bluuebunny/crossref_metadata_2025 huggingface dataset:
Xanthine Oxidoreductase Is Involved in Macrophage Foam Cell Formation and Atherosclerosis Development
Authors: Kushiyama Akifumi , Okubo Hirofumi , Sakoda Hideyuki , Kikuchi Takako , Fujishiro Midori , Sato Hirokazu , Kushiyama Sakura , Iwashita Misaki , Nishimura Fusanori , Fukushima Toshiaki , Nakatsu Yusuke , Kamata Hideaki , Kawazu Shoji , Higashi Yukihito , Kurihara Hiroki , Asano Tomoichiro
Published: November 2011

Objective—

Hyperuricemia is common in patients with metabolic syndrome. We investigated the role of xanthine oxidoreductase (XOR) in atherosclerosis development, and the effects of the XOR inhibitor allopurinol on this process.

Methods and Results—

Oral administration of allopurinol to ApoE knockout mice markedly ameliorated lipid accumulation and calcification in the aorta and aortic root. In addition, allopurinol treatment or siRNA-mediated gene knockdown of XOR suppressed transformation of J774.1 murine macrophage cells, treated with acetylated LDL or very low density lipoprotein (VLDL) into foam cells. This inhibitory effect of allopurinol was also observed in primary cultured human macrophages. In contrast, overexpression of XOR promoted transformation of J774.1 cells into foam cells. Interestingly, SR-A1, SR-B1, SR-B II, and VLDL receptors in J774.1 cells were reduced by XOR knockdown, and increased by XOR overexpression. Conversely, expressions of ABCA1 and ABCG1 were increased by XOR knockdown and suppressed by XOR overexpression. Finally, productions of inflammatory cytokines accompanied by foam cell formation were also reduced by allopurinol administration.

Conclusion—

These results strongly suggest XOR activity and/or its expression level to contribute to macrophage foam cell formation. Thus, XOR inhibitors may be useful for preventing atherosclerosis.
Apolipoprotein B-100–Containing Lipoprotein Metabolism in Subjects With Lipoprotein Lipase Gene Mutations
Authors: Ooi Esther M.M. , Russell Betsy S. , Olson Eric , Sun Sam Z. , Diffenderfer Margaret R. , Lichtenstein Alice H. , Keilson Leonard , Barrett P. Hugh R. , Schaefer Ernst J. , Sprecher Dennis L.
Published: November 2011

Objective—

We investigated the impact of lipoprotein lipase (
LPL
) gene mutations on apolipoprotein B (apoB)-100 metabolism.

Methods and Results—

We studied 3 subjects with familial LPL deficiency; 14 subjects heterozygous for the
LPL
gene mutations Gly188Glu, Trp64Stop, and Ile194Thr; and 10 control subjects. Very-low-density lipoprotein (VLDL), intermediate-density lipoprotein (IDL), and low-density lipoprotein (LDL)-apoB-100 kinetics were determined in the fed state using stable isotope methods and compartmental modeling. Compared with controls, familial LPL deficiency had markedly elevated plasma triglycerides and lower VLDL-apoB-100 fractional catabolic rate (FCR), IDL-apoB-100 FCR, VLDL-to-IDL conversion, and VLDL-apoB-100 production rate (
P
<0.01). Compared with controls, Gly188Glu had higher plasma triglyceride and VLDL- and IDL-apoB-100 concentrations and lower VLDL- and IDL-apoB-100 FCR (
P
<0.05). Plasma triglycerides were not different, but IDL-apoB-100 concentration and production rate and VLDL-to-IDL conversion were lower in Trp64Stop compared with controls (
P
<0.05). No differences between controls and Ile194Thr were observed.

Conclusion—

Our results confirm that hypertriglyceridemia is a key feature of familial LPL deficiency. This is due to impaired VLDL- and IDL-apoB-100 catabolism and VLDL-to-IDL conversion. Single-allele mutations of the
LPL
gene result in modest to elevated plasma triglycerides. The changes in plasma triglycerides and apoB-100 kinetics are attributable to the effects of the LPL genotype.
Pharmacological Suppression of Hepcidin Increases Macrophage Cholesterol Efflux and Reduces Foam Cell Formation and Atherosclerosis
Authors: Saeed Omar , Otsuka Fumiyuki , Polavarapu Rohini , Karmali Vinit , Weiss Daiana , Davis Talina , Rostad Brad , Pachura Kimberly , Adams Lila , Elliott John , Taylor W. Robert , Narula Jagat , Kolodgie Frank , Virmani Renu , Hong Charles C. , Finn Aloke V.
Published: November 2011

Objective—

We recently reported that lowering of macrophage free intracellular iron increases expression of cholesterol efflux transporters ABCA1 and ABCG1 by reducing generation of reactive oxygen species. In this study, we explored whether reducing macrophage intracellular iron levels via pharmacological suppression of hepcidin can increase macrophage-specific expression of cholesterol efflux transporters and reduce atherosclerosis.

Methods and Results—

To suppress hepcidin, increase expression of the iron exporter ferroportin, and reduce macrophage intracellular iron, we used a small molecule inhibitor of bone morphogenetic protein (BMP) signaling, LDN 193189 (LDN). LDN (10 mg/kg IP b.i.d.) was administered to mice, and its effects on atherosclerosis, intracellular iron, oxidative stress, lipid efflux, and foam cell formation were measured in plaques and peritoneal macrophages. Long-term LDN administration to apolipoprotein E−/− mice increased ABCA1 immunoreactivity within intraplaque macrophages by 3.7-fold (n=8;
P
=0.03), reduced Oil Red O–positive lipid area by 50% (n=8;
P
=0.02), and decreased total plaque area by 43% (n=8;
P
=0.001). LDN suppressed liver hepcidin transcription and increased macrophage ferroportin, lowering intracellular iron and hydrogen peroxide production. LDN treatment increased macrophage ABCA1 and ABCG1 expression, significantly raised cholesterol efflux to ApoA-1, and decreased foam cell formation. All preceding LDN-induced effects on cholesterol efflux were reversed by exogenous hepcidin administration, suggesting modulation of intracellular iron levels within macrophages as the mechanism by which LDN triggers these effects.

