bluuebunny/crossref_metadata_2025 huggingface dataset:
Repair and Neurorehabilitation Strategies for Spinal Cord Injury
Authors: Ruff Robert L. , McKerracher Lisa , Selzer Michael E.
Published: August 2009

The failure of axons in the central nervous system (CNS) to regenerate has been considered the main factor limiting recovery from spinal cord injury (SCI). Impressive gains in identification of growth‐inhibitory molecules in the CNS led to expectations that their neutralization would lead to functional regeneration. However, results of therapeutic approaches based on this assumption have been mixed. Recent data suggest that neurons differ in their ability to regenerate through similar extracellular environments, and moreover, they undergo a developmental loss of intrinsic regenerative ability. Factors mediating these intrinsic regenerative abilities include expression of (1) receptors for inhibitory molecules such as the myelin‐associated growth inhibitors and developmental guidance molecules, (2) surface molecules that permit axon adhesion to cells in the path of growth, (3) cytoskeletal proteins that mediate the mechanics of axon growth, and (4) molecules in the intracellular signaling cascades that mediate responses to chemoattractive and chemorepulsive cues. In contrast to axon development, regeneration might involve internal protrusive forces generated by microtubules, either through their own elongation or by transporting other cytoskeletal elements such as neurofilaments into the axon tip. Because of the complexity of the regenerative program, one approach will probably be insufficient to achieve functional restoration of neuronal circuits. Combination treatments will be increasingly prominent. SCI is a debilitating and costly condition that compromises pursuit of activities usually associated with an independent and productive lifestyle. This article discusses recent advances in neurorehabilitation that can improve the life quality of individuals with SCI.
Drug‐induced Movement Disorders in Children
Authors: Gilbert Donald L.
Published: August 2009

This article reviews the current state of knowledge of drug‐induced movement disorders (DIMDs) in children. The objective is to aid clinicians who treat children with medications that may induce DIMDs, as well as specialists consulted on DIMDs. As with adults, the most common agents are dopaminergic and dopamine‐blocking medications, and prescriptions for these agents have increased markedly in children. Unfortunately, most evidence‐based reviews, including those from the Cochrane Collaboration cited here, have few systematic data to analyze. Many publications are small case series. This report attempts to provide useful information, with appropriate caution and discussion of the limitations of what we know.
Task‐specific Dystonias
Authors: Torres‐Russotto Diego , Perlmutter Joel S.
Published: August 2009

Task‐specific dystonias are primary focal dystonias characterized by excessive muscle contractions producing abnormal postures during selective motor activities that often involve highly skilled, repetitive movements. Historically these peculiar postures were considered psychogenic but have now been classified as forms of dystonia. Writer's cramp is the most commonly identified task‐specific dystonia and has features typical of this group of disorders. Symptoms may begin with lack of dexterity during performance of a specific motor task with increasingly abnormal posturing of the involved body part as motor activity continues. Initially, the dystonia may manifest only during the performance of the inciting task, but as the condition progresses it may also occur during other activities or even at rest. Neurological exam is usually unremarkable except for the dystonia‐related abnormalities. Although the precise pathophysiology remains unclear, increasing evidence suggests reduced inhibition at different levels of the sensorimotor system. Symptomatic treatment options include oral medications, botulinum toxin injections, neurosurgical procedures, and adaptive strategies. Prognosis may vary depending upon body part involved and specific type of task affected. Further research may reveal new insights into the etiology, pathophysiology, natural history, and improved treatment of these conditions.
Prospects of Cell Therapy for Disorders of Myelin
Authors: Ben‐Hur Tamir , Goldman Steven A.
Published: August 2009

Recent advances in stem cell biology have raised expectations that both diseases of, and injuries to, the central nervous system may be ameliorated by cell transplantation. In particular, cell therapy has been studied for inducing efficient remyelination in disorders of myelin, including both the largely pediatric disorders of myelin formation and maintenance and the acquired demyelinations of both children and adults. Potential cell‐based treatments of two major groups of disorders include both delivery of myelinogenic replacements and mobilization of residual oligodendrocyte progenitor cells as a means of stimulating endogenous repair; the choice of modality is then predicated upon the disease target. In this review we consider the potential application of cell‐based therapeutic strategies to disorders of myelin, highlighting the promises as well as the problems and potential perils of this treatment approach.
Hereditary Episodic Ataxias
Authors: Jen Joanna C.
Published: August 2009

Hereditary episodic ataxia (EA) syndromes are rare monogenic disorders that are phenotypically and genetically heterogeneous. The number of identified EA phenotypes is expanding. So far, mutations have been identified in four genes, all coding for membrane proteins including ion channels and transporters. The study of EA has illuminated previously unrecognized but important roles of ion channels and transporters in cerebellar function. This review summarizes recent advances and focuses on practical approaches in the diagnosis and treatment of episodic ataxia.
The Estimation of Probable Maximum Precipitation
Authors: Casas M. Carmen , Rodríguez Raül , Nieto Raquel , Redaño Angel
Published: August 2009

