral sensitization to psychostimulants.
Influence of Chronic Exercise on the Amphetamine‐Induced Dopamine Release and Neurodegeneration in the Striatum of the Rat
Authors: Marques Elsa , Vasconcelos Filipa , Rolo Marta R. , Pereira Frederico C. , Silva Ana P. , Macedo Tice R. , Ribeiro Carlos F.
Published: August 2009

The aim of this study was to verify the effect of chronic exercise on the striatal dopamine (DA) outflow induced by low and high single doses of amphetamine (AMPH), and verify the existence of an exercise protective role on neurodegeneration. Adult male Sprague‐Dawley rats were randomly separated into six groups: chronic exercise, saline; chronic exercise, 5 mg kg−1AMPH; chronic exercise, 30 mg kg−1AMPH; without exercise, saline; without exercise, 5 mg kg−1AMPH; without exercise, 30 mg kg−1AMPH. Chronic exercise consisted of an 8‐week running program on a treadmill, with increasing intensity. Animals were anesthetized, placed into a stereotaxic frame and an intracerebral guide cannula implanted into the caudate–putamen. When indicated, microdialysis was performed. Dialysate samples were collected during 30‐min intervals for 6 h, before and after the intraperitonial administration of AMPH or saline solution. HPLC with electrochemical detection was used to quantify DA. Chronic exercise did not significantly change the extracellular DA basal values. Regarding the maximal DA levels in the dialysates, in the rats treated with 5 mg kg−1AMPH, there was no significant difference between groups with and without chronic exercise; on the contrary, in animals treated with 30 mg kg−1AMPH, the DA release was lower in the group with chronic exercise. Moreover, the maintenance of higher levels of DA along time in the training group suggests a diminished reuptake of DA. By using the Fluoro‐Jade C staining technique, we did not find neuronal death in any of the groups. In conclusion, these results suggest that chronic exercise leads to a diminished release and reuptake of DA after administration of a high dose of AMPH, whereas neither chronic exercise nor AMPH seems to induce neurodegeneration.
An Endocannabinoid Hypothesis of Drug Reward and Drug Addiction
Authors: Onaivi Emmanuel S.
Published: August 2009

Pharmacologic treatment of drug and alcohol dependency has largely been disappointing, and new therapeutic targets and hypotheses are needed. There is accumulating evidence indicating a central role for the previously unknown but ubiquitous endocannabinoid physiological control system (EPCS) in the regulation of the rewarding effects of abused substances. Thus an endocannabinoid hypothesis of drug reward is postulated. Endocannabinoids mediate retrograde signaling in neuronal tissues and are involved in the regulation of synaptic transmission to suppress neurotransmitter release by the presynaptic cannabinoid receptors (CB‐Rs). This powerful modulatory action on synaptic transmission has significant functional implications and interactions with the effects of abused substances. Our data, along with those from other investigators, provide strong new evidence for a role for EPCS modulation in the effects of drugs of abuse, and specifically for involvement of cannabinoid receptors in the neural basis of addiction. Cannabinoids and endocannabinoids appear to be involved in adding to the rewarding effects of addictive substances, including, nicotine, opiates, alcohol, cocaine, and BDZs. The results suggest that the EPCS may be an important natural regulatory mechanism for drug reward and a target for the treatment of addictive disorders.
Nonhuman Primate Neuroimaging and the Neurobiology of Psychostimulant Addiction
Authors: Howell Leonard L. , Murnane Kevin S.
Published: August 2009

Neuroimaging techniques have led to significant advances in our understanding of the neurobiology and treatment of drug addiction in humans. The capability to conduct parallel studies in nonhuman primates and human subjects provides a powerful translational approach to link findings in human and animal research. A significant advantage of nonhuman primate models is the ability to use drug‐naïve subjects in longitudinal designs that document the neurobiological changes that are associated with chronic drug use. Moreover, experimental therapeutics can be evaluated in subjects with well‐documented histories of drug exposure. Thein vivodistribution and pharmacokinetics of drug binding in brain have been related to the time‐course of behavioral effects associated with the addictive properties of stimulants. Importantly, the characterization of drug interactions with specific protein targets in brain has identified potential targets for medication development. Neuroimaging has proven especially useful in studying the dynamic changes in neuronal function that may be associated with environmental variables. Last, neuroimaging has been used effectively in nonhuman primates to characterize both transient and long‐lasting changes in brain chemistry associated with chronic drug exposure. Although there is some evidence to suggest neurotoxicity in humans with long histories of stimulant use, parallel studies in nonhuman primates have not identified consistent long‐term changes in such neurochemical markers. Collectively, the results of these studies of nonhuman primates have enhanced our understanding of the neurobiological basis of stimulant addiction and should have a significant impact on efforts to develop medications to treat stimulant abuse.
An Update on Substance Use and Treatment following Traumatic Brain Injury
Authors: Graham David P. , Cardon Aaron L.
Published: August 2009

