Eic1 links Mis18 with the CCAN/Mis6/Ctf19 complex to promote CENP-A assembly

Abstract: CENP-A chromatin forms the foundation for kinetochore assembly.

Abstract: Replication-independent incorporation of CENP-A at centromeres depends on its chaperone HJURP(Scm3), and Mis18 in vertebrates and fission yeast.

Abstract: The recruitment of Mis18 and HJURP(Scm3) to centromeres is cell cycle regulated.

Abstract: Vertebrate Mis18 associates with Mis18BP1(KNL2), which is critical for the recruitment of Mis18 and HJURP(Scm3).

Abstract: We identify two novel fission yeast Mis18-interacting proteins (Eic1 and Eic2), components of the Mis18 complex.

Abstract: Eic1 is essential to maintain Cnp1(CENP-A) at centromeres and is crucial for kinetochore integrity; Eic2 is dispensable.

Abstract: Eic1 also associates with Fta7(CENP-Q/Okp1), Cnl2(Nkp2) and Mal2(CENP-O/Mcm21), components of the constitutive CCAN/Mis6/Ctf19 complex.

Abstract: No Mis18BP1(KNL2) orthologue has been identified in fission yeast, consequently it remains unknown how the key Cnp1(CENP-A) loading factor Mis18 is recruited.

Abstract: Our findings suggest that Eic1 serves a function analogous to that of Mis18BP1(KNL2), thus representing the functional counterpart of Mis18BP1(KNL2) in fission yeast that connects with a module within the CCAN/Mis6/Ctf19 complex to allow the temporally regulated recruitment of the Mis18/Scm3(HJURP) Cnp1(CENP-A) loading factors.

Abstract: The novel interactions identified between CENP-A loading factors and the CCAN/Mis6/Ctf19 complex are likely to also contribute to CENP-A maintenance in other organisms.


INTRANASAL INSULIN IMPROVES COGNITION AND MODULATES β-AMYLOID IN EARLY AD

Abstract: Background: Reduced brain insulin signaling and low CSF-to-plasma insulin ratios have been observed in patients with Alzheimer disease (AD).

Abstract: Furthermore, intracerebroventricular or IV insulin administration improve memory, alter evoked potentials, and modulate neurotransmitters, possibly by augmenting low brain levels.

Abstract: After intranasal administration, insulin-like peptides follow extracellular pathways to the brain within 15 minutes.

Abstract: Objective: We tested the hypothesis that daily intranasal insulin treatment would facilitate cognition in patients with early AD or its prodrome, amnestic mild cognitive impairment (MCI).

Abstract: The proportion of verbal information retained after a delay period was the planned primary outcome measure.

Abstract: Secondary outcome measures included attention, caregiver rating of functional status, and plasma levels of insulin, glucose, β-amyloid, and cortisol.

Abstract: Methods: Twenty-five participants were randomly assigned to receive either placebo (n = 12) or 20 IU BID intranasal insulin treatment (n = 13) using an electronic atomizer, and 24 participants completed the study.

Abstract: Participants, caregivers, and all clinical evaluators were blinded to treatment assignment.

Abstract: Cognitive measures and blood were obtained at baseline and after 21 days of treatment.

Abstract: Results: Fasting plasma glucose and insulin were unchanged with treatment.

Abstract: The insulin-treated group retained more verbal information after a delay compared with the placebo-assigned group ( p = 0.0374).

Abstract: Insulin-treated subjects also showed improved attention ( p = 0.0108) and functional status ( p = 0.0410).

Abstract: Insulin treatment raised fasting plasma concentrations of the short form of the β-amyloid peptide (Aβ40; p = 0.0471) without affecting the longer isoform (Aβ42), resulting in an increased Aβ40/42 ratio ( p = 0.0207).

Abstract: Conclusions: The results of this pilot study support further investigation of the benefits of intranasal insulin for patients with Alzheimer disease, and suggest that intranasal peptide administration may be a novel approach to the treatment of neurodegenerative disorders.


Maternal alloantigens promote the development of tolerogenic fetal regulatory T cells in utero.

Abstract: As the immune system develops, T cells are selected or regulated to become tolerant of self antigens and reactive against foreign antigens.

Abstract: In mice, the induction of such tolerance is thought to be attributable to the deletion of self-reactive cells.

Abstract: Here, we show that the human fetal immune system takes advantage of an additional mechanism: the generation of regulatory T cells (Tregs) that suppress fetal immune responses.

Abstract: We find that substantial numbers of maternal cells cross the placenta to reside in fetal lymph nodes, inducing the development of CD4+CD25highFoxP3+ Tregs that suppress fetal antimaternal immunity and persist at least until early adulthood.

Abstract: These findings reveal a form of antigen-specific tolerance in humans, induced in utero and probably active in regulating immune responses after birth.


β-site amyloid precursor protein-cleaving enzyme 1(BACE1) inhibitor treatment induces Aβ5-X peptides through alternative amyloid precursor protein cleavage

Abstract: INTRODUCTION The β-secretase enzyme, β-site amyloid precursor protein-cleaving enzyme 1 (BACE1), cleaves amyloid precursor protein (APP) in the first step in β-amyloid (Aβ) peptide production.

Abstract: Thus, BACE1 is a key target for candidate disease-modifying treatment of Alzheimer's disease.

Abstract: In a previous exploratory Aβ biomarker study, we found that BACE1 inhibitor treatment resulted in decreased levels of Aβ1-34 together with increased Aβ5-40, suggesting that these Aβ species may be novel pharmacodynamic biomarkers in clinical trials.

Abstract: We have now examined whether the same holds true in humans.

Abstract: METHODS In an investigator-blind, placebo-controlled and randomized study, healthy subjects (n =18) were randomly assigned to receive a single dose of 30 mg of LY2811376 (n =6), 90 mg of LY2811376 (n =6), or placebo (n =6).

Abstract: We used hybrid immunoaffinity-mass spectrometry (HI-MS) and enzyme-linked immunosorbent assays to monitor a variety of Aβ peptides.

Abstract: RESULTS Here, we demonstrate dose-dependent changes in cerebrospinal fluid (CSF) Aβ1-34, Aβ5-40 and Aβ5-X after treatment with the BACE1-inhibitor LY2811376.

Abstract: Aβ5-40 and Aβ5-X increased dose-dependently, as reflected by two independent methods, while Aβ1-34 dose-dependently decreased.

Abstract: CONCLUSION Using HI-MS for the first time in a study where subjects have been treated with a BACE inhibitor, we confirm that CSF Aβ1-34 may be useful in clinical trials on BACE1 inhibitors to monitor target engagement.

Abstract: Since it is less hydrophobic than longer Aβ species, it is less susceptible to preanalytical confounding factors and may thus be a more stable marker.

Abstract: By independent measurement techniques, we also show that BACE1 inhibition in humans is associated with APP-processing into N-terminally truncated Aβ peptides via a BACE1-independent pathway.

Abstract: TRIAL REGISTRATION ClinicalTrials.gov NCT00838084.

Abstract: Registered: First received: January 23, 2009, Last updated: July 14, 2009, Last verified: July 2009.


Cardiovascular diseases in Chinese, Malays, and Indians in Singapore. I. Differences in mortality.

Abstract: STUDY OBJECTIVE The aim of the study was to analyse differences in mortality from the main cardiovascular diseases (ischaemic heart disease, hypertensive disease, and cerebrovascular disease) among Chinese, Malays, and Indians in Singapore.

Abstract: DESIGN

Abstract: The study was a survey using national death registration data in Singapore for the five years 1980 to 1984.

Abstract: The underlying cause of death, coded according to the ninth revision of the International Classification of Diseases, was taken for the analyses.

Abstract: SETTING The study was confined to the independent island state of Singapore, population 2.53 million (Chinese 76.5%, Malays 14.8%, Indians 6.4%, Others 2.3%).

Abstract: Death registration is thought to be complete.

Abstract: SUBJECTS All registered deaths in the age range 30-69 years during the study period were analysed by ethnic group.

Abstract: MEASUREMENT AND MAIN RESULTS Indians had higher mortality from ischaemic heart disease than the other ethnic groups in both sexes, with age-standardised relative risks of Indian v Chinese (males 3.8, females 3.4), Indian v Malay (males 1.9, females 1.6), and Malay v Chinese (males 2.0, females 2.2).

Abstract: The excess mortality in Indians declined with age.

Abstract: For hypertensive disease Malays had the highest mortality, with age-standardised relative risks of Malay v Chinese (males 3.4, females 4.4), Malay v Indian (males 2.0, females 2.5), and Indian v Chinese (males 1.6, females 1.6).

Abstract: For cerebrovascular disease there was little ethnic difference except for lower rates in Chinese females, with age-standardised relative risks of Malay v Chinese (males 1.1, females 1.9), Malay v Indian (males 1.0, females 1.1), and Indian v Chinese (males 1.1, females 1.7).

Abstract: CONCLUSIONS There are significant differences in mortality from the three main cardiovascular diseases in the different ethnic groups in Singapore.


Protein localization in electron micrographs using fluorescence nanoscopy

Abstract: A complete portrait of a cell requires a detailed description of its molecular topography: proteins must be linked to particular organelles.

Abstract: Immunocytochemical electron microscopy can reveal locations of proteins with nanometer resolution but is limited by the quality of fixation, the paucity of antibodies and the inaccessibility of antigens.

Abstract: Here we describe correlative fluorescence electron microscopy for the nanoscopic localization of proteins in electron micrographs.

Abstract: We tagged proteins with the fluorescent proteins Citrine or tdEos and expressed them in Caenorhabditis elegans, fixed the worms and embedded them in plastic.

Abstract: We imaged the tagged proteins from ultrathin sections using stimulated emission depletion (STED) microscopy or photoactivated localization microscopy (PALM).

Abstract: Fluorescence correlated with organelles imaged in electron micrographs from the same sections.

Abstract: We used these methods to localize histones, a mitochondrial protein and a presynaptic dense projection protein in electron micrographs.


The flavonoid component isorhamnetin in vitro inhibits proliferation and induces apoptosis in Eca-109 cells.

Abstract: Isorhamnetin is one member of flavonoid components which has been used in the treatment of heart disease.

Abstract: Recently the in vitro anti-cancer effect of isorhamnetin on human esophageal squamous carcinoma cell line Eca-109 was investigated in our lab.

Abstract: When Eca-109 cells were in vitro exposed to the graded doses of isorhamnetin (0-80 microg/ml) for 48 h, respectively, isorhamnetin exhibited cytostatic effect on the treated cells, with an IC(50) of 40+/-0.08 microg/ml as estimated by MTT assay.

Abstract: Inhibition on proliferation by isorhamnetin was detected by trypan blue exclusion assay, clone formation test, immunocytochemical assay of PCNA and (3)H-thymidine uptake analysis.

Abstract: Cell cycle distribution was measured by FCM.

Abstract: It was found that the viability of Eca-109 cells was significantly hampered by isorhamnetin.

Abstract: Compared with the negative control group, the treated group which was exposed to isorhamnetin had increased population in G(0)/G(1) phase from 74.6 to 84 while had a significant reduction in G(2)/M phase from 11.9 to 5.8.

Abstract: In addition to its cytostatic effect, isorhamnetin also showed stimulatory effect on apoptosis.