Conclusion—

These data suggest that pharmacological manipulation of iron homeostasis may be a promising target to increase macrophage reverse cholesterol transport and limit atherosclerosis.
Pressure-Overload–Induced Subcellular Relocalization/Oxidation of Soluble Guanylyl Cyclase in the Heart Modulates Enzyme Stimulation
Authors: Tsai Emily J. , Liu Yuchuan , Koitabashi Norimichi , Bedja Djahida , Danner Thomas , Jasmin Jean-Francois , Lisanti Michael P. , Friebe Andreas , Takimoto Eiki , Kass David A.
Published: November 2011

Rationale:

Soluble guanylyl cyclase (sGC) generates cyclic guanosine monophophate (cGMP) upon activation by nitric oxide (NO). Cardiac NO–sGC-cGMP signaling blunts cardiac stress responses, including pressure-overload–induced hypertrophy. The latter itself depresses signaling through this pathway by reducing NO generation and enhancing cGMP hydrolysis.

Objective:

We tested the hypothesis that the sGC response to NO also declines with pressure-overload stress and assessed the role of heme-oxidation and altered intracellular compartmentation of sGC as potential mechanisms.

Methods and Results:

C57BL/6 mice subjected to transverse aortic constriction (TAC) developed cardiac hypertrophy and dysfunction. NO-stimulated sGC activity was markedly depressed, whereas NO- and heme-independent sGC activation by BAY 60–2770 was preserved. Total sGCα
1
and β
1
expression were unchanged by TAC; however, sGCβ
1
subunits shifted out of caveolin-enriched microdomains. NO-stimulated sGC activity was 2- to 3-fold greater in Cav3-containing lipid raft versus nonlipid raft domains in control and 6-fold greater after TAC. In contrast, BAY 60–2770 responses were >10 fold higher in non-Cav3 domains with and without TAC, declining about 60% after TAC within each compartment. Mice genetically lacking Cav3 had reduced NO- and BAY-stimulated sGC activity in microdomains containing Cav3 for controls but no change within non–Cav3-enriched domains.

Conclusions:

Pressure overload depresses NO/heme-dependent sGC activation in the heart, consistent with enhanced oxidation. The data reveal a novel additional mechanism for reduced NO-coupled sGC activity related to dynamic shifts in membrane microdomain localization, with Cav3-microdomains protecting sGC from heme-oxidation and facilitating NO responsiveness. Translocation of sGC out of this domain favors sGC oxidation and contributes to depressed NO-stimulated sGC activity.
Multipotent Vasculogenic Pericytes From Human Pluripotent Stem Cells Promote Recovery of Murine Ischemic Limb
Authors: Dar Ayelet , Domev Hagit , Ben-Yosef Oren , Tzukerman Maty , Zeevi-Levin Naama , Novak Atara , Germanguz Igal , Amit Michal , Itskovitz-Eldor Joseph
Published: November 2011

Background—

Pericytes represent a unique subtype of microvessel-residing perivascular cells with diverse angiogenic functions and multilineage developmental features of mesenchymal stem cells. Although various protocols for derivation of endothelial and/or smooth muscle cells from human pluripotent stem cells (hPSC, either embryonic or induced) have been described, the emergence of pericytes in the course of hPSC maturation has not yet been elucidated.

Methods and Results—

We found that during hPSC development, spontaneously differentiating embryoid bodies give rise to CD105
+
CD90
+
CD73
+
CD31
−
multipotent clonogenic mesodermal precursors, which can be isolated and efficiently expanded. Isolated and propagated cells expressed characteristic pericytic markers, including CD146, NG2, and platelet-derived growth factor receptor β, but not the smooth muscle cell marker α-smooth muscle actin. Coimplantation of hPSC-derived endothelial cells with pericytes resulted in functional and rapid anastomosis to the murine vasculature. Administration of pericytes into immunodeficient mice with limb ischemia promoted significant vascular and muscle regeneration. At day 21 after transplantation, recruited hPSC pericytes were found incorporated into recovered muscle and vasculature.

Conclusions—

Derivation of vasculogenic and multipotent pericytes from hPSC can be used for the development of vasculogenic models using multiple vasculogenic cell types for basic research and drug screening and can contribute to angiogenic regenerative medicine.
Levodopa Treatment Improves Functional Recovery After Experimental Stroke
Authors: Ruscher Karsten , Kuric Enida , Wieloch Tadeusz
Published: November 2011

Background and Purpose—

Delayed treatment of patients with stroke with levodopa/benserazide contributes to enhanced functional recovery, but the mechanisms involved are poorly understood. The present study was designed to investigate if levodopa/benserazide treatment improves recovery of lost neurological function and contributes to tissue reorganization in the rat brain after stroke.