A brief overview of the different techniques used to estimate the probable maximum precipitation (PMP) is presented. As a particular case, the 1‐day PMP over Catalonia has been calculated and mapped with a high spatial resolution. For this purpose, the annual maximum daily rainfall series from 145 pluviometric stations of the Instituto Nacional de Meteorología (Spanish Weather Service) in Catalonia have been analyzed. In order to obtain values of PMP, an enveloping frequency factor curve based on the actual rainfall data of stations in the region has been developed. This enveloping curve has been used to estimate 1‐day PMP values of all the 145 stations. Applying the Cressman method, the spatial analysis of these values has been achieved. Monthly precipitation climatological data, obtained from the application of Geographic Information Systems techniques, have been used as the initial field for the analysis. The 1‐day PMP at 1 km2spatial resolution over Catalonia has been objectively determined, varying from 200 to 550 mm. Structures with wavelength longer than approximately 35 km can be identified and, despite their general concordance, the obtained 1‐day PMP spatial distribution shows remarkable differences compared to the annual mean precipitation arrangement over Catalonia.
Large‐scale Factors in Tropical and Extratropical Cyclone Transition and Extreme Weather Events
Authors: Pezza Alexandre Bernardes , Simmonds Ian
Published: August 2009

Transition mechanisms characterizing changes from hurricanes to midlatitude cyclones and vice‐versa (extratropical and tropical transition) have become a topic of increasing interest, partially because of their association with recent unusual storms that have developed in different ocean basins of both hemispheres. The aim of this work is to discuss some recent cases of transition and highly unusual hurricane developments and to address some of their wider implications for climate science. Frequently those dramatic cyclones are responsible for severe weather, potentially causing significant damage to property and infrastructure. An additional manifestation discussed here is their association with cold surges, a topic that has been very little explored in the literature. In the Southern Hemisphere, the first South Atlantic hurricane, Catarina, developed in March 2004 under very unusual large‐scale conditions. That exceptional cyclone is viewed as a case of tropical transition facilitated by a well‐developed blocking structure. A new index for monitoring tropical transition in the subtropical South Atlantic is discussed. This “South Atlantic index” is used to show that the unusual flow during and prior to Catarina's genesis can be attributed to tropical/extratropical interaction mechanisms. The “Donald Duck” case in Australia and Vince in the North Atlantic have also been examined and shown to belong to a category of hybrid‐transitioning systems that will achieve at least partial tropical transition. While clearly more research is needed on the topic of transition, as we gain further insight, it is becoming increasingly apparent that features of large‐scale circulation do play a fundamental role. A complex interaction between an extratropical transition case and an extreme summer cold surge affecting southeastern Australia is discussed as an example of wider climate implications.
Are Reanalysis Data Useful for Calculating Climate Indices over South America?
Authors: Dufek Amanda Sabatini , Ambrizzi Tércio , Da Rocha Rosmeri Porfírio
Published: August 2009

Precipitation and temperature climate indices are calculated using the National Centers for Environmental Prediction/National Center for Atmospheric Research (NCEP/NCAR) reanalysis and validated against observational data from some stations over Brazil and other data sources. The spatial patterns of the climate indices trends are analyzed for the period 1961–1990 over South America. In addition, the correlation and linear regression coefficients for some specific stations were also obtained in order to compare with the reanalysis data. In general, the results suggest that NCEP/NCAR reanalysis can provide useful information about minimum temperature and consecutive dry days indices at individual grid cells in Brazil. However, some regional differences in the climate indices trends are observed when different data sets are compared. For instance, the NCEP/NCAR reanalysis shows a reversal signal for all rainfall annual indices and the cold night index over Argentina. Despite these differences, maps of the trends for most of the annual climate indices obtained from the NCEP/NCAR reanalysis and BRANT analysis are generally in good agreement with other available data sources and previous findings in the literature for large areas of southern South America. The pattern of trends for the precipitation annual indices over the 30 years analyzed indicates a change to wetter conditions over southern and southeastern parts of Brazil, Paraguay, Uruguay, central and northern Argentina, and parts of Chile and a decrease over southwestern South America. All over South America, the climate indices related to the minimum temperature (warm or cold nights) have clearly shown a warming tendency; however, no consistent changes in maximum temperature extremes (warm and cold days) have been observed. Therefore, one must be careful before suggesting any trends for warm or cold days.
The Intra‐Americas Sea Low‐level Jet
Authors: Amador Jorge A.
Published: August 2009

A relevant climate feature of the Intra‐Americas Sea (IAS) is the low‐level jet (IALLJ) dominating the IAS circulation, both in summer and winter; and yet it is practically unknown with regard to its nature, structure, interactions with mid‐latitude and tropical phenomena, and its role in regional weather and climate. This paper updates IALLJ current knowledge and its contribution to IAS circulation–precipitation patterns and presents recent findings about the IALLJ based on firstin situobservations during Phase 3 of the Experimento Climático en las Albercas de Agua Cálida (ECAC), an international field campaign to study IALLJ dynamics during July 2001. Nonhydrostatic fifth‐generation Pennsylvania State University National Center for Atmospheric Research Mesoscale Model (MM5) simulations were compared with observations and reanalysis. Large‐scale circulation patterns of the IALLJ northern hemisphere summer and winter components suggest that trades, and so the IALLJ, are responding to land–ocean thermal contrasts during the summer season of each continent. The IALLJ is a natural component of the American monsoons as a result of the continent's approximate north–south land distribution. During warm (cold) El Niño–Southern Oscillation phases, winds associated with the IALLJ core (IALLJC) are stronger (weaker) than normal, so precipitation anomalies are positive (negative) in the western Caribbean near Central America and negative (positive) in the central IAS. During the ECAC Phase 3, strong surface winds associated with the IALLJ induced upwelling, cooling down the sea surface temperature by 1–2 °C. The atmospheric mixed layer height reached 1 km near the surface wind maximum below the IALLJC. Observations indicate that primary water vapor advection takes place in a shallow layer between the IALLJC and the ocean surface. Latent heat flux peaked below the IALLJC. Neither the reanalysis nor MM5 captured the observed thermodynamic and kinematic IALLJ structure. So far, IALLJ knowledge is based on either dynamically initialized data or simulations of global (regional) models, which implies that a more systematic and scientific approach is needed to improve it. The Intra‐Americas Study of Climate Processes is a great regional opportunity to address trough field work, modeling, and process studies, many of the IALLJ unknown features.
Cryosphere—Atmosphere Interaction Related to Variability and Change of Northern Hemisphere Annular Mode
Authors: Bojariu Roxana , García‐Herrera Ricardo , Gimeno Luis , Zhang Tingjun , Frauenfeld Oliver W.
Published: August 2009

