Poly(A)-specific ribonuclease (PARN) mediates 3′-end maturation of the telomerase RNA component

Abstract: Mutations in the PARN gene (encoding poly(A)-specific ribonuclease) cause telomere diseases including familial idiopathic pulmonary fibrosis (IPF) and dyskeratosis congenita, but how PARN deficiency impairs telomere maintenance is unclear.

Abstract: Here, using somatic cells and induced pluripotent stem cells (iPSCs) from patients with dyskeratosis congenita with PARN mutations, we show that PARN is required for the 3′-end maturation of the telomerase RNA component (TERC).

Abstract: Patient-derived cells as well as immortalized cells in which PARN is disrupted show decreased levels of TERC.

Abstract: Deep sequencing of TERC RNA 3′ termini shows that PARN is required for removal of post-transcriptionally acquired oligo(A) tails that target nuclear RNAs for degradation.

Abstract: Diminished TERC levels and the increased proportion of oligo(A) forms of TERC are normalized by restoring PARN, which is limiting for TERC maturation in cells.

Abstract: Our results demonstrate a new role for PARN in the biogenesis of TERC and provide a mechanism linking PARN mutations to telomere diseases.


Folic acid supplementation and dietary folate intake, and risk of preeclampsia

Abstract: Background/Objectives:Folic acid supplementation has been suggested to reduce the risk of preeclampsia.

Abstract: However, results from few epidemiologic studies have been inconclusive.

Abstract: We investigated the hypothesis that folic acid supplementation and dietary folate intake before conception and during pregnancy reduce the risk of preeclampsia.

Abstract: Subjects/Methods:A birth cohort study was conducted in 2010–2012 at the Gansu Provincial Maternity & Child Care Hospital in Lanzhou, China.

Abstract: A total of 10 041 pregnant women without chronic hypertension or gestational hypertension were enrolled.

Abstract: Results:Compared with nonusers, folic acid supplement users had a reduced risk of preeclampsia (OR=0.61, 95% CI: 0.43–0.87).

Abstract: A significant dose–response of duration of use was observed among women who used folic acid supplemention during pregnancy only (P-trend=0.007).

Abstract: The reduced risk associated with folic acid supplement was similar for mild or severe preeclampsia and for early- or late-onset preeclampsia, although the statistical significant associations were only observed for mild (OR=0.50, 95% CI: 0.30–0.81) and late-onset (OR=0.60, 95% CI: 0.42–0.86) preeclampsia.

Abstract: The reduced risk associated with dietary folate intake during pregnancy was only seen for severe preeclampsia (OR=0.52, 95% CI: 0.31–0.87, for the highest quartile of dietary folate intake compared with the lowest).Conclusions:Our study results suggest that folic acid supplementation and higher dietary folate intake during pregnancy reduce the risk of preeclampsia.

Abstract: Future studies are needed to confirm the associations.


Model-based Analysis of ChIP-Seq (MACS)

Abstract: We present Model-based Analysis of ChIP-Seq data, MACS, which analyzes data generated by short read sequencers such as Solexa's Genome Analyzer.

Abstract: MACS empirically models the shift size of ChIP-Seq tags, and uses it to improve the spatial resolution of predicted binding sites.

Abstract: MACS also uses a dynamic Poisson distribution to effectively capture local biases in the genome, allowing for more robust predictions.

Abstract: MACS compares favorably to existing ChIP-Seq peak-finding algorithms, and is freely available.


Familial obesity and leanness.

Abstract: Using the Princeton School District Family Study cohort, our specific aim was to estimate the prevalence of suspected familial ponderosity and leanness, to provide empirical risk estimates for the proportion of probands' first-degree relatives who were similarly affected, and to estimate the contributions of diseases, drugs and caloric intake to relative obesity and leanness.

Abstract: We studied 379 probands, 125 whites and 52 blacks from a random recall group, 147 whites and 55 blacks from a hyperlipidemic recall group.

Abstract: Suspected familial obesity and leanness were arbitrarily identified in those kindreds with at least two first-degree relatives in the same Quetelet index decile as the proband, top or bottom respectively.

Abstract: Suspected familial obesity was observed in 2.4 percent and 6 percent respectively of random and hyperlipidemic recall group whites.

Abstract: Suspected familial leanness was identified in 2.4 percent and 1.4 percent of random and hyperlipidemic recall whites and in 3.8 percent of randomly recalled blacks.

Abstract: Approximately twice as many as expected white first-degree relatives of top Quetelet index decile probands themselves had top decile Quetelet indices; approximately three times as many as expected first-degree relatives of bottom decile Quetelet index probands themselves had bottom decile Quetelet indices.

Abstract: Nineteen percent and 31 percent of top decile Quetelet index white probands from random and hyperlipidemic recall groups came from families where at least two other first-degree relatives were similarly obese; 18 percent and 20 percent of white random and hyperlipidemic recall group probands with bottom decile Quetelet indices had suspected familial leanness.

Abstract: Nearly all subjects with familial obesity or leanness had no overt metabolic or pharmacological explanations for their body habitus.

Abstract: Within-family clustering of hypertension was common in kindreds with suspected familial obesity and was absent in kindreds with suspected familial leanness.

Abstract: Marked within-family clustering of both obesity and leanness is useful diagnostically; therapeutic intervention to reduce obesity, to be most effective, should be family-wide in the many kindreds which share familial obesity.


In-hospital switching of oral P2Y12 inhibitor treatment in patients with acute coronary syndrome undergoing percutaneous coronary intervention: prevalence, predictors and short-term outcome.

Abstract: BACKGROUND P2Y12 inhibitor switching has appeared in clinical practice as a consequence of prasugrel and ticagrelor availability, apart from clopidogrel, for use in patients with acute coronary syndrome (ACS) undergoing percutaneous coronary intervention (PCI).