Methods—

Male Wistar rats were subjected to transient occlusion of the middle cerebral artery (120 minutes) and treated with levodopa (1, 5, and 20 mg/kg)/benserazide (15 mg/kg) or saline for 12 consecutive days starting on Day 2 after transient occlusion of the middle cerebral artery. Infarct volume was determined and sensorimotor function was assessed using the rotating pole test, a 28-point neuroscore, and a cylinder test on Days 2, 7, and 14 after transient occlusion of the middle cerebral artery. The spatiotemporal expression pattern of dopamine-1 and dopamine-2 receptors and the dopamine- and cAMP-regulated neuronal phosphoprotein in reactive astrocytes were analyzed in the ischemic hemisphere as well as in cultured astrocytes.

Results—

Treatment with levodopa/benserazide significantly improved the recovery of sensorimotor function after transient occlusion of the middle cerebral artery without affecting the infarct volume. In addition, we found that different subpopulations of glial fibrillary acidic protein-positive astrocytes in the peri-infarct area express dopamine-1 receptors and dopamine-2 receptors as well as dopamine- and cAMP-regulated neuronal phosphoprotein.

Conclusions—

Our results strongly corroborate the concept of recovery enhancing actions of levodopa treatment after stroke. Also, astrocytes in the peri-infarct area may contribute to the dopamine enhanced recovery mechanisms.
Starving Where People Can See: The 1939 Bootheel Sharecroppers' Demonstration
Authors: White Ann Folino
Published: November 2011

Beginning on 10 January 1939, Missouri sharecropping families—1,300 women, men, and children—lived in makeshift shelters and out in the open on US highways 60 and 61. Their strategy of in situ exhibition manipulated conventions of minstrelsy through which media recognized them, successfully shaming landowners as violators of the moral economy.
(Gob Squad's) Revolution Now! Or Never?
Authors: Woolf Brandon
Published: November 2011

Gob Squad—love children of the contemporary European performance scene—have “occupied” the Berlin Volksbühne for their newest action: Revolution Now! So let the Molotov cocktails fly: “this revolution will be broadcast live!”
SKRIP (Screech) Orkestra: Speaking Truth
Authors: Malpede Karen
Published: November 2011

Skrip (Screech) Orkestra produces punk-noir theatre events orchestrated by a band of appealing performer-songsters from the republics of the former-Yugoslavia. This new generation of Balkan artists speaks, sings, and dances their harsh but enlivening vision of postwar existence.
Structural Integrity of the Prefrontal Cortex Modulates Electrocortical Sensitivity to Reward
Authors: Parvaz Muhammad A. , Konova Anna B. , Tomasi Dardo , Volkow Nora D. , Goldstein Rita Z.
Published: November 2011

The P300 is a known ERP component assessing stimulus value, including the value of a monetary reward. In parallel, the incentive value of reinforcers relies on the PFC, a major cortical projection region of the mesocortical reward pathway. Here we show a significant positive correlation between P300 response to money (vs. no money) with PFC gray matter volume in the OFC, ACC, and dorsolateral and ventrolateral PFC in healthy control participants. In contrast, individuals with cocaine use disorders showed compromises in both P300 sensitivity to money and PFC gray matter volume in the ventrolateral PFC and OFC and their interdependence. These results document for the first time the importance of gray matter structural integrity of subregions of PFC to the reward-modulated P300 response.
Persistent Neuronal Firing in Primary Somatosensory Cortex in the Absence of Working Memory of Trial-specific Features of the Sample Stimuli in a Haptic Working Memory Task
Authors: Wang Liping , Li Xianchun , Hsiao Steven S. , Bodner Mark , Lenz Fred , Zhou Yong-Di
Published: November 2011

Previous studies suggested that primary somatosensory (SI) neurons in well-trained monkeys participated in the haptic–haptic unimodal delayed matching-to-sample (DMS) task. In this study, 585 SI neurons were recorded in monkeys performing a task that was identical to that in the previous studies but without requiring discrimination and active memorization of specific features of a tactile or visual memorandum. A substantial number of those cells significantly changed their firing rate in the delay compared with the baseline, and some of them showed differential delay activity. These firing changes are similar to those recorded from monkeys engaged in active (working) memory. We conclude that the delay activity is not necessarily only observed as was generally thought in the situation of active memorization of different features between memoranda after those features have been actively discriminated. The delay activity observed in this study appears to be an intrinsic property of SI neurons and suggests that there exists a neural network in SI (the primary sensory cortex) for haptic working memory no matter whether the difference in features of memoranda needs to be memorized in the task or not. Over 400 SI neurons were also recorded in monkeys well-trained to discriminate two memoranda in the haptic–haptic DMS task for comparison of delay firing of SI neurons between the two different working memory tasks used in this study. The similarity observed in those two situations suggests that working memory uses already-existing memory apparatus by activating it temporarily. Our data also suggest that, through training (repetitive exposure to the stimulus), SI neurons may increase their involvement in the working memory of the memorandum.
Integrative Analysis Strategies for Mixed Data Sources
Authors: Bazeley Pat
Published: November 2011

The approach taken to integration of diverse data sources and analytical approaches in mixed methods studies is a crucial feature of those studies. Models of integration in analysis range from discussing separately generated results from different components or phases of a study together as part of the conclusion, through synthesis of data from these different components, to combination of data sources or conversion of data types to build a blended set of results. Although different models of integration are appropriate for different research settings and purposes, an overcautious approach to integration can generate invalid or weakened conclusions through a failure to consider all available information together. Strategies for making the most of opportunities to integrate process and variable data in analysis to build strong and useful conclusions are identified and illustrated through reference to a variety of mixed methods studies, including several with a focus on transition to school.
From Autonomous Areas to Non- Autonomous Areas
Authors: Paik Wooyeal , Ham Myungsik
Published: November 2011

