The Somatic Genomic Landscape of Glioblastoma

Abstract: We describe the landscape of somatic genomic alterations based on multidimensional and comprehensive characterization of more than 500 glioblastoma tumors (GBMs).

Abstract: We identify several novel mutated genes as well as complex rearrangements of signature receptors, including EGFR and PDGFRA.

Abstract: TERT promoter mutations are shown to correlate with elevated mRNA expression, supporting a role in telomerase reactivation.

Abstract: Correlative analyses confirm that the survival advantage of the proneural subtype is conferred by the G-CIMP phenotype, and MGMT DNA methylation may be a predictive biomarker for treatment response only in classical subtype GBM.

Abstract: Integrative analysis of genomic and proteomic profiles challenges the notion of therapeutic inhibition of a pathway as an alternative to inhibition of the target itself.

Abstract: These data will facilitate the discovery of therapeutic and diagnostic target candidates, the validation of research and clinical observations and the generation of unanticipated hypotheses that can advance our molecular understanding of this lethal cancer.


Radioiodine treatment of multinodular non-toxic goitre.

Abstract: OBJECTIVE To investigate the long term effect of radioactive iodine on thyroid function and size in patients with non-toxic multinodular goitre.

Abstract: DESIGN Consecutive patients with multinodular non-toxic goitre selected for radioactive iodine treatment and followed for a minimum of 12 months (median 48 months) after an intended dose of 3.7 MBq/g thyroid tissue corrected to a 100% uptake of iodine-131 in 24 hours.

Abstract: PATIENTS 69 patients with a growing multinodular non-toxic goitre causing local compression symptoms or cosmetic inconveniences.

Abstract: The treatment was chosen because of a high operative risk, previous thyroidectomy, or refusal to be operated on.

Abstract: MAIN OUTCOME MEASUREMENTS Standard thyroid function variables and ultrasonically determined thyroid volume before treatment as well as 1, 2, 3, 6, and 12 months after treatment and then once a year.

Abstract: RESULTS 56 patients were treated with a single dose of 131I, 12 with two doses, and one with four doses.

Abstract: In 45 patients treated with one dose and remaining euthyroid the median thyroid volume was reduced from 73 (interquartile range 50-106) ml to 29 (23-48) ml at 24 months in the 39 patients in whom this was measured during follow up.

Abstract: The median reduction was 40 (22-48) ml (60% reduction, p < 0.0001), half of which occurred within three months.

Abstract: Patients treated with two doses as well as those developing hypothyroidism and hyperthyroidism had a significant reduction in thyroid volume.

Abstract: Eleven patients developed hypothyroidism (cumulative five year risk 22%, 95% confidence interval 4.8% to 38.4%).

Abstract: Side effects were few: three cases of hyperthyroidism and two cases of radiation thyroiditis.

Abstract: Only one patient was dissatisfied with the result; she was referred for operation six months after treatment.

Abstract: CONCLUSIONS A substantial reduction in thyroid volume accompanied by a low incidence of hypothyroidism and few side effects makes the use of radioactive iodine an attractive alternative to surgery in selected cases of non-toxic multinodular goitre.


Alterations in microRNA-124 and AMPA receptors contribute to social behavioral deficits in frontotemporal dementia

Abstract: Neurodegenerative diseases, such as frontotemporal dementia (FTD), are often associated with behavioral deficits, but the underlying anatomical and molecular causes remain poorly understood.

Abstract: Here we show that forebrain-specific expression of FTD-associated mutant CHMP2B in mice causes several age-dependent neurodegenerative phenotypes, including social behavioral impairments.

Abstract: The social deficits were accompanied by a change in AMPA receptor (AMPAR) composition, leading to an imbalance between Ca(2+)-permeable and Ca(2+)-impermeable AMPARs.

Abstract: Expression of most AMPAR subunits was regulated by the brain-enriched microRNA miR-124, whose abundance was markedly decreased in the superficial layers of the cerebral cortex of mice expressing the mutant CHMP2B.

Abstract: We found similar changes in miR-124 and AMPAR levels in the frontal cortex and induced pluripotent stem cell-derived neurons from subjects with behavioral variant FTD.

Abstract: Moreover, ectopic miR-124 expression in the medial prefrontal cortex of mutant mice decreased AMPAR levels and partially rescued behavioral deficits.

Abstract: Knockdown of the AMPAR subunit Gria2 also alleviated social impairments.

Abstract: Our results identify a previously undescribed mechanism involving miR-124 and AMPARs in regulating social behavior in FTD and suggest a potential therapeutic avenue.


Activated AMPK boosts the Nrf2/HO-1 signaling axis—A role for the unfolded protein response

Abstract: In light of the emerging interplay between redox and metabolic signaling pathways we investigated the potential cross talk between nuclear factor E2-related factor 2 (Nrf2) and AMP-activated kinase (AMPK), central regulators of the cellular redox and energy balance, respectively.

Abstract: Making use of xanthohumol (XN) as an activator of both the AMPK and the Nrf2 signaling pathway we show that AMPK exerts a positive influence on Nrf2/heme oxygenase (HO)-1 signaling in mouse embryonic fibroblasts.

Abstract: Genetic ablation and pharmacological inhibition of AMPK blunts Nrf2-dependent HO-1 expression by XN already at the mRNA level.

Abstract: XN leads to AMPK activation via interference with mitochondrial function and activation of liver kinase B1 as upstream AMPK kinase.

Abstract: The subsequent AMPK-mediated enhancement of the Nrf2/HO-1 response does not depend on inhibition of the mammalian target of rapamycin, inhibition of glycogen synthase kinase 3β, or altered abundance of Nrf2 (total and nuclear).

Abstract: However, reduced endoplasmic reticulum stress was identified and elaborated as a step in the AMPK-augmented Nrf2/HO-1 response.

Abstract: Overall, we shed more light on the hitherto incompletely understood cross talk between the LKB1/AMPK and the Nrf2/HO-1 axis revealing for the first time involvement of the unfolded protein response as an additional player and suggesting tight cooperation between signaling pathways controlling cellular redox, energy, or protein homeostasis.


The IDEAL recommendations and urological innovation

Abstract: Like other branches of surgery, Urology has encountered major challenges in aligning the research processes by which new interventions are assessed with the principles of Evidence-Based Medicine.

Abstract: This article explains the IDEAL framework and recommendations and illustrates how they might affect the evaluation of current controversial urological procedures.

Abstract: From an inside perspective, we provide an overview of the efforts of the IDEAL Working Group to date with special emphasis on the field of Urology.

Abstract: There are clear differences between drugs and interventions in the natural history of innovations.

Abstract: Since the conventional framework for conducting trials of new treatments is largely based on the former, the evaluation of surgical innovations using the same template can encounter significant problems.

Abstract: Difficulties in performing randomized controlled trials of surgical techniques and the persistence of the case series as an important feature of the scientific literature have been the two most controversial aspects of this mismatch between the subject of research and the methodology used.

Abstract: The IDEAL framework provides a description of the process of innovation and development for surgical trials, and the associated recommendations provide a suggested alternative approach to developing study designs, which are appropriate for the specific problems of new techniques.

Abstract: IDEAL provides a new framework for surgical innovation that was developed with broad stakeholder input from the surgical community and is expected to have a transformative impact on the way that urologists perform clinical research.


Prolonged conservative care versus early surgery in patients with sciatica caused by lumbar disc herniation: two year results of a randomised controlled trial.

Abstract: OBJECTIVES To evaluate the effects of early lumbar disc surgery compared with prolonged conservative care for patients with sciatica over two years of follow-up.

Abstract: DESIGN Randomised controlled trial.

Abstract: SETTING Nine Dutch hospitals.

Abstract: PARTICIPANTS 283 patients with 6-12 weeks of sciatica.

Abstract: INTERVENTIONS Early surgery or an intended six months of continued conservative treatment, with delayed surgery if needed.

Abstract: MAIN OUTCOME MEASURES Scores from Roland disability questionnaire for sciatica, visual analogue scale for leg pain, and Likert self rating scale of global perceived recovery.

Abstract: RESULTS Of the 141 patients assigned to undergo early surgery, 125 (89%) underwent microdiscectomy.

Abstract: Of the 142 patients assigned to conservative treatment, 62 (44%) eventually required surgery, seven doing so in the second year of follow-up.

Abstract: There was no significant overall difference between treatment arms in disability scores during the first two years (P=0.25).

Abstract: Improvement in leg pain was faster for patients randomised to early surgery, with a significant difference between "areas under the curves" over two years (P=0.05).

Abstract: This short term benefit of early surgery was no longer significant by six months and continued to narrow between six months and 24 months.

Abstract: Patient satisfaction decreased slightly between one and two years for both groups.

Abstract: At two years 20% of all patients reported an unsatisfactory outcome.

Abstract: CONCLUSIONS Early surgery achieved more rapid relief of sciatica than conservative care, but outcomes were similar by one year and these did not change during the second year.

Abstract: TRIAL REGISTRY ISRCT

Abstract: No 26872154.


Human papillomavirus testing for the detection of high-grade cervical intraepithelial neoplasia and cancer: final results of the POBASCAM randomised controlled trial.

Abstract: BACKGROUND Human papillomavirus (HPV) testing is more sensitive for the detection of high-grade cervical lesions than is cytology, but detection of HPV by DNA screening in two screening rounds 5 years apart has not been assessed.

Abstract: The aim of this study was to assess whether HPV DNA testing in the first screen decreases detection of cervical intraepithelial neoplasia (CIN) grade 3 or worse, CIN grade 2 or worse, and cervical cancer in the second screening.

Abstract: METHODS In this randomised trial, women aged 29-56 years participating in the cervical screening programme in the Netherlands were randomly assigned to receive HPV DNA (GP5+/6+-PCR method) and cytology co-testing or cytology testing alone, from January, 1999, to September, 2002.

Abstract: Randomisation (in a 1:1 ratio) was done with computer-generated random numbers after the cervical specimen had been taken.

Abstract: At the second screening 5 years later, HPV DNA and cytology co-testing was done in both groups; researchers were masked to the patient's assignment.

Abstract: The primary endpoint was the number of CIN grade 3 or worse detected.

Abstract: Analysis was done by intention to screen.

Abstract: The trial is now finished and is registered, number ISRCTN20781131.

Abstract: FINDINGS 22,420 women were randomly assigned to the intervention group and 22 518 to the control group; 19 999 in the intervention group and 20,106 in the control group were eligible for analysis at the first screen.

Abstract: At the second screen, 19 579 women in the intervention group and 19,731 in the control group were eligible, of whom 16,750 and 16,743, respectively, attended the second screen.

Abstract: In the second round, CIN grade 3 or worse was less common in the intervention group than in the control group (88 of 19 579 in the intervention group vs 122 of 19,731 in the control group; relative risk 0·73, 95% CI 0·55-0·96; p=0·023).

Abstract: Cervical cancer was also less common in the intervention group than in the control group (four of 19 579 in the intervention group vs 14 of 19,731; 0·29, 0·10-0·87; p=0·031).