Abstract: Typical apoptotic morphology such as condensation and fragmentation of nuclei and blebbing membrane of the apoptotic cells could be observed through transmission electron microscope.

Abstract: Moreover, the sharp increase in apoptosis rate between the control and treated group were detected by FCM from 6.3 to 16.3.

Abstract: To explore the possible molecular mechanisms that underlie the growth inhibition and apoptosis stimulatory effects of isorhamnetin, the expressions of six proliferation- and death-related genes were detected by FCM.

Abstract: Expressions of bcl-2, c-myc and H-ras were downregulated whereas Bax, c-fos and p53 were upregulated.

Abstract: However, the in vivo experiments were required to further confirm the anti-cancer effects of isorhamnetin.

Abstract: In conclusion, isorhamnetin appears to be a potent drug against esophageal cancer due to its in vitro potential to not only inhibit proliferation but also induce apoptosis of Eca-109 cells.


Arachidonic Acid in the Diabetic Rat Kidney

Abstract: In the rat isolated perfused kidney, arachidonic acid elicits cyclooxygenase-dependent vasoconstriction through activation of PGH2/TxA2 receptors; responses are enhanced in kidneys from diabetic rats.

Abstract: This study examined the roles of cyclooxygenase-1/cyclooxygenase-2 in the enhanced renal vasoconstrictor effect of arachidonic acid in streptozotocin-diabetic rats.

Abstract: Release of 20-HETE was also determined, as this eicosanoid has been reported to elicit cyclooxygenase-dependent vasoconstriction.

Abstract: We confirmed that vasoconstrictor responses to arachidonic acid were enhanced in the diabetic rat kidney associated with a 2-fold-greater increase in the release of 6-ketoPGF1alpha, which was used as an index of cyclooxygenase activity.

Abstract: One and three micrograms of arachidonic acid increased perfusion pressure by 85+/-37 and 186+/-6 mm Hg, respectively, in diabetic rat kidneys compared with 3+/-1 and 17+/-8 mm Hg, respectively, in control rat kidneys.

Abstract: Inhibition of both cyclooxygenase isoforms with indomethacin (10 micromol/L) abolished the vasoconstrictor response to arachidonic acid in both diabetic and control rat kidneys, whereas inhibition of cyclooxygenase-2 with nimesulide (5 micromol/L) reduced perfusion pressure responses to 1 and 3 microg arachidonic acid only in the diabetic rat kidney to 15+/-8 and 108+/-26 mm Hg, respectively, consistent with a 3-fold increase in the renal cortical expression of cyclooxygenase-2.

Abstract: 20-HETE release from the diabetic rat kidney was reduced almost 6-fold and was not increased in response to arachidonic acid.

Abstract: These results demonstrate that the renal vasoconstrictor effect of arachidonic acid is solely dependent on cyclooxygenase activity, with no evidence for a contribution from 20-HETE; in the diabetic rat, cyclooxygenase-2 activity contributes to the renal vasoconstrictor effect of arachidonic acid.


Two sequence motifs from HIF-1alpha bind to the DNA-binding site of p53.

Abstract: There is evidence that hypoxia-inducible factor-1alpha (HIF-1alpha) interacts with the tumor suppressor p53.

Abstract: To characterize the putative interaction, we mapped the binding of the core domain of p53 (p53c) to an array of immobilized HIF-1alpha-derived peptides and found two peptide-sequence motifs that bound to p53c with micromolar affinity in solution.

Abstract: One sequence was adjacent to and the other coincided with the two proline residues of the oxygen-dependent degradation domain (P402 and P564) that act as switches for the oxygen-dependent regulation of HIF-1alpha.

Abstract: The binding affinity was independent of the hydroxylation state of P564.

Abstract: We found from NMR spectroscopy that these sequence motifs bind to the DNA-binding site of p53c.

Abstract: Because the two sequences are homologous and separated by 120 residues, and one is in a largely unstructured transactivation domain, we speculate that each sequence motif in HIF-1alpha binds to a different subunit of the p53 tetramer, leading to very tight binding.

Abstract: The binding data support the proposal that p53 provides a route for the degradation in hypoxic tumor cells of HIF-1alpha that is not hydroxylated at the two proline residues.


A functionally characterized test set of human induced pluripotent stem cells

Abstract: Human induced pluripotent stem cells (iPSCs) present exciting opportunities for studying development and for in vitro disease modeling.

Abstract: However, reported variability in the behavior of iPSCs has called their utility into question.

Abstract: We established a test set of 16 iPSC lines from seven individuals of varying age, sex and health status, and extensively characterized the lines with respect to pluripotency and the ability to terminally differentiate.

Abstract: Under standardized procedures in two independent laboratories, 13 of the iPSC lines gave rise to functional motor neurons with a range of efficiencies similar to that of human embryonic stem cells (ESCs).

Abstract: Although three iPSC lines were resistant to neural differentiation, early neuralization rescued their performance.

Abstract: Therefore, all 16 iPSC lines passed a stringent test of differentiation capacity despite variations in karyotype and in the expression of early pluripotency markers and transgenes.

Abstract: This iPSC and ESC test set is a robust resource for those interested in the basic biology of stem cells and their applications.


Differential regulation of MAP kinase activation by a novel splice variant of human MAP kinase phosphatase-2.

Abstract: MAP kinase phosphatase-2 (MKP-2) is a member of the family of dual specificity phosphatases that functions to inactivate the ERK and JNK MAP kinase signalling pathways.

Abstract: Here, we identify a novel human MKP-2 variant (MKP-2-S) lacking the MAP kinase binding site but retaining the phosphatase catalytic domain.

Abstract: Endogenous MKP-2-S transcripts and proteins were found in PC3 prostate and MDA-MB-231 breast cancer cells and also human prostate biopsies.

Abstract: Cellular transfection of MKP-2-S gave rise to a nuclear protein of 33kDa which displayed phosphatase activity comparable to the formerly described long form of MKP-2 (MKP-2-L).

Abstract: Due to its lack of a kinase interacting motif (KIM), MKP-2-S did not bind to JNK or ERK; MKP-2-L bound ERK and to a lesser extent JNK.

Abstract: Protein turnover of adenoviral expressed MKP-2-S was accelerated relative to MKP-2-L, with a greater susceptibility to proteosomal-mediated degradation.

Abstract: MKP-2-S retained its ability to deactivate JNK in a similar manner as MKP-2-L and was an effective inhibitor of LPS-stimulated COX-2 induction.

Abstract: However, unlike MKP-2-L, MKP-2-S was unable to reverse serum-induced ERK activation or significantly inhibit endothelial cell proliferation.

Abstract: These findings reveal the occurrence of a novel splice variant of MKP-2 which is unable to bind ERK and may be significant in the dysregulation of MAP kinase activity in certain disease states, particularly in breast and prostate cancers.


Myeloperoxidase acts as a profibrotic mediator of atrial fibrillation

Abstract: Observational clinical and ex vivo studies have established a strong association between atrial fibrillation and inflammation.

Abstract: However, whether inflammation is the cause or the consequence of atrial fibrillation and which specific inflammatory mediators may increase the atria's susceptibility to fibrillation remain elusive.

Abstract: Here we provide experimental and clinical evidence for the mechanistic involvement of myeloperoxidase (MPO), a heme enzyme abundantly expressed by neutrophils, in the pathophysiology of atrial fibrillation.

Abstract: MPO-deficient mice pretreated with angiotensin II (AngII) to provoke leukocyte activation showed lower atrial tissue abundance of the MPO product 3-chlorotyrosine, reduced activity of matrix metalloproteinases and blunted atrial fibrosis as compared to wild-type mice.

Abstract: Upon right atrial electrophysiological stimulation, MPO-deficient mice were protected from atrial fibrillation, which was reversed when MPO was restored.

Abstract: Humans with atrial fibrillation had higher plasma concentrations of MPO and a larger MPO burden in right atrial tissue as compared to individuals devoid of atrial fibrillation.

Abstract: In the atria, MPO colocalized with markedly increased formation of 3-chlorotyrosine.

Abstract: Our data demonstrate that MPO is a crucial prerequisite for structural remodeling of the myocardium, leading to an increased vulnerability to atrial fibrillation.


Knowledge and use of secondary contraception among patients requesting termination of pregnancy.

Abstract: The results of a survey of 769 patients attending the St. James's University Fertility Control Clinic, England, for abortion services showed that patients seeing general practitioners were less knowledgeable than those attending specialist clinics.

Abstract: There was a demonstrated need for counseling on pill and condom use and protection against sexually transmitted diseases.

Abstract: Knowledge of postcoital methods was also found to be lacking.

Abstract: The survey was conducted between April 1, 1991, and January 31, 1992.

Abstract: Respondents included minorities such as Afro-Caribbean (8%) and Asian (9%).

Abstract: 307 of the cases were using a less effective form of contraception at the time of conception, usually a change from the pill to condoms.

Abstract: Of the 171 people reporting failure of contraception, 93 noted a split or leaking condom; 13, a condom falling off during intercourse; 32, inconsistent use of condoms;l 32, forgetting to take contraceptive pills or using antibiotics with the pill; and 1, a late injection of medroxyprogesterone acetate.

Abstract: 45 of the 309 people who had conceived while using condoms recognized a potential condom failure, and only 20 attempted any emergency contraceptive method such as the postcoital pill.

Abstract: Only 30% of the 171 patients with recognized condom failure and 12% of the 210 who had not used any contraception had adequate knowledge of the existence, timing, and source of postcoital pills; i.e., 20% of 381.

Abstract: Only 2% of the 171 nd 2% of the 381 patients, had knowledge of postcoital insertion of an intrauterine contraceptive device.

Abstract: Given the choice between and unplanned pregnancy and postcoital contraceptive, most (718 out of 769) preferred using postcoital contraception.

Abstract: Contraceptive information was given to 501 by a general practitioner, to 102 by a community family planning clinic, and 163 had no medical advice.

Abstract: There was a range of knowledge of postcoital contraceptive methods.

Abstract: Knowledge of how to deal with forgotten pills, severe vomiting, severe diarrhea, and concurrent antibiotic treatment among the 422 patients who had ever used the combined pill also was variable.

Abstract: 19% of the 372 patients treated by general practitioners knew 4 correct answers, but 50% of the 50 patients in community family planning clinics answered correctly 4 times.

Abstract: Differences could not be explained by other demographic characteristics.


Domestication and Divergence of Saccharomyces cerevisiae Beer Yeasts

Abstract: Whereas domestication of livestock, pets, and crops is well documented, it is still unclear to what extent microbes associated with the production of food have also undergone human selection and where the plethora of industrial strains originates from.

Abstract: Here, we present the genomes and phenomes of 157 industrial Saccharomyces cerevisiae yeasts.

Abstract: Our analyses reveal that today's industrial yeasts can be divided into five sublineages that are genetically and phenotypically separated from wild strains and originate from only a few ancestors through complex patterns of domestication and local divergence.

Abstract: Large-scale phenotyping and genome analysis further show strong industry-specific selection for stress tolerance, sugar utilization, and flavor production, while the sexual cycle and other phenotypes related to survival in nature show decay, particularly in beer yeasts.