Since the start of the market reform in 1978, approximately 30 million of China’s rural ethnic minority people have moved to China’s industrializing coastal cities. This extraordinary migration wave has included in the past twenty years more than 60% of the two million Korean Chinese. How has this migration of Korean Chinese influenced the political and economic power structures in the autonomous areas and the new non-autonomous areas? This article argues that (a) the decrease in the Korean population in established autonomous areas has weakened the minority’s autonomous power. The higher level governments of the autonomous areas have pursued more comprehensive and Han-majority-oriented macroeconomic policies—such as the abolition or merger of smaller Korean autonomous areas and the diminution of the political and economic power of larger Korean autonomous areas—rather than minority-protective policies. (b) The increasing Korean population in new migratory areas motivates these minority migrants to establish governmental and non-governmental organizations to further their political representation in their areas of settlement.
Accent, listening assessment and the potential for a shared-L1 advantage: A DIF perspective
Authors: Harding Luke
Published: November 2011

This paper reports on an investigation of the potential for a shared-L1 advantage on an academic English listening test featuring speakers with L2 accents. Two hundred and twelve second-language listeners (including 70 Mandarin Chinese L1 listeners and 60 Japanese L1 listeners) completed three versions of the University Test of English as a Second Language (UTESL) listening sub-test which featured an Australian English-accented speaker, a Japanese-accented speaker and a Mandarin Chinese-accented speaker. Differential item functioning (DIF) analyses were conducted on data from the tests which featured L2-accented speakers using two methods of DIF detection – the standardization procedure and the Mantel-Haenszel procedure – with candidates matched for ability on the test featuring the Australian English-accented speaker. Findings showed that Japanese L1 listeners were advantaged on a small number of items on the test featuring the Japanese-accented speaker, but these were balanced by items which favoured non-Japanese L1 listeners. By contrast, Mandarin Chinese L1 listeners were clearly advantaged across several items on the test featuring a Mandarin Chinese L1 speaker. The implications of these findings for claims of bias are discussed with reference to the role of speaker accent in the listening construct.
Misuse of Prescribed Stimulant Medication for ADHD and Associated Patterns of Substance Use: Preliminary Analysis Among College Students
Authors: Sepúlveda Dalissa R. , Thomas Lisl M. , McCabe Sean Esteban , Cranford James A. , Boyd Carol J. , Teter Christian J.
Published: November 2011

Objectives: To explore the prevalence and characteristics associated with college students who misuse their prescribed stimulants for attention-deficit hyperactivity disorder (ADHD) and examine diversion and substance use behaviors as a function of misuse. Methods: Cohort of 55 past-year prescribed stimulant users was identified from a random sample (n = 1738) at a large Midwestern research university following the self-administration of a web-based survey. An index was created to assess misuse of prescribed stimulants (i.e., Misuse Index). Results: Of 55 college students who reported past-year use of prescribed stimulants for ADHD, 22 (40%) endorsed at least one item on the misuse index. The most frequently endorsed misuse items were used too much (36%), self-reported misuse (19%), and intentionally used with alcohol or other drugs (19%). Misusers of prescribed stimulant medication were more likely to report cigarette smoking (p = 0.022), binge drinking (p = 0.022), illicit use of cocaine (p = 0.032), and screen positive on the Drug Abuse Screening test (DAST-10) criteria (p = 0.002). The bivariate odds ratio for the DAST-10 findings was 8.4 (95% CI: 2.0-34.6). Diversion of prescribed stimulants was common (36%) and occurred more frequently among stimulant misusers (57%; p = 0.008). Conclusion: There is a strong relationship between misuse of prescribed stimulants for ADHD and substance use behaviors, as well as other deleterious behaviors such as diversion. These findings suggest the need for close screening, assessment, and therapeutic monitoring of medication use in the college population.
A framework for GPU accelerated deformable object modeling
Authors: Shahingohar Aria , Eagleson Roy
Published: November 2011

We have developed a framework that uses multicore CPUs and GPUs found on personal computers to accelerate the computations needed for a class of deformable object modeling algorithms. In recent years there has been a growing interest in using deformable objects in computer applications such as animation, video games, garment CAD, and surgical simulation. Deformable object modeling is quite expensive computationally. However, since most of the related calculations can be parallelized, we have developed a framework that utilizes NVIDIA’s CUDA technology to accelerate a set of deformable object modeling algorithms by transferring their core computations to the GPU. Our results show that frame rates can be improved more than 20 times using GPU compared with using a multicore CPU. In addition, we have developed a method called Local Shape Matching which is an extension to the Shape Matching method. Using this new method we have achieved fast and robust simulations whose implementations have good numerical stability.
Integrin-αvβ3 regulates thrombopoietin-mediated maintenance of hematopoietic stem cells
Authors: Umemoto Terumasa , Yamato Masayuki , Ishihara Jun , Shiratsuchi Yoshiko , Utsumi Mika , Morita Yohei , Tsukui Hiroko , Terasawa Masao , Shibata Takehiko , Nishida Kohji , Kobayashi Yoshiro , Petrich Brian G. , Nakauchi Hiromitsu , Eto Koji , Okano Teruo
Published: November 2011