Abstract: In the baseline round, detection of CIN grade 3 or worse did not differ significantly between groups (171 of 19 999 vs 150 of 20,106; 1·15, 0·92-1·43; p=0·239) but was significantly more common in women with normal cytology (34 of 19,286 vs 12 of 19,373; 2·85, 1·47-5·49; p=0·001).

Abstract: Furthermore, significantly more cases of CIN grade 2 or worse were detected in the intervention group than in the control group (267 of 19 999 vs 215 of 20,106; 1·25, 1·05-1·50; p=0·015).

Abstract: In the second screen, fewer HPV16-positive CIN grade 3 or worse were detected in the intervention group than in the control group (17 of 9481 vs 35 of 9354; 0·48, 0·27-0·85; p=0·012); detection of non-HPV16-positive CIN grade 3 or worse did not differ between groups (25 of 9481 vs 25 of 9354; 0·99, 0·57-1·72; p=1·00).

Abstract: The cumulative detection of CIN grade 3 or worse and CIN grade 2 or worse did not differ significantly between study arms, neither for the whole study group (CIN grade 3 or worse: 259 of 19 999 vs 272 of 20,106; 0·96, 0·81-1·14, p=0·631; CIN grade 2 or worse: 427 of 19 999 vs 399 of 20,106; 1·08, 0·94-1·24; p=0·292), nor for subgroups of women invited for the first time (CIN grade 3 or worse in women aged 29-33 years: 102 of 3139 vs 105 of 3128; 0·97, 0·74-1·27; CIN grade 2 or worse in women aged 29-33 years: 153 of 3139 vs 151 of 3128; 1·01, 0·81-1·26; CIN grade 3 or worse in women aged 34-56 years: 157 of 16,860 vs 167 of 16 978; 0·95, 0·76-1·18; CIN grade 2 or worse in women aged 34-56 years: 274 of 16,860 vs 248 of 16 978; 1·11, 0·94-1·32).

Abstract: INTERPRETATION Implementation of HPV DNA testing in cervical screening leads to earlier detection of clinically relevant CIN grade 2 or worse, which when adequately treated, improves protection against CIN grade 3 or worse and cervical cancer.

Abstract: Early detection of high-grade cervical legions caused by HPV16 was a major component of this benefit.

Abstract: Our results lend support to the use of HPV DNA testing for all women aged 29 years and older.

Abstract: FUNDING Zorg Onderzoek Nederland (Netherlands Organisation for Health Research and Development).


Epigenetic therapy activates type I interferon signaling in murine ovarian cancer to reduce immunosuppression and tumor burden.

Abstract: Ovarian cancer is the most lethal of all gynecological cancers, and there is an urgent unmet need to develop new therapies.

Abstract: Epithelial ovarian cancer (EOC) is characterized by an immune suppressive microenvironment, and response of ovarian cancers to immune therapies has thus far been disappointing.

Abstract: We now find, in a mouse model of EOC, that clinically relevant doses of DNA methyltransferase and histone deacetylase inhibitors (DNMTi and HDACi, respectively) reduce the immune suppressive microenvironment through type I IFN signaling and improve response to immune checkpoint therapy.

Abstract: These data indicate that the type I IFN response is required for effective in vivo antitumorigenic actions of the DNMTi 5-azacytidine (AZA).

Abstract: Through type I IFN signaling, AZA increases the numbers of CD45+ immune cells and the percentage of active CD8+ T and natural killer (NK) cells in the tumor microenvironment, while reducing tumor burden and extending survival.

Abstract: AZA also increases viral defense gene expression in both tumor and immune cells, and reduces the percentage of macrophages and myeloid-derived suppressor cells in the tumor microenvironment.

Abstract: The addition of an HDACi to AZA enhances the modulation of the immune microenvironment, specifically increasing T and NK cell activation and reducing macrophages over AZA treatment alone, while further increasing the survival of the mice.

Abstract: Finally, a triple combination of DNMTi/HDACi plus the immune checkpoint inhibitor α-PD-1 provides the best antitumor effect and longest overall survival, and may be an attractive candidate for future clinical trials in ovarian cancer.


IGF2 is parentally imprinted during human embryogenesis and in the Beckwith–Wiedemann syndrome

Abstract: The phenomenon of parental imprinting involves the preferential expression of one parental allele of a subset of chromosomal genes and has so far only been documented in the mouse.

Abstract: We show here, by exploiting sequence polymorphisms in exon nine of the human insulin–like growth factor 2 (IGF2) gene, that only the paternally–inherited allele is active in embryonic and extra–embryonic cells from first trimester pregnancies.

Abstract: In addition, only the paternal allele is expressed in tissues from a patient who suffered from Beckwith–Wiedemann syndrome.

Abstract: Thus the parental imprinting of IGF2 appears to be evolutionarily conserved from mouse to man and has implications for the generation of the Beckwith–Wiedemann syndrome.


Safety and reliability of Radio Frequency Identification Devices in Magnetic Resonance Imaging and Computed Tomography

Abstract: BACKGROUND Radio Frequency Identification (RFID) devices are becoming more and more essential for patient safety in hospitals.

Abstract: The purpose of this study was to determine patient safety, data reliability and signal loss wearing on skin RFID devices during magnetic resonance imaging (MRI) and computed tomography (CT) scanning.

Abstract: METHODS Sixty RFID tags of the type I-Code SLI, 13.56 MHz, ISO 18000-3.1 were tested: Thirty type 1, an RFID tag with a 76 x 45 mm aluminum-etched antenna and 30 type 2, a tag with a 31 x 14 mm copper-etched antenna.

Abstract: The signal loss, material movement and heat tests were performed in a 1.5 T and a 3 T MR system.

Abstract: For data integrity, the tags were tested additionally during CT scanning.

Abstract: Standardized function tests were performed with all transponders before and after all imaging studies.

Abstract: RESULTS There was no memory loss or data alteration in the RFID tags after MRI and CT scanning.

Abstract: Concerning heating (a maximum of 3.6 degrees C) and device movement (below 1 N/kg) no relevant influence was found.

Abstract: Concerning signal loss (artifacts 2 - 4 mm), interpretability of MR images was impaired when superficial structures such as skin, subcutaneous tissues or tendons were assessed.

Abstract: CONCLUSIONS Patients wearing RFID wristbands are safe in 1.5 T and 3 T MR scanners using normal operation mode for RF-field.

Abstract: The findings are specific to the RFID tags that underwent testing.


LXR-Dependent Gene Expression Is Important for Macrophage Survival and the Innate Immune Response

Abstract: The liver X receptors (LXRs) are nuclear receptors with established roles in the regulation of lipid metabolism.

Abstract: We now show that LXR signaling not only regulates macrophage cholesterol metabolism but also impacts antimicrobial responses.

Abstract: Mice lacking LXRs are highly susceptible to infection with the intracellular bacteria Listeria monocytogenes (LM).

Abstract: Bone marrow transplant studies point to altered macrophage function as the major determinant of susceptibility.

Abstract: LXR-null macrophages undergo accelerated apoptosis when challenged with LM and exhibit defective bacterial clearance in vivo.

Abstract: These defects result, at least in part, from loss of regulation of the antiapoptotic factor SPalpha, a direct target for regulation by LXRalpha.

Abstract: Expression of LXRalpha or SPalpha in macrophages inhibits apoptosis in the setting of LM infection.

Abstract: Our results demonstrate that LXR-dependent gene expression plays an unexpected role in innate immunity and suggest that common nuclear receptor pathways mediate macrophage responses to modified lipoproteins and intracellular pathogens.


Alterations on peripheral B cell subsets following an acute uncomplicated clinical malaria infection in children

Abstract: BACKGROUND The effects of Plasmodium falciparum on B-cell homeostasis have not been well characterized.

Abstract: This study investigated whether an episode of acute malaria in young children results in changes in the peripheral B cell phenotype.

Abstract: METHODS Using flow-cytofluorimetric analysis, the B cell phenotypes found in the peripheral blood of children aged 2-5 years were characterized during an episode of acute uncomplicated clinical malaria and four weeks post-recovery and in healthy age-matched controls.

Abstract: RESULTS There was a significant decrease in CD19+ B lymphocytes during acute malaria.

Abstract: Characterization of the CD19+ B cell subsets in the peripheral blood based on expression of IgD and CD38 revealed a significant decrease in the numbers of naive 1 CD38-IgD+ B cells while there was an increase in CD38+IgD- memory 3 B cells during acute malaria.

Abstract: Further analysis of the peripheral B cell phenotype also identified an expansion of transitional CD10+CD19+ B cells in children following an episode of acute malaria with up to 25% of total CD19+ B cell pool residing in this subset.

Abstract: CONCLUSION Children experiencing an episode of acute uncomplicated clinical malaria experienced profound disturbances in B cell homeostasis.


SETD2: an epigenetic modifier with tumor suppressor functionality

Abstract: In the past decade important progress has been made in our understanding of the epigenetic regulatory machinery.

Abstract: It has become clear that genetic aberrations in multiple epigenetic modifier proteins are associated with various types of cancer.

Abstract: Moreover, targeting the epigenome has emerged as a novel tool to treat cancer patients.

Abstract: Recently, the first drugs have been reported that specifically target SETD2-negative tumors.

Abstract: In this review we discuss the studies on the associated protein, Set domain containing 2 (SETD2), a histone modifier for which mutations have only recently been associated with cancer development.

Abstract: Our review starts with the structural characteristics of SETD2 and extends to its corresponding function by combining studies on SETD2 function in yeast, Drosophila, Caenorhabditis elegans, mice, and humans.

Abstract: SETD2 is now generally known as the single human gene responsible for trimethylation of lysine 36 of Histone H3 (H3K36).

Abstract: H3K36me3 readers that recruit protein complexes to carry out specific processes, including transcription elongation, RNA processing, and DNA repair, determine the impact of this histone modification.

Abstract: Finally, we describe the prevalence of SETD2-inactivating mutations in cancer, with the highest frequency in clear cell Renal Cell Cancer, and explore how SETD2-inactivation might contribute to tumor development.


Updated energy budgets for neural computation in the neocortex and cerebellum.

Abstract: The brain's energy supply determines its information processing power, and generates functional imaging signals.

Abstract: The energy use on the different subcellular processes underlying neural information processing has been estimated previously for the grey matter of the cerebral and cerebellar cortex.

Abstract: However, these estimates need reevaluating following recent work demonstrating that action potentials in mammalian neurons are much more energy efficient than was previously thought.

Abstract: Using this new knowledge, this paper provides revised estimates for the energy expenditure on neural computation in a simple model for the cerebral cortex and a detailed model of the cerebellar cortex.

Abstract: In cerebral cortex, most signaling energy (50%) is used on postsynaptic glutamate receptors, 21% is used on action potentials, 20% on resting potentials, 5% on presynaptic transmitter release, and 4% on transmitter recycling.

Abstract: In the cerebellar cortex, excitatory neurons use 75% and inhibitory neurons 25% of the signaling energy, and most energy is used on information processing by non-principal neurons: Purkinje cells use only 15% of the signaling energy.

Abstract: The majority of cerebellar signaling energy use is on the maintenance of resting potentials (54%) and postsynaptic receptors (22%), while action potentials account for only 17% of the signaling energy use.