Abstract: Together, these results shed light on the origins, evolutionary history, and phenotypic diversity of industrial yeasts and provide a resource for further selection of superior strains.

Abstract: PAPERCLIP.


The ascendancy of Amblyomma americanum as a vector of pathogens affecting humans in the United States.

Abstract: Until the 1990s, Amblyomma americanum was regarded primarily as a nuisance species, but a tick of minor importance as a vector of zoonotic pathogens affecting humans.

Abstract: With the recent discoveries of Ehrlichia chaffeensis, Ehrlichia ewingii, and "Borrelia lonestari," the public health relevance of lone star ticks is no longer in question.

Abstract: During the next 25 years, the number of cases of human disease caused by A. americanum-associated pathogens will probably increase.

Abstract: Based on current trajectories and historic precedents, the increase will be primarily driven by biological and environmental factors that alter the geographic distribution and intensity of transmission of zoonotic pathogens.

Abstract: Sociologic and demographic changes that influence the likelihood of highly susceptible humans coming into contact with infected lone star ticks, in addition to advances in diagnostic capabilities and national surveillance efforts, will also contribute to the anticipated increase in the number of recognized cases of disease.


Biallelic mutations in PALB2 cause Fanconi anemia subtype FA-N and predispose to childhood cancer

Abstract: PALB2 was recently identified as a nuclear binding partner of BRCA2.

Abstract: Biallelic BRCA2 mutations cause Fanconi anemia subtype FA-D1 and predispose to childhood malignancies.

Abstract: We identified pathogenic mutations in PALB2 (also known as FANCN) in seven families affected with Fanconi anemia and cancer in early childhood, demonstrating that biallelic PALB2 mutations cause a new subtype of Fanconi anemia, FA-N, and, similar to biallelic BRCA2 mutations, confer a high risk of childhood cancer.


Long tandem arrays of complex repeat units in Chironomus telomeres.

Abstract: A cloned 340-bp DNA fragment excised by EcoRI from the Chironomus pallividittatus genome has been localized to the telomeres by in situ hybridization as well as to connectives between telomeres.

Abstract: No hybridization was observed in other regions of the chromosomes.

Abstract: Another cloned EcoRI fragment, 525 bp long has also been studied.

Abstract: This represents a partial duplication of the 340-bp sequence.

Abstract: Genomic blot hybridization experiments show that the 340-bp sequence is a representative monomeric unit of tandemly repeated arrays which account for 1.2% of the Chironomus genome, on average 300 kb per telomere.

Abstract: The repeat unit contains two types of subrepeats each present twice per repeat unit.

Abstract: Northern blot hybridization experiments show that the telomere-associated sequences are transcribed into a discrete RNA species approximately 20 kb in size.

Abstract: The evolution of this telomere-associated DNA is discussed.


Localization of Na+ channel isoforms at the atrioventricular junction and atrioventricular node in the rat.

Abstract: BACKGROUND The electrical activity of the atrioventricular node (AVN) is functionally heterogeneous, but how this relates to distinct cell types and the 3-dimensional structure of the AVN is unknown.

Abstract: To address this, we have studied the expression of Na(V)1.5 and other Na+ channel isoforms in the AVN.

Abstract: METHODS AND RESULTS The rat AVN was identified by Masson's trichrome staining together with immunolabeling of marker proteins: connexin40, connexin43, desmoplakin, atrial natriuretic peptide, and hyperpolarization-activated and cyclic nucleotide-gated channel 4.

Abstract: Na+ channel expression was investigated with immunohistochemistry with isoform-specific Na+ channel antibodies.

Abstract: Na(V)1.1 was distributed in a similar manner to Na(V)1.5.

Abstract: Na(V)1.2 was not detected.

Abstract: Na(V)1.3 labeling was present in nerve fibers and cell bodies (but not myocytes) and was abundant in the penetrating atrioventricular (AV) bundle and the common bundle but was much less abundant in other regions.

Abstract: Na(V)1.5 labeling was abundant in the atrial and ventricular myocardium and the left bundle branch.

Abstract: Na(V)1.5 labeling was absent in the open node, penetrating AV bundle, AV ring bundle, and common bundle but present at a reduced level in the inferior nodal extension and transitional zone.

Abstract: Na(V)1.6 was not detected.

Abstract: CONCLUSIONS Our findings provide molecular evidence of multiple electrophysiological cell types at the AV junction.

Abstract: Impaired AV conduction as a result of mutations in or loss of Na(V)1.5 must be the result of impaired conduction in the AVN inputs (inferior nodal extension and transitional zone) or output (bundle branches) rather than the AVN itself (open node and penetrating AV bundle).


Follicular B cell trafficking within the spleen actively restricts humoral immune responses.

Abstract: Follicular (FO) and marginal zone (MZ) B cells are maintained in distinct locations within the spleen, but the genetic basis for this separation is still enigmatic.

Abstract: We now report that B cell sequestration requires lineage-specific regulation of migratory receptors by the transcription factor Klf2.

Abstract: Moreover, using gene-targeted mice we show that altered splenic B cell migration confers a significant in vivo gain-of-function phenotype to FO B cells, including the ability to quickly respond to MZ-associated antigens and pathogens in a T cell-dependent manner.

Abstract: This work demonstrates that in wild-type animals, naive FO B cells are actively removed from the MZ, thus restricting their capacity to respond to blood-borne pathogens.


Interleukin-2 and inflammation induce distinct transcriptional programs that promote the differentiation of effector cytolytic T cells.

Abstract: Interleukin(IL)-2 and inflammation regulate effector and memory cytolytic T-lymphocyte (CTL) generation during infection.

Abstract: We demonstrate a complex interplay between IL-2 and inflammatory signals during CTL differentiation.

Abstract: IL-2 stimulation induced the transcription factor eomesodermin (Eomes), upregulated perforin (Prf1) transcription, and repressed re-expression of memory CTL markers Bcl6 and IL-7Ralpha.

Abstract: Binding of Eomes and STAT5 to Prf1 cis-regulatory regions correlated with transcriptional initiation (increased recruitment of RNA polymerase II to the Prf1 promoter).

Abstract: Inflammation (CpG, IL-12) enhanced expression of IL-2Ralpha and the transcription factor T-bet, but countered late Eomes and perforin induction while preventing IL-7Ralpha repression by IL-2.

Abstract: After infection of mice with lymphocytic choriomeningitis virus, IL-2Ralpha-deficient effector CD8(+) T cells expressed more Bcl6 but less perforin and granzyme B, formed fewer KLRG-1(+) and T-bet-expressing CTL, and killed poorly.

Abstract: Thus, inflammation influences both effector and memory CTL differentiation, whereas persistent IL-2 stimulation promotes effector at the expense of memory CTL development.


Neighbourhood risk factors for tuberculosis in Hong Kong.

Abstract: BACKGROUND Tuberculosis (TB) has been reported to be associated with poverty, especially in developing countries.

Abstract: Hong Kong is one of the few industrialised areas with a high incidence of TB where previous reports on the effect of poverty at neighbourhood level have been conflicting.

Abstract: OBJECTIVE To examine the spatial distribution of TB and its association with neighbourhood risk factors.

Abstract: METHOD A total of 17 294 TB cases notified from 2005 to 2007 were mapped down to the District Council Constituency Area (DCCA) level, and were indirectly standardised by age and sex using 2006 census population data.

Abstract: The standardised TB ratio was correlated with neighbourhood risk factors classified by family, ethnicity, economic and environmental domains.

Abstract: RESULTS The indirect age- and sex-standardised ratio demonstrated a spatially varied pattern, and was significantly associated with all neighbourhood factors on univariate analysis.

Abstract: Only marital status, place of birth and low household income were independently associated with the standardised TB ratio on multivariate analysis.

Abstract: CONCLUSION Despite the virtual elimination of absolute poverty by a well-developed social assistance scheme, low household income in the neighbourhood was significantly associated with TB, independently of place of birth, marital status and other risk factors.


Genomic stability and tumour suppression by the APC/C cofactor Cdh1

Abstract: The anaphase promoting complex or cyclosome (APC/C) is a ubiquitin protein ligase that, together with Cdc20 or Cdh1, targets cell-cycle proteins for degradation.

Abstract: APC/C–Cdh1 specifically promotes protein degradation in late mitosis and G1.

Abstract: Mutant embryos lacking Cdh1 die at E9.5–E10.5 due to defects in the endoreduplication of trophoblast cells and placental malfunction.

Abstract: This lethality is prevented when Cdh1 is expressed in the placenta.

Abstract: Cdh1-deficient cells proliferate inefficiently and accumulate numeric and structural chromosomal aberrations, indicating that Cdh1 contributes to the maintenance of genomic stability.

Abstract: Cdh1 heterozygous animals show increased susceptibility to spontaneous tumours, suggesting that Cdh1 functions as a haploinsufficient tumour suppressor.

Abstract: These heterozygous mice also show several defects in behaviour associated with increased proliferation of stem cells in the nervous system.

Abstract: These results indicate that Cdh1 is required for preventing unscheduled proliferation of specific progenitor cells and protecting mammalian cells from genomic instability.


Developing primary palliative care.

Abstract: Although 65% of people with cancer want to die at home, only about 30% are successful in doing so.1,2 A government committed to choice for patients must improve this figure.3 Developing palliative care services in primary care is essential for realising the expectations of dying people.

Abstract: Such services could also offer important opportunities for extending supportive humane care at an earlier stage, and to people not only with cancer but with chronic obstructive pulmonary disease, motor neurone disease, and cardiac failure, for example, who also often have palliative care needs.

Abstract: Primary care professionals have the potential and ability to provide end of life care for most patients, given adequate training, resources, and, when needed, specialist advice.4,5 They share common values with palliative care specialists—holistic, patient centred care, delivered in the context of families and friends.6 However, until recently, apart from Macmillan general practitioners and nurse facilitators, few comprehensive workforce initiatives have been undertaken in primary care that focus on end of life care.

Abstract: Many cancer patients and their carers experience existential distress long before they die.7 Recognising and alleviating such suffering is important, but it often goes unrecognised or is overlooked by services focusing on the terminal phase of illnesses.

Abstract: Primary care teams may know patients over long periods of time.

Abstract: They can readily identify patients from cancer and chronic disease registers who might benefit from an early palliative care approach.

Abstract: Such patients could be identified by clinicians asking one simple question of themselves: “Would I be surprised if my patient were to die in the next 12 months?”8 By identifying such patients proactively we could deliver, simultaneously, active treatment and patient centred supportive care, through a team with whom many patients have a valued long term relationship.

Abstract: Palliative care services need to be extended to patients with non-malignant conditions who have comparable concerns to and in some cases even greater unmet needs than cancer patients.9 Progress by palliative medicine specialists is hampered by issues such as uncertainty about the most effective models of care, lack of non-cancer expertise, and concerns about pressure on specialist services.

Abstract: General practitioners and community nurses can lead the way in providing a palliative care approach for patients with terminal organ failure illness.

Abstract: The first step in such an approach is for the goals of care to be discussed and agreed.

Abstract: Management plans are adjusted accordingly.

Abstract: Effective control of symptoms and maintaining quality of life are prioritised.

Abstract: In the light of these important opportunities it is regrettable that the new general medical services contract has not prioritised palliative care.