Traumatic brain injury (TBI) is a leading cause of mortality and morbidity among young adults. Substance abusers constitute a disproportionate percentage of these patients. A history of substance abuse predicts increased disability, poorer prognosis, and delayed recovery. While consensus in the literature indicates that substance‐abuse rates decline following injury, conflicting literature shows a significant history of brain injury in addicts. We reviewed the literature on substance abuse after TBI to explore the state of knowledge on TBI as a risk factor for substance abuse. While recent reviews regarding substance abuse in TBI patients concur that substance‐abuse rates decline even after mild TBI, an emerging literature suggests mild TBI may cause subtle impairments in cognitive, executive, and decision‐making functions that are often poorly recognized in early diagnosis and treatment. When combined with difficulties in psychosocial adjustment and coping skills, these impairments may increase the risk for chronic substance abuse in a subset of TBI patients. Preliminary results from veterans indicate these patterns hold in a combat‐related post‐traumatic stress disorder population with TBI. This increasingly prevalent combination presents a specific challenge in rehabilitation. While this comorbidity presents a challenge for the successful treatment and rehabilitation of both disorders, there is sparse evidence to recommend any specific treatment strategy for these individuals. Mild TBI and substance abuse are bidirectionally related both for risks and treatment. Further understanding the neuropsychiatric pathology and different effects of different types of injuries will likely improve the implementation of effective treatments for each of these two conditions.
MicroRNAs and Their Emerging Roles in Immunology
Authors: Pauley Kaleb M. , Chan Edward K.L.
Published: August 2009

MicroRNAs (miRNAs) are evolutionarily conserved small noncoding RNAs that post‐transcriptionally regulate gene expression by targeting specific messenger RNAs (mRNAs) for degradation or translational repression. Recent evidence indicates that miRNA‐mediated gene regulation is critical for normal cellular functions, and as much as one‐third of human mRNAs may be miRNA targets. Emerging evidence suggests that miRNAs play a key role in the regulation of immunological functions including innate and adaptive immune responses, development and differentiation of immune cells, and the prevention of autoimmunity. Here, we review the mechanisms of miRNA maturation and function, the roles of several miRNAs in immunological functions, and the involvement of miRNAs in disease pathogenesis.
Retinoic Acid in the Immune System
Authors: Pino‐Lagos Karina , Benson Micah J. , Noelle Randolph J.
Published: August 2009

On occasion, emerging scientific fields intersect and great discoveries result. In the last decade, the discovery of regulatory T cells (Treg) in immunity has revolutionized our understanding of how the immune system is controlled. Intersecting the rapidly emerging field of Treg function, has been the discovery that retinoic acid (RA) controls both the homing and differentiation of Treg. Instantly, the wealth and breadth of knowledge of the molecular basis for RA action, its receptors, and how it controls cellular differentiation can and will be exploited to understand its profound effects on Treg. Historically, vitamin A deprivation and repletion and RA agonists have been shown to profoundly affect immunity. Now these findings can be interpreted in light of the revelations that RA controls leukocyte homing and Treg function.
Mast Cells
Authors: Rao Kavitha N. , Brown Melissa A.
Published: August 2009

Mast cells were discovered more than 100 years ago and until recently, have been considered renegades of the host with the sole purpose of perpetuating allergy. The discovery of mast cell–deficient mice that could be reconstituted with mast cells (the so called “mast cell knock‐in” mice) has allowed the study of the in vivo functions of mast cells and revealed several new facets of these cells. It is now evident that mast cells have a much broader impact on many physiological and pathologic processes. Mast cells, particularly through their dynamic interaction with the nervous system, have been implicated in wound healing, tissue remodeling, and homeostasis. Perhaps the most progress has been made in our understanding of the role of mast cells in immunity outside the realm of allergy, and host defense. Mast cells play critical roles in both innate and adaptive immunity, including immune tolerance. Greater insight into mast cell biology has prompted studies probing the additional consequences of mast cell dysfunction, which reveal a central role for mast cells in the pathogenesis of autoimmune disorders, cardiovascular disorders, and cancer. Here, we review recent developments in the study of mast cells, which present a complex picture of mast cell functions.
Neurological Aspects of Obstructive Sleep Apnea
Authors: Broderick Meredith , Guilleminault Christian
Published: August 2009

Obstructive sleep apnea is often regarded as a structural disorder causing narrowing of the airway. This article reviews the neurological aspects of obstructive sleep apnea, including the upper airway reflex, cortical arousal thresholds, and motor function as they pertain to the pathophysiology of disease. We also discuss the relationship of obstructive sleep apnea to other neurological diseases.
Mitochondrial Encephalopathy, Lactic Acidosis, and Strokelike Episodes
Authors: Sproule Douglas M. , Kaufmann Petra
Published: August 2009