Abstract: METHODS In the context of the GReek AntiPlatelet REgistry (GRAPE) we assessed the prevalence, predictive factors and short-term outcome of in-hospital P2Y12 inhibitor switching in 1794 ACS patients undergoing PCI.

Abstract: RESULTS Switching occurred in 636 (35.5%) patients of which in the form of clopidogrel to a novel agent, novel agent to clopidogrel and between prasugrel and ticagrelor in 574 (90.4%), 34 (5.3%) and 27 (4.3%) patients, respectively.

Abstract: Presentation to non PCI-capable hospital, bivalirudin use, age ≥75 years (inverse predictor), and regional trends emerged as predictive factors of switching to a novel agent.

Abstract: At combined in-hospital and one-month follow-up, propensity matched pairs analysis showed no differences in major adverse cardiovascular (MACE) or bleeding events between switching from clopidogrel to a novel agent vs novel agent constant administration.

Abstract: More Bleeding Academic Research Consortium type 1, type 2 and any type events and fewer MACE were seen when switching from clopidogrel to a novel agent vs only clopidogrel administration (23.7%, 3.8%, 30.6%, 1.2% vs 8.9%, 1.2%, 12.0%, 3.8% with P < .001, P = .03, P < .001 and P = .03 respectively).

Abstract: CONCLUSIONS In a real-life experience with contemporary antiplatelet treatment in ACS patients undergoing PCI, in-hospital switching represents common clinical practice.

Abstract: Clinical factors and regional practice differences seem to affect this strategy's choice, while switching to a novel agent may be associated with higher risk of bleeding.


Structural basis of allosteric and synergistic activation of AMPK by furan-2-phosphonic derivative C2 binding.

Abstract: The metabolic stress-sensing enzyme AMP-activated protein kinase (AMPK) is responsible for regulating metabolism in response to energy supply and demand.

Abstract: Drugs that activate AMPK may be useful in the treatment of metabolic diseases including type 2 diabetes.

Abstract: We have determined the crystal structure of AMPK in complex with its activator 5-(5-hydroxyl-isoxazol-3-yl)-furan-2-phosphonic acid (C2), revealing two C2-binding sites in the γ-subunit distinct from nucleotide sites.

Abstract: C2 acts synergistically with the drug A769662 to activate AMPK α1-containing complexes independent of upstream kinases.

Abstract: Our results show that dual drug therapies could be effective AMPK-targeting strategies to treat metabolic diseases.


Validity of age at menarche self-reported in adulthood.

Abstract: OBJECTIVE To test the validity of age at menarche self-reported in adulthood and examine whether socioeconomic position, education, experience of gynaecological events and psychological symptoms influence the accuracy of recall.

Abstract: DESIGN Prospective birth cohort study.

Abstract: SETTING England, Scotland and Wales.

Abstract: PARTICIPANTS 1050 women from the Medical Research Council National Survey of Health and Development, with two measures of age at menarche, one recorded in adolescence and the other self-reported at age 48 years.

Abstract: RESULTS By calculating the limits of agreement, kappa statistic and Pearson's correlation coefficients (r), we found that the validity of age at menarche self-reported in middle age compared with that recorded in adolescence was moderate (kappa = 0.35, r = 0.66, n = 1050).

Abstract: Validity was improved by categorising age at menarche into three groups: early, normal and late (kappa = 0.43).

Abstract: Agreement was influenced by educational level and having had a stillbirth or miscarriage.

Abstract: CONCLUSIONS The level of validity shown in this study throws some doubt on whether it is justifiable to use age at menarche self-reported in middle age.

Abstract: It is likely to introduce error and bias, and researchers should be aware of these limitations and use such measures with caution.


Seroepidemiologic survey of captive Old-World primates for antibodies to human and simian retroviruses, and isolation of a lentivirus from sooty mangabeys (Cercocebus atys).

Abstract: Sera from 526 Old-World monkeys and apes, representing 50 species and 20 genera and living in US zoos and vivaria, were screened for antibodies to HTLV-I, HTLV-III/LAV, and simian-AIDS retrovirus, type I (SRV-I).

Abstract: Sera were screened initially by ELISA, and ELISA-positive sera, as well as ELISA-negative sera from cage contacts, were further tested by Western blotting.

Abstract: A large number of false-positive and a small number of false-negative ELISA sera were identified.

Abstract: Although most true positive reactions were directed to a single retrovirus, a number of individuals from 4 species were positive for more than one retrovirus.

Abstract: Specific seroreactivity to HTLV-I was found in 39/526 (7%) animals of 15 species.

Abstract: True positive reactions to SRV-I were found in 21/516 (4%) animals, including talapoins and 2 species of macaques.

Abstract: Specific serologic reactions to HTLV-III/LAV were detected in 23/526 (4%) monkeys.

Abstract: Many of the HTLV-III/LAV seropositive animals were from one mixed-species zoo exhibit, containing sooty mangabeys, mandrills, Kolb's guenons, and talapoins.

Abstract: A type D virus was isolated from the blood of 3/10 SRV-I antibody-positive Tonkeana macaques, but from none of 11 seropositive talapoins.

Abstract: A lentivirus was isolated from the blood of 4/7 HTLV-III/LAV seropositive sooty mangabeys, but not from seropositive talapoins in the same exhibit or from 2 seropositive colobus from another zoo.

Abstract: The sooty mangabey lentivirus produced generalized lymphadenopathy, leukopenia, and decreased levels of T4 lymphocytes in 2 experimentally infected rhesus macaques.


Coronary plaque disruption.

Abstract: Coronary atherosclerosis is by far the most frequent cause of ischemic heart disease, and plaque disruption with superimposed thrombosis is the main cause of the acute coronary syndromes of unstable angina, myocardial infarction, and sudden death.1 2 3 4 5 Therefore, for event-free survival, the vital question is not why atherosclerosis develops but rather why, after years of indolent growth, it suddenly becomes complicated by life-threatening thrombosis.