The Northern Hemisphere annular mode, also known as the Arctic Oscillation/North Atlantic Oscillation (AO/NAO) is a dominant atmospheric mode in the Northern Hemisphere winter that influences climate fluctuations from the eastern seaboard of the United States to Siberia and from the Arctic to the subtropical Atlantic. After almost a century of scientific investigation, the fundamental mechanisms determining the evolution of the AO/NAO are not yet completely understood. The ocean is favored as the most likely forcing of atmospheric variability, given the time scales of oceanic circulation and its large heat capacity. Our analyses of snow cover, soil temperatures, zonal winds, and geopotential heights identify the effect of land–atmosphere interaction over Eurasia on Northern Hemisphere atmospheric circulation, explaining the predictive signal that links fluctuations of April–October snow cover with the following winter AO/NAO phases.
Comparative Analysis of the Islet Proteome between NOD/Lt and ALR/Lt Mice
Authors: Yang Ping , Li Manyu , Guo Dehuang , Gong Feili , Adam Bao‐Ling , Atkinson Mark A. , Wang Cong‐Yi
Published: August 2009

Although alloxan‐resistant (ALR) mice share 70% of the NOD genome, ALR islets are remarkably resistant to oxidative stress and autoimmunity. Using 2D PAGE comparative analysis, we have characterized 55 proteins that are differentially expressed between the NOD and ALR islet proteome. Ten proteins were found to be highly expressed in the NOD islets. Typically, proteins associated with pancreatic inflammation and autoimmunity, such as amylase and carboxypeptidase, are highly expressed in the NOD islets. Forty‐five proteins showed significantly higher expression in the ALR islets. Among these, 30 are proteins implicated in the regulation of intracellular stress including heat‐shock proteins, disulfide isomerase–associated proteins, ROS detoxification enzymes, and apoptotic regulators. Our results clearly demonstrate that the 30% unique ALR genome encodes protective determinants expressed at islet levels, which render the islets of this strain of mice resistant to oxidative stress and autoimmunity.
HLA Class I Supertypes in Type 1 Diabetic Children in an Urban Children's Hospital
Authors: Antal Zoltan , Jarchum Irene , DiLorenzo Teresa P.
Published: August 2009

Type 1 diabetes is an autoimmune disorder characterized by progressive destruction of insulin‐secreting β cells of the pancreas, in which CD8+ T cells play a critical role. The diversity in the HLA alleles expressed among various racial and ethnic groups leads to great variability in antigen presentation and recognition by CD8+ T cells in the context of MHC class I molecules. To date, studies aimed at identifying disease‐relevant antigenic epitopes have focused on using mice transgenic for HLA‐A*0201, a common allele, particularly among Caucasians. We present HLA class I typing data from 88 children with type 1 diabetes at the Children's Hospital at Montefiore, where the patient population is ethnically diverse, but largely minority. When categorized into the HLA supertypes A2, A3, B7, and C1, 77% of those studied have alleles belonging to at least one supertype, and of these patients, 65% do not belong to the A2 supertype, which is the supertype represented by the HLA‐A*0201 allele. These results support the need for studies using HLA transgenic mice expressing MHC molecules representative of a variety of HLA supertypes, particularly when searching for antigenic epitopes applicable for study among largely urban, minority pediatric populations.
Autoreactive T Cell Response in CD25‐Negative Fraction of Peripheral Blood Mononuclear Cells in Established Type 1 Diabetes
Authors: Moriyama Hiroaki , Kotani Reiko , Katsuta Atsumi , Kameno Mami , Arai Takashi , Okumachi Yasuyo , Kishi Minoru , Yamada Katsumi , Yasuda Hisafumi , Hara Kenta , Yokono Koichi , Nagata Masao
Published: August 2009

CD4+CD25+ T cells (Tregs) play a critical role in maintaining dominant peripheral tolerance, and pathogenic autoreactive T cells may be frequent in the CD25‐negative fraction of peripheral blood mononuclear cells (PBMCs) from patients with autoimmune disease. We therefore investigated whether T cell autoimmune responses to recombinant GAD65 can be detected by the use of ELISPOT assay in the CD25‐negative fraction of PMBCs from Japanese type 1 diabetes (T1D) patients. The frequency of CD4+CD25+ T cells was not different among patients with newly developed T1D, established T1D, and healthy controls. The CD25 positive cell–depleted fraction was obtained by negative selection with antihuman CD25 magnetic beads, reducing the number of CD4+CD25+ T cells from 4–5% to less than 1%. In whole PBMC fraction, there was a significant elevation of IFN‐γ spots in PBMCs from recently diagnosed patients with T1D (P < 0.05), whereas the number of IFN‐γ spots from patients with established T1D was not significant. In the CD25‐negative fraction, unlike whole PBMCs, we observed the significant IFN‐γ spots to GAD65 in the fraction from patients with established T1D (P < 0.05), but not in those with recently diagnosed disease. The phenomena were not observed for IL‐4 spots. Our data suggest a possible role of Tregs maintaining dominant peripheral tolerance in T1D and application of further improved T cell assay detecting autoimmunity even in established T1D.
Induction of Apoptosis of β Cells of the Pancreas by Advanced Glycation End‐Products, Important Mediators of Chronic Complications of Diabetes Mellitus
Authors: Lim Minsu , Park Leejin , Shin Geewook , Hong Hekyung , Kang Incheol , Park Yongsoo
Published: August 2009