Since the initial description almost 25 years ago, the syndrome of mitochondrial encephalopathy, lactic acidosis, and strokelike episodes (MELAS) has been a useful model to study the complex interplay of factors that define mitochondrial disease. This syndrome, most commonly caused by an A‐to‐G transition mutation at position 3243 of the mitochondrial genome, is typified by characteristic neurological manifestations including seizures, encephalopathy, and strokelike episodes, as well as other frequent secondary manifestations including short stature, cognitive impairment, migraines, depression, cardiomyopathy, cardiac conduction defects, and diabetes mellitus. In this review, we discuss the history, pathogenesis, clinical features, and diagnostic and management strategies of mitochondrial disease in general and of MELAS in particular. We explore features of mitochondrial genetics, including the concepts of heteroplasmy, mitotic segregation, and threshold effect, as a basis for understanding the variability and complicated inheritance patterns seen with this group of diseases. We also describe systemic manifestations of MELAS‐associated mutations, including cardiac, renal, endocrine, gastrointestinal, and endothelial abnormalities and pathology, as well as the hypothetical role of derangements to COX enzymatic function in driving the unique pathology and clinical manifestations of MELAS. Although therapeutic options for MELAS and other mitochondrial diseases remain limited, and recent trials have been disappointing, we also consider current and potential therapeutic modalities.
Stroke Treatment
Authors: Cronin C.A. , Weisman C.J. , Llinas R.H.
Published: August 2009

For acute stroke patients who arrive at the hospital within 3 h of symptom onset, the focus of care involves screening for eligibility to receive intravenous tissue plasminogen activator. The publication of the National Institute of Neurological Disorders and Stroke recombinant tissue–type plasminogen activator (tPA, or alteplase) study in 1995 (Marler, J.R. 1995, New England Journal of Medicine333: 1581–1587) spurred protocol changes, which continue to evolve, throughout the health care system in an effort to streamline the patient through the Emergency Medical System. The need to expedite patient evaluation involving emergency department, laboratory, radiology, and clinical neurology testing is clear and has been a focus of many stroke centers. For some patients, intravenous thrombolysis within 3 h has a dramatic effect on outcome. However, that is not the only course of action for acute stroke patients. This article will review some of the effective treatments for stroke patients beyond the first 3 h of their care.
Cervical Arterial Dissection
Authors: Menon Ranjith K. , Norris John W.
Published: August 2009

The increasing use and safety of noninvasive imaging in recent years has revealed the surprising frequency of dissection of the carotid and vertebral arteries (cervical arterial dissection [CAD]) as a cause of ischemic and hemorrhagic stroke. This review is an overview of current concepts and practice of patients with CAD, but our ideas are constantly evolving with new discoveries from neurovascular imaging and medical and surgical management in this area.
Impacts of Climate Change and Variability on European Agriculture
Authors: Orlandini Simone , Nejedlik Pavol , Eitzinger Josef , Alexandrov Vesselin , Toulios Leonidas , Calanca Pierluigi , Trnka Miroslav , Olesen Jørgen E.
Published: August 2009

Climate plays a fundamental role in agriculture because of to its influence on production. All processes are regulated by specific climatic requirements. Furthermore, European agriculture, based on highly developed farming techniques, is mainly oriented to high quality food production that is more susceptible to meteorological hazards. These hazards can modify environment–genotype interactions, which can affect the quality of production. The COST 734 Action (Impacts of Climate Change and Variability on European Agriculture), launched in 2006, is composed of 28 signature countries and is funded by the European Commission. The main objective of the Action is the evaluation of possible impacts arising from climate change and variability on agriculture and the assessment of critical thresholds for various European areas. The Action will concentrate on four different tasks: agroclimatic indices and simulation models, including review and assessment of tools used to relate climate and agricultural processes; evaluation of the current trends of agroclimatic indices and model outputs, including remote sensing; developing and assessing future regional and local scenarios of agroclimatic conditions; and risk assessment and foreseen impacts on agriculture. The work will be carried out by respective Working Groups. This paper presents the results of the analysis of the first phase of inventory activity. Specific questionnaires were disseminated among COST 734 countries to collect information on climate change analysis, studies, and impact at the European level. The results were discussed with respect to their spatial distribution in Europe and to identify possible common long‐ and short‐term strategies for adaptation.
Humanized Mice for the Study of Type 1 Diabetes and Beta Cell Function
Authors: King Marie , Pearson Todd , Rossini Aldo A. , Shultz Leonard D. , Greiner Dale L.
Published: August 2009

Our understanding of the basic biology of diabetes has been guided by observations made using animal models, particularly rodents. However, humans are not mice, and outcomes predicted by murine studies are not always representative of actual outcomes in the clinic. In particular, investigators studying diabetes have relied heavily on mouse and rat models of autoimmune type 1‐like diabetes, and experimental results using these models have not been representative of many of the clinical trials in type 1 diabetes. In this article, we describe the availability of new models of humanized mice for the study of three areas of diabetes. These include the use of humanized mice for the study of (1) human islet stem and progenitor cells, (2) human islet allograft rejection, and (3) human immunity and autoimmunity. These humanized mouse models provide an important preclinical bridge between in vitro studies and rodent models and the translation of discoveries in these model systems to the clinic.
Diabetogenic Effects of the Most Prevalent Enteroviruses in Finnish Sewage
Authors: Klemola Paivi , Kaijalainen Svetlana , Ylipaasto Petri , Roivainen Merja
Published: August 2009