Abstract: The composition and vulnerability of plaque rather than its volume or the consequent severity of stenosis produced have emerged as being the most important determinants for the development of the thrombus-mediated acute coronary syndromes; lipid-rich and soft plaques are more dangerous than collagen-rich and hard plaques because they are more unstable and rupture-prone and highly thrombogenic after disruption.6 This review will explore potential mechanisms responsible for the sudden conversion of a stable atherosclerotic plaque to an unstable and life-threatening atherothrombotic lesion—an event known as plaque fissuring, rupture, or disruption.7 8   Atherosclerosis is the result of a complex interaction between blood elements, disturbed flow, and vessel wall abnormality, involving several pathological processes: inflammation, with increased endothelial permeability, endothelial activation, and monocyte recruitment9 10 11 12 13 14 ; growth, with smooth muscle cell (SMC) proliferation, migration, and matrix synthesis15 16 ; degeneration, with lipid accumulation17 18 ; necrosis, possibly related to the cytotoxic effect of oxidized lipid19 ; calcification/ossification, which may represent an active rather than a dystrophic process20 21 ; and thrombosis, with platelet recruitment and fibrin formation.1 22 23 Thrombotic factors may play a role early during atherogenesis, but a flow-limiting thrombus does not develop until mature plaques are present, which is why thrombosis often is classified as a complication rather than a genuine component of atherosclerosis.

Abstract: ### Mature Plaques: Atherosis and Sclerosis   As the name atherosclerosis implies, mature …


p16INK4A is a robust in vivo biomarker of cellular aging in human skin.

Abstract: The cell-cycle regulating gene, p16INK4A, encoding an inhibitor of cyclin-dependent kinases 4 and 6, is considered to play an important role in cellular aging and in premature senescence.

Abstract: Although there is an age-dependent increase of p16INK4A expression in human fibroblast senescence in vitro, no data are available regarding the age dependency of p16INK4A in vivo.

Abstract: To determine whether p16INK4A expression in human skin correlates with donor age, p16INK4A expression was analyzed by immunohistochemistry as well as the expression of the p16INK4A repressor BMI1.

Abstract: Samples from the age groups 0-20, 21-70, and 71-95 years were selected from a bank of healthy human skin.

Abstract: We show that the number of p16INK4A positive cells is significantly higher in elderly individuals compared to the younger age groups.

Abstract: The number of p16INK4A positive cells was found to be increased in both epidermis and dermis, compartments with strictly different proliferative activities.

Abstract: BMI1 gene expression was significantly down-regulated with increasing donor age, whereas no striking age differences were observed for Ki67.

Abstract: In immunofluorescence co-expression studies, Ki67-positive cells were negative for p16INK4A and BMI1-expressing cells also stained negatively for Ki67.

Abstract: In conclusion, we provide for the first time evidence that p16INK4A expression directly correlates with chronological aging of human skin in vivo.

Abstract: p16INK4A therefore is a biomarker for human aging in vivo.

Abstract: The data reported here suggest a model for changes in regulatory gene expression that drive aging in human skin.


hPrimpol1/CCDC111 is a human DNA primase-polymerase required for the maintenance of genome integrity.

Abstract: Prim-pol is a recently identified DNA primase-polymerase belonging to the archaeao-eukaryotic primase (AEP) superfamily.

Abstract: Here, we characterize a previously unrecognized prim-pol in human cells, which we designate hPrimpol1 (human primase-polymerase 1).

Abstract: hPrimpol1 possesses primase and DNA polymerase activities in vitro, interacts directly with RPA1 and is recruited to sites of DNA damage and stalled replication forks in an RPA1-dependent manner.

Abstract: Cells depleted of hPrimpol1 display increased spontaneous DNA damage and defects in the restart of stalled replication forks.

Abstract: Both RPA1 binding and the primase activity of hPrimpol1 are required for its cellular function during DNA replication.

Abstract: Our results indicate that hPrimpol1 is a novel factor involved in the response to DNA replication stress.


Functional proteomics identifies miRNAs to target a p27/Myc/phospho-Rb signature in breast and ovarian cancer

Abstract: The myc oncogene is overexpressed in almost half of all breast and ovarian cancers, but attempts at therapeutic interventions against myc have proven to be challenging.

Abstract: Myc regulates multiple biological processes, including the cell cycle, and as such is associated with cell proliferation and tumor progression.

Abstract: We identified a protein signature of high myc, low p27 and high phospho-Rb significantly correlated with poor patient survival in breast and ovarian cancers.

Abstract: Screening of a miRNA library by functional proteomics in multiple cell lines and integration of data from patient tumors revealed a panel of five microRNAs (miRNAs) (miR-124, miR-365, miR-34b*, miR-18a and miR-506) as potential tumor suppressors capable of reversing the p27/myc/phospho-Rb protein signature.

Abstract: Mechanistic studies revealed an RNA-activation function of miR-124 resulting in direct induction of p27 protein levels by binding to and inducing transcription on the p27 promoter region leading to a subsequent G1 arrest.

Abstract: Additionally, in vivo studies utilizing a xenograft model demonstrated that nanoparticle-mediated delivery of miR-124 could reduce tumor growth and sensitize cells to etoposide, suggesting a clinical application of miRNAs as therapeutics to target the functional effect of myc on tumor growth.


The potential role of miRNAs 21 and 199-a in early diagnosis of hepatocellular carcinoma.

Abstract: BACKGROUND Hepatocellular carcinoma (HCC) is regarded as one of the most common malignancies and among the leading causes of cancer death among the whole world.

Abstract: The most urgent needs are to find sensitive markers for early diagnosis for HCC.

Abstract: MicroRNAs (miRNAs) are reported as a group of small non-coding RNAs that can function as endogenous RNA interference to regulate expression of the targeted genes.