Abstract: By day, other developments to achieve the quality indicators are taking precedence.

Abstract: By night and at weekends, the new unscheduled care services (which are responsible for providing care for 75% of the hours in the week) are even less well configured than previous out of hours provision to facilitate dying at home.

Abstract: Such services specialise in dealing with acute emergencies and, as such, often struggle to meet the medical, nursing, and social care needs of dying people and their families.

Abstract: These changes will greatly affect care for dying people and may increase the number of hospital admissions.

Abstract: However, one important initiative is gaining momentum within primary care.

Abstract: The Gold Standards Framework is a resource for organising proactive palliative care in the community and is supported by funding from the Cancer Services Collaborative, Macmillan Cancer Relief, and the National Lottery.10 The framework provides a detailed guide to providing holistic, patient centred care and thereby facilitates effective care in the community.

Abstract: Other recently initiated mechanisms for developing primary palliative care include the training of general practitioners with a special interest in palliative care and the new end of life initiative in England to improve palliative care provision by generalists and to share examples of good practice.

Abstract: To support such developments it is essential that primary palliative care is supported by an adequate academic base.11 This is admittedly a challenging arena in which to undertake research, but progress has been made in recent years in developing conceptual models and research architectures for studying end of life issues.

Abstract: Now we need to build on this work to ensure that the understanding and insights gleaned can be translated into effective interventions.

Abstract: Every person with a progressive illness has a right to palliative care.12 Patients desire a reassuring professional presence in the face of death.

Abstract: General practitioners and community nurses are trusted by patients and are in a position to provide effective, equitable, and accessible palliative care.

Abstract: This will happen only if they have adequate time and resources and work in a system that encourages such care.

Abstract: Patients who receive holistic support in the community may be less likely to require expensive admission to hospital and often futile treatments at the end of their lives.


The effects of self-administering emergency contraception.

Abstract: BACKGROUND Emergency postcoital contraception prevents pregnancy, but it must be prescribed by a doctor and taken within 72 hours of intercourse.

Abstract: It has been proposed that emergency contraception be made available without a prescription.

Abstract: We undertook a study to learn how women might behave if given a supply of emergency contraceptive pills to keep at home.

Abstract: METHODS We assigned 553 women to be given a replaceable supply of hormonal emergency contraceptive pills to take home (the treatment group) and 530 women to use emergency contraception obtained by visiting a doctor (the control group).

Abstract: The frequency of use of emergency contraception, the use of other contraceptives, and the incidence of unwanted pregnancy were determined in both groups of women one year later.

Abstract: RESULTS The results for 549 women in the treatment group and 522 women in the control group were available for analysis.

Abstract: Three hundred seventy-nine of the women in the treatment group (69 percent) and 326 of the women in the control group (62 percent) contributed detailed information at follow-up.

Abstract: One hundred eighty of the women in the treatment group (47 percent) used emergency contraception at least once.

Abstract: Among those who returned the study questionnaire, 98 percent used emergency contraception correctly.

Abstract: There were no serious adverse effects.

Abstract: Eighty-seven women in the control group (27 percent) used emergency contraception at least once (P<0.001 for the comparison with the treatment group).

Abstract: The women in the treatment group were not more likely to use emergency contraception repeatedly.

Abstract: Their use of other methods of contraception was no different from that of the women in the control group.

Abstract: There were 18 unintended pregnancies in the treatment group and 25 in the control group (relative risk, 0.7; 95 percent confidence interval, 0.4 to 1.2).

Abstract: CONCLUSIONS Making emergency contraception more easily obtainable does no harm and may reduce the rate of unwanted pregnancies.


A coactivator of pre-mRNA splicing.

Abstract: The nuclear matrix antigen recognized by the monoclonal antibody (mAb) B1C8 is a novel serine (S) and arginine (R)-rich protein associated with splicing complexes and is named here SRm160 (SR-related matrix protein of 160 kD).

Abstract: SRm160 contains multiple SR repeats, but unlike proteins of the SR family of splicing factors, lacks an RNA recognition motif.

Abstract: SRm160 and a related protein SRm300 (the 300-kD nuclear matrix antigen recognized by mAb B4A11) form a complex that is required for the splicing of specific pre-mRNAs.

Abstract: The SRm160/300 complex associates with splicing complexes and promotes splicing through interactions with SR family proteins.

Abstract: Binding of SRm160/300 to pre-mRNA is normally also dependent on U1 snRNP and is stabilized by U2 snRNP.

Abstract: Thus, SRm160/300 forms multiple interactions with components bound directly to important sites within pre-mRNA.

Abstract: The results suggest that a complex of the nuclear matrix proteins SRm160 and SRm300 functions as a coactivator of pre-mRNA splicing.


Comparison of pioglitazone vs glimepiride on progression of coronary atherosclerosis in patients with type 2 diabetes: the PERISCOPE randomized controlled trial.

Abstract: CONTEXT No antidiabetic regimen has demonstrated the ability to reduce progression of coronary atherosclerosis.

Abstract: Commonly used oral glucose-lowering agents include sulfonylureas, which are insulin secretagogues, and thiazolidinediones, which are insulin sensitizers.

Abstract: OBJECTIVE To compare the effects of an insulin sensitizer, pioglitazone, with an insulin secretagogue, glimepiride, on the progression of coronary atherosclerosis in patients with type 2 diabetes.

Abstract: DESIGN, SETTING, AND PARTICIPANTS Double-blind, randomized, multicenter trial at 97 academic and community hospitals in North and South America (enrollment August 2003-March 2006) in 543 patients with coronary disease and type 2 diabetes.

Abstract: INTERVENTIONS A total of 543 patients underwent coronary intravascular ultrasonography and were randomized to receive glimepiride, 1 to 4 mg, or pioglitazone, 15 to 45 mg, for 18 months with titration to maximum dosage, if tolerated.

Abstract: Atherosclerosis progression was measured by repeat intravascular ultrasonography examination in 360 patients at study completion.

Abstract: MAIN OUTCOME MEASURE Change in percent atheroma volume (PAV) from baseline to study completion.

Abstract: RESULTS Least squares mean PAV increased 0.73% (95% CI, 0.33% to 1.12%) with glimepiride and decreased 0.16% (95% CI, -0.57% to 0.25%) with pioglitazone(P = .002).

Abstract: An alternative analysis imputing values for noncompleters based on baseline characteristics showed an increase in PAV of 0.64% (95% CI, 0.23% to 1.05%) for glimepiride and a decrease of 0.06% (-0.47% to 0.35%) for pioglitazone (between-group P = .02).

Abstract: Mean (SD) baseline HbA(1c) levels were 7.4% (1.0%) in both groups and declined during treatment an average 0.55% (95% CI, -0.68% to -0.42%) with pioglitazone and 0.36% (95% CI, -0.48% to -0.24%) with glimepiride (between-group P = .03).

Abstract: In the pioglitazone group, compared with glimepiride, high-density lipoprotein levels increased 5.7 mg/dL (95% CI, 4.4 to 7.0 mg/dL; 16.0%) vs 0.9 mg/dL (95% CI, -0.3 to 2.1 mg/dL; 4.1%), and median triglyceride levels decreased 16.3 mg/dL (95% CI, -27.7 to -11.0 mg/dL; 15.3%) vs an increase of 3.3 mg/dL (95% CI, -10.7 to 11.7 mg/dL; 0.6%) (P < .001 for both comparisons).

Abstract: Median fasting insulin levels decreased with pioglitazone and increased with glimepiride (P < .001).

Abstract: Hypoglycemia was more common in the glimepiride group and edema, fractures, and decreased hemoglobin levels occurred more frequently in the pioglitazone group.

Abstract: CONCLUSION In patients with type 2 diabetes and coronary artery disease, treatment with pioglitazone resulted in a significantly lower rate of progression of coronary atherosclerosis compared with glimepiride.

Abstract: TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT00225277.


Staphylococcus aureus RNAIII coordinately represses the synthesis of virulence factors and the transcription regulator Rot by an antisense mechanism.

Abstract: RNAIII is the intracellular effector of the quorum-sensing system in Staphylococcus aureus.

Abstract: It is one of the largest regulatory RNAs (514 nucleotides long) that are known to control the expression of a large number of virulence genes.

Abstract: Here, we show that the 3' domain of RNAIII coordinately represses at the post-transcriptional level, the expression of mRNAs that encode a class of virulence factors that act early in the infection process.

Abstract: We demonstrate that the 3' domain acts primarily as an antisense RNA and rapidly anneals to these mRNAs, forming long RNA duplexes.

Abstract: The interaction between RNAIII and the mRNAs results in repression of translation initiation and triggers endoribonuclease III hydrolysis.

Abstract: These processes are followed by rapid depletion of the mRNA pool.

Abstract: In addition, we show that RNAIII and its 3' domain mediate translational repression of rot mRNA through a limited number of base pairings involving two loop-loop interactions.

Abstract: Since Rot is a transcriptional regulatory protein, we proposed that RNAIII indirectly acts on many downstream genes, resulting in the activation of the synthesis of several exoproteins.

Abstract: These data emphasize the multitude of regulatory steps affected by RNAIII and its 3' domain in establishing a network of S. aureus virulence factors.


Human Cytomegalovirus Latency-Associated Proteins Elicit Immune-Suppressive IL-10 Producing CD4+ T Cells

Abstract: Human cytomegalovirus (HCMV) is a widely prevalent human herpesvirus, which, after primary infection, persists in the host for life.

Abstract: In healthy individuals, the virus is well controlled by the HCMV-specific T cell response.

Abstract: A key feature of this persistence, in the face of a normally robust host immune response, is the establishment of viral latency.

Abstract: In contrast to lytic infection, which is characterised by extensive viral gene expression and virus production, long-term latency in cells of the myeloid lineage is characterised by highly restricted expression of viral genes, including UL138 and LUNA.

Abstract: Here we report that both UL138 and LUNA-specific T cells were detectable directly ex vivo in healthy HCMV seropositive subjects and that this response is principally CD4⁺ T cell mediated.

Abstract: These UL138-specific CD4⁺ T cells are able to mediate MHC class II restricted cytotoxicity and, importantly, show IFNγ effector function in the context of both lytic and latent infection.

Abstract: Furthermore, in contrast to CDCD4⁺ T cells specific to antigens expressed solely during lytic infection, both the UL138 and LUNA-specific CD4⁺ T cell responses included CD4⁺ T cells that secreted the immunosuppressive cytokine cIL-10.

Abstract: We also show that cIL-10 expressing CD4⁺ T-cells are directed against latently expressed US28 and UL111A.

Abstract: Taken together, our data show that latency-associated gene products of HCMV generate CD4⁺ T cell responses in vivo, which are able to elicit effector function in response to both lytic and latently infected cells.

Abstract: Importantly and in contrast to CD4⁺ T cell populations, which recognise antigens solely expressed during lytic infection, include a subset of cells that secrete the immunosuppressive cytokine cIL-10.

Abstract: This suggests that HCMV skews the T cell responses to latency-associated antigens to one that is overall suppressive in order to sustain latent carriage in vivo.


Pamidronate induced anti-proliferative, apoptotic, and anti-migratory effects in hepatocellular carcinoma.