Throughout life, one's blood supply depends on sustained division of hematopoietic stem cells (HSCs) for self-renewal and differentiation. Within the bone marrow microenvironment, an adhesion-dependent or -independent niche system regulates HSC function. Here we show that a novel adhesion-dependent mechanism via integrin-β3 signaling contributes to HSC maintenance. Specific ligation of β3-integrin on HSCs using an antibody or extracellular matrix protein prevented loss of long-term repopulating (LTR) activity during ex vivo culture. The actions required activation of αvβ3-integrin “inside-out” signaling, which is dependent on thrombopoietin (TPO), an essential cytokine for activation of dormant HSCs. Subsequent “outside-in” signaling via phosphorylation of Tyr747 in the β3-subunit cytoplasmic domain was indispensable for TPO-dependent, but not stem cell factor-dependent, LTR activity in HSCs in vivo. This was accompanied with enhanced expression of Vps72, Mll1, and Runx1, 3 factors known to be critical for maintaining HSC activity. Thus, our findings demonstrate a mechanistic link between β3-integrin and TPO in HSCs, which may contribute to maintenance of LTR activity in vivo as well as during ex vivo culture.
Fetal hemoglobin levels and morbidity in untransfused patients with β-thalassemia intermedia
Authors: Musallam Khaled M. , Sankaran Vijay G. , Cappellini Maria Domenica , Duca Lorena , Nathan David G. , Taher Ali T.
Published: November 2011

To evaluate the association between fetal hemoglobin (HbF) levels and morbidity in β-thalassemia intermedia (TI), we analyzed data from 63 untransfused patients who had also never received HbF induction therapy. Patient records were reviewed for any history of 10 predefined morbidities. Laboratory measurements for markers of ineffective erythropoiesis were also obtained. The mean age of patients was 32.1 years, 47.6% were males, and the median HbF level was 37.2%. HbF levels correlated positively with total hemoglobin, yet negatively with growth differentiation factor-15 and non–transferrin-bound iron levels. Median HbF levels were significantly lower in patients with the majority of evaluated morbidities than in those without. There was a strong negative adjusted linear correlation between the HbF level and the total number of morbidities (R2 = 0.825, P < .001). The HbF threshold of 63.7% had 95.5% sensitivity and 100% specificity for ensuring absence of morbidity. There exists a strong association between HbF levels and morbidity in the subset of untransfused patients with TI.
The adaptor protein Shc integrates growth factor and ECM signaling during postnatal angiogenesis
Authors: Sweet Daniel Timothy , Chen Zhongming , Wiley David M. , Bautch Victoria L. , Tzima Ellie
Published: November 2011

Angiogenesis requires integration of cues from growth factors, extracellular matrix (ECM) proteins, and their receptors in endothelial cells. In the present study, we show that the adaptor protein Shc is required for angiogenesis in zebrafish, mice, and cell-culture models. Shc knockdown zebrafish embryos show defects in intersegmental vessel sprouting in the trunk. Shc flox/flox; Tie2-Cre mice display reduced angiogenesis in the retinal neovascularization model and in response to VEGF in the Matrigel plug assay in vivo. Functional studies reveal a model in which Shc is required for integrin-mediated spreading and migration specifically on fibronectin, as well as endothelial cell survival in response to VEGF. Mechanistically, Shc is required for activation of the Akt pathway downstream of both integrin and VEGF signaling, as well as for integration of signals from these 2 receptors when cells are grown on fibronectin. Therefore, we have identified a unique mechanism in which signals from 2 critical angiogenic signaling axes, integrins and VEGFR-2, converge at Shc to regulate postnatal angiogenesis.
Evaluation of High Resolution Melting analysis as an alternate tool to screen for risk alleles associated with small kidneys in Indian newborns
Authors: Raghavendra Ashwini , Siji Annes , Sridhar TS , Phadke Kishore , Vasudevan Anil
Published: November 2011

Background

Single nucleotide polymorphisms (SNPs) are the most common forms of sequence variations in the human genome. They contribute to the human phenotypic spectrum and are associated with variations in response to pathogens, drugs and vaccines. Recently, SNPs in three human genes involved in kidney development (RET, PAX2 and ALDH1A2) have been reported to be associated with variation in renal size and function. These known SNPs could potentially be used in the clinic as markers for identifying babies who may have smaller kidneys and permit close follow up for early detection of hypertension and acquired renal dysfunction. The aim of this study was to evaluate the use of High Resolution Melting technique (HRM) as a tool for detecting the known SNPs in these three genes in comparison to sequencing which is the gold standard.

Methods

High resolution melting analysis was performed on 75 DNA samples that were previously sequenced for the known polymorphisms in RET (rs1800860), PAX2 (rs11190688) and ALDH1A2 (rs7169289) genes. The SNPs were G > A transitions in RET and PAX2 and A > G in ALDH1A2 gene. A blinded assessment was performed on these samples for evaluation of the HRM technique as compared to sequencing.

Results

Each variant had a unique melt curve profile that was reproducible. The shift in melting temperature (Tm) allowed visual discrimination between the homozygous alleles (major and minor) in all three genes. The shape of the melting curve as compared to the major allele homozygous curve allowed the identification of the heterozygotes in each of the three SNPs. For validation, HRM was performed on 25 samples for each of the three SNPs. The results were compared with the sequencing results and 100% correct identification of the samples was obtained for RET, PAX2, and ALDA1H2 gene.

Conclusion

High Resolution Melting analysis is a simple, rapid and cost effective technique that could be used in a large population to identify babies with the risk alleles. These high risk children could be followed up for early detection of hypertension and acquired renal dysfunction.
Fusion primer and nested integrated PCR (FPNI-PCR): a new high-efficiency strategy for rapid chromosome walking or flanking sequence cloning
Authors: Wang Zhen , Ye Shafei , Li Jingjing , Zheng Bo , Bao Manzhu , Ning Guogui
Published: November 2011

Background

The advent of genomics-based technologies has revolutionized many fields of biological enquiry. However, chromosome walking or flanking sequence cloning is still a necessary and important procedure to determining gene structure. Such methods are used to identify T-DNA insertion sites and so are especially relevant for organisms where large T-DNA insertion libraries have been created, such as rice and Arabidopsis. The currently available methods for flanking sequence cloning, including the popular TAIL-PCR technique, are relatively laborious and slow.