Common enterovirus infections appear to initiate or facilitate the pathogenetic processes leading to type 1 diabetes (T1D) and also sometimes precipitate the clinical disease. We have recently demonstrated that (1) enterovirus‐positive islet cells were seen on postmortem pancreatic specimens of several T1D patients but not in the corresponding samples of nondiabetic controls, and (2) several different enteroviruses can be associated with T1D. Enterovirus infections are transmitted from person to person by fecal–oral or respiratory routes, which means that infections usually start from the respiratory or gastrointestinal mucosa. Regardless of the clinical symptoms of the disease, viral replication continues in the submucosal lymphatic tissue for several weeks, up to a couple of months, and during that time the virus is excreted into the feces and translocated to the environment. Monitoring of sewage samples for enteroviruses can be used as a tool in epidemiologic studies of enterovirus. Finland has successfully used environmental control data in poliovirus surveillance for decades. About 24 samples have been collected annually from the Helsinki region, which covers about 20% of the population. In the present study, we have reanalyzed the sewage samples of the years 1993–2004 for nonpolio enteroviruses by inoculating them into five different continuous cell lines known to cover a wide range of serotypes. Isolated strains were identified by RT‐PCR and VP1 sequencing. The most commonly detected serotypes were coxsackie B viruses (CBV1–5) and echoviruses (E6, 7, 11, 25, 30). Diabetogenic effects of the most prevalent enterovirus serotypes were studied in primary human β cells.
SlC30A8 Is a Major Target of Humoral Autoimmunity in Type 1 Diabetes and a Predictive Marker in Prediabetes
Authors: Wenzlau Janet M. , Moua Ong , Sarkar Suparna A. , Yu Liping , Rewers Marian , Eisenbarth George S. , Davidson Howard W. , Hutton John C.
Published: August 2009

Type 1A diabetes (T1D) results from autoimmunity targeted at a limited number of molecules that are expressed in the pancreatic β cell. Putative novel autoantigen candidates were identified from microarray expression profiling of human and rodent islet cells. The highest ranking candidate was Slc30A8 (zinc transporter 8; ZnT8), which was screened by radioimmunoprecipitation assays against new‐onset T1D and prediabetic sera. Such assays detected 63% of subjects with new‐onset diabetes, but fewer than 2% of controls, 3% of those with type 2 diabetes, and 10% of patients with other autoimmune disorders. ZnT8 autoantibodies were found, however, in 26% of T1D subjects previously classified as autoantibody‐negative on the basis of existing markers (GADA, IA2 A, IAA, and ICA). We conclude that SLC30A8 provides an important additional and independent predictive marker for T1D.
Different KIRs Confer Susceptibility and Protection to Adults with Latent Autoimmune Diabetes in Latvian and Asian Indian Populations
Authors: Shastry Arun , Sedimbi Saikiran K. , Rajalingam Raja , Rumba Ingrida , Kanungo Alok , Sanjeevi C.B.
Published: August 2009

KIRs (killer Ig‐like receptors) expressed on natural killer (NK) cells are an important component of innate (and adaptive) immunity. They are either activatory or inhibitory, and certain KIRs are known to interact with specific motifs of HLA Class I molecules, which is very crucial in determining whether a cell is targeted to lysis or otherwise. Latent autoimmune diabetes in adults (LADA) is a slowly progressive form of autoimmune diabetes, with an adult onset (>30 years). Because autoantibodies and autoimmunity involved are involved in the etiology of LADA, KIRs might play an important role in conferring susceptibility to or protection against the disease. The purpose of this study was to identify killer immunoglobulin‐like receptor (KIR) genes, which are associated with susceptibility to and protection against type 1 diabetes in Latvian and Asian Indian patients with LADA. KIR and HLA‐C ligand genotyping was performed using PCR‐SSP in LADA patients from Latvia (n= 45) with age‐ and sex‐matched controls (n= 92) and from India (n= 86) with controls (n= 98). Results showed that in Latvian patients with LADA, KIRs 2DL1, 2DS2, and 2DS4 were associated with susceptibility and KIR 2DS5 with protection. In Asian Indian LADA patients, KIRs 2DL5 and 3DL1 were associated with susceptibility and KIRs 2DS1 and 2DS3 with protection. Stratification analyses for KIRs that bind to HLA‐C1 and C2 were performed. We concluded that KIRs are important in conferring susceptibility (or protection) to adult patients with LADA in both our study populations. However the KIR genes (and their HLA‐C ligands) conferring susceptibility or protection in these two populations differ, showing a role of ethnicity in disease susceptibility.
Overview of Circulating Nucleic Acids in Plasma/Serum
Authors: Butt Asif N. , Swaminathan R.
Published: August 2009

The existence of circulating nucleic acids in plasma and serum (CNAPS) was first described almost six decades ago. However, the prognostic and diagnostic utility of this circulating DNA/RNA has only really begun to be appreciated in the last decade. Earlier studies concentrated mainly on investigations concerned with fetal medicine and oncology, and significant progress was made in both specialities. More recently the field of enquiry has extended further, and attention has turned to other pathologic states, including trauma, sepsis, myocardial infarction, stroke, transplantation, diabetes mellitus, and hematologic disorders. In some of these studies, mitochondrial as well as genomic DNA and tissue‐specific mRNA have been analyzed, either quantitatively or qualitatively or both, and have been shown to be modified in the presence of disease. While there is tremendous potential for CNAPS as a clinical modality, many of the emerging studies seem to be confined to a few dedicated labs. Therefore, additional independent studies are necessary in some cases in order to show reproducibility, which will further consolidate the field. Despite this shortcoming, and with the evidently increasing number of applications for CNAPS, it is highly likely that routine testing, as described, will become reality within the next 5 to 10 years.
Stability of β‐Actin mRNA in Plasma
Authors: Holford N.C. , Sandhu H.S. , Thakkar H. , Butt A.N. , Swaminathan R.
Published: August 2009