Abstract: BACKGROUND/AIMS The small GTPase of Ras and Rho families are widely involved in human tumorgenesis and metastasis.

Abstract: It has recently been reported that pamidronate inhibits the mevalonate pathway, which is required for the prenylation of the small GTPase.

Abstract: We demonstrated a possible beneficial use of pamidronate in the treatment of hepatocellular carcinoma (HCC).

Abstract: METHODS The effect of pamidronate on cell proliferation was analyzed with five hepatoma cell lines using MTT assay.

Abstract: Apoptosis was evaluated by staining with DAPI and a histon ELISA assay.

Abstract: A cell migration assay was performed using the Modified Boyden Chamber.

Abstract: To analyze anti-proliferation effect of pamidronate in vivo, tumor volumes were monitored with the intraperitoneal injection of pamidronate after subcutaneous inoculation of PLC/PRF/5 cells into nude mice.

Abstract: RESULTS Pamidronate inhibited cell growth for all hepatoma cell lines.

Abstract: The amount of membrane associated Ras and phosphorylated extracellular signal-regulated kinase 2 (ERK 2) were reduced after pamidronate treatment.

Abstract: Pamidronate increased apoptosis and cleavage of Caspase-3, and -9.

Abstract: Pamidronate suppressed membrane associated RhoA and cell motility.

Abstract: In vivo, tumor volumes were significantly suppressed by pamidronate at three weeks (P<0.03).

Abstract: CONCLUSIONS We conclude that pamidronate has therapeutic potential in inducing anti-proliferative, apoptotic, and anti-migratory effects in HCC.


Defective regulation of adipose tissue autophagy in obesity

Abstract: Objectives:

Abstract: Autophagy is a highly regulated process that has an important role in the control of a wide range of cellular functions, such as organelle recycling, nutrient availability and tissue differentiation.

Abstract: A recent study has shown an increased autophagic activity in the adipose tissue of obese subjects, and a role for autophagy in obesity-associated insulin resistance was proposed.

Abstract: Body mass reduction is the most efficient approach to tackle insulin resistance in over-weight subjects; however, the impact of weight loss in adipose tissue autophagy is unknown.

Abstract: Subjects:Adipose tissue autophagy was evaluated in mice and humans.

Abstract: Results:First, a mouse model of diet-induced obesity and diabetes was maintained on a 15-day, 40% caloric restriction.

Abstract: At baseline, markers of autophagy were increased in obese mice as compared with lean controls.

Abstract: Upon caloric restriction, autophagy increased in the lean mice, whereas it decreased in the obese mice.

Abstract: The reintroduction of ad libitum feeding was sufficient to rapidly reduce autophagy in the lean mice and increase autophagy in the obese mice.

Abstract: In the second part of the study, autophagy was evaluated in the subcutaneous adipose tissue of nine obese-non-diabetic and six obese-diabetic subjects undergoing bariatric surgery for body mass reduction.

Abstract: Specimens were collected during the surgery and approximately 1 year later.

Abstract: Markers of systemic inflammation, such as tumor necrosis factor-1α, interleukin (IL)-6 and IL-1β were evaluated.

Abstract: As in the mouse model, human obesity was associated with increased autophagy, and body mass reduction led to an attenuation of autophagy in the adipose tissue.

Abstract: Conclusion:Obesity and caloric overfeeding are associated with the defective regulation of autophagy in the adipose tissue.

Abstract: The studies in obese-diabetic subjects undergoing improved metabolic control following calorie restriction suggest that autophagy and inflammation are regulated independently.


Recovery of visual response of injured adult rat optic nerves treated with transglutaminase.

Abstract: Failure of axons of the central nervous system in adult mammals to regenerate spontaneously after injury is attributed in part to inhibitory molecules associated with oligodendrocytes.

Abstract: Regeneration of central nervous system axons in fish is correlated with the presence of a transglutaminase.

Abstract: This enzyme dimerizes interleukin-2, and the product is cytotoxic to oligodendrocytes in vitro.

Abstract: Application of this nerve-derived transglutaminase to rat optic nerves, in which the injury had caused the loss of visual evoked potential response to light, promoted the recovery of that response within 6 weeks after injury.

Abstract: Transmission electron microscopy analysis revealed the concomitant appearance of axons in the distal stump of the optic nerve.


Cell survival during complete nutrient deprivation depends on lipid droplet-fueled β-oxidation of fatty acids.

Abstract: Cells exposed to stress of different origins synthesize triacylglycerols and generate lipid droplets (LD), but the physiological relevance of this response is uncertain.

Abstract: Using complete nutrient deprivation of cells in culture as a simple model of stress, we have addressed whether LD biogenesis has a protective role in cells committed to die.

Abstract: Complete nutrient deprivation induced the biogenesis of LD in human LN18 glioblastoma and HeLa cells and also in CHO and rat primary astrocytes.

Abstract: In all cell types, death was associated with LD depletion and was accelerated by blocking LD biogenesis after pharmacological inhibition of Group IVA phospholipase A2 (cPLA2α) or down-regulation of ceramide kinase.

Abstract: Nutrient deprivation also induced β-oxidation of fatty acids that was sensitive to cPLA2α inhibition, and cell survival in these conditions became strictly dependent on fatty acid catabolism.

Abstract: These results show that, during nutrient deprivation, cell viability is sustained by β-oxidation of fatty acids that requires biogenesis and mobilization of LD.


Molecular and cytogenetic changes involved in the immortalization of nasopharyngeal epithelial cells by telomerase.

Abstract: Nasopharyngeal carcinoma (NPC) is a common disease in Hong Kong and southern provinces of China.

Abstract: EBV infection is believed to play a critical role in the development of NPC.

Abstract: Previous studies on the transformation mechanism of EBV genes were mostly performed in either NPC or nonnasopharyngeal epithelial cells which may not be representative of premalignant nasopharyngeal epithelial cells.

Abstract: Establishment of a representative cell system would greatly facilitate the elucidation of the role of EBV infection in the development of NPC.

Abstract: Using telomerase alone, we were able to establish an immortalized nasopharyngeal epithelial cell line from primary nonmalignant nasopharyngeal biopsies.

Abstract: The telomerase-immortalized nasopharyngeal epithelial cells are largely diploid in karyotype.

Abstract: Interestingly, this newly immortalized nasopharyngeal epithelial cell line, referred as NP460hTert, harbors genetic alterations previously identified in premalignant and malignant nasopharyngeal epithelial cells.

Abstract: These include inactivation of p16 by homozygous deletion of the p16(INK4A) locus and downregulation of RASSF1A expression.

Abstract: The deletion of the p16(INK4A) locus appears to be the most crucial event for the immortalization of nasopharyngeal epithelial cells by telomerase and precedes RASSF1A downregulation.

Abstract: In addition, detailed analysis of the cytogenetic changes by conventional cytogenetics, spectral karyotyping (SKY) and array-based CGH revealed a gain of a 17q21-q25 fragment on 11p15 chromosome in all NP460hTert cells which occurred before deletion of the p16(INK4A) locus.

Abstract: Gain of 17q has been previously reported in NPC.

Abstract: In addition, activation of NF-kappaB was observed in immortalized NP460hTert cells at the later population doublings, and may play a role in the survival of immortalized NP epithelial cells.

Abstract: Id1 which is commonly expressed in various human cancers, including NPC, was also upregulated in the immortalized NP460hTert cells.

Abstract: Thus, the establishment of an immortalized nasopharyngeal epithelial cell line harboring common genetic alterations present in premalignant and cancerous nasopharyngeal epithelial cells may provide a valuable cell system to examine for early events involved in NPC carcinogenesis, particularly in elucidating the role of EBV infection in NPC development.


Cellular nucleic acid binding protein suppresses tumor cell metastasis and induces tumor cell death by downregulating heterogeneous ribonucleoprotein K in fibrosarcoma cells.

Abstract: BACKGROUND Cellular nucleic acid binding protein (CNBP) has been implicated in vertebrate craniofacial development and in myotonic dystrophy type 2 (DM2) and sporadic inclusion body myositis (sIBM) human diseases by controlling cell proliferation and survival to mediate neural crest expansion.

Abstract: CNBP has been found to bind single-stranded nucleic acid and promote rearrangements of nucleic acid secondary structure in an ATP-independent manner, acting as a nucleic acid chaperone.

Abstract: METHODS A variety of methods were used, including cell viability assays, wound-scratch assays, chemotaxis assays, invasion assays, circular dichroic (CD) spectroscopy, NMR spectroscopy, chromatin immunoprecipitation, expression and purification of recombinant human CNBP, electrophoretic mobility shift assay (EMSA), surface plasmon resonance (SPR), fluorescence resonance energy transfer (FRET) analyses, luciferase reporter assay, Western blotting, and isothermal titration calorimetry (ITC).

Abstract: RESULTS Up-regulation of CNBP induced human fibrosarcoma cell death and suppressed fibrosarcoma cell motility and invasiveness.

Abstract: It was found that CNBP transcriptionally down-regulated the expression of heterogeneous ribonucleoprotein K (hnRNP K) through its conversion of a G-rich sequence into G-quadruplex in the promoter of hnRNP K. G-quadruplex stabilizing ligand tetra-(N-methyl-4-pyridyl) porphyrin (TMPyP4) could interact with and stabilize the G-quadruplex, resulting in downregulation of hnRNP K transcription.

Abstract: CONCLUSIONS CNBP overexpression caused increase of cell death and suppression of cell metastasis through its induction of G-quadruplex formation in the promoter of hnRNP K resulting in hnRNP K down-regulation.

Abstract: GENERAL SIGNIFICANCE The present result provided a new solution for controlling hnRNP K expression, which should shed light on new anticancer drug design and development.


The DHR96 nuclear receptor regulates xenobiotic responses in Drosophila.

Abstract: Exposure to xenobiotics such as plant toxins, pollutants, or prescription drugs triggers a defense response, inducing genes that encode key detoxification enzymes.

Abstract: Although xenobiotic responses have been studied in vertebrates, little effort has been made to exploit a simple genetic system for characterizing the molecular basis of this coordinated transcriptional response.

Abstract: We show here that approximately 1000 transcripts are significantly affected by phenobarbital treatment in Drosophila.

Abstract: We also demonstrate that the Drosophila ortholog of the human SXR and CAR xenobiotic receptors, DHR96, plays a role in this response.

Abstract: A DHR96 null mutant displays increased sensitivity to the sedative effects of phenobarbital and the pesticide DDT as well as defects in the expression of many phenobarbital-regulated genes.

Abstract: Metabolic and stress-response genes are also controlled by DHR96, implicating its role in coordinating multiple response pathways.

Abstract: This work establishes a new model system for defining the genetic control of xenobiotic stress responses.


A primate virus generates transformed human cells by fusion

Abstract: Amodel that explains both the origin and sporadic nature of cancer argues that cancer cells are a chance result of events that cause genomic and epigenetic variability.

Abstract: The prevailing view is that these events are mutations that affect chromosome segregation or stability.

Abstract: However, genomic and epigenetic variability is also triggered by cell fusion, which is often caused by viruses.

Abstract: Yet, cells fused by viruses are considered harmless because they die.

Abstract: We provide evidence that a primate virus uses both viral and exosomal proteins involved in cell fusion to produce transformed proliferating human cells.