Results

Here, we report a simple and effective fusion primer and nested integrated PCR method (FPNI-PCR) for the identification and cloning of unknown genomic regions flanked known sequences. In brief, a set of universal primers was designed that consisted of various 15-16 base arbitrary degenerate oligonucleotides. These arbitrary degenerate primers were fused to the 3' end of an adaptor oligonucleotide which provided a known sequence without degenerate nucleotides, thereby forming the fusion primers (FPs). These fusion primers are employed in the first step of an integrated nested PCR strategy which defines the overall FPNI-PCR protocol. In order to demonstrate the efficacy of this novel strategy, we have successfully used it to isolate multiple genomic sequences namely, 21 orthologs of genes in various species of Rosaceace, 4 MYB genes of Rosa rugosa, 3 promoters of transcription factors of Petunia hybrida, and 4 flanking sequences of T-DNA insertion sites in transgenic tobacco lines and 6 specific genes from sequenced genome of rice and Arabidopsis.

Conclusions

The successful amplification of target products through FPNI-PCR verified that this novel strategy is an effective, low cost and simple procedure. Furthermore, FPNI-PCR represents a more sensitive, rapid and accurate technique than the established TAIL-PCR and hiTAIL-PCR procedures.
Empowerment in outpatient care for patients with chronic kidney disease - from the family member's perspective
Authors: Nygårdh Annette , Wikby Kerstin , Malm Dan , Ahlstrom Gerd
Published: November 2011

Background

Family members of persons with pre-dialysis chronic kidney disease may experience feelings of vulnerability and insecurity as the disease follows its course. Against this background, the aim of the present study was to explore empowerment in outpatient care as experienced by these family members.

Methods

An inductive approach for qualitative data analysis was chosen. The study sample comprised 12 family members of pre-dialysis patients at an outpatient kidney clinic. Two interviews with each family member were subjected to content analysis to gain an understanding of empowerment from the family members' perspective.

Results

Having strength to assume the responsibility was the main theme that emerged from the following five sub-themes: Being an involved participant, Having confirming encounters, Trusting in health-care staff, Comprehending through knowledge, and Feeling left out. Four of these five sub-themes were positive. The fifth subtheme illuminated negative experience, indicating the absence of empowerment.

Conclusions

Family members' experience of empowerment is dependent on their ability to assume the responsibility for a relative with chronic kidney disease when needed. The findings emphasise the need for a family perspective and the significance of a supportive environment for family members of persons in outpatient care.
A mass spectrometric strategy for absolute quantification of Plasmodium falciparum proteins of low abundance
Authors: Southworth Paul M , Hyde John E , Sims Paul FG
Published: November 2011

Selected reaction monitoring mass spectrometry has been combined with the use of an isotopically labelled synthetic protein, made up of proteotypic tryptic peptides selected from parasite proteins of interest. This allows, for the first time, absolute quantification of proteins from Plasmodium falciparum. This methodology is demonstrated to be of sufficient sensitivity to quantify, even within whole cell extracts, proteins of low abundance from the folate pathway as well as more abundant "housekeeping" proteins.
Lack of association between apolipoprotein C3 gene polymorphisms and risk of coronary heart disease in a Han population in East China
Authors: Yu Juan , Huang Jingjing , Liang Yan , Qin Baodong , He Su , Xiao Jing , Wang Huimin , Zhong Renqian
Published: November 2011

Background

Several polymorphisms in the apolipoprotein C3 (APOC3) gene have been found association with hypertriglyceridemia(HTG), but the link with coronary heart disease(CHD) risk between ethnicities was still controversial. Among them, reseachers paid more attentions to the promoter polymorphisms T-455C and C-482T because both of them located in insulin-responsive element (IRE) and insulin was thought to exert its action by down-regulating APOC3 gene expression. The aim of this study was to investigate the association of the two polymorphisms of APOC3 with CHD in a Han population in East China.

Methods

TaqMan SNP Genotyping Assays were carried out to detect the genotypes of APOC3 gene, including the T-455C and C-482T, in 286 subjects with CHD and 325 controls without CHD. The levels of serum lipid profiles were also detected by biochemical methods.

Results

There was no difference of genotype frequencies and allele frequencies between the CHD population and the controls(P > 0.05). Compared with the most common genotype -455TT or -482CC, the variants had neither significantly increased CHD risk, nor the lipid variables showed any statistically relevant differences in the research population. The adjusted OR of CHD were 5.67 [0.27-18.74] and 0.75 [0.20-2.73] in carriers of the APOC3 -455C and -482T variants, respectively(P > 0.05). There was also no significant difference in APOC3 haplotype distribution in CHD and controls, but there was a strong linkage disequilibrium between T-455C and C-482T with D' = 0.9293, 0.8881, respectively(P < 0.0001).

Conclusions

Our data did not support a relationship between the two polymorphisms of APOC3 gene and risk of CHD in the Han population in East China.
Diagnosis of tuberculosis: the experience at a specialized diagnostic laboratory
Authors: Mashta Anita , Mishra Pooja , Philipose Sonia , Tamilzhalagan S , Mahmud Hanif , Bhaskar Sangeeta , Upadhyay Pramod
Published: November 2011

This work describes the experience at a tuberculosis clinical laboratory where relatively new TB diagnosis technologies; nucleic acid detection of two target strands, IS6110 and devR, by PCR and microscopic observation drug susceptibility (MODS) were used. The LJ culture was the gold standard. This evaluation was done from August 2007 to July 2009 on 463 sputum samples of tuberculosis suspects at a specialized tuberculosis clinic in Delhi, India.