It has long been thought that mRNA is labile and easily prone to degradation. However, a recent study demonstrated that GAPDH mRNA in cell‐free plasma may remain stable up to 24 hours after blood collection. As there are no other independent studies, we attempted to reproduce the findings of that study. In our study, blood was collected from a healthy male volunteer into Vacutainer tubes containing EDTA. Blood samples were placed on ice and plasma separated by double‐centrifugation at times 0, 1, 2, and 5 hours after blood collection. mRNA was extracted from four aliquots of the blood sample by means of the QIAamp Viral RNA kit. Extracted mRNA was converted to cDNA by reverse transcription before real time quantitative PCR measurement of the housekeeping β‐actin gene. Plasma β‐actin mRNA at 2 hours (0.012; 0.0031–0.0297, median and range) was significantly lower (P= 0.022) than at 0 hours (0.12; 0.057–0.165) (P= 0.016). The levels decreased further at 5 hours (0.0037; 0.0024–0.011) (P= 0.004). The results show that plasma β‐actin mRNA levels decrease with time after blood collection and that this is likely to be due to degradation in vitro.
Clinical Relevance of Circulating Nucleosomes in Cancer
Authors: Holdenrieder Stefan , Nagel Dorothea , Schalhorn Andreas , Heinemann Volker , Wilkowski Ralf , Von Pawel Joachim , Raith Hannelore , Feldmann Knut , Kremer Andreas E. , Müller Susanne , Geiger Sandra , Hamann Gerhard F. , Seidel Dietrich , Stieber Petra
Published: August 2009

Nucleosomes, complexes of DNA and histone proteins, are released during cell death into the blood circulation. Elevated serum and plasma levels have been found in various forms of cancer, but also in autoimmune diseases and acute situations such as stroke, trauma, and during sepsis. Here, the clinical relevance of circulating nucleosomes for diagnosis, staging, prognosis, and therapeutic monitoring of cancer is reviewed. Several studies have shown that levels of nucleosomes are significantly higher in serum and plasma of cancer patients in comparison to healthy controls. However, because of elevations of nucleosome levels in patients with benign diseases relevant for differential diagnosis, they are not suitable for cancer diagnosis. Concerning tumor staging, nucleosome levels correlate with tumor stage and presence of metastases in gastrointestinal cancer, but not in other tumor types. Prognostic value of circulating nucleosomes is found in lung cancer in univariate analyses, but not in multivariate analyses. Circulating nucleosomes are most informative for the monitoring of cytotoxic therapy. Strongly decreasing levels are mainly found in patients with remission of disease, whereas constantly high or increasing values are associated with progressive disease during chemo‐ and radiotherapy. In addition, therapy outcome is already indicated by the nucleosomal course during the first week of chemo‐ and radiotherapy in patients with lung, pancreatic, and colorectal cancer as well as in hematologic malignancies. Despite their non‐tumor‐specificity, kinetics of nucleosomes are valuable markers for the early estimation of therapeutic efficacy and may be helpful to adapting early cancer therapy in the future.
Isolation and Extraction of Circulating Tumor DNA from Patients with Small Cell Lung Cancer
Authors: Board Ruth E. , Williams Victoria S. , Knight Lucy , Shaw Jacqueline , Greystoke Alastair , Ranson Malcolm , Dive Caroline , Blackhall Fiona H. , Hughes Andrew
Published: August 2009

There is no consensus on the optimal protocol for isolation of circulating tumor DNA. We report our comparison of several extraction methods and variables that may affect yield, quality, and contamination of tumor DNA. DNA was extracted from the plasma and serum of five healthy volunteers by means of four different commercially available kits and DNA yield was quantified by real‐time PCR. DNA was extracted using the optimum kit from the plasma and serum of an additional 10 healthy volunteers and 10 patients with small cell lung cancer (SCLC) to compare yield and DNA fragment size in plasma versus serum and in those with SCLC versus controls. Time to sample processing was also examined. We found that DNA yield was greatest using the QIAamp Viral Spin Kit. Delayed time to processing led to increased DNA concentrations in serum, but not plasma. The plasma DNA concentration in SCLC patients was significantly higher than in healthy volunteers (24.5 ng/mL versus 5.1 ng/mL, P= 0.002). In contrast, there was no significant difference in serum DNA concentrations between controls and patients that may be explained by the wide variability and range of DNA concentrations in serum. A significantly higher proportion of longer fragments (272 bp/60 bp) was observed in the plasma DNA extracted from patients with SCLC than in healthy controls (13% versus 8%, P= 0.04). There was absence of DNA fragments of 512 bp in healthy control plasma, but faint bands were observed in serum, which is thought to be due to cellular contamination. We conclude that plasma is a more reliable source of tumor DNA then serum and have optimized a robust procedure for plasma tumor DNA isolation that can be applied to translational research studies.
The Origin of Elevated Levels of Circulating DNA in Blood Plasma of Premature Neonates
Authors: Tuaeva N. O. , Abramova Z. I. , Sofronov V. V.
Published: August 2009