Abstract: Although normal cells indeed fail to proliferate after fusion, expression of an oncogene or a mutated tumor suppressor p53 in just one of the fusion partners is sufficient to produce heterogeneous progeny.

Abstract: We also show that this virus can produce viable oncogenically transformed cells by fusing cells that are otherwise destined to die.

Abstract: Therefore, we argue that viruses can contribute to carcinogenesis by fusing cells.


TAA1-Mediated Auxin Biosynthesis Is Essential for Hormone Crosstalk and Plant Development

Abstract: Plants have evolved a tremendous ability to respond to environmental changes by adapting their growth and development.

Abstract: The interaction between hormonal and developmental signals is a critical mechanism in the generation of this enormous plasticity.

Abstract: A good example is the response to the hormone ethylene that depends on tissue type, developmental stage, and environmental conditions.

Abstract: By characterizing the Arabidopsis wei8 mutant, we have found that a small family of genes mediates tissue-specific responses to ethylene.

Abstract: Biochemical studies revealed that WEI8 encodes a long-anticipated tryptophan aminotransferase, TAA1, in the essential, yet genetically uncharacterized, indole-3-pyruvic acid (IPA) branch of the auxin biosynthetic pathway.

Abstract: Analysis of TAA1 and its paralogues revealed a link between local auxin production, tissue-specific ethylene effects, and organ development.

Abstract: Thus, the IPA route of auxin production is key to generating robust auxin gradients in response to environmental and developmental cues.


A research agenda for public health workforce development.

Abstract: In the past decades, public health research has focused on categorical rather than cross-cutting or systems issues.

Abstract: Little research has been carried out on the infrastructure required to support public health programs.

Abstract: This article describes the results of an interactive process to develop a research agenda for public health workforce development to inform all those with stakes in the public health system.

Abstract: This research is defined as a multidisciplinary field of inquiry, both basic and applied, that examines the workforce in terms of costs, quality, accessibility, delivery, organization, financing, and outcomes of public health services to increase knowledge and understanding of the relationships among workforce and structure, processes, and effects of public health services.

Abstract: A logic model and five priority research areas resulted from meetings of expert panels during 2000 to 2003.

Abstract: Innovative public and private partnerships will be required to advance cross-cutting and systems-focused research.


Parallel processing in the mammalian retina

Abstract: Our eyes send different 'images' of the outside world to the brain — an image of contours (line drawing), a colour image (watercolour painting) or an image of moving objects (movie).

Abstract: This is commonly referred to as parallel processing, and starts as early as the first synapse of the retina, the cone pedicle.

Abstract: Here, the molecular composition of the transmitter receptors of the postsynaptic neurons defines which images are transferred to the inner retina.

Abstract: Within the second synaptic layer — the inner plexiform layer — circuits that involve complex inhibitory and excitatory interactions represent filters that select 'what the eye tells the brain'.


Oxymetholone Therapy of Fanconi Anemia Suppresses Osteopontin Transcription and Induces Hematopoietic Stem Cell Cycling

Abstract: Androgens are widely used for treating Fanconi anemia (FA) and other human bone marrow failure syndromes, but their mode of action remains incompletely understood.

Abstract: Aged Fancd2(-/-) mice were used to assess the therapeutic efficacy of oxymetholone (OXM) and its mechanism of action.

Abstract: Eighteen-month-old Fancd2(-/-) mice recapitulated key human FA phenotypes, including reduced bone marrow cellularity, red cell macrocytosis, and peripheral pancytopenia.

Abstract: As in humans, chronic OXM treatment significantly improved these hematological parameters and stimulated the proliferation of hematopoietic stem and progenitor cells.

Abstract: RNA-Seq analysis implicated downregulation of osteopontin as an important potential mechanism for the drug's action.

Abstract: Consistent with the increased stem cell proliferation, competitive repopulation assays demonstrated that chronic OXM therapy eventually resulted in stem cell exhaustion.

Abstract: These results expand our knowledge of the regulation of hematopoietic stem cell proliferation and have direct clinical implications for the treatment of bone marrow failure.


A point mutation in KINDLIN3 ablates activation of three integrin subfamilies in humans

Abstract: Monogenic deficiency diseases provide unique opportunities to define the contributions of individual molecules to human physiology and to identify pathologies arising from their dysfunction.

Abstract: Here we describe a deficiency disease in two human siblings that presented with severe bleeding, frequent infections and osteopetrosis at an early age.

Abstract: These symptoms are consistent with but more severe than those reported for people with leukocyte adhesion deficiency III (LAD-III).

Abstract: Mechanistically, these symptoms arose from an inability to activate the integrins expressed on hematopoietic cells, including platelets and leukocytes.

Abstract: Immortalized lymphocyte cell lines isolated from the two individuals showed integrin activation defects.

Abstract: Several proteins previously implicated in integrin activation, including Ras-associated protein-1 (RAP1) and calcium and diacylglycerol-regulated guanine nucleotide exchange factor-1 (CALDAG-GEF1), were present and functional in these cell lines.

Abstract: The genetic basis for this disease was traced to a point mutation in the coding region of the KINDLIN3 (official gene symbol FERMT3) gene.

Abstract: When wild-type KINDLIN-3 was expressed in the immortalized lymphocytes, their integrins became responsive to activation signals.

Abstract: These results identify a genetic disease that severely compromises the health of the affected individuals and establish an essential role of KINDLIN-3 in integrin activation in humans.

Abstract: Furthermore, allogeneic bone marrow transplantation was shown to alleviate the symptoms of the disease.


Fasting Activation of AgRP Neurons Requires NMDA Receptors and Involves Spinogenesis and Increased Excitatory Tone

Abstract: AgRP neuron activity drives feeding and weight gain whereas that of nearby POMC neurons does the opposite.

Abstract: However, the role of excitatory glutamatergic input in controlling these neurons is unknown.

Abstract: To address this question, we generated mice lacking NMDA receptors (NMDARs) on either AgRP or POMC neurons.

Abstract: Deletion of NMDARs from AgRP neurons markedly reduced weight, body fat and food intake whereas deletion from POMC neurons had no effect.

Abstract: Activation of AgRP neurons by fasting, as assessed by c-Fos, Agrp and Npy mRNA expression, AMPA receptor-mediated EPSCs, depolarization and firing rates, required NMDARs.

Abstract: Furthermore, AgRP but not POMC neurons have dendritic spines and increased glutamatergic input onto AgRP neurons caused by fasting was paralleled by an increase in spines, suggesting fasting induced synaptogenesis and spinogenesis.

Abstract: Thus glutamatergic synaptic transmission and its modulation by NMDARs play key roles in controlling AgRP neurons and determining the cellular and behavioral response to fasting.


Prevalence and Causes of Blindness and Low Vision in Southern Sudan

Abstract: Background Blindness and low vision are thought to be common in southern Sudan.

Abstract: However, the magnitude and geographical distribution are largely unknown.

Abstract: We aimed to estimate the prevalence of blindness and low vision, identify the main causes of blindness and low vision, and estimate targets for blindness prevention programs in Mankien payam (district), southern Sudan.


Origin of nitrite and nitrate in nasal and exhaled breath condensate and relation to nitric oxide formation.

Abstract: BACKGROUND Raised concentrations of nitrate and nitrite have been found in exhaled breath condensate (EBC) in airway disease, and it has been postulated that this reflects increased nitric oxide (NO) metabolism.

Abstract: However, the chemical and anatomical origin of nitrate and nitrite in the airways has not yet been sufficiently studied.

Abstract: METHODS The fraction of exhaled NO at an exhalation flow rate of 50 ml/s (FE(NO)) and nitrite and nitrate in EBC, nasal condensate, and saliva were measured in 17 tracheostomised and 15 non-tracheostomised subjects, all of whom were non-smokers without respiratory disease.

Abstract: Tracheal and oral samples were taken from the tracheostomised subjects and nasal (during velum closure) and oral samples from the non-tracheostomised subjects.

Abstract: Measurements were performed before and after sodium nitrate ingestion (10 mg/kg) and use of antibacterial mouthwash (chlorhexidine 0.2%).

Abstract: RESULTS In tracheostomised subjects oral FE(NO) increased by 90% (p<0.01) while tracheal FE(NO) was not affected 60 minutes after nitrate ingestion.

Abstract: Oral EBC nitrite levels were increased 23-fold at 60 minutes (p<0.001) whereas the nitrite levels in tracheal EBC showed only a minor increase (fourfold, p<0.05).

Abstract: Nitrate was increased the same amount in oral and tracheal EBC at 60 minutes (2.5-fold, p<0.05).

Abstract: In non-tracheostomised subjects oral FE(NO) and EBC nitrite increased after nitrate ingestion and after chlorhexidine mouthwash they approached baseline levels again (p<0.001).

Abstract: Nasal NO, nitrate, and nitrite were not affected by nitrate intake or mouthwash.

Abstract: At baseline, mouthwash with deionised water did not affect nitrite in oral EBC or saliva, whereas significant reductions were seen after antibacterial mouthwash (p<0.05 and p<0.001, respectively).

Abstract: CONCLUSIONS Besides the salivary glands, plasma nitrate is taken up by the lower airways but not the nasal airways.

Abstract: Nitrate levels in EBC are thus influenced by dietary intake.

Abstract: Nitrate is reduced to nitrite by bacterial activity which takes place primarily in the oropharyngeal tract of healthy subjects.

Abstract: Only oropharyngeal nitrite seems to contribute to exhaled NO in non-inflamed airways, and there is also a substantial contribution of nitrite from the oropharyngeal tract during standard collection of EBC.


AZD9150, a next-generation antisense oligonucleotide inhibitor of STAT3 with early evidence of clinical activity in lymphoma and lung cancer

Abstract: Next-generation sequencing technologies have greatly expanded our understanding of cancer genetics.

Abstract: Antisense technology is an attractive platform with the potential to translate these advances into improved cancer therapeutics, because antisense oligonucleotide (ASO) inhibitors can be designed on the basis of gene sequence information alone.

Abstract: Recent human clinical data have demonstrated the potent activity of systemically administered ASOs targeted to genes expressed in the liver.

Abstract: We describe the preclinical activity and initial clinical evaluation of a class of ASOs containing constrained ethyl modifications for targeting the gene encoding the transcription factor STAT3, a notoriously difficult protein to inhibit therapeutically.

Abstract: Systemic delivery of the unformulated ASO, AZD9150, decreased STAT3 expression in a broad range of preclinical cancer models and showed antitumor activity in lymphoma and lung cancer models.

Abstract: AZD9150 preclinical activity translated into single-agent antitumor activity in patients with highly treatment-refractory lymphoma and non–small cell lung cancer in a phase 1 dose-escalation study.


Evidence of a pluripotent human embryonic stem cell line derived from a cloned blastocyst.

Abstract: Somatic cell nuclear transfer (SCNT) technology has recently been used to generate animals with a common genetic composition.

Abstract: In this study, we report the derivation of a pluripotent embryonic stem (ES) cell line (SCNT-hES-1) from a cloned human blastocyst.