Plasma homocysteine is a risk factor for recurrent vascular events in young patients with an ischaemic stroke or TIA

Abstract: Young patients with an ischaemic stroke or transient ischaemic attack (TIA) often have no vascular risk factors.

Abstract: Hyperhomocysteinaemia is an established risk factor for stroke in elderly patients but it is uncertain whether it is also important for the prognosis of young ischaemic stroke and TIA patients.

Abstract: We examined the possible effect of the plasma homocysteine level on the risk of recurrent vascular events in patients between 18 and 45 years of age.

Abstract: The study population consisted of 161 consecutive patients with a recent cerebral infarction or TIA.

Abstract: Data on the primary event and the homocysteine level were collected retrospectively from hospital records.

Abstract: General practitioners and patients were contacted by telephone to record vascular events and the type of medication used during the follow–up period.

Abstract: Vascular events included cerebral infarction, TIA, pulmonary embolism, venous thrombosis, myocardial infarction and peripheral arterial disease.

Abstract: A Kaplan- Meier curve showed a dose effect relationship between event-free survival time and tertiles of the homocysteine level (Log rank statistic 5.91; p = 0.05).

Abstract: The Cox hazard ratio, after adjustment for homocysteine lowering treatment, was 1.7 (95 % CI, 1.1 to 2.8) for any vascular outcome event, 1.9 (95% CI, 1.1 to 3.0) for arterial outcome events and 1.8 (95 % CI, 1.1 to 2.9) for cerebral outcome events.

Abstract: In spite of our small number of outcome events we found a significant association at the 95% confidence level between homocysteine level and the risk of recurrent vascular events in young patients with an ischaemic stroke or TIA.

Abstract: The association is of the same magnitude as in elderly people.


LDL inhibits the mediation of cholesterol efflux from macrophage foam cells by apoA-I-containing lipoproteins. A putative mechanism for foam cell formation.

Abstract: Although the accumulation of cholesterol in macrophages appears to be an initial step in atherogenesis, low-density lipoprotein (LDL), a major risk factor for atherosclerosis, does not promote cholesterol accumulation in macrophages in its native form.

Abstract: On the other hand, apolipoprotein (apo) A-I-containing lipoprotein removes cholesterol from cholesterol-loaded macrophages (foam cells) and prevents cholesterol from accumulating in the cells.

Abstract: We examined the effect of LDL on cholesterol removal by two species of apoA-I-containing lipoproteins, one containing only apoA-I (LpA-I) and the other containing apoA-I and apoA-II (LpA-I/A-II).

Abstract: When foam cells were incubated with LpA-I or LpA-I/A-II, cellular cholesterol mass was reduced.

Abstract: In contrast, when LDL was added, the cholesterol-reducing capacities of these lipoproteins were dose-dependently inhibited by LDL.

Abstract: In the presence of LDL, LpA-I and LpA-I/A-II removed free cholesterol preferentially from LDL rather than from the plasma membrane of foam cells.

Abstract: In addition, a fair amount of cellular cholesterol was directly moved to LDL rather than to LpA-I or LpA-I/A-II.

Abstract: The cellular cholesterol that moved to LDL was completely compensated for by the cholesterol influx from LDL to foam cells.

Abstract: Thus, net cholesterol efflux (a combination of influx and efflux) from foam cells was inhibited by LDL.

Abstract: These results, taken together, indicate that LDL may accelerate foam cell formation by inhibiting cholesterol removal from the cells and that elevated levels of plasma LDL may become a risk factor for atherosclerosis by inhibiting the function of LpA-I and LpA-I/A-II at the cellular level.


Sugar-sweetened beverages, weight gain, and incidence of type 2 diabetes in young and middle-aged women.

Abstract: CONTEXT Sugar-sweetened beverages like soft drinks and fruit punches contain large amounts of readily absorbable sugars and may contribute to weight gain and an increased risk of type 2 diabetes, but these relationships have been minimally addressed in adults.

Abstract: OBJECTIVE To examine the association between consumption of sugar-sweetened beverages and weight change and risk of type 2 diabetes in women.

Abstract: DESIGN, SETTING, AND PARTICIPANTS Prospective cohort analyses conducted from 1991 to 1999 among women in the Nurses' Health Study II.

Abstract: The diabetes analysis included 91,249 women free of diabetes and other major chronic diseases at baseline in 1991.

Abstract: The weight change analysis included 51,603 women for whom complete dietary information and body weight were ascertained in 1991, 1995, and 1999.

Abstract: We identified 741 incident cases of confirmed type 2 diabetes during 716,300 person-years of follow-up.

Abstract: MAIN OUTCOME MEASURES Weight gain and incidence of type 2 diabetes.

Abstract: RESULTS Those with stable consumption patterns had no difference in weight gain, but weight gain over a 4-year period was highest among women who increased their sugar-sweetened soft drink consumption from 1 or fewer drinks per week to 1 or more drinks per day (multivariate-adjusted means, 4.69 kg for 1991 to 1995 and 4.20 kg for 1995 to 1999) and was smallest among women who decreased their intake (1.34 and 0.15 kg for the 2 periods, respectively) after adjusting for lifestyle and dietary confounders.

Abstract: Increased consumption of fruit punch was also associated with greater weight gain compared with decreased consumption.

Abstract: After adjustment for potential confounders, women consuming 1 or more sugar-sweetened soft drinks per day had a relative risk [RR] of type 2 diabetes of 1.83 (95% confidence interval [CI], 1.42-2.36; P<.001 for trend) compared with those who consumed less than 1 of these beverages per month.

Abstract: Similarly, consumption of fruit punch was associated with increased diabetes risk (RR for > or =1 drink per day compared with <1 drink per month, 2.00; 95% CI, 1.33-3.03; P =.001).

Abstract: CONCLUSION Higher consumption of sugar-sweetened beverages is associated with a greater magnitude of weight gain and an increased risk for development of type 2 diabetes in women, possibly by providing excessive calories and large amounts of rapidly absorbable sugars.


Pancreatic stellate cells: partners in crime with pancreatic cancer cells.

Abstract: Pancreatic stellate cells (PSC) produce the stromal reaction in pancreatic cancer, but their role in cancer progression is not fully elucidated.

Abstract: We examined the influence of PSCs on pancreatic cancer growth using (a) an orthotopic model of pancreatic cancer and (b) cultured human PSCs (hPSC) and human pancreatic cancer cell lines MiaPaCa-2 and Panc-1.

Abstract: Athymic mice received an intrapancreatic injection of saline, hPSCs, MiaPaCa-2 cells, or hPSCs + MiaPaCa-2.

Abstract: After 7 weeks, tumor size, metastases, and tumor histology were assessed.

Abstract: In vitro studies assessed the effect of cancer cell secretions on PSC migration and the effect of hPSC secretions on cancer cell proliferation, apoptosis, and migration.

Abstract: Possible mediators of the effects of hPSC secretions on cancer cell proliferation were examined using neutralizing antibodies.

Abstract: Compared with mice receiving MiaPaCa-2 cells alone, mice injected with hPSCs + MiaPaCa-2 exhibited (a) increased tumor size and regional and distant metastasis, (b) fibrotic bands (desmoplasia) containing activated PSCs within tumors, and (c) increased tumor cell numbers.

Abstract: In vitro studies showed that, in the presence of pancreatic cancer cells, PSC migration was significantly increased.

Abstract: Furthermore, hPSC secretions induced the proliferation and migration, but inhibited the apoptosis, of MiaPaCa-2 and Panc-1 cells.

Abstract: The proliferative effect of hPSC secretions on pancreatic cancer cells was inhibited in the presence of neutralizing antibody to platelet-derived growth factor.

Abstract: Our studies indicate a significant interaction between pancreatic cancer cells and stromal cells (PSCs) and imply that pancreatic cancer cells recruit stromal cells to establish an environment that promotes cancer progression.


Preoperative statin treatment reduces systemic inflammatory response and myocardial damage in cardiac surgery.

Abstract: OBJECTIVE To determine if preoperative statin treatment is associated with a reduction in systemic inflammatory response (SIR) and myocardial damage markers following cardiac surgery with cardiopulmonary bypass (CPB).

Abstract: METHODS We study a prospective cohort of 138 patients who underwent coronary and valvular surgery with CPB.

Abstract: We differentiate two study groups: patients with (group A, n=72) or without (group B, n=66) statins.

Abstract: Plasma levels of pro-inflammatory interleukins (tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, IL-8 and IL-2R), creatine phosphokinase (CPK), CPK-MB and troponin I were measured before and 1, 6, 24 and >72 h after surgery.

Abstract: RESULTS The baseline, operative and postoperative morbidity and mortality characteristics were similar for both the groups.

Abstract: Group A had significantly lower postoperative levels of IL-6 than group B at 6h (68.8+/-5 pg ml(-1) vs 108.9+/-108 pg ml(-1), p=0.01), 24h (71.7+/-7 pg ml(-1) vs 110.4+/-106 pg ml(-1), p=0.01) and before hospital discharge (21.6+/-12 pg ml(-1) vs 32.8+/-27 pg ml(-1), p=0.005), as well as significantly lower average IL-6 levels in the first 24h following surgery (71.8+/-5 pgml(-1) vs 112.8+/-82 pg ml(-1), p=0.002).

Abstract: The postoperative CPK-MB at 24h (19.7+/-23 ng ml(-1) vs 33.1+/-32 ng ml(-1), p=0.02) and troponin I levels at the end of the intervention (2.2+/-2.2 ng ml(-1) vs 3.3+/-3.1 ng ml(-1), p=0.03) and at 24h (4.1+/-3.5 ng ml(-1) vs 6.6+/-8 ng ml(-1), p=0.04) were also significantly lower in the group treated with statins prior to surgery.

Abstract: CONCLUSIONS Preoperative treatment with statins is associated with a lower biochemical parameters of SIR and myocardial damage following cardiac surgery with CPB, regardless of it being coronary bypass grafting (CABG) or valvular surgery.


MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets.

Abstract: We present the latest version of the Molecular Evolutionary Genetics Analysis (Mega) software, which contains many sophisticated methods and tools for phylogenomics and phylomedicine.

Abstract: In this major upgrade, Mega has been optimized for use on 64-bit computing systems for analyzing larger datasets.

Abstract: Researchers can now explore and analyze tens of thousands of sequences in Mega The new version also provides an advanced wizard for building timetrees and includes a new functionality to automatically predict gene duplication events in gene family trees.

Abstract: The 64-bit Mega is made available in two interfaces: graphical and command line.

Abstract: The graphical user interface (GUI) is a native Microsoft Windows application that can also be used on Mac OS X. The command line Mega is available as native applications for Windows, Linux, and Mac OS X. They are intended for use in high-throughput and scripted analysis.

Abstract: Both versions are available from www.megasoftware.net free of charge.


DNA replication fidelity

Abstract: DNA replication fidelity is a key determinant of genome stability and is central to the evolution of species and to the origins of human diseases.

Abstract: Here we review our current understanding of replication fidelity, with emphasis on structural and biochemical studies of DNA polymerases that provide new insights into the importance of hydrogen bonding, base pair geometry, and substrate-induced conformational changes to fidelity.