Abstract: This study was conducted to detect the serum and tissue expression of miR 21 and miR 199-a to be applied as early detectors for HCC.

Abstract: METHODS A total of 40 serum and tissue samples (17 samples from chronic hepatitis and 23 samples from HCC patients) were collected.

Abstract: The levels of the two mature miRNAs (miR-21 and miR-199-a) were detected by real time quantitative reverse-transcriptase PCR (RT-qPCR) in sera and tissues of chronic hepatitis and HCC patients.

Abstract: Besides, miR-21 and miR-199-a levels in relation to clinical and pathological factors were explored.

Abstract: RESULTS We found that the expression of serum miR-21 was distinctly increased in HCC compared with chronic hepatitis (P<0.001).

Abstract: miR 199-a was distinctly decreased in HCC compared with chronic hepatitis (P<0.001).

Abstract: In addition, median of miR 21 was increased in malignant when compared to adjacent non-malignant tissues without significant differences (P=0.191) while miR 199-a was significantly decreased in malignant when compared to adjacent nonmalignant tissues (P<0.001).

Abstract: ROC analysis showed that miR-21 and miR-199-a might be potential biomarkers for HCC.

Abstract: CONCLUSIONS In conclusion, the expression of miR-21 was significantly up-regulated and miR-199-a was significantly down regulated in serum of patients with HCC.

Abstract: Due to their reasonable sensitivity and specificity for disease progression, miR-21 and miR-199-a could be used as potential circulating biomarkers for HCC.


Tex10 Coordinates Epigenetic Control of Super-Enhancer Activity in Pluripotency and Reprogramming.

Abstract: Super-enhancers (SEs) are large clusters of transcriptional enhancers that are co-occupied by multiple lineage-specific transcription factors driving expression of genes that define cell identity.

Abstract: In embryonic stem cells (ESCs), SEs are highly enriched for the core pluripotency factors Oct4, Sox2, and Nanog.

Abstract: In this study, we sought to dissect the molecular control mechanism of SE activity in pluripotency and reprogramming.

Abstract: Starting from a protein interaction network surrounding Sox2, we identified Tex10 as a key pluripotency factor that plays a functionally significant role in ESC self-renewal, early embryo development, and reprogramming.

Abstract: Tex10 is enriched at SEs in a Sox2-dependent manner and coordinates histone acetylation and DNA demethylation at SEs.

Abstract: Tex10 activity is also important for pluripotency and reprogramming in human cells.

Abstract: Our study therefore highlights Tex10 as a core component of the pluripotency network and sheds light on its role in epigenetic control of SE activity for cell fate determination.


Value of whole brain re-irradiation for brain metastases--single centre experience.

Abstract: AIMS There is controversy in published studies regarding the role of repeat whole brain radiation (WBRT) for previously irradiated brain metastases.

Abstract: The aim of our retrospective study was to document the practice at Princess Margaret Hospital with respect to the re-irradiation of patients with progressive or recurrent brain metastatic disease after initial WBRT.

Abstract: MATERIALS AND METHODS A comprehensive computerised database was used to identify patients treated for brain metastases with more than one course of WBRT between 1997 and 2003.

Abstract: Seventy-two patients were treated with WBRT for brain metastases and retreated with WBRT at a later date.

Abstract: The records of these patients were reviewed.

Abstract: RESULTS The median age was 56.5 years.

Abstract: The most common primary sites were lung (51 patients) and breast (17 patients).

Abstract: The most frequent dose used for the initial radiotherapy was 20 Gy/5 fractions (62 patients).

Abstract: The most common doses of re-irradiation were 25 Gy/10 fractions (22 patients), 20 Gy/10 fractions (12 patients), 15 Gy/5 fractions (11 patients) and 20 Gy/8 fractions (10 patients).

Abstract: Thirty-one per cent of patients experienced a partial clinical response after re-irradiation, as judged by follow-up clinical notes; 27% remained stable; 32% deteriorated after re-irradiation.

Abstract: Patients who had Eastern Cooperative Oncology Group performance status 0-1 at the time of retreatment lived longer.

Abstract: In responders, the mean duration of response was 5.1 months.

Abstract: The median survival after re-irradiation was 4.1 months.

Abstract: One patient was reported as having memory impairment and pituitary insufficiency after 5 months of progression-free survival.

Abstract: CONCLUSION Repeat radiotherapy may be a useful treatment in carefully selected patients.

Abstract: With increased survival and better systemic options for patients with metastatic disease, more patients may be candidates for consideration of repeat WBRT for recurrent brain metastases, but prospective studies are needed to more clearly document their outcomes.


Bisphosphonates in oncology: rising stars or fallen heroes.

Abstract: The introduction of bisphosphonates in oncology has dramatically changed the management of patients with metastatic bone disease.

Abstract: In this manuscript, we thoroughly scrutinize the available body of clinical trials supporting the use of bisphosphonates in this setting and review new and ongoing research.

Abstract: Additionally, we summarize the data showing the benefits of bisphosphonate use in the prevention of treatment-induced bone loss and the intriguing emerging evidence on the antitumor potential of some of these agents when used in the adjuvant setting.

Abstract: Finally, we address the need for a careful consideration of potential benefits of bisphosphonate therapy and the risk for osteonecrosis of the jaw, a recently recognized late-toxicity of their use.


IL-33, a recently identified interleukin-1 gene family member, is expressed in human adipocytes.

Abstract: Inflammation occurs in adipose tissue in obesity.

Abstract: We have examined whether IL-33, a recently identified IL-1 gene family member, and its associated receptors are expressed in human adipocytes.

Abstract: IL-33, IL-1RL1 and IL-1RAP gene expression was observed in human visceral white fat, in preadipocytes and in adipocytes (SGBS cells).

Abstract: Treatment with TNFalpha for 24h induced a 6-fold increase in IL-33 mRNA level in preadipocytes and adipocytes.