The relationship between the concentration of cell‐free DNA (cfDNA) and the number of proliferating/apoptotic lymphocytes in peripheral blood of premature newborns of different gestation age and full‐term newborns was determined. The experiments were performed using fluorescent spectrophotometry (with Hoechst 33342), flow cytometry, and microscopy (Feulgen staining of lymphocytes). It was determined that the lymphocyte population of premature newborns may consist of 4.6% of proliferating and 22.1% apoptotic cells. For full‐term newborns, the percentage was 2.5% and 2.9%, respectively. A direct correlation between the concentration of extracellular DNA and the number of proliferating lymphocytes of full‐term newborns was ascertained (r= 0.400; P < 0.05). For premature newborns, the concentration of extracellular DNA correlated both with proliferating lymphocytes and apoptotic cells. The results show that premature birth causes the induction in lymphocytes of both apoptosis and proliferation that are accompanied by an increased extracellular DNA concentration in the blood of newborn babies.
Dichlorodiphenyldichloroethylene Exposure during the First Trimester of Pregnancy Alters the Anal Position in Male Infants
Authors: Torres‐Sanchez Luisa , Zepeda Monica , Cebrián Mariano E. , Belkind‐Gerson Jaime , Garcia‐Hernandez Rosa M. , Belkind‐Valdovinos Uri , López‐Carrillo Lizbeth
Published: August 2009

Anogenital distance (AGD) at birth is regarded as a useful measurement that reflects the prenatal androgenic status in rodents. However, the impact of xenoantiandrogens on human development is largely unknown. The aim of this study was to evaluate the potential antiandrogenic impact of prenatal DDT metabolites (p,p′‐DDE and p,p′‐DDT) exposure on infant AGD, using a non‐age–dependent anal position index (API). As part of an ongoing perinatal cohort study on the effects of organochlorine pesticides in children's neurodevelopment, we conducted a cross‐sectional study in 71 infants (37 males and 34 females). Maternal serum levels of DDT metabolites (p,p′‐DDE and p,p′‐DDT) before and during each trimester of pregnancy were determined by electron capture gas–liquid chromatography. During postnatal home visits at 3, 6, and 12 or 18 months of age, the children's weight and API were evaluated. Multiple lineal regression models were used to estimate the potential endocrine disruptor activity of prenatal p,p′‐DDE exposure. Boys had significantly higher API values than girls (0.6 versus 0.5; P < 0.001). Only among boys, a doubling increase of maternal p,p′‐DDE serum levels during the first trimester of pregnancy, were associated with a significant reduction of API (β=−0.02; P= 0.02). No effect of p,p′‐DDT on AGD was observed. Evidence of the effect of prenatal p,p′‐DDE on external genital differentiation is scarce and not consistent in the literature. Further studies are needed to confirm a hormonal disruptive effect on the development of external genitalia, due not only to p,p′‐DDE but also due to other antiandrogenic persistent compounds.
Monitoring of DNA Damage in Individuals Exposed to Petroleum Hydrocarbons in Ecuador
Authors: Paz‐y‐Miño César , López‐Cortés Andrés , Arévalo Melissa , Sánchez María Eugenia
Published: August 2009

Currently, it is known that several chemical agents used or generated by the oil industry are classified as mutagens and/or carcinogens. Among these we have gasoline, diesel, butane gas, styrene, benzene, chloroform, and others. Studies have verified that these chemicals have effects in fertility (abortions, sterility); produce various upheavals, such as dizziness, nausea, muscular pain; and produce chromosomal damage at the DNA level, which in the long or medium run, can develop into cancer and leukemia. The genetic damage in exposed individuals was measured by means of the comet test, chromosomal alterations test, and the study of the CYP 1A1 and MSH2 genes. These methods were applied to determine the genotoxicity of hydrocarbons and their residue in human beings. When conducting these tests on the blood samples of individuals exposed to hydrocarbons (workers of oil companies) and of a control population of the area of study and Quito, it was found that, in effect, the exposed individuals presented a greater amount of damage at the DNA level as well as at the chromosomal level than the individuals from the control populations (P< 0.001). Thus, it can be determined that populations that are exposed to hydrocarbons are susceptible to developing genetic damage. Therefore, risk groups can be determined in certain zones where the oil impact has been greater.
Bioremediation of Heavy Metal–Contaminated Soils by Sulfate‐Reducing Bacteria
Authors: Jiang Wei , Fan Wenhong
Published: August 2009