We herein report cytotoxicity of advanced glycation end‐products (AGEs) on pancreatic β cells. AGEs stimulated reactive oxygen species (ROS) generation but did not arrest proliferation of the INS‐1 cell line. Pancreatic β cell lines or primary cultured islets possess a receptor for AGE (RAGE), and its expression increased after AGE treatment. TUNEL staining and FACS analysis using annexin V/PI antibodies showed that apoptosis increased in INS‐1 cells or primary cultured islets when incubated with BSA conjugated with glyceraldehyde (AGE2) or glucoaldehyde (AGE3), compared with those conjugated with glucose (AGE1). Reaction of INS‐1 cells to Ki67, which is a cellular marker for proliferation, was also increased after AGE treatment. The ability of primary cultured islets to secrete insulin was retained even after AGE treatment under either low or high glucose conditions. The antiserum against RAGE partially prevented AGE‐induced cellular events. Treatment of β cells with the antioxidant metallothionein results in a significant reduction in pathologic changes. AGEs might be able to induce apoptosis as well as proliferation of pancreatic β cell lines or primary cultured islets. Moreover, antibody array showed that RAD51 and RAD52 were significantly decreased in AGE2‐treated INS‐1 cells. AGEs might inhibit homologous DNA recombination for repairing DNA of INS‐1 cells damaged by ROS generation. It might be suggested that treatment of AGEs resulted in ROS production and apoptosis through their receptor on pancreatic β cells. AGEs might deteriorate function of pancreatic β cells in patients with long‐term hyperglycemia.
Tobacco Exposure and Diabetes‐Related Autoantibodies in Children
Authors: Johansson AnnaKarin , Hermansson Göran , Ludvigsson Johnny ,   for the ABIS Study Group
Published: August 2009

Passive smoking has decreased in recent years (“increased hygiene”). Less environmental tobacco smoke (ETS) gives increased hygiene that, if the hygiene hypothesis is true, in turn might give more autoimmune diseases. The presence of auto antibodies is considered to be an early indicator of type 1 diabetes (T1D). Because tobacco exposure may influence the immune system, we analyzed the relation between passive smoking and development of autoantibodies. A subsample (n= 8794) of the children in the ABIS study was used for this analysis. The parents answered questionnaires on smoking from pregnancy and onwards, and blood samples from the children aged 2.5–3 years were analyzed for GADA and IA‐2A. Results showed that there was no significant difference in the prevalence of GADA or IA‐2A (>95 percentile) between tobacco‐exposed and nonexposed children. It was concluded that passive smoking does not seem to influence development of diabetes‐related autoantibodies early in life.
Conserved Epitopes in the Protein Tyrosine Phosphatase Family of Diabetes Autoantigens
Authors: Liu Y. , Wenzlau J.M. , Yu L. , Patel C. , Eisenbarth G.S. , Hutton J.C. , Davidson H.W.
Published: August 2009

IA2 and phogrin are important targets of humoral and cell‐mediated autoimmunity in type 1 diabetes in man. They belong to a conserved subfamily of transmembrane protein tyrosine phosphatases (PTPs) associated with the regulatory pathway of secretion. To examine potential cross‐reactivity between PTP family members we tested sera from T1D patients for reactivity to IA2, and the Drosophila (FLYDA) and C. elegans (IDA) orthologs using radioimmunoprecipitation assays of 35S Met–labeled in vitro translated products of the cytosolic domains of these proteins. Approximately 80% of sera reacted with at least one probe. Of these, 82.5% showed reactivity to human IA2, 74.1% to FLYDA, and 33.7% to IDA. The majority of sera that bound FLYDA and/or IDA also recognized IA2. This raises the possibility that in some cases reactivity to IA2 may have arisen by molecular mimicry.
From Type 1, through LADA, to Type 2 Diabetes: A Continuous Spectrum?
Authors: Lin J. , Zhou Z.G. , Wang J.P. , Zhang C. , Huang G.
Published: August 2009