Abstract: The SCNT-hES-1 cells displayed typical ES cell morphology and cell surface markers and were capable of differentiating into embryoid bodies in vitro and of forming teratomas in vivo containing cell derivatives from all three embryonic germ layers in severe combined immunodeficient mice.

Abstract: After continuous proliferation for more than 70 passages, SCNT-hES-1 cells maintained normal karyotypes and were genetically identical to the somatic nuclear donor cells.

Abstract: Although we cannot completely exclude the possibility that the cells had a parthenogenetic origin, imprinting analyses support a SCNT origin of the derived human ES cells.


Ultrarapid drug metabolism: PCR-based detection of CYP2D6 gene duplication.

Abstract: The enzyme debrisoquine 4-hydroxylase (CYP2D6), which metabolizes many widely used drugs, is highly polymorphic.

Abstract: The activity of the enzyme ranges between subjects from ultrafast to a complete absence.

Abstract: Therefore, metabolic capacity varies, producing intersubject differences in therapeutic efficacy and side effects at standard recommended doses.

Abstract: Up to 7% of Caucasians may demonstrate ultrarapid drug metabolism (UM) because of inherited alleles with multiplicate functional CYP2D6 genes, causing an increased amount of enzyme to be expressed.

Abstract: Identification of UM subjects is of potential clinical importance for adjustment of doses in drug therapy, as well as to avoid misidentification of noncompliance.

Abstract: In our study, we tested recently designed PCR assays for the detection of the UM genotype.

Abstract: We found a 3.5% prevalence of UMs carrying duplicate active CYP2D6 genes in a population consisting of 202 psychiatric patients.


The Actions of Synaptically Released Zinc at Hippocampal Mossy Fiber Synapses

Abstract: Zn2+ is present at high concentrations in the synaptic vesicles of hippocampal mossy fibers.

Abstract: We have used Zn2+ chelators and the mocha mutant mouse to address the physiological role of Zn2+ in this pathway.

Abstract: Zn2+ is not involved in the unique presynaptic plasticities observed at mossy fiber synapses but is coreleased with glutamate from these synapses, both spontaneously and with electrical stimulation, where it exerts a strong modulatory effect on the NMDA receptors.

Abstract: Zn2+ tonically occupies the high-affinity binding site of NMDA receptors at mossy fiber synapses, whereas the lower affinity voltage-dependent Zn2+ binding site is occupied during action potential driven-release.

Abstract: We conclude that Zn2+ is a modulatory neurotransmitter released from mossy fiber synapses and plays an important role in shaping the NMDA receptor response at these synapses.


Association between folate intake from different food sources in Norway and homocysteine status in a dietary intervention among young male adults.

Abstract: The aim of the present investigation was to study the effect of a dietary intervention which combined nutrition information with increased availability of vegetables, fruits and wholegrain bread.

Abstract: The effect of the intervention was determined by changes in the intake of vegetables, fruits, wholegrain bread and estimated nutrients.

Abstract: Furthermore, the study investigated whether changes in relative contribution from different food sources of folate were related to changes in the concentration of plasma total homocysteine (p-tHcy).

Abstract: The 5-month intervention study included 376 male recruits from the Norwegian National Guard, Vaernes (intervention group) and 105 male recruits from the Norwegian National Guard, Heggelia (control group).

Abstract: The study resulted in an increase in the total consumption of vegetables, fruits, berries and juice (P < 0.001) and of wholegrain bread (P < 0.001).

Abstract: The participants in the intervention group showed a higher increase in the intake of dietary fibre (P < 0.001) and folate (P < 0.001) compared with the control group.

Abstract: The relative contribution of folate intake from fruits, vegetables and wholegrain bread was higher in the intervention group compared with the control group (P < 0.001 for all).

Abstract: The increased intake of folate from wholegrain bread was inversely associated with a reduced concentration of p-tHcy (P = 0.017).

Abstract: In summary, the dietary intervention resulted in an increased intake of vegetables, fruits and wholegrain bread and a subsequent increase in folate intake from these food components.

Abstract: Reduction in the concentration of p-tHcy was significantly related to an increased folate intake due to an increased consumption of wholegrain bread.


Beta interferon controls West Nile virus infection and pathogenesis in mice.

Abstract: Studies with mice lacking the common plasma membrane receptor for type I interferon (IFN-αβR(-)(/)(-)) have revealed that IFN signaling restricts tropism, dissemination, and lethality after infection with West Nile virus (WNV) or several other pathogenic viruses.

Abstract: However, the specific functions of individual IFN subtypes remain uncertain.

Abstract: Here, using IFN-β(-)(/)(-) mice, we defined the antiviral and immunomodulatory function of this IFN subtype in restricting viral infection.

Abstract: IFN-β(-)(/)(-) mice were more vulnerable to WNV infection than wild-type mice, succumbing more quickly and with greater overall mortality, although the phenotype was less severe than that of IFN-αβR(-)(/)(-) mice.

Abstract: The increased susceptibility of IFN-β(-)(/)(-) mice was accompanied by enhanced viral replication in different tissues.

Abstract: Consistent with a direct role for IFN-β in control of WNV replication, viral titers in ex vivo cultures of macrophages, dendritic cells, fibroblasts, and cerebellar granule cell neurons, but not cortical neurons, from IFN-β(-)(/)(-) mice were greater than in wild-type cells.

Abstract: Although detailed immunological analysis revealed no major deficits in the quality or quantity of WNV-specific antibodies or CD8(+) T cells, we observed an altered CD4(+) CD25(+) FoxP3(+) regulatory T cell response, with greater numbers after infection.

Abstract: Collectively, these results suggest that IFN-β controls WNV pathogenesis by restricting infection in key cell types and by modulating T cell regulatory networks.


Frontrunners of T cell activation: Initial, localized Ca2+ signals mediated by NAADP and the type 1 ryanodine receptor

Abstract: The activation of T cells is the fundamental on switch for the adaptive immune system.

Abstract: Ca2+ signaling is essential for T cell activation and starts as initial, short-lived, localized Ca2+ signals.

Abstract: The second messenger nicotinic acid adenine dinucleotide phosphate (NAADP) forms rapidly upon T cell activation and stimulates early Ca2+ signaling.

Abstract: We developed a high-resolution imaging technique using multiple fluorescent Ca2+ indicator dyes to characterize these early signaling events and investigate the channels involved in NAADP-dependent Ca2+ signals.

Abstract: In the first seconds of activation of either primary murine T cells or human Jurkat cells with beads coated with an antibody against CD3, we detected Ca2+ signals with diameters close to the limit of detection and that were close to the activation site at the plasma membrane.

Abstract: In Jurkat cells in which the ryanodine receptor (RyR) was knocked down or in primary T cells from RyR1−/− mice, either these early Ca2+ signals were not detected or the number of signals was markedly reduced.

Abstract: Local Ca2+ signals observed within 20 ms upon microinjection of Jurkat cells with NAADP were also sensitive to RyR knockdown.

Abstract: In contrast, TRPM2 (transient receptor potential channel, subtype melastatin 2), a potential NAADP target channel, was not required for the formation of initial Ca2+ signals in primary T cells.

Abstract: Thus, through our high-resolution imaging method, we characterized early Ca2+ release events in T cells and obtained evidence for the involvement of RyR and NAADP in such signals.


A missense LAMB2 mutation causes congenital nephrotic syndrome by impairing laminin secretion.

Abstract: Laminin β2 is a component of laminin-521, which is an important constituent of the glomerular basement membrane (GBM).

Abstract: Null mutations in laminin β2 (LAMB2) cause Pierson syndrome, a severe congenital nephrotic syndrome with ocular and neurologic defects.

Abstract: In contrast, patients with LAMB2 missense mutations, such as R246Q, can have less severe extrarenal defects but still exhibit congenital nephrotic syndrome.

Abstract: To investigate how such missense mutations in LAMB2 cause proteinuria, we generated three transgenic lines of mice in which R246Q-mutant rat laminin β2 replaced the wild-type mouse laminin β2 in the GBM.

Abstract: These transgenic mice developed much less severe proteinuria than their nontransgenic Lamb2-deficient littermates; the level of proteinuria correlated inversely with R246Q-LAMB2 expression.

Abstract: At the onset of proteinuria, expression and localization of proteins associated with the slit diaphragm and foot processes were normal, and there were no obvious ultrastructural abnormalities.

Abstract: Low transgene expressors developed heavy proteinuria, foot process effacement, GBM thickening, and renal failure by 3 months, but high expressors developed only mild proteinuria by 9 months.

Abstract: In vitro studies demonstrated that the R246Q mutation results in impaired secretion of laminin.

Abstract: Taken together, these results suggest that the R246Q mutation causes nephrotic syndrome by impairing secretion of laminin-521 from podocytes into the GBM; however, increased expression of the mutant protein is able to overcome this secretion defect and improve glomerular permselectivity.


Interleukin-33 prevents apoptosis and improves survival after experimental myocardial infarction through ST2 signaling.

Abstract: BACKGROUND ST2 is an interleukin (IL)-1 receptor family member with membrane-bound (ST2L) and soluble (sST2) isoforms, and sST2 is a biomarker for poor outcome in patients with myocardial infarction (MI).

Abstract: IL-33, the recently discovered ligand for ST2, activates nuclear factor kappaB and thus may regulate apoptotic cell death.

Abstract: We tested the hypothesis that IL-33 is cardioprotective after MI through ST2 signaling.

Abstract: METHODS AND RESULTS IL-33 protected cultured cardiomyocytes from hypoxia-induced apoptosis, and this cardioprotection was partially inhibited by sST2.

Abstract: IL-33 induced expression of the antiapoptotic factors XIAP, cIAP1, and survivin.

Abstract: To define the cardioprotective role of IL-33 in vivo, we performed a blinded and randomized study of ischemia/reperfusion in rats.

Abstract: IL-33 reduced cardiomyocyte apoptosis, suppressed caspase-3 activity, and increased expression of IAP family member proteins.

Abstract: IL-33 decreased both infarct and fibrosis volumes at 15 days; furthermore, both echocardiographic and hemodynamic studies revealed that IL-33 improved ventricular function.

Abstract: To determine whether cardioprotection by IL-33 is mediated through ST2 signaling, a randomized and blinded study of ST2(-/-) versus wild-type littermate mice was performed in 98 mice subjected to MI.

Abstract: At 4 weeks after MI, IL-33 reduced ventricular dilation and improved contractile function in wild-type mice but not in ST2(-/-) mice.

Abstract: Finally, IL-33 improved survival after MI in wild-type but not in ST2(-/-) mice.

Abstract: CONCLUSIONS IL-33 prevents cardiomyocyte apoptosis and improves cardiac function and survival after MI through ST2 signaling.


The impact of open versus closed format ICU admission practices on the outcome of high risk surgical patients: a cohort analysis

Abstract: BACKGROUND In the year 2000, the organizational structure of the ICU in the Zaandam Medical Centre (ZMC) changed from an open to a closed format ICU.

Abstract: The objective of this study was to evaluate the effect of this organizational change on outcome in high risk surgical patients.

Abstract: METHODS The medical records of all consecutive high risk surgical patients admitted to the ICU from 1996 to 1998 (open format) and from 2003 to 2005 (closed format), were reviewed.

Abstract: High-risk patients were defined according to the Identification of Risk in Surgical patients (IRIS) score.