None of the tests we evaluated can accurately detect the presence or absence of Mycobacterium tuberculosis in all the samples and smear microscopy was found to be the most reliable assay in this study.

The PCR assay could detect down to 2 pg of H37Rv DNA. Sensitivity, specificity was 0.40, 0.60 and 0.19, 0.81 for smear positive (n = 228) and negative samples (n = 235) respectively. In the MODS assay, sensitivity, specificity of 0.48, 0.52 and 0.38, 0.76 was observed for smear positive and negative samples. Sputum smear microscopy had sensitivity of 0.77 and specificity of 0.70.
Area of Wharton's jelly as an estimate of the thickness of the umbilical cord and its relationship with estimated fetal weight
Authors: Barbieri Cristiane , Cecatti Jose G , Surita Fernanda G , Costa Maria L , Marussi Emilio F , Costa Jose V
Published: November 2011

Background

To build a reference curve for the area of Wharton's jelly (WJ) in low-risk pregnancies from 13 to 40 weeks and to assess its relationship with estimated fetal weight (EFW).

Methods

2,189 low-risk pregnancies had the area of WJ estimated by ultrasound and the 10th, 50th and 90th percentiles calculated using a third-degree polynomial regression procedure. EFW by ultrasound was correlated with the measurement of the area of WJ.

Results

The area of WJ increased according to gestational age (R2 = 0.64), stabilizing from the 32nd week onwards. There was a significant linear correlation between area of WJ and EFW up to 26 weeks (R = 0.782) and after that 5t remained practically constant (R = 0.047).

Conclusion

The area of WJ increases according to gestational age, with a trend to stabilize at around 32 weeks of gestation. It is also linearly correlated with EFW only up to 26 weeks of gestation.
Drosophila G-protein-coupled receptor kinase 2 regulates cAMP-dependent Hedgehog signaling
Authors: Cheng Shuofei , Maier Dominic , Hipfner David R.
Published: November 2011

G-protein-coupled receptor kinases (GRKs) play a conserved role in Hedgehog (Hh) signaling. In several systems, GRKs are required for efficient Hh target gene expression. Their principal target appears to be Smoothened (Smo), the intracellular signal-generating component of the pathway and a member of the G-protein-coupled receptor (GPCR) protein family. In Drosophila, a GRK called Gprk2 is needed for internalization and downregulation of activated Smo, consistent with the typical role of these kinases in negatively regulating GPCRs. However, Hh target gene activation is strongly impaired in gprk2 mutant flies, indicating that Gprk2 must also positively regulate Hh signaling at some level. To investigate its function in signaling, we analyzed several different readouts of Hh pathway activity in animals or cells lacking Gprk2. Surprisingly, although target gene expression was impaired, Smo-dependent activation of downstream components of the signaling pathway was increased in the absence of Gprk2. This suggests that Gprk2 does indeed play a role in terminating Smo signaling. However, loss of Gprk2 resulted in a decrease in cellular cAMP concentrations to a level that was limiting for Hh target gene activation. Normal expression of target genes was restored in gprk2 mutants by stimulating cAMP production or activating the cAMP-dependent Protein kinase A (Pka). Our results suggest that direct regulation of Smo by Gprk2 is not absolutely required for Hh target gene expression. Gprk2 is important for normal cAMP regulation, and thus has an indirect effect on the activity of Pka-regulated components of the Hh pathway, including Smo itself.
The role of the histone H2A ubiquitinase Sce in Polycomb repression
Authors: Gutiérrez Luis , Oktaba Katarzyna , Scheuermann Johanna C. , Gambetta Maria Cristina , Ly-Hartig Nga , Müller Jürg
Published: November 2011

Polycomb group (PcG) proteins exist in multiprotein complexes that modify chromatin to repress transcription. Drosophila PcG proteins Sex combs extra (Sce; dRing) and Posterior sex combs (Psc) are core subunits of PRC1-type complexes. The Sce:Psc module acts as an E3 ligase for monoubiquitylation of histone H2A, an activity thought to be crucial for repression by PRC1-type complexes. Here, we created an Sce knockout allele and show that depletion of Sce results in loss of H2A monoubiquitylation in developing Drosophila. Genome-wide profiling identified a set of target genes co-bound by Sce and all other PRC1 subunits. Analyses in mutants lacking individual PRC1 subunits reveals that these target genes comprise two distinct classes. Class I genes are misexpressed in mutants lacking any of the PRC1 subunits. Class II genes are only misexpressed in animals lacking the Psc-Su(z)2 and Polyhomeotic (Ph) subunits but remain stably repressed in the absence of the Sce and Polycomb (Pc) subunits. Repression of class II target genes therefore does not require Sce and H2A monoubiquitylation but might rely on the ability of Psc-Su(z)2 and Ph to inhibit nucleosome remodeling or to compact chromatin. Similarly, Sce does not provide tumor suppressor activity in larval tissues under conditions in which Psc-Su(z)2, Ph and Pc show such activity. Sce and H2A monoubiquitylation are therefore only crucial for repression of a subset of genes and processes regulated by PRC1-type complexes. Sce synergizes with the Polycomb repressive deubiquitinase (PR-DUB) complex to repress transcription at class I genes, suggesting that H2A monoubiquitylation must be appropriately balanced for their transcriptional repression.
Seneca’s Presence in Pliny’s Epistle 1. 12
Authors: Tzounakas Spyridon
Published: November 2011