Abstract: Time-course studies with adipocytes showed that the increase in IL-33 mRNA with TNFalpha was maximal (>55-fold) at 12h.

Abstract: This response was markedly different to IL-1beta (peak mRNA increase at 2h; 5.4-fold) and 1L-18 (peak mRNA increase at 6h; >1500-fold).

Abstract: Exposure of adipocytes to hypoxia (1% O(2), 24h) did not alter IL-33 mRNA level; in preadipocytes, however, there was a 3-fold increase.

Abstract: Human adipocytes and preadipocytes express IL-33, but the various IL-1 family members exhibit major differences in responsiveness to TNFalpha.


Imaging techniques for assaying lymphocyte activation in action

Abstract: Imaging techniques have greatly improved our understanding of lymphocyte activation.

Abstract: Technical advances in spatial and temporal resolution and new labelling tools have enabled researchers to directly observe the activation process.

Abstract: Consequently, research using imaging approaches to study lymphocyte activation has expanded, providing an unprecedented level of cellular and molecular detail in the field.

Abstract: As a result, certain models of lymphocyte activation have been verified, others have been revised and yet others have been replaced with new concepts.

Abstract: In this article, we review the current imaging techniques that are used to assess lymphocyte activation in different contexts, from whole animals to single molecules, and discuss the advantages and potential limitations of these methods.


Extending healthy life span--from yeast to humans.

Abstract: When the food intake of organisms such as yeast and rodents is reduced (dietary restriction), they live longer than organisms fed a normal diet.

Abstract: A similar effect is seen when the activity of nutrient-sensing pathways is reduced by mutations or chemical inhibitors.

Abstract: In rodents, both dietary restriction and decreased nutrient-sensing pathway activity can lower the incidence of age-related loss of function and disease, including tumors and neurodegeneration.

Abstract: Dietary restriction also increases life span and protects against diabetes, cancer, and cardiovascular disease in rhesus monkeys, and in humans it causes changes that protect against these age-related pathologies.

Abstract: Tumors and diabetes are also uncommon in humans with mutations in the growth hormone receptor, and natural genetic variants in nutrient-sensing pathways are associated with increased human life span.

Abstract: Dietary restriction and reduced activity of nutrient-sensing pathways may thus slow aging by similar mechanisms, which have been conserved during evolution.

Abstract: We discuss these findings and their potential application to prevention of age-related disease and promotion of healthy aging in humans, and the challenge of possible negative side effects.


Hospital and 1-year survival of patients admitted to intensive care units with acute exacerbation of chronic obstructive pulmonary disease.

Abstract: OBJECTIVE To describe outcomes and identify variables associated with hospital and 1-year survival for patients admitted to an intensive care unit (ICU) with an acute exacerbation of chronic obstructive pulmonary disease (COPD).

Abstract: DESIGN Prospective, multicenter, inception cohort study.

Abstract: SETTING Forty-two ICUs at 40 US hospitals.

Abstract: PATIENTS A total of 362 admissions for COPD exacerbation selected from the Acute Physiology and Chronic Health Evaluation (APACHE) III database of 17,440 ICU admissions.

Abstract: MEASUREMENTS AND RESULTS Hospital mortality for the 362 admissions was 24%.

Abstract: For the 167 patients aged 65 years or older, mortality was 30% at hospital discharge, 41% at 90 days, 47% at 180 days, and 59% at 1 year.

Abstract: Median survival for all patients was 224 days, and median survival for the patients who died within 1 year was 30.5 days.

Abstract: On multiple regression analysis, variables associated with hospital mortality included age, severity of respiratory and nonrespiratory organ system dysfunction, and hospital length of stay before ICU admission.

Abstract: Development of nonrespiratory organ system dysfunction was the major predictor of hospital mortality (60% of total explanatory power) and 180-day outcomes (54% of explanatory power).

Abstract: Respiratory physiological variables (respiratory rate, serum pH, PaCO2, PaO2, and alveolar-arterial difference in partial pressure of oxygen [PAO2-PaO2]) indicative of advanced dysfunction were more strongly associated with 180-day mortality rates (22% of explanatory power) than hospital death rates (4% of explanatory power).

Abstract: After controlling for severity of illness, mechanical ventilation at ICU admission was not associated with either hospital mortality or subsequent survival.

Abstract: CONCLUSIONS Patients with COPD admitted to an ICU for an acute exacerbation have a substantial hospital mortality (24%).

Abstract: For patients aged 65 years or older, mortality doubles in 1 year from 30% to 59%.

Abstract: Hospital and longer-term mortality is closely associated with development of nonrespiratory organ system dysfunction; severity of the underlying respiratory function substantially influences mortality following hospital discharge.

Abstract: The need for mechanical ventilation at ICU admission did not influence either short- or long-term outcomes.

Abstract: Physicians should be aware of these relationships when making treatment decisions or evaluating new therapies.


Kindlin-3 is essential for integrin activation and platelet aggregation

Abstract: Integrin-mediated platelet adhesion and aggregation are essential for sealing injured blood vessels and preventing blood loss, and excessive platelet aggregation can initiate arterial thrombosis, causing heart attacks and stroke.

Abstract: To ensure that platelets aggregate only at injury sites, integrins on circulating platelets exist in a low-affinity state and shift to a high-affinity state (in a process known as integrin activation or priming) after contacting a wounded vessel.

Abstract: The shift is mediated through binding of the cytoskeletal protein Talin to the β subunit cytoplasmic tail.

Abstract: Here we show that platelets lacking the adhesion plaque protein Kindlin-3 cannot activate integrins despite normal Talin expression.

Abstract: As a direct consequence, Kindlin-3 deficiency results in severe bleeding and resistance to arterial thrombosis.

Abstract: Mechanistically, Kindlin-3 can directly bind to regions of β-integrin tails distinct from those of Talin and trigger integrin activation.