This study aims to explore whether diabetes mellitus from T1DM, through LADA, to T2DM presents a continuous spectrum in terms of HLA‐DQ genetic background. We recruited 223 patients with autoimmune T1DM, 215 with LADA, 206 with T2DM, and 228 nondiabetic controls, and then defined their HLA‐DQA1 and –DQB1 genotypes and haplotypes. T1DM patients were divided into two groups depending on age of onset of disease: juvenile‐onset (JO; before the age of 20) and adult‐onset (AO; after the age of 20). LADA patients were sorted according to the GADA titer: the LADA1 group had titers higher than 175 U/mL, whereas the LADA2 group had lower titers. The susceptible haplotypes of T1DM were DQA1*03‐DQB1*0303, DQA1*03‐DQB1*0401, and DQA1*05‐DQB1*0201. The protective haplotype was DQA1*0102‐DQB1*0602. The frequency of DQA1*03‐DQB1*0303 in JO, AO, LADA1, LADA2, T2DM, and control groups were 38.2%, 34.2%, 25.3%, 18.9%, 17.5%, and 16.5%, respectively. The frequencies of DQA1*05‐DQB1*0201 were 21.2%, 15.0%, 12.7%, 4.6%, 3.6% and 3.3%; the frequencies of DQA1*03‐DQB1*0401 were 11.3%, 9.4%, 11.3%, 5.4%, 4.4% and 3.3%; and the frequencies of DQA1*0102‐DQB1*0602 were 1.4%, 1.7%, 0.7%, 4.6%, 7.0%, and 5.3%. The linear‐by‐linear association showed that the frequency of DQA1*03‐DQB1*0303, DQA1*05‐DQB1*0201, and DQA1*03‐DQB1*0401 presented a decremental tendency in JO, AO, LADA1, LADA2, T2DM, and control groups. The preliminary data demonstrated that the susceptible haplotypes of the HLA‐DQ gene present a continuous spectrum from typical T1DM, through LADA, to T2DM, which deserves further investigation.
Apoptotic Non–β Cells Suppress β Cell Antigen–Reactive T Cells and Induce β Cell Antigen–Specific Regulatory T Cells
Authors: Xia Chang‐Qing , Qiu Yushi , Peng Ruihua , Lo‐Dauer Jeannette , Clare‐Salzler Michael J.
Published: August 2009

Steady‐state cell apoptosis plays an important role in maintenance of self‐tolerance. Based on this notion, the use of apoptotic cells to restore self‐tolerance to β cell antigens is a rational approach to type 1 diabetes (T1D) prevention. Our previous study demonstrated that transfusion of apoptotic β cells induced immune tolerance to β cell antigens in NOD mice. However, concerned about the limited β cell source for future clinical applications, we attempted in the present study to develop a more practical approach for T1D prevention using apoptotic non–β cells. We found that UVB‐irradiation‐induced apoptotic NOD splenic stromal cells significantly suppressed β cell antigen‐specific T cell proliferation in vitro and in vivo. Furthermore, TCR‐transgenic CD4+ T cells primed by the antigens to which they were specific in the presence of UVB‐irradiated stromal cells were rendered unresponsive to the antigen restimulation, a result that was partially attributed to the induced IL‐10‐producing regulatory T cells. Of more interest, transfusion of UVB‐irradiated stromal cells appeared to induce β cell antigen–responding IL‐10‐producing regulatory T cells in vivo. Most importantly, transfusion of UVB‐irradiated stromal cells effectively prevented T1D in NOD mice, which is consistent with these findings. This study suggests that it is possible to use apoptotic non–β cells such as peripheral blood mononuclear cells to induce β cell antigen–specific tolerance, thereby preventing T1D in humans.
Developing a Novel Model System to Target Insulin‐Reactive CD8 T Cells
Authors: Scott Gwen S. , Fishman Sigal , Margalit Alon , Siew L. Khai , Chapman Stephen , Wen Li , Gross Gideon , Wong F. Susan
Published: August 2009

CD8 T cells play an important role in autoimmune diabetes development, and therefore removing these cells may protect against disease. To test this, we designed a novel method using engineered cells (InsCD3‐ζ) to target insulin‐specific CD8 T cells. Insulin‐reactive target cells were cultured with InsCD3‐ζ CD8 T cells and cytotoxicity was assessed. Activated, but not naïve, InsCD3‐ζ CD8 T cells readily killed insulin‐reactive target CD8 T cells. This approach to immunotarget relevant pathogenic CD8 T cells may be a therapeutic option to delay or prevent type 1 diabetes.
What Will It Take to Get Therapies Approved for Type 1 Diabetes?
Authors: Fleming Alexander
Published: August 2009

The development of therapies for T1D has been neglected in favor of efforts in advancing therapies for the larger T2D population. Pharmaceutical companies have also been deterred by lack of clarity around the regulatory expectations for such therapies. The prospects for therapy for new‐onset T1D have brightened in some respects because of convergence among regulators and clinical experts in views about how these therapies should be assessed. The most important consensus is that the primary efficacy end point for treatments directed at the underlying autoimmune cause of T1D should be endogenous insulin secretion, as reflected by standardized C‐peptide measurements. Most T1D therapeutic development efforts are directed at new‐onset disease, which represents a small proportion of the entire T1D population. A major deficiency in T1D therapeutic development is the lack of activity in advancing therapies for people with established T1D, a population that far outnumbers those with new‐onset disease. Complete remission of new‐onset or established T1D will almost certainly require a combination of two or more therapies to address the underlying cause of the disease and restore normal insulin function.
Evaluation of Cytokine Production from Peripheral Blood Mononuclear Cells of Type 1 Diabetic Patients
Authors: Foss‐Freitas Maria Cristina , Foss Norma Tiraboschi , Rassi Diane Meyre , Donadi Eduardo Antônio , Foss Milton C.
Published: August 2009

This study aims to evaluate the production of cytokines, tumor necrosis factor (TNF), and interleukin 10 (IL‐10) in peripheral blood mononuclear cells (PBMCs) from type 1 diabetic (T1D) patients by means of intracellular staining, flow cytometry, and ELISA and to correlate it with inadequate (IN) and adequate (A) metabolic controls. We studied 28 patients with T1D and 20 healthy individuals (C) paired by sex and age. T1D patients were divided in patients with IN and A metabolic control. PBMC cultures were stimulated with LPS to evaluate TNF or were stimulated with PMA/ionomycin or concanavalin A to evaluate IL‐10. The TNF levels in supernatant of stimulated cultures, evaluated by ELISA, of diabetic patients were similar to those of healthy individuals, although the percentage of CD 33+ cells that were positive for TNF was higher in the T1D IN group compared to the T1D A group (P= 0.01). Similarly, the IL‐10 levels evaluated by ELISA in stimulated cultures of T1D patients were not different from those in the control group; moreover, the percentage of CD3+ cells positive for intracellular IL‐10 were higher in the T1D IN group compared to C groups (P= 0.007). The increased levels of cytokines in T1D IN diabetic patients, with reduction in the A group, suggests that hyperglycemia stimulates an inflammatory state that can result in a deficient immune cellular response. The data suggest that assessment by intracellular staining seems to be more accurate than the ELISA technique in evaluating diabetic patients.
Update on Worldwide Efforts to Prevent Type 1 Diabetes
Authors: Skyler Jay S. ,   for the Type 1 Diabetes TrialNet Study Group
Published: August 2009