Many sewage irrigation areas in China have lost their agricultural productivity because they are polluted by cadmium (Cd). Use of sulfate‐reducing bacteria (SRB) may have a positive ecological effect on the bioremediation of heavy‐metal Cd‐contaminated soils. A five‐step Tessier sequential extraction procedure to analyze the geochemical speciation of metal Cd and investigate the bioremediation efficiency by SRB was adopted. The results indicate that the SRB can reduce the concentrations of the exchangeable fraction of Cd, which can be easily absorbed by plants. The removal is approximately 70%, which demonstrates bioremediation efficiency. In the contaminated soils before bioremediation, the exchangeable fraction of Cd is the dominate carrier and the fraction of Fe‐Mn oxides is next in importance, accounting for 45–60% and 35–55% of the total Cd, respectively. After adding the SRB into the water‐logged soils, the concentration of different geochemical speciation of Cd was determined every 7 days for 33 days. During this period, the total concentrations of Cd did not change, while the form of Cd changed significantly. The concentrations of the exchangeable fraction of Cd decreased significantly, which shows that the Cd bioavailability in the soils markedly decreased. The concentrations of the Fe‐Mn oxides fraction of Cd rose conspicuously, ranging from 60% to 120%; the concentrations of the carbonate fraction of Cd and organic matter fraction of Cd both increased slightly. The residue fraction of Cd did not change. The scope of oxidation–reduction potential was from 150 mv to −450 mv, and the rate of reduction of sulfate decreased with the increase of total Cd concentration.
Management of Endocrine Manifestations and the Use of Mitotane As a Chemotherapeutic Agent for Adrenocortical Carcinoma
Authors: Veytsman Irina , Nieman Lynnette , Fojo Tito
Published: August 2009

Adrenal cortical carcinoma (ACC) is a rare malignancy in which patients have poor overall 5-year survival. Patients with ACC can present with symptoms of hormone excess, including Cushing's syndrome, virilization, feminization, or—less frequently—hypertension with hypokalemia. In many patients with ACC, advanced disease at presentation precludes surgery or is followed by local relapse or distant metastatic disease that cannot be managed surgically. In these instances, chemotherapy is often tried, but its limited efficacy all too often leaves the problem of persistent hormonal excess. Physicians who treat patients with ACC and severe hypercortisolism should recognize that uncontrolled hormone production is a malignant disease, which has severe consequences that require aggressive management. Because chemotherapy benefits only a small percentage of patients, steroidogenesis inhibitors, including mitotane, ketoconazole, metyrapone, and etomidate, should be used singly or in combination even as chemotherapy is administered. Diligent management with frequent adjustments is required, especially in patients with chemotherapy-refractory tumors that continue to grow. In the absence of randomized, controlled trials, adjuvant use of mitotane remains controversial, although the authors of a recent case-control study argue for its use. Despite difficulty administering effective doses, most clinicians agree that mitotane should be used if the tumor cannot be removed surgically or should be used as adjuvant therapy if there is a high likelihood of recurrence. The option of long-term monotherapy is restricted to patients who tolerate mitotane and either experience a clinical response or are at high risk for recurrence. Recommendations are provided to help manage patients with this difficult disease and to improve the quality of their lives.
Risk Assessment Among Prostate Cancer Patients Receiving Primary Androgen Deprivation Therapy
Authors: Cooperberg Matthew R. , Hinotsu Shiro , Namiki Mikio , Ito Kazuto , Broering Jeanette , Carroll Peter R. , Akaza Hideyuki
Published: August 2009

Purpose

Prostate cancer epidemiology has been marked overall by a downward risk migration over time. However, in some populations, both in the United States and abroad, many men are still diagnosed with high-risk and/or advanced disease. Primary androgen deprivation therapy (PADT) is frequently offered to these patients, and disease risk prediction is not well-established in this context. We compared risk features between large disease registries from the United States and Japan, and aimed to build and validate a risk prediction model applicable to PADT patients.

Methods

Data were analyzed from 13,740 men in the United States community-based Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE) registry and 19,265 men in the Japan Study Group of Prostate Cancer (J-CaP) database, a national Japanese registry of men receiving androgen deprivation therapy. Risk distribution was compared between the two datasets using three well-described multivariable instruments. A novel instrument (Japan Cancer of the Prostate Risk Assessment [J-CAPRA]) was designed and validated to be specifically applicable to PADT patients, and more relevant to high-risk patients than existing instruments.

Results

J-CaP patients are more likely than CaPSURE patients to be diagnosed with high-risk features; 43% of J-CaP versus 5% of CaPSURE patients had locally advanced or metastatic disease that could not be stratified with the standard risk assessment tools. J-CAPRA—scored 0 to 12 based on Gleason score, prostate-specific antigen level, and clinical stage—predicts progression-free survival among PADT patients in J-CaP with a c-index of 0.71, and cancer-specific survival among PADT patients in CaPSURE with a c-index of 0.84.