Abstract: We have therefore identified Kindlin-3 as a novel and essential element for platelet integrin activation in hemostasis and thrombosis.


Opening of compacted chromatin by early developmental transcription factors HNF3 (FoxA) and GATA-4.

Abstract: The transcription factors HNF3 (FoxA) and GATA-4 are the earliest known to bind the albumin gene enhancer in liver precursor cells in embryos.

Abstract: To understand how they access sites in silent chromatin, we assembled nucleosome arrays containing albumin enhancer sequences and compacted them with linker histone.

Abstract: HNF3 and GATA-4, but not NF-1, C/EBP, and GAL4-AH, bound their sites in compacted chromatin and opened the local nucleosomal domain in the absence of ATP-dependent enzymes.

Abstract: The ability of HNF3 to open chromatin is mediated by a high affinity DNA binding site and by the C-terminal domain of the protein, which binds histones H3 and H4.

Abstract: Thus, factors that potentiate transcription in development are inherently capable of initiating chromatin opening events.


N- and E-cadherins in Xenopus are specifically required in the neural and non-neural ectoderm, respectively, for F-actin assembly and morphogenetic movements.

Abstract: Transmembrane cadherins are calcium-dependent intercellular adhesion molecules.

Abstract: Recently, they have also been shown to be sites of actin assembly during adhesive contact formation.

Abstract: However, the roles of actin assembly on transmembrane cadherins during development are not fully understood.

Abstract: We show here, using the developing ectoderm of the Xenopus embryo as a model, that F-actin assembly is a primary function of both N-cadherin in the neural ectoderm and E-cadherin in the non-neural (epidermal) ectoderm, and that each cadherin is essential for the characteristic morphogenetic movements of these two tissues.

Abstract: However, depletion of N-cadherin and E-cadherin did not cause dissociation in these tissues at the neurula stage, probably owing to the expression of C-cadherin in each tissue.

Abstract: Depletion of each of these cadherins is not rescued by the other, nor by the expression of C-cadherin, which is expressed in both tissues.

Abstract: One possible reason for this is that each cadherin is expressed in a different domain of the cell membrane.

Abstract: These data indicate the combinatorial nature of cadherin function, the fact that N- and E-cadherin play primary roles in F-actin assembly in addition to roles in cell adhesion, and that this function is specific to individual cadherins.

Abstract: They also show how cell adhesion and motility can be combined in morphogenetic tissue movements that generate the form and shape of the embryonic organs.


Mutation of the fumarase gene in two siblings with progressive encephalopathy and fumarase deficiency.

Abstract: We report an inborn error of the tricarboxylic acid cycle, fumarase deficiency, in two siblings born to first cousin parents.

Abstract: They presented with progressive encephalopathy, dystonia, leucopenia, and neutropenia.

Abstract: Elevation of lactate in the cerebrospinal fluid and high fumarate excretion in the urine led us to investigate the activities of the respiratory chain and of the Krebs cycle, and to finally identify fumarase deficiency in these two children.

Abstract: The deficiency was profound and present in all tissues investigated, affecting the cytosolic and the mitochondrial fumarase isoenzymes to the same degree.

Abstract: Analysis of fumarase cDNA demonstrated that both patients were homozygous for a missense mutation, a G-955-->C transversion, predicting a Glu-319-->Gln substitution.

Abstract: This substitution occurred in a highly conserved region of the fumarase cDNA.

Abstract: Both parents exhibited half the expected fumarase activity in their lymphocytes and were found to be heterozygous for this substitution.

Abstract: The present study is to our knowledge the first molecular characterization of tricarboxylic acid deficiency, a rare inherited inborn error of metabolism in childhood.


Adult Neurogenesis Is Sustained by Symmetric Self-Renewal and Differentiation.

Abstract: Somatic stem cells have been identified in multiple adult tissues.

Abstract: Whether self-renewal occurs symmetrically or asymmetrically is key to understanding long-term stem cell maintenance and generation of progeny for cell replacement.

Abstract: In the adult mouse brain, neural stem cells (NSCs) (B1 cells) are retained in the walls of the lateral ventricles (ventricular-subventricular zone [V-SVZ]).

Abstract: The mechanism of B1 cell retention into adulthood for lifelong neurogenesis is unknown.

Abstract: Using multiple clonal labeling techniques, we show that the vast majority of B1 cells divide symmetrically.

Abstract: Whereas 20%-30% symmetrically self-renew and can remain in the niche for several months before generating neurons, 70%-80% undergo consuming divisions generating progeny, resulting in the depletion of B1 cells over time.

Abstract: This cellular mechanism decouples self-renewal from the generation of progeny.

Abstract: Limited rounds of symmetric self-renewal and consuming symmetric differentiation divisions can explain the levels of neurogenesis observed throughout life.


A common missense variant in NUDT15 confers susceptibility to thiopurine-induced leukopenia

Abstract: Thiopurine therapy, commonly used in autoimmune conditions, can be complicated by life-threatening leukopenia.

Abstract: This leukopenia is associated with genetic variation in TPMT (encoding thiopurine S-methyltransferase).

Abstract: Despite a lower frequency of TPMT mutations in Asians, the incidence of thiopurine-induced leukopenia is higher in Asians than in individuals of European descent.

Abstract: Here we performed an Immunochip-based 2-stage association study in 978 Korean subjects with Crohn's disease treated with thiopurines.

Abstract: We identified a nonsynonymous SNP in NUDT15 (encoding p.

Abstract: Arg139Cys) that was strongly associated with thiopurine-induced early leukopenia (odds ratio (OR) = 35.6; Pcombined = 4.88 × 10−94).

Abstract: In Koreans, this variant demonstrated sensitivity and specificity of 89.4% and 93.2%, respectively, for thiopurine-induced early leukopenia (in comparison to 12.1% and 97.6% for TPMT variants).