In his exitus letter on the death of Corellius Rufus (Ep. 1. 12), Pliny attempts to present his dead friend with Stoic characteristics. Not only does Corellius follow the Stoic view on suicide in the case of an incurable disease, but also he is implicitly compared to the Stoic sapiens. This is greatly facilitated by allusions to Seneca’s Epistulae Morales, and in particular to epistle 85, where the sapiens is described and dolor is presented as indifferent to the pursuit of virtus. These allusions allow Pliny to both praise Corellius and at the same time express his objections to the prose consolationes of his predecessors.
MANAGEMENT OF ENDOCRINE DISEASE: Beneficial effect of real-time continuous glucose monitoring system on glycemic control in type 1 diabetic patients: systematic review and meta-analysis of randomized trials
Authors: Szypowska A , Ramotowska A , Dżygało K , Golicki D
Published: November 2011

Objective

Real-time continuous glucose monitoring (RT-CGM) provides detailed information on glucose patterns and trends, thus allowing the patients to manage their diabetes more effectively.

Design

The aim of this study was to explore the potential beneficial effects of the use of RT-CGM on diabetes management compared with self blood glucose measurement (SBGM) in patients with type 1 diabetes mellitus (T1DM), by means of a systematic review and meta-analysis of randomized controlled trials (RCTs).

Methods

MEDLINE, EMBASE, and the Cochrane Library were searched through by two independent investigators for RCTs concerning the use of RT-CGM in patients with T1DM. Only studies with a similar insulin regimen in the experimental and control groups were included in the analysis.

Results

Seven RCTs (n=948) met the inclusion criteria. Combined data from all studies showed better HbA1c reduction in subjects using RT-CGM compared with those using SBGM (mean difference (MD) −0.25; 95% confidence interval (95% CI): from −0.34 to −0.17; P<0.001). Patients treated with insulin pump and RT-CGM had a lower HbA1c level compared with subjects managed with insulin pump and SBGM (four RCTs, n=497; MD −0.26; 95% CI: from −0.43 to −0.10; P=0.002). The benefits of applying RT-CGM were not associated with an increasing rate of major hypoglycemic episodes. The use of RT-CGM for over 60–70% of time was associated with a significant lowering of HbA1c.

Conclusions

RT-CGM is more beneficial than SBGM in reducing HbA1c in patients with type 1 diabetes. Further studies are needed to evaluate the efficacy of this system in the pediatric population, especially in very young children.
MECHANISMS IN ENDOCRINOLOGY: Recent advances in cardiovascular aspects of polycystic ovary syndrome
Authors: Sathyapalan T , Atkin S L
Published: November 2011

Polycystic ovary syndrome (PCOS) has been associated with increased cardiovascular risk (CVR) markers, but population studies have not clarified whether there is an increase in cardiovascular morbidity and mortality. Four different PCOS phenotypes resulted from the Rotterdam criteria that may differ in their CVR potential, thus introducing further complexity. This has led to studies using surrogate CVR markers including biomarkers in blood and imaging such as flow-mediated vasodilatation. In PCOS, both peripheral and central insulin resistance (IR) have been shown. Weight loss has been shown to improve IR and visceral fat, while insulin sensitizer therapies with metformin or thiazolidinediones improve IR and endothelial dysfunction. IR is also found in non-alcoholic fatty liver disease that in turn is very common in PCOS; studies have suggested that IR may be improved by treatment with metformin and omega-3 fish oils. PCOS patients have a more dyslipidemic phenotype that is worse in ‘classical PCOS’ associated with a higher CVR. Studies with atorvastatin and simvastatin have reported a decrease in the lipid parameters and an improvement in CVR indices including IR, but it is unclear whether this is due to their lipid-lowering action or a pleiotropic effect of the statin. In this expert opinion review, the relevant literature published during the last 2 years was considered. It focuses on some recent important data that has emerged while also exposing the gaps that remain in our knowledge that need to be addressed.
Molecular characterization of Trypanosoma cruzi Mexican strains and their behavior in the mouse experimental model
Authors: Gómez-Hernández César , Rezende-Oliveira Karine , Nascentes Gabriel Antônio Nogueira , Batista Lara Rocha , Kappel Henrique Borges , Martinez-Ibarra José Alejandro , Trujillo Contreras Francisco , Lages-Silva Eliane , Ramírez Luis Eduardo
Published: November 2011

INTRODUCTION: For a long time, the importance of Chagas disease in Mexico, where many regarded it as an exotic malady, was questioned. Considering the great genetic diversity among isolates of Trypanosoma cruzi, the importance of this biological characterization, and the paucity of information on the clinical and biological aspects of Chagas disease in Mexico, this study aimed to identify the molecular and biological characterization of Trypanosoma cruzi isolates from different endemic areas of this country, especially of the State of Jalisco. METHODS: Eight Mexican Trypanosoma cruzi strains were biologically and genetically characterized (PCR specific for Trypanosoma cruzi, multiplex-PCR, amplification of space no transcript of the genes of the mini-exon, amplification of polymorphic regions of the mini-exon, classification by amplification of intergenic regions of the spliced leader genes, RAPD - (random amplified polymorphic DNA). RESULTS: Two profiles of parasitaemia were observed, patent (peak parasitaemia of 4.6×10(6) to 10(7) parasites/mL) and subpatent. In addition, all isolates were able to infect 100% of the animals. The isolates mainly displayed tropism for striated (cardiac and skeletal) muscle. PCR amplification of the mini-exon gene classified the eight strains as TcI. The RAPD technique revealed intraspecies variation among isolates, distinguishing strains isolated from humans and triatomines and according to geographic origin. CONCLUSIONS: The Mexican T. cruzi strains are myotrophic and belong to group TcI.