Abstract: These studies also reveal polymerase interactions with the DNA minor groove at and upstream of the active site that influence nucleotide selectivity, the efficiency of exonucleolytic proofreading, and the rate of forming errors via strand misalignments.

Abstract: We highlight common features that are relevant to the fidelity of any DNA synthesis reaction, and consider why fidelity varies depending on the enzymes, the error, and the local sequence environment.


Viable neutrophils release mitochondrial DNA to form neutrophil extracellular traps

Abstract: Neutrophil extracellular traps (NETs) represent extracellular structures able to bind and kill microorganisms.

Abstract: It is believed that they are generated by neutrophils undergoing cell death, allowing these dying or dead cells to kill microbes.

Abstract: We show that, following priming with granulocyte/macrophage colony-stimulating factor (GM-CSF) and subsequent short-term toll-like receptor 4 (TLR4) or complement factor 5a (C5a) receptor stimulation, viable neutrophils are able to generate NETs.

Abstract: Strikingly, NETs formed by living cells contain mitochondrial, but no nuclear, DNA.

Abstract: Pharmacological or genetic approaches to block reactive oxygen species (ROS) production suggested that NET formation is ROS dependent.

Abstract: Moreover, neutrophil populations stimulated with GM-CSF and C5a showed increased survival compared with resting neutrophils, which did not generate NETs.

Abstract: In conclusion, mitochondrial DNA release by neutrophils and NET formation do not require neutrophil death and do also not limit the lifespan of these cells.


ATP-driven exchange of histone H2AZ variant catalyzed by SWR1 chromatin remodeling complex.

Abstract: The conserved histone variant H2AZ has an important role in the regulation of gene expression and the establishment of a buffer to the spread of silent heterochromatin.

Abstract: How histone variants such as H2AZ are incorporated into nucleosomes has been obscure.

Abstract: We have found that Swr1, a Swi2/Snf2-related adenosine triphosphatase, is the catalytic core of a multisubunit, histone-variant exchanger that efficiently replaces conventional histone H2A with histone H2AZ in nucleosome arrays.

Abstract: Swr1 is required for the deposition of histone H2AZ at specific chromosome locations in vivo, and Swr1 and H2AZ commonly regulate a subset of yeast genes.

Abstract: These findings define a previously unknown role for the adenosine triphosphate-dependent chromatin remodeling machinery.


FACT Disrupts Nucleosome Structure by Binding H2A-H2B with Conserved Peptide Motifs.

Abstract: FACT, a heterodimer of Spt16 and Pob3, is an essential histone chaperone.

Abstract: We show that the H2A-H2B binding activity that is central to FACT function resides in short acidic regions near the C termini of each subunit.

Abstract: Mutations throughout these regions affect binding and cause correlated phenotypes that range from mild to lethal, with the largest individual contributions unexpectedly coming from an aromatic residue and a nearby carboxylate residue within each domain.

Abstract: Spt16 and Pob3 bind overlapping sites on H2A-H2B, and Spt16-Pob3 heterodimers simultaneously bind two H2A-H2B dimers, the same stoichiometry as the components of a nucleosome.

Abstract: An Spt16:H2A-H2B crystal structure explains the biochemical and genetic data, provides a model for Pob3 binding, and implies a mechanism for FACT reorganization that we confirm biochemically.

Abstract: Moreover, unexpected similarity to binding of ANP32E and Swr1 with H2A.Z-H2B reveals that diverse H2A-H2B chaperones use common mechanisms of histone binding and regulating nucleosome functions.


Role of AMP-activated protein kinase in mechanism of metformin action.

Abstract: Metformin is a widely used drug for treatment of type 2 diabetes with no defined cellular mechanism of action.

Abstract: Its glucose-lowering effect results from decreased hepatic glucose production and increased glucose utilization.

Abstract: Metformin's beneficial effects on circulating lipids have been linked to reduced fatty liver.

Abstract: AMP-activated protein kinase (AMPK) is a major cellular regulator of lipid and glucose metabolism.

Abstract: Here we report that metformin activates AMPK in hepatocytes; as a result, acetyl-CoA carboxylase (ACC) activity is reduced, fatty acid oxidation is induced, and expression of lipogenic enzymes is suppressed.

Abstract: Activation of AMPK by metformin or an adenosine analogue suppresses expression of SREBP-1, a key lipogenic transcription factor.

Abstract: In metformin-treated rats, hepatic expression of SREBP-1 (and other lipogenic) mRNAs and protein is reduced; activity of the AMPK target, ACC, is also reduced.

Abstract: Using a novel AMPK inhibitor, we find that AMPK activation is required for metformin's inhibitory effect on glucose production by hepatocytes.

Abstract: In isolated rat skeletal muscles, metformin stimulates glucose uptake coincident with AMPK activation.

Abstract: Activation of AMPK provides a unified explanation for the pleiotropic beneficial effects of this drug; these results also suggest that alternative means of modulating AMPK should be useful for the treatment of metabolic disorders.


Interferon alfa therapy for chronic hepatitis B in children: a multinational randomized controlled trial.

Abstract: BACKGROUND & AIMS Treatment of chronic hepatitis B with interferon alfa is not approved in children.

Abstract: The aim of this study was to evaluate the safety and efficacy of interferon alfa (IFN-alpha) in children with chronic hepatitis B and increased transaminase levels.

Abstract: METHODS Children were given either IFN-alpha2b (6 megaunits/m2 thrice weekly for 24 weeks) or no treatment.

Abstract: Clearance of markers of viral replication was evaluated 24 weeks after therapy and after 48 weeks of observation in controls.

Abstract: RESULTS Of 149 children enrolled, 144 were evaluable (70 treated and 74 controls).

Abstract: Serum hepatitis B e antigen and viral DNA became negative in 26% of treated children and 11% of controls (P < 0.05).

Abstract: Serum aminotransferase levels normalized and liver histology improved among responders.

Abstract: Hepatitis B surface antigen became undetectable in 10% of treated patients and 1% of controls.

Abstract: Female gender and interferon treatment were the only significant predictors of response.

Abstract: Ethnic origin, baseline aminotransferase level, initial DNA levels, and histology did not correlate with response.

Abstract: Most adverse reactions were mild or moderate, and dose was reduced in 24% of children.

Abstract: CONCLUSIONS In children with chronic hepatitis B, INF-alpha promotes loss of viral replication markers and surface antigen and improves aminotransferases and histology.


ImmTACs for targeted cancer therapy: Why, what, how, and which.

Abstract: Overcoming immunosuppression and activating a cytotoxic T cell response has the potential to halt the progression of cancer and, in some circumstances, eradicate it.

Abstract: Designing therapeutic interventions that achieve this goal has proven challenging, but now a greater understanding of the complexities of immune responses is beginning to produce some notable breakthroughs.

Abstract: ImmTACs (immune-mobilising monoclonal TCRs against cancer) are a new class of bispecific reagents, based on soluble monoclonal T cell receptors, which have been engineered to possess extremely high affinity for cognate tumour antigen.

Abstract: In this way, ImmTACs overcome the problem of low affinity tumour-specific T cells imposed by thymic selection and provide access to the large number of antigens presented as peptide-HLA complexes.

Abstract: Once bound to tumour cells the anti-CD3 effector end of the ImmTAC drives recruitment of polyclonal T cells to the tumour site, leading to a potent redirected T cell response and tumour cell destruction.

Abstract: Extensive in vitro testing coupled with promising early clinical data has provided an enhanced appreciation of ImmTAC function in vivo and indicates their potential therapeutic benefit in terms of a durable response and ultimately the breaking of T cell tolerance.

Abstract: This review introduces ImmTACs in the context of immunotherapy, and outlines their design, construction and mechanism of action, as well as examining target selection and aspects of preclinical safety testing.


Angiopoietin-like protein 3 supports the activity of hematopoietic stem cells in the bone marrow niche.

Abstract: The physiologic roles of angiopoietin-like proteins (Angptls) in the hematopoietic system remain unknown.

Abstract: Here we show that hematopoietic stem cells (HSCs) in Angptl3-null mice are decreased in number and quiescence.

Abstract: HSCs transplanted into Angptl3-null recipient mice exhibited impaired repopulation.

Abstract: Bone marrow sinusoidal endothelial cells express high levels of Angptl3 and are adjacent to HSCs.

Abstract: Importantly, bone marrow stromal cells or endothelium deficient in Angptl3 have a significantly decreased ability to support the expansion of repopulating HSCs.

Abstract: Angptl3 represses the expression of the transcription factor Ikaros, whose unregulated overexpression diminishes the repopulation activity of HSCs.

Abstract: Angptl3, as an extrinsic factor, thus supports the stemness of HSCs in the bone marrow niche.


Subsequent chemotherapy reverses acquired tyrosine kinase inhibitor resistance and restores response to tyrosine kinase inhibitor in advanced non-small-cell lung cancer

Abstract: BACKGROUND Patients with advanced or metastatic non-small cell lung cancer (NSCLC) can develop acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors (TKIs) erlotinib and gefitinib.

Abstract: Here, we report the successful treatment with alternating chemotherapy and TKIs of two cases of advanced NSCLC who developed resistance to TKI.

Abstract: CASE PRESENTATION Two patients with advanced or metastatic NSCLC were treated with palliative chemotherapy followed by erlotinib/gefitinib.

Abstract: When TKI therapy failed, two cycles of chemotherapy were provided, which were followed by re-challenge with erlotinib or gefitinib.

Abstract: CONCLUSION NSCLC patients with acquired TKI resistance should be managed aggressively whenever possible.

Abstract: Subsequent chemotherapy and target treatment is one of the reasonable choices for those with an initial dramatic clinical response with erlotinib/gefitinib treatment.

Abstract: Further studies are warranted to substantiate the association of erlotinib /gefitinib treatment with the efficacy of NSCLC patients with acquired TKI failure.


Epigenetic Therapy Ties MYC Depletion to Reversing Immune Evasion and Treating Lung Cancer

Abstract: Combining DNA-demethylating agents (DNA methyltransferase inhibitors [DNMTis]) with histone deacetylase inhibitors (HDACis) holds promise for enhancing cancer immune therapy.

Abstract: Herein, pharmacologic and isoform specificity of HDACis are investigated to guide their addition to a DNMTi, thus devising a new, low-dose, sequential regimen that imparts a robust anti-tumor effect for non-small-cell lung cancer (NSCLC).

Abstract: Using in-vitro-treated NSCLC cell lines, we elucidate an interferon α/β-based transcriptional program with accompanying upregulation of antigen presentation machinery, mediated in part through double-stranded RNA (dsRNA) induction.

Abstract: This is accompanied by suppression of MYC signaling and an increase in the T cell chemoattractant CCL5.

Abstract: Use of this combination treatment schema in mouse models of NSCLC reverses tumor immune evasion and modulates T cell exhaustion state towards memory and effector T cell phenotypes.

Abstract: Key correlative science metrics emerge for an upcoming clinical trial, testing enhancement of immune checkpoint therapy for NSCLC.


The Role of PPARγ in Advanced Glycation End Products-Induced Inflammatory Response in Human Chondrocytes

Abstract: OBJECTIVE Advances made in the past ten years highlight the notion that peroxisome proliferator-activated receptors gamma (PPARγ) has protective properties in the pathophysiology of osteoarthritis (OA).