Abstract: Although rare, this SNP was also strongly associated with thiopurine-induced leukopenia in subjects with inflammatory bowel disease of European descent (OR = 9.50; P = 4.64 × 10−4).

Abstract: Thus, NUDT15 is a pharmacogenetic determinant for thiopurine-induced leukopenia in diverse populations.


Induction of cell cycle entry eliminates human leukemia stem cells in a mouse model of AML

Abstract: Cancer stem cells have been proposed to be important for initiation, maintenance and recurrence of various malignancies, including acute myeloid leukemia (AML).

Abstract: We have previously reported that CD34+CD38− human primary AML stem cells residing in the endosteal region of the bone marrow are relatively chemotherapy resistant.

Abstract: Using a NOD/SCID/IL2rγnull mouse model of human AML, we now show that the AML stem cells in the endosteal region are cell cycle quiescent and that these stem cells can be induced to enter the cell cycle by treatment with granulocyte colony-stimulating factor (G-CSF).

Abstract: In combination with cell cycle-dependent chemotherapy, G-CSF treatment significantly enhances induction of apoptosis and elimination of human primary AML stem cells in vivo.

Abstract: The combination therapy leads to significantly increased survival of secondary recipients after transplantation of leukemia cells compared with chemotherapy alone.


Reconstitution of the immune system after hematopoietic stem cell transplantation in humans

Abstract: Hematopoietic stem cell transplantation is associated with a severe immune deficiency.

Abstract: As a result, the patient is at high risk of infections.

Abstract: Innate immunity, including epithelial barriers, monocytes, granulocytes, and NK cells recovers within weeks after transplantation.

Abstract: By contrast, adaptive immunity recovers much slower.

Abstract: B- and T-cell counts normalize during the first months after transplantation, but in particular, T-cell immunity may remain impaired for years.

Abstract: During the last decade, much of the underlying mechanisms have been identified.

Abstract: These insights may provide new therapies to accelerate recovery.


Vision tests in the mouse: Functional phenotyping with electroretinography.

Abstract: Electroretinography (ERG) is an established diagnostic technique in clinical ophthalmology and provides objective information about retinal function.

Abstract: This technique is also applied in basic research, where animal models of hereditary retinopathies have significantly contributed to our understanding of the composition of ERG responses in general and how retinal degenerative pathologies alter retinal function specifically.

Abstract: Indeed, electrophysiologic assessment of transgenic mice, which are genetically engineered to mimic human mutations that lead to retinal diseases, can be well compared with clinical data.

Abstract: Furthermore, limitations on examinations (e.g. length of measurement, range of light intensity) are much less of a concern when assessing mice compared to human patients.

Abstract: In order to measure and analyze retinal responses properly, several important aspects have to be considered.

Abstract: This paper focuses on these aspects, and shows exemplary ERG data which were obtained from normal wild-type mice and from transgenic mice with specific functional properties, namely Rho-/- (rod opsin knockout, cone function only), and Cnga3-/- (cone CNG channel deficient, rod function only) to illustrate rod and cone system contributions to ERG responses.


Cyclin-dependent kinase 2 is essential for meiosis but not for mitotic cell division in mice

Abstract: We targeted the locus encoding the cyclin-dependent kinase 2 (CDK2) by homologous recombination in mouse embryonic stem (ES) cells.

Abstract: Embryonic fibroblasts lacking CDK2 proliferate normally and become immortal after continuous passage in culture.

Abstract: Elimination of a conditional Cdk2 allele in immortal cells does not have a significant effect on proliferation.

Abstract: Cdk2−/− mice are viable and survive for up to two years, indicating that CDK2 is also dispensable for proliferation and survival of most cell types.

Abstract: But CDK2 is essential for completion of prophase I during meiotic cell division in male and female germ cells, an unforeseen role for this cell cycle kinase.


Capsaicin-sensitive sensory nerve fibers contribute to the generation and maintenance of skeletal fracture pain.

Abstract: Although skeletal pain can have a marked impact on a patient's functional status and quality of life, relatively little is known about the specific populations of peripheral nerve fibers that drive non-malignant bone pain.

Abstract: In the present report, neonatal male Sprague-Dawley rats were treated with capsaicin or vehicle and femoral fracture was produced when the animals were young adults (15-16 weeks old).

Abstract: Capsaicin treatment, but not vehicle, resulted in a significant (>70%) depletion in the density of calcitonin-gene related peptide positive (CGRP(+)) sensory nerve fibers, but not 200 kDa neurofilament H positive (NF200(+)) sensory nerve fibers in the periosteum.

Abstract: The periosteum is a thin, cellular and fibrous tissue that tightly adheres to the outer surface of all but the articulated surface of bone and appears to play a pivotal role in driving fracture pain.

Abstract: In animals treated with capsaicin, but not vehicle, there was a 50% reduction in the severity, but no change in the time course, of fracture-induced skeletal pain-related behaviors as measured by spontaneous flinching, guarding and weight bearing.

Abstract: These results suggest that both capsaicin-sensitive (primarily CGRP(+) C-fibers) and capsaicin-insensitive (primarily NF200(+) A-delta fibers) sensory nerve fibers participate in driving skeletal fracture pain.

Abstract: Skeletal pain can be a significant impediment to functional recovery following trauma-induced fracture, osteoporosis-induced fracture and orthopedic surgery procedures such as knee and hip replacement.

Abstract: Understanding the specific populations of sensory nerve fibers that need to be targeted to inhibit the generation and maintenance of skeletal pain may allow the development of more specific mechanism-based therapies that can effectively attenuate acute and chronic skeletal pain.


Surgical therapy for dystonia.

Abstract: Surgical treatments for dystonia have been available since the early 20th century, but have improved in their efficacy to adversity ratio through a combination of technologic advances and better understanding of the role of the basal ganglia in dystonia.

Abstract: The word "dystonia" describes a phenotype of involuntary movement that may manifest from a variety of conditions.