This paper reviews worldwide efforts to interdict the type 1 diabetes (T1D) disease process, during the stage of evolution of the disease prior to the time of disease onset. The goal of intervention before disease onset is to arrest immune destruction and thus prevent or delay clinical disease. In this regard, there have been several large‐scale multicenter randomized controlled clinical trials designed to prevent T1D. These have tested nicotinamide, parenteral insulin, oral insulin, nasal insulin, and the elimination of cow's milk from infant feeding.
Type 1 Diabetes versus Type 2 Diabetes with Onset in Persons Younger than 20 Years of Age
Authors: Unnikrishnan A. G. , Bhatia Eesh , Bhatia Vijayalakshmi , Bhadada Sanjay Kumar , Sahay Rakesh Kumar , Kannan Arun , Kumaravel V. , Sarma Dipti , Ganapathy Bantwal , Thomas Nihal , John Mathew , Jayakumar R. V. , Kumar Harish , Nair Vasantha , Sanjeevi Carani B.
Published: August 2009

Type 1 diabetes (T1D) is the most common form of diabetes in children in Western countries. There have been no large studies of childhood diabetes from India. We undertook the MEDI study (Multicenter Survey of Early Onset Diabetes in India) to assess the proportion of various subtypes of diabetes among the young subjects presenting to the endocrinology divisions of seven large teaching hospitals in different regions of India. In addition, we compared the clinical features of T1D and type 2 diabetes (T2D) in Indian subjects. Patients with onset of disease at younger than 20 years of age were included in this study. Six hundred and three subjects (603) were studied of whom 535 subjects (89%) had T1D, 36 (6%) had T2D, 18 (3%) had diabetes related to tropical pancreatitis or other forms of chronic pancreatitis, while other subtypes accounted for the rest. Compared to those with T2D, subjects with T1D were younger, had a lower C‐peptide level, higher prevalence of ketosis, lower prevalence of acanthosis nigricans, and lower LDL and triglyceride levels. When compared with that of T2D, a higher proportion of patients with T1D were positive for GAD‐65 and IA‐2 antibodies, and this difference was statistically significant for GAD‐65 antibodies. Overall, this large multicenter study showed that T1D is the commonest form of diabetes in childhood. T2D is the next most common kind, while chronic pancreatitis‐related diabetes is uncommon.
The Risk Conferred by HLA‐DR and DQ for Type 1 Diabetes in 0–35‐Years Age Group are Different in Different Regions of Sweden
Authors: Sanjeevi Carani B. , Sedimbi Saikiran K. , Landin‐Olsson Mona , Kockum Ingrid , Lernmark Åke ,   on behalf of the Swedish Childhood Diabetes and the Diabetes Incidence in Sweden Study Groups
Published: August 2009

HLA DR4‐DQ8 and DR3‐DQ2 haplotypes account for 89% of newly diagnosed cases of type 1 diabetes (T1D) in Sweden. The presence of a single copy of DQ6 confers protection. The aim of the present study is to evaluate whether the risk conferred by high risk HLA DR and DQ to T1D is similar in all regions of Sweden and see whether there are any significant regional differences. The subjects comprised 799 consecutively diagnosed T1D patients and 585 age‐, sex‐, and geography‐matched healthy controls in the age group 0–35 years. HLA typing for high‐risk haplotypes was previously performed using PCR‐SSOP and RFLP. The results showed that HLA DR3‐DR4 gave an odds ratio of 8.14 for the whole of Sweden. However, when the study group was divided into six geographical regions, subjects from Stockholm had the highest OR, followed by those from Lund, Linköping, Gothenburg, Umeå, and Uppsala. Absolute protection was conferred by the presence of DQ6 in subjects from the Linköping region, but varied in the other regions. The frequency of DR3 and DQ2, DR4 and DQ8, DR15, and DQ6 in patients showed high linkage for each region, but were different between regions. In conclusion: The risk conferred by high‐risk HLA varies in different regions for a homogenous population in Sweden. The results highlight the important role played by the various environmental factors in the precipitation of T1D.
Circulating Nucleic Acids in the Assessment of Endogenous Growth Hormone Production
Authors: Thakkar H. , Butt A.N. , Powrie J. , Holt R. , Swaminathan R.
Published: August 2009