Conclusion

The novel J-CAPRA is the first risk instrument developed and validated for patients undergoing PADT. It is applicable to those with both localized and advanced disease, and performs well in diverse populations.
Clinical Features and Outcome of Patients With Non–Small-Cell Lung Cancer Who Harbor EML4-ALK
Authors: Shaw Alice T. , Yeap Beow Y. , Mino-Kenudson Mari , Digumarthy Subba R. , Costa Daniel B. , Heist Rebecca S. , Solomon Benjamin , Stubbs Hannah , Admane Sonal , McDermott Ultan , Settleman Jeffrey , Kobayashi Susumu , Mark Eugene J. , Rodig Scott J. , Chirieac Lucian R. , Kwak Eunice L. , Lynch Thomas J. , Iafrate A. John
Published: August 2009

Purpose

The EML4-ALK fusion oncogene represents a novel molecular target in a small subset of non–small-cell lung cancers (NSCLC). To aid in identification and treatment of these patients, we examined the clinical characteristics and treatment outcomes of patients who had NSCLC with and without EML4-ALK.

Patients and Methods

Patients with NSCLC were selected for genetic screening on the basis of two or more of the following characteristics: female sex, Asian ethnicity, never/light smoking history, and adenocarcinoma histology. EML4-ALK was identified by using fluorescent in situ hybridization for ALK rearrangements and was confirmed by immunohistochemistry for ALK expression. EGFR and KRAS mutations were determined by DNA sequencing.

Results

Of 141 tumors screened, 19 (13%) were EML4-ALK mutant, 31 (22%) were EGFR mutant, and 91 (65%) were wild type (WT/WT) for both ALK and EGFR. Compared with the EGFR mutant and WT/WT cohorts, patients with EML4-ALK mutant tumors were significantly younger (P < .001 and P = .005) and were more likely to be men (P = .036 and P = .039). Patients with EML4-ALK–positive tumors, like patients who harbored EGFR mutations, also were more likely to be never/light smokers compared with patients in the WT/WT cohort (P < .001). Eighteen of the 19 EML4-ALK tumors were adenocarcinomas, predominantly the signet ring cell subtype. Among patients with metastatic disease, EML4-ALK positivity was associated with resistance to EGFR tyrosine kinase inhibitors (TKIs). Patients in the EML4-ALK cohort and the WT/WT cohort showed similar response rates to platinum-based combination chemotherapy and no difference in overall survival.

Conclusion

EML4-ALK defines a molecular subset of NSCLC with distinct clinical characteristics. Patients who harbor this mutation do not benefit from EGFR TKIs and should be directed to trials of ALK-targeted agents.
The use of the defence style questionnaire in major depressive and panic disorders: A comprehensive meta‐analysis
Authors: Calati Raffaella. , Oasi Osmano. , De Ronchi Diana. , Serretti Alessandro.
Published: August 2009

Purpose. The issue of defence mechanisms is of great importance in clinical practice. The aim of this meta‐analysis is to compare different defence styles (Mature, Neurotic, and Immature), assessed using the three‐factor defence style questionnaire (DSQ), in major depressive disorder (MDD), and panic disorder (PD) in order to evaluate potential differences in defence profiles among these disorders.

Methods. We assessed all publications listed in PubMed, PsycINFO, ISI, and Lilacs databases focusing on DSQ defence styles in MDD and PD. To be included studies had to be published in English, to include psychiatric patients, to provide defence style mean scores, and to employ the three‐factor DSQ versions. Data were entered into the Cochrane Collaboration Review Manager Software and analysed by RevMan version 4.2.

Results. MDD patients reported significantly lower scores in Mature style and both MDD and PD patients reported significantly higher scores in Neurotic and Immature styles.

Conclusions. A dissimilarity between depressive and anxious profiles emerged, since the former profile was characterized by low Mature and high Neurotic and Immature scores while the latter by high Neurotic and Immature scores only. These results could indicate the use of specific strategies in clinical and psychotherapeutic management of patients with these diagnoses. Nevertheless, present results should be considered with caution because of the high heterogeneity of the studies and some weaknesses in the psychometric properties of the DSQ.
Freud's Uncanny Egypt: Prolegomena
Authors: Bergstein Mary
Published: August 2009

Egypt in all its guises—biblical, ancient, and modern—was a place of uncanny "otherness" in Freud's visual and historical imagination. This mental construct was formed from both hieroglyphic writing and Freud's collection of small-scale statuary. Uncanny visual representations of Egypt were also found in books and photographs in his archaeological library, as well as in wood engravings in the Philippson Bible, which he encountered for the first time in early childhood. Manifest subject matter associated with Egypt included mummies, dancing-girls, works of ancient art, and romantic desert landscapes. The present reflections on Freud's multifaceted fascination with Egypt are offered as prolegomena to open prospects for further research.
The Magic of the Manifest: Freud's Egyptian Dream Book
Authors: O'Donoghue Diane
Published: August 2009

Textual evidence shows that dream interpretation preceded Freud by several millennia, but his method was unprecedented. In asserting that the dream's remembered, or manifest, content was merely a disguise for censored, latent meaning, he associated manifest content with notions of animism and magical thinking that characterized colonialist attitudes toward non-European cultures. This paper examines Freud's investment in devaluing dream imagery and relates his distrust at once to the dream books that originated in the classical period and to those that, with pretensions to an ancient lineage, were popular in his time. In examining the anti-Semitism present in dream books of both periods, the paper identifies ways in which Freud's theories evolved as a cultural construction rather than being, as he claimed, the "discovery" of innate psychic functioning.