Abstract: Parameters studied were: mortality, morbidity, ICU length of stay (LOS) and hospital LOS.

Abstract: RESULTS Mortality of ICU patients was 25.7% in the open format group and 15.8% in the closed format group (p = 0.01).

Abstract: Morbidity decreased from 48.6% to 46.1% (p = 0.6).

Abstract: The average length of hospital stay was 17 days in the open format group, and 21 days in the closed format group (p = 0.03).

Abstract: CONCLUSIONS High risk surgical patients in the ICU are patients that have undergone complex and often extensive surgery.

Abstract: These patients are in need of specialized treatment and careful monitoring for maximum safety and optimal care.

Abstract: Our results suggest that closed format is a more favourable setting than open format to minimize the effects of high risk surgery, and to warrant safe outcome in this patient group.


Induction therapy with autologous mesenchymal stem cells in living-related kidney transplants: a randomized controlled trial.

Abstract: CONTEXT Antibody-based induction therapy plus calcineurin inhibitors (CNIs) reduce acute rejection rates in kidney recipients; however, opportunistic infections and toxic CNI effects remain challenging.

Abstract: Reportedly, mesenchymal stem cells (MSCs) have successfully treated graft-vs-host disease.

Abstract: OBJECTIVE To assess autologous MSCs as replacement of antibody induction for patients with end-stage renal disease who undergo ABO-compatible, cross-match-negative kidney transplants from a living-related donor.

Abstract: DESIGN, SETTING, AND PATIENTS One hundred fifty-nine patients were enrolled in this single-site, prospective, open-label, randomized study from February 2008-May 2009, when recruitment was completed.

Abstract: INTERVENTION Patients were inoculated with marrow-derived autologous MSC (1-2 x 10(6)/kg) at kidney reperfusion and two weeks later.

Abstract: Fifty-three patients received standard-dose and 52 patients received low-dose CNIs (80% of standard); 51 patients in the control group received anti-IL-2 receptor antibody plus standard-dose CNIs.

Abstract: MAIN OUTCOME MEASURES The primary measure was 1-year incidence of acute rejection and renal function (estimated glomerular filtration rate [eGFR]); the secondary measure was patient and graft survival and incidence of adverse events.

Abstract: RESULTS Patient and graft survival at 13 to 30 months was similar in all groups.

Abstract: After 6 months, 4 of 53 patients (7.5%) in the autologous MSC plus standard-dose CNI group (95% CI, 0.4%-14.7%; P = .04) and 4 of 52 patients (7.7%) in the low-dose group (95% CI, 0.5%-14.9%; P = .046) compared with 11 of 51 controls (21.6%; 95% CI, 10.5%-32.6%) had biopsy-confirmed acute rejection.

Abstract: None of the patients in either autologous MSC group had glucorticoid-resistant rejection, whereas 4 patients (7.8%) in the control group did (95% CI, 0.6%-15.1%; overall P = .02).

Abstract: Renal function recovered faster among both MSC groups showing increased eGFR levels during the first month after surgery than the control group.

Abstract: Patients receiving standard-dose CNI had a mean difference of 6.2 mL/min per 1.73 m(2) (95% CI, 0.4-11.9; P=.04) and those in the low-dose CNI of 10.0 mL/min per 1.73 m(2) (95% CI, 3.8-16.2; P=.002).

Abstract: Also, during the 1-year follow-up, combined analysis of MSC-treated groups revealed significantly decreased risk of opportunistic infections than the control group (hazard ratio, 0.42; 95% CI, 0.20-0.85, P=.02)   CONCLUSION Among patients undergoing renal transplant, the use of autologous MSCs compared with anti-IL-2 receptor antibody induction therapy resulted in lower incidence of acute rejection, decreased risk of opportunistic infection, and better estimated renal function at 1 year.

Abstract: TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT00658073.


Cerebral cortex assembly: generating and reprogramming projection neuron diversity

Abstract: The mammalian cerebral cortex is responsible for the highest levels of associative, cognitive and motor functions.

Abstract: In the central nervous system (CNS) the cortex stands as a prime example of extreme neuronal diversity, broadly classified into excitatory projection neurons (PNs) and inhibitory interneurons (INs).

Abstract: We review here recent progress made in understanding the strategies and mechanisms that shape PN diversity during embryogenesis, and discuss how PN classes may be maintained, postnatally, for the life of the organism.

Abstract: In addition, we consider the intriguing possibility that PNs may be amenable to directed reprogramming of their class-specific features to allow enhanced cortical plasticity in the adult.


Mechanical modulation of receptor-ligand interactions at cell-cell interfaces.

Abstract: Cell surface receptors have been extensively studied because they initiate and regulate signal transduction cascades leading to a variety of functional cellular outcomes.

Abstract: An important class of immune receptors (e.g., T-cell antigen receptors) whose ligands are anchored to the surfaces of other cells remain poorly understood.

Abstract: The mechanism by which ligand binding initiates receptor phosphorylation, a process termed "receptor triggering", remains controversial.

Abstract: Recently, direct measurements of the (two-dimensional) receptor-ligand complex lifetimes at cell-cell interface were found to be smaller than (three-dimensional) lifetimes in solution but the underlying mechanism is unknown.

Abstract: At the cell-cell interface, the receptor-ligand complex spans a short intermembrane distance (15 nm) compared to long surface molecules (LSMs) whose ectodomains span >40 nm and these LSMs include phosphatases (e.g., CD45) that dephosphorylate the receptor.

Abstract: It has been proposed that size-based segregation of LSMs from a receptor-ligand complex is a mechanism of receptor triggering but it is unclear whether the mechanochemistry supports such small-scale segregation.

Abstract: Here we present a nanometer-scale mathematical model that couples membrane elasticity with the compressional stiffness and lateral mobility of LSMs.

Abstract: We find robust supradiffusive segregation of LSMs from a single receptor-ligand complex.

Abstract: The model predicts that LSM redistribution will result in a time-dependent tension on the complex leading to a decreased two-dimensional lifetime.

Abstract: Interestingly, the model predicts a nonlinear relationship between the three- and two-dimensional lifetimes, which can enhance the ability of receptors to discriminate between similar ligands.


High-performance neuroprosthetic control by an individual with tetraplegia.

Abstract: BACKGROUND Paralysis or amputation of an arm results in the loss of the ability to orient the hand and grasp, manipulate, and carry objects, functions that are essential for activities of daily living.

Abstract: Brain-machine interfaces could provide a solution to restoring many of these lost functions.

Abstract: We therefore tested whether an individual with tetraplegia could rapidly achieve neurological control of a high-performance prosthetic limb using this type of an interface.

Abstract: METHODS We implanted two 96-channel intracortical microelectrodes in the motor cortex of a 52-year-old individual with tetraplegia.

Abstract: Brain-machine-interface training was done for 13 weeks with the goal of controlling an anthropomorphic prosthetic limb with seven degrees of freedom (three-dimensional translation, three-dimensional orientation, one-dimensional grasping).

Abstract: The participant's ability to control the prosthetic limb was assessed with clinical measures of upper limb function.

Abstract: This study is registered with ClinicalTrials.gov, NCT01364480.

Abstract: FINDINGS The participant was able to move the prosthetic limb freely in the three-dimensional workspace on the second day of training.

Abstract: After 13 weeks, robust seven-dimensional movements were performed routinely.

Abstract: Mean success rate on target-based reaching tasks was 91·6% (SD 4·4) versus median chance level 6·2% (95% CI 2·0-15·3).

Abstract: Improvements were seen in completion time (decreased from a mean of 148 s [SD 60] to 112 s [6]) and path efficiency (increased from 0·30 [0·04] to 0·38 [0·02]).

Abstract: The participant was also able to use the prosthetic limb to do skilful and coordinated reach and grasp movements that resulted in clinically significant gains in tests of upper limb function.

Abstract: No adverse events were reported.

Abstract: INTERPRETATION With continued development of neuroprosthetic limbs, individuals with long-term paralysis could recover the natural and intuitive command signals for hand placement, orientation, and reaching, allowing them to perform activities of daily living.

Abstract: FUNDING Defense Advanced Research Projects Agency, National Institutes of Health, Department of Veterans Affairs, and UPMC Rehabilitation Institute.


Phase locking and multiple oscillating attractors for the coupled mammalian clock and cell cycle.

Abstract: Daily synchronous rhythms of cell division at the tissue or organism level are observed in many species and suggest that the circadian clock and cell cycle oscillators are coupled.

Abstract: For mammals, despite known mechanistic interactions, the effect of such coupling on clock and cell cycle progression, and hence its biological relevance, is not understood.

Abstract: In particular, we do not know how the temporal organization of cell division at the single-cell level produces this daily rhythm at the tissue level.

Abstract: Here we use multispectral imaging of single live cells, computational methods, and mathematical modeling to address this question in proliferating mouse fibroblasts.

Abstract: We show that in unsynchronized cells the cell cycle and circadian clock robustly phase lock each other in a 1:1 fashion so that in an expanding cell population the two oscillators oscillate in a synchronized way with a common frequency.

Abstract: Dexamethasone-induced synchronization reveals additional clock states.

Abstract: As well as the low-period phase-locked state there are distinct coexisting states with a significantly higher period clock.

Abstract: Cells transition to these states after dexamethasone synchronization.

Abstract: The temporal coordination of cell division by phase locking to the clock at a single-cell level has significant implications because disordered circadian function is increasingly being linked to the pathogenesis of many diseases, including cancer.


Moderate effect of artemisinin-based combination therapy on transmission of Plasmodium falciparum.

Abstract: Background.

Abstract: Artemisinin-based combination therapy (ACT) reduces microscopically confirmed gametocytemia and mosquito infection.

Abstract: However, molecular techniques have recently revealed high prevalences of submicroscopic gametocytemia.

Abstract: Our objective here was to determine the effect of sulfadoxine-pyrimethamine (SP) monotherapy and treatment with SP plus amodiaquine (AQ), SP plus artesunate (AS), and artemether-lumefantrine (AL; Coartem) on submicroscopic gametocytemia and infectiousness.

Abstract: Methods.

Abstract: Kenyan children (n=528) 6 months-10 years of age were randomized to 4 treatment arms.

Abstract: Gametocytemia was determined by both microscopy and Pfs25 RNA-based quantitative nucleic acid sequence-based amplification (Pfs25 QT-NASBA).

Abstract: Transmission was determined by membrane-feeding assays.

Abstract: Results.

Abstract: Gametocyte prevalence, as determined by Pfs25 QT-NASBA, was 89.4% (219/245) at enrollment and decreased after treatment with SP plus AS, SP plus AQ, and AL.

Abstract: Membrane-feeding assays for a group of randomly selected children revealed that the proportion of infectious children was as much as 4-fold higher than expected when based on microscopy.

Abstract: ACT did not significantly reduce the proportion of infectious children but did reduce the proportion of infected mosquitoes.

Abstract: Conclusions.

Abstract: Submicroscopic gametocytemia is common after treatment and contributes considerably to mosquito infection.

Abstract: Our findings should be interpreted in the context of transmission intensity, but the effect of ACT on malaria transmission appears to be moderate and restricted to the duration of gametocyte carriage and the proportion of mosquitoes that are infected by carriers.