Abstract: The aim of this study was to define the roles of PPARγ in AGEs-induced inflammatory response in human chondrocytes.

Abstract: METHODS Primary human chondrocytes were stimulated with AGEs in the presence or absence of neutralizing antibody against RAGE (anti-RAGE), MAPK specific inhibitors and PPARγ agonist pioglitazone.

Abstract: The expression of IL-1, MMP-13, TNF-α, PPARγ, nuclear NF-κB p65 and cytosol IκBα was determined by western blotting and real-time PCR.

Abstract: RESULTS AGEs could enhance the expression of IL-1, TNF-α, and MMP-13, but the level of PPARγ was decreased in a time- and dose-dependent manner, which was inhibited by anti-RAGE, SB203580 (P38 MAPK specific inhibitor) and SP600125 (a selective inhibitor of JNK).

Abstract: PPARγ agonist pioglitazone could inhibit the effects of AGEs-induced inflammatory response and PPARγ down-regulation.

Abstract: In human chondrocytes, AGEs could induce cytosol IκBα degradation and increase the level of nuclear NF-κB p65, which was inhibited by PPARγ agonist pioglitazone.

Abstract: CONCLUSIONS In primary human chondrocytes, AGEs could down-regulate PPARγ expression and increase the inflammatory mediators, which could be reversed by PPARγ agonist pioglitazone.

Abstract: Activation of RAGE by AGEs triggers a cascade of downstream signaling, including MAPK JNK/ p38, PPARγ and NF-κB. Taken together, PPARγ could be a potential target for pharmacologic intervention in the treatment of OA.


Surgical versus medical treatment with cyclooxygenase inhibitors for symptomatic patent ductus arteriosus in preterm infants.

Abstract: BACKGROUND Patent ductus arteriosus (PDA) with significant left to right shunt in preterm infants increases morbidity and mortality.

Abstract: Early closure of the ductus arteriosus may be achieved pharmacologically using cyclooxygenase inhibitors or by surgery.

Abstract: The efficacy of both treatment modalities is well established.

Abstract: However, the preferred initial treatment of a symptomatic PDA in a preterm infant, surgical ligation or treatment with indomethacin, has not been well established.

Abstract: OBJECTIVES To compare the effect of surgical ligation of PDA vs. medical treatment with cyclooxygenase inhibitors (using indomethacin, ibuprofen, or mefenamic acid), each used as the initial treatment, on neonatal mortality in preterm infants with a symptomatic PDA.

Abstract: SEARCH STRATEGY The standard search strategy of the Cochrane Neonatal Review Group was used.

Abstract: This included search of electronic databases: Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 2, 2007), MEDLINE (1966 - July 2007), CINAHL (1982 - July 2007), EMBASE (1980 - July 2007); and hand search of abstracts of Pediatric Academic Societies annual meetings published in Pediatric Research (1990 - April 2002) or on line from May 2002 -July 2007.

Abstract: No language restrictions were applied.

Abstract: SELECTION CRITERIA All trials 1) using randomized or quasi-randomized patient allocation, 2) in preterm infants < 37 weeks gestational age or low-birth-weight infants (< 2500 grams) with symptomatic PDA in the neonatal period (< 28 days) and 3) comparing surgical ligation with medical treatment with cyclooxygenase inhibitors, each used as the initial treatment for closure of PDA.

Abstract: DATA COLLECTION AND ANALYSIS Assessment of methodological quality and extraction of data for included trials was undertaken independently by the authors.

Abstract: RevMan 4.1 was used for analysis of the data.

Abstract: MAIN RESULTS Only one study, trial B in the report of Gersony 1983, was found eligible.

Abstract: No additional studies were identified in the literature searches performed in July 2007.

Abstract: The trial compared the effect of surgical ligation of PDA vs. medical treatment with indomethacin, each used as the primary treatment.

Abstract: No trials comparing surgery to other cyclooxygenase inhibitors (ibuprofen, mefenamic acid) were found.

Abstract: Trial B of Gersony 1983 enrolled 154 infants.

Abstract: The study found no statistically significant difference between surgical closure and indomethacin treatment in mortality during hospital stay, chronic lung disease, other bleeding, necrotizing enterocolitis, sepsis, creatinine level, or intraventricular hemorrhage.

Abstract: There was a statistically significant increase in the surgical group in incidence of pneumothorax [RR 2.68 (95% CI 1.45, 4.93); RD 0.25 (95% CI 0.11, 0.38); NNH 4 (95% CI 3, 9)] and retinopathy of prematurity stage III and IV [RR 3.80 (95% CI 1.12, 12.93); RD 0.11 (95% CI 0.02, 0.20), NNH 9 (95% CI 5, 50] compared to the indomethacin group.

Abstract: There was as expected a statistically significant decrease in failure of ductal closure rate in the surgical group as compared to the indomethacin group: [RR 0.04 (95% CI 0.01, 0.27); RD -0.32 (95% CI -0.43, -0.21), NNT 3 (95% CI 2, 4)].

Abstract: AUTHORS' CONCLUSIONS The data regarding net benefit/harm are insufficient to make a conclusion as to whether surgical ligation or medical treatment with indomethacin is preferred as initial treatment for symptomatic PDA in preterm infants.

Abstract: It should be noted that three recent observational studies indicated an increased risk for one or more of the following outcomes associated with PDA ligation; chronic lung disease, retinopathy of prematurity and neurosensory impairment .

Abstract: It is possible that the duration of the "waiting-time" and transport to another facility with surgical capacity to have the PDA ligated could adversely affect outcomes, as could the perioperative care.


The Effect of Collinearity on Parameter Estimates in Nonlinear Mixed Effect Models

Abstract: Purpose.

Abstract: To demonstrate how correlations among predictor variables in a population pharmacokinetic model affect the ability to discern which covariates should enter into the structural pharmacokinetic model.

Abstract: Methods.

Abstract: Monte Carlo simulation was used to generate multiple-dose concentration-time data similar to that seen in a Phase III clinical trial.

Abstract: The drugs' pharmacokinetics were dependent on two covariates.

Abstract: Five data sets were simulated with increasing correlation between the two covariates.

Abstract: All data sets were analyzed using NONMEM both with and without inclusion of the covariates in the structural pharmacokinetic model.

Abstract: Summary measures for ill-conditioning and sensitivity analysis were used to examine how increasing correlation among covariates affects the accuracy and precision of the parameter estimates.

Abstract: Results.

Abstract: When covariates were included in the structural pharmacokinetic model and the correlation between covariates increased, the standard error of the parameter estimates increased and the value of parameter estimates themselves became increasingly biased.

Abstract: When the correlation between predictor variables was 0.75, the standard errors of the parameter estimates were too large to declare statistical significance.

Abstract: Conclusions.

Abstract: Correlations among predictor variables greater than 0.5 when entered into the model simultaneously should be a warning to researchers because the (1) the accuracy of the parameter estimates themselves may be biased and (2) the precision of the estimates may be inflated due to ill-conditioning.


An analysis of age and gender influences on the relative risk for suicide and psychotropic drug self-poisoning.

Abstract: Psychiatric illness is a significant risk factor for both attempted and completed suicide and psychotropic medications account for 80% of all drug overdoses involving prescription medications.

Abstract: One challenge facing clinicians is to balance the benefit of treatment against the risk of drug overdose.

Abstract: The aim of the present study was to compare the age and gender distribution of patients prescribed psychotropic drugs with patients attempting and completing suicide with these drugs.

Abstract: Data were obtained from the Australian census and studies of general practitioner prescribing, patients who committed suicide or presented with self-poisoning within a defined geographic area.

Abstract: The characteristics of these populations were compared to calculate odds ratios for attempting or completing suicide with psychotropic drugs, before and after correction for rates of prescription, in different age and gender groups.

Abstract: The odds ratios (ORs) for self-poisoning were higher for those aged less than 45 years and yet this group was least likely to be prescribed psychotropic drugs.

Abstract: Men had a much higher rate of completed suicide using more lethal methods.

Abstract: The ORs for self-poisoning and suicide with psychotropic drugs, after correction for prescription rates, for those aged 15 to 24 years were 11.1 and 1.7, respectively.

Abstract: Those aged 25 to 44 years had ORs of 4.9 and 4.3, and, by contrast, those over 75 years had ORs of 0.03 and 0.

Abstract: Women were slightly more likely to poison themselves with psychotropic drugs (OR 1.2).

Abstract: However, the situation reversed after correction for prescription rates (OR 0.69).

Abstract: It is concluded that greater caution should be exercised in prescribing for those under 45 years of age, given their relatively higher risk of drug overdose, and that the least toxic compounds should be used.

Abstract: The risk (of self-poisoning) among the elderly may have been overstated, so that some patients may have been denied the benefit of adequate treatment.


Cell death and immune privilege.

Abstract: The host response to pathogens involves complex inflammatory responses and immune reactions.

Abstract: While these are central to host defense and vital to clearing infections, they are often accompanied by injury to surrounding tissue.

Abstract: Most organ systems can tolerate these responses without permanent consequences.

Abstract: However, there are sites that limit the spread of inflammation because it can threaten organ function.

Abstract: The most prominent examples of these are the eye, brain, and reproductive organs (testis, ovary), where even minor bouts of inflammation can have long-term consequences for the survival of the organism.

Abstract: In these organs immune responses either do not proceed, or proceed in a manner different from other areas; thus, they are called "immunologically privileged.

Abstract: Here a functioning immune response can be the culprit that leads to disease.


Niche-Independent Symmetrical Self-Renewal of a Mammalian Tissue Stem Cell

Abstract: Pluripotent mouse embryonic stem (ES) cells multiply in simple monoculture by symmetrical divisions.

Abstract: In vivo, however, stem cells are generally thought to depend on specialised cellular microenvironments and to undergo predominantly asymmetric divisions.

Abstract: Ex vivo expansion of pure populations of tissue stem cells has proven elusive.

Abstract: Neural progenitor cells are propagated in combination with differentiating progeny in floating clusters called neurospheres.

Abstract: The proportion of stem cells in neurospheres is low, however, and they cannot be directly observed or interrogated.

Abstract: Here we demonstrate that the complex neurosphere environment is dispensable for stem cell maintenance, and that the combination of fibroblast growth factor 2 (FGF-2) and epidermal growth factor (EGF) is sufficient for derivation and continuous expansion by symmetrical division of pure cultures of neural stem (NS) cells.

Abstract: NS cells were derived first from mouse ES cells.

Abstract: Neural lineage induction was followed by growth factor addition in basal culture media.

Abstract: In the presence of only EGF and FGF-2, resulting NS cells proliferate continuously, are diploid, and clonogenic.

Abstract: After prolonged expansion, they remain able to differentiate efficiently into neurons and astrocytes in vitro and upon transplantation into the adult brain.

Abstract: Colonies generated from single NS cells all produce neurons upon growth factor withdrawal.

Abstract: NS cells uniformly express morphological, cell biological, and molecular features of radial glia, developmental precursors of neurons and glia.

Abstract: Consistent with this profile, adherent NS cell lines can readily be established from foetal mouse brain.