There is growing concern about the use of recombinant human growth hormone (rhGH) by individuals taking part in competitive sports. Although rhGH is banned by the international organizations, the detection of GH doping is difficult. We postulated that rhGH will suppress endogenous GH production, which can be assessed by the measurement of mRNA for GH and growth hormone–releasing hormone (GHRH). In order to prove this concept, we undertook a pilot study to examine whether circulating nucleic acids are useful in the detection of endogenous GH production. Blood samples were collected into PAXgene tubes from 37 healthy controls and 12 acromegalic patients. RNA was extracted from the samples, cDNA was obtained, and the quantities of mRNA for GH and GHRH were measured using real‐time PCR. In acromegalic patients, median mRNA concentration for GHRH (corrected for β‐actin mRNA) was 30.7 times lower than in controls (median delta CT) value of −0.128 versus 3.927, P < 0.001). There was a significant correlation between serum IGF‐1 SD score and mRNA for GHRH (r= 0.407). In acromegalic patients, mRNA for GH was significantly higher than in controls (median values of −4.694 versus −0.044, P < 0.05). As GH production is known to decline with age, we also examined mRNA for GH and GHRH according to age subgroups. Both markers were significantly lower in the older age group (>50 years) compared to the younger age group (<34 years). These results show that mRNA for GH and GHRH can be detected in the peripheral circulation and raises the possibility of using these markers in the detection of exogenously administered GH.
Effect of Cell‐Free DNA of Patients with Cardiomyopathy and rDNA on the Frequency of Contraction of Electrically Paced Neonatal Rat Ventricular Myocytes in Culture
Authors: Bulicheva N. , Fidelina O. , Mkrtumova N. , Neverova M. , Bogush A. , Bogush M. , Roginko O. , Veiko N.
Published: August 2009

There is evidence that infarcted myocardium contributes to the increase of cell‐free DNA levels (cfDNA). We studied the effect of different human DNA fragments on the rate of contraction of the electrically paced neonatal rat ventricular myocytes in culture (spontaneously hypertensive line SHR). AT‐rich fragments of the human satellite 3 tandem repeat (1q12 region) at a concentration of 1 ng/mL increase the frequency of cardiomyocyte contractions by 2–2.5 times. GC‐rich fragments of the rDNA at 0.5 ng/mL decrease the cardiomyocyte contraction frequency by 1.5–2 times. The serum cfDNA of patients with acute myocardial infarction decreases the frequency of the contraction of cardiomyocytes proportionally to the amount of the rDNA fragments it contains. The rDNA effect is similar to the E. coli DNA effect and is presumably being realized through TLR9. In the presence of the inhibitor of these receptors, rDNA operates the same way as the fragment of the satellite 3—increasing the frequency of the cardiomyocyte contractions. Accumulation of the GC‐rich fragments of the cfDNA in the blood of the AMI patients might have an influence on the contractile function of myocardial cells.
Detection of Mutated K‐ras DNA in Urine, Plasma, and Serum of Patients with Colorectal Carcinoma or Adenomatous Polyps
Authors: Su Ying‐Hsiu , Wang Mengjun , Brenner Dean E. , Norton Pamela A. , Block Timothy M.
Published: August 2009

Our previous studies demonstrated that urine contains DNA derived from the circulation and that this DNA originated, in part, from organ sites and tumors distal to the urinary tract. To explore the potential use of DNA from urine as compared to other body fluids as a source for circulating DNA for cancer detection, the DNA concentration and the frequency of detection of mutated Kristin‐ras (K‐ras) DNA in serum, plasma, and urine were examined. The concentration of DNA in the urine was similar to that in the serum, but the DNA concentration in plasma was significantly lower than in either urine or serum (P < 0.05). When DNA derived from 10 μL of body fluid was used in each mutation assay, the detection frequency of mutated K‐ras DNA was comparable among serum, plasma, and urine. However, when DNA derived from 200 μL of body fluid was used, the incidence of detecting mutated K‐ras DNA in urine was significant higher (95%) than in either serum (35%) or plasma (40%) (P < 0.0005), suggesting that inhibitory factors in serum/plasma may be more limiting than in urine. The use and practicality of urine as a source of circulating DNA for cancer detection are discussed.
Arrayed Primer Extension for the Noninvasive Prenatal Diagnosis of β‐Thalassemia Based on Detection of Single Nucleotide Polymorphisms
Authors: Papasavva Thessalia , Kalikas Ioannis , Kyrri Andreanni , Kleanthous Marina
Published: August 2009

β‐Thalassemia is one of the most common autosomal recessive single‐gene disorders in Cyprus. Development of a noninvasive prenatal diagnostic (NIPD) assay for β‐thalassemia is based mostly on the detection of paternally inherited single nucleotide polymorphisms (SNPs) using the arrayed primer extension (APEX) method. Eleven SNPs with high degree of heterozygosity in the Cypriot population were selected and analyzed on 34 families and the informative SNPs were determined. The APEX assay was used on maternal plasma of seven families using the informative SNPs; paternal allele of the fetus was noninvasively detected in five families.
Binding and Penetration of Methylated DNA into Primary and Transformed Human Cells
Authors: Skvortsova Tatyana E. , Vlassov Valentin V. , Laktionov Pavel P.
Published: August 2009

Tumor progression was shown to be accompanied by the circulation of aberrantly methylated DNA both in the plasma and at the surface of blood cells. As the methylated DNA of tumor suppression genes in blood is a carrier of potentially harmful genetic information, we have studied the stability of methylated DNA fragments in the extracellular space, their binding to the cell surface, and their penetration into cells. It has been shown that the methylated fragments of the RARβ2 gene are more stable and penetrate into the primary and transformed cells at least twice as efficiently as the unmethylated fragments. The data obtained demonstrate the prevalence of methylated DNA in intracellular traffic compared to the same sequences of unmethylated DNA and, thus, a high transformation potential of methylated DNA.
Integrity of Cell‐Free Plasma DNA in Patients with Lung Cancer and Nonmalignant Lung Disease
Authors: Schmidt Bernd , Weickmann Sabine , Witt Christian , Fleischhacker Michael
Published: August 2009