Abstract: Similar NS cells can be generated from human ES cells and human foetal brain.

Abstract: The extrinsic factors EGF plus FGF-2 are sufficient to sustain pure symmetrical self-renewing divisions of NS cells.

Abstract: The resultant cultures constitute the first known example of tissue-specific stem cells that can be propagated without accompanying differentiation.

Abstract: These homogenous cultures will enable delineation of molecular mechanisms that define a tissue-specific stem cell and allow direct comparison with pluripotent ES cells.


A Bivalent Chromatin Structure Marks Key Developmental Genes in Embryonic Stem Cells

Abstract: The most highly conserved noncoding elements (HCNEs) in mammalian genomes cluster within regions enriched for genes encoding developmentally important transcription factors (TFs).

Abstract: This suggests that HCNE-rich regions may contain key regulatory controls involved in development.

Abstract: We explored this by examining histone methylation in mouse embryonic stem (ES) cells across 56 large HCNE-rich loci.

Abstract: We identified a specific modification pattern, termed "bivalent domains," consisting of large regions of H3 lysine 27 methylation harboring smaller regions of H3 lysine 4 methylation.

Abstract: Bivalent domains tend to coincide with TF genes expressed at low levels.

Abstract: We propose that bivalent domains silence developmental genes in ES cells while keeping them poised for activation.

Abstract: We also found striking correspondences between genome sequence and histone methylation in ES cells, which become notably weaker in differentiated cells.

Abstract: These results highlight the importance of DNA sequence in defining the initial epigenetic landscape and suggest a novel chromatin-based mechanism for maintaining pluripotency.


Ovarian Surface Epithelium in Patients with Severe Ovarian Infertility: A Potential Source of Cells Expressing Markers of Pluripotent/Multipotent Stem Cells

Abstract: The aim of this study was to confirm the presence of stem cells in the ovarian surface epithelium of patients with premature ovarian failure and no mature follicles and oocytes.

Abstract: In these patients, small round cells of unknown origin expressing SOX-2 marker of pluripotency were observed among the epithelial cells just after the ovarian surface epithelium scraping.

Abstract: These cells were an integral part of the ovarian surface epithelium.

Abstract: When the scraped cells were cultured in a medium with added follicular fluid to provide some ovarian niche, primitive oocyte-like cells and typical round-shaped cell clusters positively stained on alkaline phosphatase, and markers of pluripotency, such as SOX-2 and SSEA-4, were developed.

Abstract: These markers were expressed early and also later in the culture.

Abstract: Single oocyte-like cells expressed genes OCT4A, SOX-2, NANOG, NANOS, STELLA, CD9, LIN28, KLF4, GDF3, and MYC, characteristic for pluripotent stem cells.

Abstract: The results of this study confirmed the presence of putative stem cells in the ovarian surface epithelium of these patients and provided some basis to create a stem cell line in the future.


Structural Homeostasis: Compensatory Adjustments of Dendritic Arbor Geometry in Response to Variations of Synaptic Input

Abstract: As the nervous system develops, there is an inherent variability in the connections formed between differentiating neurons.

Abstract: Despite this variability, neural circuits form that are functional and remarkably robust.

Abstract: One way in which neurons deal with variability in their inputs is through compensatory, homeostatic changes in their electrical properties.

Abstract: Here, we show that neurons also make compensatory adjustments to their structure.

Abstract: We analysed the development of dendrites on an identified central neuron (aCC) in the late Drosophila embryo at the stage when it receives its first connections and first becomes electrically active.

Abstract: At the same time, we charted the distribution of presynaptic sites on the developing postsynaptic arbor.

Abstract: Genetic manipulations of the presynaptic partners demonstrate that the postsynaptic dendritic arbor adjusts its growth to compensate for changes in the activity and density of synaptic sites.

Abstract: Blocking the synthesis or evoked release of presynaptic neurotransmitter results in greater dendritic extension.

Abstract: Conversely, an increase in the density of presynaptic release sites induces a reduction in the extent of the dendritic arbor.

Abstract: These growth adjustments occur locally in the arbor and are the result of the promotion or inhibition of growth of neurites in the proximity of presynaptic sites.

Abstract: We provide evidence that suggest a role for the postsynaptic activity state of protein kinase A in mediating this structural adjustment, which modifies dendritic growth in response to synaptic activity.

Abstract: These findings suggest that the dendritic arbor, at least during early stages of connectivity, behaves as a homeostatic device that adjusts its size and geometry to the level and the distribution of input received.

Abstract: The growing arbor thus counterbalances naturally occurring variations in synaptic density and activity so as to ensure that an appropriate level of input is achieved.


Nutrition and physical activity guidelines for cancer survivors.

Abstract: Cancer survivors are often highly motivated to seek information about food choices, physical activity, and dietary supplements to improve their treatment outcomes, quality of life, and overall survival.

Abstract: To address these concerns, the American Cancer Society (ACS) convened a group of experts in nutrition, physical activity, and cancer survivorship to evaluate the scientific evidence and best clinical practices related to optimal nutrition and physical activity after the diagnosis of cancer.

Abstract: This report summarizes their findings and is intended to present health care providers with the best possible information with which to help cancer survivors and their families make informed choices related to nutrition and physical activity.

Abstract: The report discusses nutrition and physical activity guidelines during the continuum of cancer care, briefly highlighting important issues during cancer treatment and for patients with advanced cancer, but focusing largely on the needs of the population of individuals who are disease free or who have stable disease following their recovery from treatment.

Abstract: It also discusses select nutrition and physical activity issues such as body weight, food choices, food safety, and dietary supplements; issues related to selected cancer sites; and common questions about diet, physical activity, and cancer survivorship.


The role of protein clearance mechanisms in organismal ageing and age-related diseases.

Abstract: The ability to maintain a functional proteome, or proteostasis, declines during the ageing process.

Abstract: Damaged and misfolded proteins accumulate with age, impairing cell function and tissue homeostasis.

Abstract: The accumulation of damaged proteins contributes to multiple age-related diseases such as Alzheimer's, Parkinson's or Huntington's disease.

Abstract: Damaged proteins are degraded by the ubiquitin-proteasome system or through autophagy-lysosome, key components of the proteostasis network.

Abstract: Modulation of either proteasome activity or autophagic-lysosomal potential extends lifespan and protects organisms from symptoms associated with proteostasis disorders, suggesting that protein clearance mechanisms are directly linked to ageing and age-associated diseases.


A distinctive DNA damage response in human hematopoietic stem cells reveals an apoptosis-independent role for p53 in self-renewal.

Abstract: Highly regenerative tissues such as blood must possess effective DNA damage responses (DDR) that balance long-term regeneration with protection from leukemogenesis.

Abstract: Hematopoietic stem cells (HSCs) sustain life-long blood production, yet their response to DNA damage remains largely unexplored.

Abstract: We report that human HSCs exhibit delayed DNA double-strand break rejoining, persistent gammaH2AX foci, and enhanced p53- and ASPP1-dependent apoptosis after gamma-radiation compared to progenitors.

Abstract: p53 inactivation or Bcl-2 overexpression reduced radiation-induced apoptosis and preserved in vivo repopulating HSC function.

Abstract: Despite similar protection from irradiation-induced apoptosis, only Bcl-2-overexpressing HSCs showed higher self-renewal capacity, establishing that intact p53 positively regulates self-renewal independently from apoptosis.

Abstract: The reduced self-renewal of HSCs with inactivated p53 was associated with increased spontaneous gammaH2AX foci in secondary transplants of HSCs.

Abstract: Our data reveal distinct physiological roles of p53 that together ensure optimal HSC function: apoptosis regulation and prevention of gammaH2AX foci accumulation upon HSC self-renewal.


Behavioral characterization of neuropeptide Y knockout mice.

Abstract: An extensive behavioral characterization was conducted with mice lacking the gene for neuropeptide Y (NPY) including response to 24 and 48 h fast and challenge with small molecule antagonists of NPY receptors implicated in mediating the feeding effects of NPY (i.e., Y1 and Y5).

Abstract: In addition, wildtype (WT) and NPY knockout (KO) mice were tested in locomotor monitors, elevated plus maze, inhibitory avoidance, acoustic startle, prepulse inhibition, and hot plate assays.

Abstract: One of the major findings was that the NPY KO mice have a reduced food intake relative to WT controls in response to fasting.

Abstract: Also, based on data from the behavioral models, the NPY KO mice may have an anxiogenic-like phenotype, and appear to be hypoalgesic in the hot plate paradigm.

Abstract: The data from these studies provide further evidence of involvement of NPY in energy balance, anxiety, and possibly nociception.


Epigenetic-induced repression of microRNA-205 is associated with MED1 activation and a poorer prognosis in localized prostate cancer

Abstract: Deregulation of microRNA (miRNA) expression can have a critical role in carcinogenesis.

Abstract: Here we show in prostate cancer that miRNA-205 (miR-205) transcription is commonly repressed and the MIR-205 locus is hypermethylated.

Abstract: LOC642587, the MIR-205 host gene of unknown function, is also concordantly inactivated.

Abstract: We show that miR-205 targets mediator 1 (MED1, also called TRAP220 and PPARBP) for transcriptional silencing in normal prostate cells, leading to reduction in MED1 mRNA levels, and in total and active phospho-MED1 protein.

Abstract: Overexpression of miR-205 in prostate cancer cells negatively affects cell viability, consistent with a tumor suppressor function.

Abstract: We found that hypermethylation of the MIR-205 locus was strongly related with a decrease in miR-205 expression and an increase in MED1 expression in primary tumor samples (n=14), when compared with matched normal prostate (n=7).

Abstract: An expanded patient cohort (tumor n=149, matched normal n=30) also showed significant MIR-205 DNA methylation in tumors compared with normal, and MIR-205 hypermethylation is significantly associated with biochemical recurrence (hazard ratio=2.005, 95% confidence interval (1.109, 3.625), P=0.02), in patients with low preoperative prostate specific antigen.

Abstract: In summary, these results suggest that miR-205 is an epigenetically regulated tumor suppressor that targets MED1 and may provide a potential biomarker in prostate cancer management.


Th2 cytokine response in Major Depressive Disorder patients before treatment

Abstract: In Major Depressive Disorder (MDD), the neuroendocrine and immune systems interactions are impaired.

Abstract: We investigated the pro/anti-inflammatory Th1/Th2 cytokine balance in MDD patients and in non-depressed control group.

Abstract: The MDD subjects showed higher levels of cortisol and TNF-alpha, increased CD3+CD8+ and NK percentages, diminished B cell counts and no significant variations in CD3+CD4+ lymphocyte.

Abstract: Moreover, higher levels of IL-4 and IL-13 (Th2) and significantly lower measurements of IL-2 and IFN-gamma (Th1) cytokines were also observed in the MDD group.

Abstract: Overall, we propose that all these changes could be related to the elevated cortisol levels seen in the MDD patients.

Abstract: Further studies are necessary to explore these findings and its implication in future therapeutic approach of MDD patients.


CDD: conserved domains and protein three-dimensional structure

Abstract: CDD, the Conserved Domain Database, is part of NCBI's Entrez query and retrieval system and is also accessible via http://www.ncbi.nlm.nih.gov/Structure/cdd/cdd.shtml.

Abstract: CDD provides annotation of protein sequences with the location of conserved domain footprints and functional sites inferred from these footprints.

Abstract: Pre-computed annotation is available via Entrez, and interactive search services accept single protein or nucleotide queries, as well as batch submissions of protein query sequences, utilizing RPS-BLAST to rapidly identify putative matches.

Abstract: CDD incorporates several protein domain and full-length protein model collections, and maintains an active curation effort that aims at providing fine grained classifications for major and well-characterized protein domain families, as supported by available protein three-dimensional (3D) structure and the published literature.

Abstract: To this date, the majority of protein 3D structures are represented by models tracked by CDD, and CDD curators are characterizing novel families that emerge from protein structure determination efforts.


Extensive diversity of Ig-superfamily proteins in the immune system of insects.

Abstract: The extensive somatic diversification of immune receptors is a hallmark of higher vertebrates.

Abstract: However, whether molecular diversity contributes to immune protection in invertebrates is unknown.

Abstract: We present evidence that Drosophila immune-competent cells have the potential to express more than 18,000 isoforms of the immunoglobulin (Ig)-superfamily receptor Down syndrome cell adhesion molecule (Dscam).

Abstract: Secreted protein isoforms of Dscam were detected in the hemolymph, and hemocyte-specific loss of Dscam impaired the efficiency of phagocytic uptake of bacteria, possibly due to reduced bacterial binding.

Abstract: Importantly, the molecular diversity of Dscam transcripts generated through a mechanism of alternative splicing is highly conserved across major insect orders, suggesting an unsuspected molecular complexity of the innate immune system of insects.


Preparing the outbreak assistance laboratory network in the Netherlands for the detection of the influenza virus A(H1N1) variant.

Abstract: BACKGROUND Late April 2009, human infection with variant influenza virus A(H1N1)v emerged in the Northern Americas posing a threat that this virus may become the next pandemic influenza virus.

Abstract: OBJECTIVES To prepare laboratories for surge capacity for molecular diagnosis of patients suspected for A(H1N1)v infection in the Netherlands.

Abstract: STUDY DESIGN A panel of 10 blinded specimens containing seasonal A(H1N1) or A(H3N2), or A/Netherlands/602/2009(H1N1)v influenza virus, or negative control was distributed to the outbreak assistance laboratories (OAL) together with influenza virus A (M-gene), swine influenza virus A (NP-gene) and influenza virus A(H1N1)v (H1v-gene) specific primers and probes and protocol (CDC Atlanta, USA).

Abstract: Laboratories were asked to implement and test this protocol.

Abstract: RESULTS All OAL were able to detect A(H1N1)v using the CDC M-gene reagents, the majority with similar sensitivity as the in-house M-gene based assays.

Abstract: RT-PCRs used in routine diagnostic setting in the OAL specifically designed to detect H1, H3, or NS1 from seasonal influenza A viruses, did not or at very low level cross-react with A(H1N1)v.

Abstract: The CDC swine NP-gene and H1v-gene RT-PCRs showed somewhat reduced sensitivity compared to the CDC and in-house M-gene RT-PCRs.

Abstract: In contrast, in-house developed A(H1N1)v specific H1v-gene and N1v-gene RT-PCRs showed equal sensitivity to CDC and in-house M-gene RT-PCRs.

Abstract: CONCLUSIONS The Dutch OAL are prepared for detection and specific identification of A(H1N1)v, although some level of cross-reactivity was observed with seasonal influenza viruses.

Abstract: Additionally, M-gene based generic influenza A virus detection is recommended to be able to detect emerging influenza A viruses in routine settings.


Averting Obesity and Type 2 Diabetes in India through Sugar-Sweetened Beverage Taxation: An Economic-Epidemiologic Modeling Study

Abstract: BACKGROUND Taxing sugar-sweetened beverages (SSBs) has been proposed in high-income countries to reduce obesity and type 2 diabetes.

Abstract: We sought to estimate the potential health effects of such a fiscal strategy in the middle-income country of India, where there is heterogeneity in SSB consumption, patterns of substitution between SSBs and other beverages after tax increases, and vast differences in chronic disease risk within the population.

Abstract: METHODS AND FINDINGS Using consumption and price variations data from a nationally representative survey of 100,855 Indian households, we first calculated how changes in SSB price alter per capita consumption of SSBs and substitution with other beverages.

Abstract: We then incorporated SSB sales trends, body mass index (BMI), and diabetes incidence data stratified by age, sex, income, and urban/rural residence into a validated microsimulation of caloric consumption, glycemic load, overweight/obesity prevalence, and type 2 diabetes incidence among Indian subpopulations facing a 20% SSB excise tax.

Abstract: The 20% SSB tax was anticipated to reduce overweight and obesity prevalence by 3.0% (95% CI 1.6%-5.9%) and type 2 diabetes incidence by 1.6% (95% CI 1.2%-1.9%) among various Indian subpopulations over the period 2014-2023, if SSB consumption continued to increase linearly in accordance with secular trends.

Abstract: However, acceleration in SSB consumption trends consistent with industry marketing models would be expected to increase the impact efficacy of taxation, averting 4.2% of prevalent overweight/obesity (95% CI 2.5-10.0%) and 2.5% (95% CI 1.0-2.8%) of incident type 2 diabetes from 2014-2023.

Abstract: Given current consumption and BMI distributions, our results suggest the largest relative effect would be expected among young rural men, refuting our a priori hypothesis that urban populations would be isolated beneficiaries of SSB taxation.

Abstract: Key limitations of this estimation approach include the assumption that consumer expenditure behavior from prior years, captured in price elasticities, will reflect future behavior among consumers, and potential underreporting of consumption in dietary recall data used to inform our calculations.

Abstract: CONCLUSION Sustained SSB taxation at a high tax rate could mitigate rising obesity and type 2 diabetes in India among both urban and rural subpopulations.


Acute myopericarditis after multiple vaccinations in an adolescent: case report and review of the literature.

Abstract: We report a case of postvaccination acute myopericarditis in an adolescent.

Abstract: The patient presented with acute chest pain, diffuse ST-segment elevation, and elevated cardiac enzyme levels.

Abstract: Cardiac MRI was consistent with acute myocarditis.

Abstract: He recovered within a few days with nonsteroidal antiinflammatory treatment and remains clinically stable, with improvement of MRI findings at the 10-week follow-up.

Abstract: Postvaccination cases of myopericarditis reported in the pediatric literature are also reviewed.


Cardiovascular protection with antihypertensive drugs in dialysis patients: systematic review and meta-analysis.

Abstract: Epidemiological studies demonstrate that a lower blood pressure and decline in blood pressure over months or years are associated with higher mortality in dialysis patients.

Abstract: In contrast, randomized, controlled trials lack power to establish benefits of antihypertensive therapy.

Abstract: Patients on long-term dialysis participating in randomized, controlled trials and receiving antihypertensive drug therapy were the subject of this meta-analysis.

Abstract: Outcomes assessed were the hazard ratio of cardiovascular events and all-cause mortality in treated group compared with controls.

Abstract: Among 1202 patients who we identified in 5 studies, the overall benefit of antihypertensive therapy compared with the control or placebo group had a combined hazard ratio for cardiovascular events of 0.69 (95% CI: 0.56 to 0.84) using a fixed-effects model and 0.62 (95% CI: 0.45 to 0.86) using a random-effects model.

Abstract: In a sensitivity analysis, we found that the hypertensive group had a pooled hazard ratio of 0.49 (95% CI: 0.35 to 0.67), but when normotensives were included in the trial, lesser cardiovascular protection was seen (pooled hazard ratio of 0.86 [95% CI: 0.67 to 1.12]).

Abstract: Test for heterogeneity between hypertensive and "normotensive-included" groups was significant (P<0.006).

Abstract: Similar results were seen for risk ratio for death and cardiovascular events.

Abstract: There was evidence of publication bias based on Egger's test and funnel plot.

Abstract: Randomized trials suggested a benefit of antihypertensive therapy among hemodialysis patients.

Abstract: Adequately powered randomized trials are required to confirm these observations, especially among those with hypertension.


Protoplasmic astrocytes in CA1 stratum radiatum occupy separate anatomical domains.

Abstract: Protoplasmic astrocytes are increasingly thought to interact extensively with neuronal elements in the brain and to influence their activity.

Abstract: Recent reports have also begun to suggest that physiologically, and perhaps functionally, diverse forms of these cells may be present in the CNS.

Abstract: Our current understanding of astrocyte form and distribution is based predominantly on studies that used the astrocytic marker glial fibrillary acidic protein (GFAP) and on studies using metal-impregnation techniques.

Abstract: The prevalent opinion, based on studies using these methods, is that astrocytic processes overlap extensively and primarily share the underlying neuropil.

Abstract: However, both of these techniques have serious shortcomings for visualizing the interactions among these structurally complex cells.

Abstract: In the present study, intracellular injection combined with immunohistochemistry for GFAP show that GFAP delineates only approximately 15% of the total volume of the astrocyte.

Abstract: As a result, GFAP-based images have led to incorrect conclusions regarding the interaction of processes of neighboring astrocytes.

Abstract: To investigate these interactions in detail, groups of adjacent protoplasmic astrocytes in the CA1 stratum radiatum were injected with fluorescent intracellular tracers of distinctive emissive wavelengths and analyzed using three-dimensional (3D) confocal analysis and electron microscopy.

Abstract: Our findings show that protoplasmic astrocytes establish primarily exclusive territories.

Abstract: The knowledge of how the complex morphology of protoplasmic astrocytes affects their 3D relationships with other astrocytes, oligodendroglia, neurons, and vasculature of the brain should have important implications for our understanding of nervous system function.


Leptin-receptor-expressing mesenchymal stromal cells represent the main source of bone formed by adult bone marrow.

Abstract: Studies of the identity and physiological function of mesenchymal stromal cells (MSCs) have been hampered by a lack of markers that permit both prospective identification and fate mapping in vivo.

Abstract: We found that Leptin Receptor (LepR) is a marker that highly enriches bone marrow MSCs.

Abstract: Approximately 0.3% of bone marrow cells were LepR(+)

Abstract: , 10% of which were CFU-Fs, accounting for 94% of bone marrow CFU-Fs.

Abstract: LepR(+) cells formed bone, cartilage, and adipocytes in culture and upon transplantation in vivo.

Abstract: LepR(+) cells were Scf-GFP(+), Cxcl12-DsRed(high), and Nestin-GFP(low), markers which also highly enriched CFU-Fs, but negative for Nestin-CreER and NG2-CreER, markers which were unlikely to be found in CFU-Fs.

Abstract: Fate-mapping showed that LepR(+) cells arose postnatally and gave rise to most bone and adipocytes formed in adult bone marrow, including bone regenerated after irradiation or fracture.

Abstract: LepR(+) cells were quiescent, but they proliferated after injury.

Abstract: Therefore, LepR(+) cells are the major source of bone and adipocytes in adult bone marrow.


High-resolution sperm typing of meiotic recombination in the mouse MHC Ebeta gene.

Abstract: Meiotic crossovers detected by pedigree analysis in the mouse MHC cluster into hotspots.