In several papers an increased quantity of cell‐free plasma DNA as well as the presence of long DNA fragments in cell‐free plasma and serum has been described. We isolated cell‐free DNA from plasma, serum, and bronchial lavage supernatants from 33 lung cancer patients and 27 patients with a benign lung disease. The DNA was amplified by real‐time PCR, and the quantity as well as the DNA integrity was determined. We did not find significant differences between the patient populations. Our results led us to conclude that this method is not useful in a diagnostic setting and is not able to differentiate between lung cancer patients and patients with a benign lung disease.
An Improved Method for the Isolation of Free‐Circulating Plasma DNA and Cell‐Free DNA from Other Body Fluids
Authors: Kirsch Christoph , Weickmann Sabine , Schmidt Bernd , Fleischhacker Michael
Published: August 2009

Cell‐free DNA is of increasing interest in different fields, such as cancer research, prenatal diagnosis, and the diagnosis of nonmalignant diseases. The quantity of free‐circulating DNA in plasma, serum, and other body fluids is usually low and its isolation is still a challenge. Here we evaluate the application of the new commercially available NucleoSpin Plasma XS Kit for the isolation of cell‐free DNA from plasma, serum, and cell‐free bronchial lavage samples. The NucleoSpin columns proved superior to the QIAamp system in terms of DNA yield, purity, and retrieval of small DNA fragments. The procedure is fast and easy to standardize and might be valuable for laboratories working in the field of CNAPS.
Fetal Sex Identification in Maternal Plasma by Means of Short Tandem Repeats on Chromosome X
Authors: Vecchione Gennaro , Tomaiuolo Michela , Sarno Michelina , Colaizzo Donatella , Petraroli Rosella , Matteo Maria , Greco Pantaleo , Grandone Elvira , Margaglione Maurizio
Published: August 2009

Analysis of fetal DNA in maternal plasma has recently been introduced as a new method for noninvasive prenatal diagnosis. In the majority of cases, the Y chromosome–specific sequences are commonly used as a fetus‐specific marker with a high risk of false‐negative cases. We attempted to develop a sensitive and reliable X chromosome short tandem repeat (STR) multiplex PCR amplification system that is suitable for the amplification of short‐sized templates of free fetal DNA. Because of specific characteristics of fetal DNA in maternal plasma, cell‐free fetal DNA is smaller than corresponding maternal DNA, and so we selected 10 X‐STR loci in which the allele size was 250 bp. In addition, fetal sex was also investigated using the amelogenin gene in the same multiplex assay. Twenty‐six women were enrolled in the study. Eight of 26 total fetuses analyzed were male and 18 were female. In the whole sample, X‐STRs were informative with a mean of 4.84 ± 1.43. A mean of 2.67 ± 1.28 X‐STR markers per sample (range 1–5) of paternally inherited fetal alleles were detected in pregnant women carrying a female fetus. In all cases, blind determination of fetal sex by means of the identification of amelogenin and X‐STR markers was confirmed by fetal karyotyping. This study showed that this noninvasive technique is a reliable and accurate tool to investigate free fetal DNA in pregnancies within the first trimester and could be widely used in clinical research and diagnosis.
Methylation‐Specific Sequencing of GSTP1 Gene Promoter in Circulating/Extracellular DNA from Blood and Urine of Healthy Donors and Prostate Cancer Patients
Authors: Bryzgunova Olga E. , Morozkin Evgeniy S. , Yarmoschuk Sergey V. , Vlassov Valentin V. , Laktionov Pavel P.
Published: August 2009

Hypermethylated promoters of cancer‐related genes represent convenient targets for early cancer diagnosis and monitoring based on circulating/extracellular DNA (cir/exDNA) from human blood and urine. The frequency of detection of methylated tumor suppressor genes in plasma or urine samples is usually lower than in the samples of tumor tissue because of a low concentration of target DNA and potential polymorphism of cirDNA methylation. Sequencing of the methylated cir/exDNA of tumor suppression genes provides information about methylation of the cirDNA originating from the tumor cells, which is necessary for optimization of cancer diagnosis. In this work, by sequencing chemically converted cir/exDNA, we have studied the cytosine methylation profile of GSTP1 gene promoter (1001–1302, X08058) in the pool of cir/exDNA from the blood and urine of healthy men, prostate cancer (PCa) patients, and patients with benign prostatic hyperplasia (BPH). We demonstrated that the data on cir/exDNA methylation could be obtained from sequencing of the cir/exDNA from blood and urine. The DNA isolated from blood plasma and the eluates of blood cells and urine of each patient were characterized by the same methylation profile of the GSTP1 gene. The profile of GSTP1 gene methylation in the extracellular DNA of PCa patients differs from the profiles characteristic of healthy donors and patients with BPH.
A New Y Chromosome Marker for Noninvasive Fetal Gender Determination
Authors: Vainer Olga B. , Katokhin Aleksey V. , Kustov Sergey M. , Vlassov Valentin V. , Laktionov Pavel P.
Published: August 2009

We have developed a real‐time PCR assay for early fetal gender diagnostics based on the analysis of the Y chromosome–specific multicopy DAZ sequence in the circulating DNA from maternal blood. One fluorescent probe and two pairs of primers used simultaneously in one PCR reaction make it possible to detect as little as 1.5 × 10−6 ng of DNA isolated from male plasma. Fetal DNA was found both in the plasma and at the surface of cells from mother's blood. The TaqMan real‐time PCR assay of the multicopy DAZ sequence in mother's blood detected correctly the fetal gender in the tested group of pregnant women.