Abstract: To explore the properties of hotspots, we subjected the class II E(beta) gene to high-resolution sperm crossover analysis.

Abstract: We confirm the presence of a highly localized hotspot 1.0-1.6 kb wide in the second intron of E(beta) and show that it is flanked by DNA which is almost completely recombinationally inert.

Abstract: Mice heterozygous for haplotype s and another MHC haplotype show major haplotype-dependant variation in crossover rate but always the same hotspot, even in crosses including the highly diverged p haplotype.

Abstract: Crossovers in reciprocal orientations occur at similar rates but show different distributions across the hotspot, with the position of centre points in the two orientations shifted on average by 400 bp.

Abstract: This asymmetry results in crossover products showing biased gene conversion in favour of hotspot markers from the non-initiating haplotype, and supports the double-strand break repair model of recombination, with haplotype s as the most efficient crossover initiator.

Abstract: The detailed behaviour of the E(beta) hotspot, including evidence for highly localized recombination initiation, is strikingly similar to human hotspots.


Long-term survival of beta thalassemia major patients treated with hematopoietic stem cell transplantation compared with survival with conventional treatment.

Abstract: Allogeneic hematopoietic stem cell transplantation (HSCT) in thalassemia remains a challenge.

Abstract: We reported a single-centre case-control study of a large cohort of 516 children and adult patients treated with HSCT or blood transfusion support and iron chelation therapy; 258 patients (median age 12, range 1-45) underwent sibling (67%) or unrelated (33%) HSCT; 97 patients were adults (age ≥ 16 years).

Abstract: The median follow-up after HSCT was 11 years (range 1-30).

Abstract: The conditioning regimen was busulfan (80.6%) or treosulfan-based (19.4%).

Abstract: A cohort of 258 age-sex matched conventionally treated (CT) patients was randomly selected.

Abstract: In transplanted patients the 30-year overall survival (OS) and thalassemia-free survival (TFS) were 82.6 ± 2.7% and 77.8 ± 2.9%, compared to the OS of 85.3 ± 2.7% in CT patients (P = NS); The incidence of grade II-IV acute and chronic graft versus host disease (GvHD) was 23.6% and 12.9% respectively.

Abstract: The probability of rejection was 6.9%.

Abstract: Transplant-related mortality (TRM) (13.8%) was similar to the probability of dying of cardiovascular events in CT patients (12.2%).

Abstract: High-risk Pesaro score (class 3) was associated with lower OS (OR = 1.99, 95% C.I.=1.31-3.03) and TFS (OR = 1.54, 95% C.I.=1.12-2.12).

Abstract: In adult patients, the 23-years OS and TFS after HSCT were 70 ± 5% and 67.3 ± 5%, compared to 71.2 ± 5% of OS in CT (P = NS).

Abstract: Finally, treosulfan was associated with lower risk of acute GvHD (P = .004; OR = 0.28, 95% C.I.=0.12-0.67).

Abstract: In conclusion, the 30-year survival rate of ex-thalassemia patients after HSCT was similar to that expected in CT thalassemia patients, with the vast majority of HSCT survivors cured from thalassemia.


Global analysis of ATM polymorphism reveals significant functional constraint.

Abstract: ATM, the gene that is mutated in ataxia-telangiectasia, is associated with cerebellar degeneration, abnormal proliferation of small blood vessels, and cancer.

Abstract: These clinically important manifestations have stimulated interest in defining the sequence variation in the ATM gene.

Abstract: Therefore, we undertook a comprehensive survey of sequence variation in ATM in diverse human populations.

Abstract: The protein-encoding exons of the gene (9,168 bp) and the adjacent intron and untranslated sequences (14,661 bp) were analyzed in 93 individuals from seven major human populations.

Abstract: In addition, the coding sequence was analyzed in one chimpanzee, one gorilla, one orangutan, and one Old World monkey.

Abstract: In human ATM, 88 variant sites were discovered by denaturing high-performance liquid chromatography, which is 96%-100% sensitive for detection of DNA sequence variation.

Abstract: ATM was compared to 14 other autosomal genes for nucleotide diversity.

Abstract: The noncoding regions of ATM had diversity values comparable to other genes, but the coding regions had very low diversity, especially in the last 29% of the protein sequence.

Abstract: A test of the neutral evolution hypothesis, through use of the Hudson/Kreitman/Aguadé statistic, revealed that this region of the human ATM gene was significantly constrained relative to that of the orangutan, the Old World monkey, and the mouse, but not relative to that of the chimpanzee or the gorilla.

Abstract: ATM displayed extensive linkage disequilibrium, consistent with suppression of meiotic recombination at this locus.

Abstract: Seven haplotypes were defined.

Abstract: Two haplotypes accounted for 82% of all chromosomes analyzed in all major populations; two others carrying the same D126E missense polymorphism accounted for 33% of chromosomes in Africa but were never observed outside of Africa.

Abstract: The high frequency of this polymorphism may be due either to a population expansion within Africa or to selective pressure.


Ablation and regeneration of tolerance-inducing medullary thymic epithelial cells after cyclosporine, cyclophosphamide, and dexamethasone treatment.

Abstract: Immunosuppressive drugs and cytotoxic chemotherapy agents are designed to kill or suppress autoreactive, alloaggressive, or hyperinflammatory T cells, or disseminated malignancies.

Abstract: However, they also cause severe immunological side effects ranging from interrupted thymopoiesis and general immunodeficiency to, paradoxically, autoimmunity.

Abstract: Consistent with the cross-talk between thymocytes and stromal cells, we now show that these common therapeutic agents have major effects on murine thymic epithelial cells (TEC), crucially required to rebuild immunity posttreatment.

Abstract: We show that the immunosuppressant cyclosporine A, which has been linked to a thymus-dependent autoimmune syndrome in some patients, causes extensive loss of autoimmune regulator (Aire(+)) tolerance-inducing MHC class II(high) medullary TEC (mTEC(high)).

Abstract: Post-cyclosporine A, Aire expression was restored within 7 days.

Abstract: Full recovery of the mTEC(high) subset occurred within 10 days and was linked to a decrease in a relatively resistant MHC class II(low) mTEC subset (mTEC(low)), consistent with a previously described precursor-product relationship.

Abstract: Cyclophosphamide and dexamethasone caused more extensive ablation of thymocytes and stromal cells but again severely depleted tolerance-inducing mTEC(high).

Abstract: Together, these data show that Aire(+) mTECs are highly sensitive to damage and that mTEC regeneration follows a conserved pattern regardless of the treatment regimen used.


GATA-3 regulates hematopoietic stem cell maintenance and cell-cycle entry.

Abstract: Maintaining hematopoietic stem cell (HSC) quiescence is a critical property for the life-long generation of blood cells.

Abstract: Approximately 75% of cells in a highly enriched long-term repopulating HSC (LT-HSC) pool (Lin(-)Sca1(+)c-Kit(hi)CD150(+)CD48(-)) are quiescent, with only a small percentage of the LT-HSCs in cycle.

Abstract: Transcription factor GATA-3 is known to be vital for the development of T cells at multiple stages in the thymus and for Th2 differentiation in the peripheral organs.

Abstract: Although it is well documented that GATA-3 is expressed in HSCs, a role for GATA-3 in any prethymic progenitor cell has not been established.

Abstract: In the present study, we show that Gata3-null mutant mice generate fewer LT-HSCs and that fewer Gata3-null LT-HSCs are in cycle.

Abstract: Furthermore, Gata3 mutant hematopoietic progenitor cells fail to be recruited into an increased cycling state after 5-fluorouracil-induced myelosuppression.

Abstract: Therefore, GATA-3 is required for the maintenance of a normal number of LT-HSCs and for their entry into the cell cycle.


CpG island methylator phenotype predicts progression of malignant melanoma.

Abstract: PURPOSE The CpG island methylator phenotype (CIMP) may be associated with development of malignancy through coordinated inactivation of tumor suppressor and tumor-related genes (TRG) and methylation of multiple noncoding, methylated-in-tumor (MINT) loci.

Abstract: These epigenetic changes create a distinct CIMP pattern that has been linked to recurrence and survival in gastrointestinal cancers.

Abstract: Because epigenetic inactivation of TRGs also has been shown in malignant melanoma, we hypothesized the existence of a clinically significant CIMP in cutaneous melanoma progression.

Abstract: EXPERIMENTAL DESIGN The methylation status of the CpG island promoter region of TRGs related to melanoma pathophysiology (WIF1, TFPI2, RASSF1A, RARbeta2, SOCS1, and GATA4) and a panel of MINT loci (MINT1, MINT2, MINT3, MINT12, MINT17, MINT25, and MINT31) in primary and metastatic tumors of different clinical stages (n=122) was assessed.

Abstract: RESULTS Here, we show an increase in hypermethylation of the TRGs WIF1, TFPI2, RASSF1A, and SOCS1 with advancing clinical tumor stage.

Abstract: Furthermore, we find a significant positive association between the methylation status of MINT17, MINT31, and TRGs.

Abstract: The methylation status of MINT31 is associated with disease outcome in stage III melanoma.

Abstract: CONCLUSIONS These findings show the significance of a CIMP pattern that is associated with advancing clinical stage of malignant melanoma.

Abstract: Future prospective large-scale studies may determine if CIMP-positive primary melanomas are at high risk of metastasis or recurrence.


SLC1A5 mediates glutamine transport required for lung cancer cell growth and survival.

Abstract: PURPOSE We have previously identified solute-linked carrier family A1 member 5 (SLC1A5) as an overexpressed protein in a shotgun proteomic analysis of stage I non-small cell lung cancer (NSCLC) when compared with matched controls.

Abstract: We hypothesized that overexpression of SLC1A5 occurs to meet the metabolic demand for lung cancer cell growth and survival.

Abstract: EXPERIMENTAL DESIGN To test our hypothesis, we first analyzed the protein expression of SLC1A5 in archival lung cancer tissues by immunohistochemistry and immunoblotting (N = 98) and in cell lines (N = 36).

Abstract: To examine SLC1A5 involvement in amino acid transportation, we conducted kinetic analysis of l-glutamine (Gln) uptake in lung cancer cell lines in the presence and absence of a pharmacologic inhibitor of SLC1A5, gamma-l-Glutamyl-p-Nitroanilide (GPNA).

Abstract: Finally, we examined the effect of Gln deprivation and uptake inhibition on cell growth, cell-cycle progression, and growth signaling pathways of five lung cancer cell lines.

Abstract: RESULTS Our results show that (i) SLC1A5 protein is expressed in 95% of squamous cell carcinomas (SCC), 74% of adenocarcinomas (ADC), and 50% of neuroendocrine tumors; (ii) SLC1A5 is located at the cytoplasmic membrane and is significantly associated with SCC histology and male gender; (iii) 68% of Gln is transported in a Na(+)-dependent manner, 50% of which is attributed to SLC1A5 activity; and (iv) pharmacologic and genetic targeting of SLC1A5 decreased cell growth and viability in lung cancer cells, an effect mediated in part by mTOR signaling.

Abstract: CONCLUSIONS These results suggest that SLC1A5 plays a key role in Gln transport controlling lung cancer cells' metabolism, growth, and survival.