Abstract: Dystonia may affect only certain regions of the body or may be generalized.

Abstract: It appears to be critical to determine whether the etiology underlying the dystonia is "primary" (ie, occurring from a genetic or idiopathic origin) or "secondary" (ie, occurring as a result of structural, metabolic, or neurodegenerative disorders).

Abstract: Secondary dystonias are far more common than primary dystonias.

Abstract: Primary dystonias respond well to pallidotomy or deep brain stimulation of the internal segment of the globus pallidum, whereas secondary dystonias appear to respond partially at best.

Abstract: Limited historic and current data suggest that the thalamus may be a promising target for the treatment of secondary dystonias, but more careful, prospective, randomized studies are needed.

Abstract: Combinations of bilateral targets are possible with the current technology of DBS, but not widely used due to surgical morbidity and expense.

Abstract: This article reviews the surgical treatment of dystonia from past to present, with a focus on separating the outcomes for primary versus secondary and generalized versus cervical dystonia.


MEG3 imprinted gene contribution in tumorigenesis.

Abstract: Maternally expressed gene 3 (MEG3) is a maternally expressed imprinted gene representing a large noncoding RNA in which microRNAs (miRNAs) and small nucleolar RNAs are also hosted.

Abstract: It is capable of interacting with cyclic AMP, p53, murine double minute 2 (MDM2) and growth differentiation factor 15 (GDF15) playing a role in cell proliferation control.

Abstract: MEG3 expression is under epigenetic control, and aberrant CpG methylation has been observed in several types of cancer.

Abstract: Moreover, gene copy number loss has been reported as additional mechanism associated with tumorigenesis.

Abstract: MEG3 deletion seems to upregulate the paternally expressed genes and on the other hand downregulate the expression of downstream maternally expressed genes and tumor suppressor miRNAs, although there are conflicting data on the topic.

Abstract: MEG3 could represent a tumor suppressor gene located in chromosome 14q32 and its association with tumorigenesis is growing every day.


Uptake of rosuvastatin by isolated rat hepatocytes: comparison with pravastatin.

Abstract: The liver is the target organ for the lipid-regulating effect of rosuvastatin, a new 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, and liver-selective uptake of this drug is therefore a desirable property.

Abstract: The uptake kinetics of rosuvastatin were investigated and compared with those of pravastatin using isolated rat hepatocytes.

Abstract: Uptake for both drugs involved both active transport and passive diffusion processes.

Abstract: The Michaelis constant (K(m)) of uptake rate for rosuvastatin (9.17 micro M) was approximately half that for pravastatin (16.5 micro M).

Abstract: However, the maximum uptake rate (V(max)) and carrier-mediated uptake clearance (V(max)/K(m)) of rosuvastatin were significantly (p < 0.01) greater than those of pravastatin, and a larger contribution of carrier-mediated uptake clearance to total uptake clearance was shown for rosuvastatin (contribution ratio 0.903 versus pravastatin 0.654).

Abstract: Sodium and chloride ions did not play a significant role in the uptake of rosuvastatin and pravastatin, but the uptake of both drugs was inhibited both by depletion of cellular ATP and by organic anions such as bromosulfophthalein.

Abstract: Rosuvastatin competitively inhibited the uptake of pravastatin, with an inhibition constant (K(i)) (2.75 micro M) relatively similar to its K(m).

Abstract: The results suggest that an organic anion transport protein is the main mediator of the hepatic uptake of rosuvastatin and pravastatin, which occurs in an ATP-dependent manner.

Abstract: Our results indicated that rosuvastatin was taken up by the hepatocytes via the same transport systems as pravastatin, but with a greater affinity and efficiency than pravastatin.


Junctophilin-4, a component of the endoplasmic reticulum-plasma membrane junctions, regulates Ca2+ dynamics in T cells.

Abstract: Orai1 and stromal interaction molecule 1 (STIM1) mediate store-operated Ca(2+) entry (SOCE) in immune cells.

Abstract: STIM1, an endoplasmic reticulum (ER) Ca(2+) sensor, detects store depletion and interacts with plasma membrane (PM)-resident Orai1 channels at the ER-PM junctions.

Abstract: However, the molecular composition of these junctions in T cells remains poorly understood.

Abstract: Here, we show that junctophilin-4 (JP4), a member of junctional proteins in excitable cells, is expressed in T cells and localized at the ER-PM junctions to regulate Ca(2+) signaling.

Abstract: Silencing or genetic manipulation of JP4 decreased ER Ca(2+) content and SOCE in T cells, impaired activation of the nuclear factor of activated T cells (NFAT) and extracellular signaling-related kinase (ERK) signaling pathways, and diminished expression of activation markers and cytokines.

Abstract: Mechanistically, JP4 directly interacted with STIM1 via its cytoplasmic domain and facilitated its recruitment into the junctions.

Abstract: Accordingly, expression of this cytoplasmic fragment of JP4 inhibited SOCE.

Abstract: Furthermore, JP4 also formed a complex with junctate, a Ca(2+)-sensing ER-resident protein, previously shown to mediate STIM1 recruitment into the junctions.

Abstract: We propose that the junctate-JP4 complex located at the junctions cooperatively interacts with STIM1 to maintain ER Ca(2+) homeostasis and mediate SOCE in T cells.


The PI3K inhibitor arsenal: choose your weapon!

Abstract: Owing to its widespread activation in inflammation and cancer, a growing appreciation of the therapeutic potential of inhibitors of the phosphoinositide 3-kinase (PI3K) pathway has stimulated intense interest in compounds with suitable pharmacological profiles.

Abstract: These are primarily directed toward PI3K itself.

Abstract: However, as class I PI3Ks are also essential for a range of normal physiological processes, broad spectrum PI3K inhibition could be poorly tolerated.

Abstract: In recent years, patents describing a new generation of PI3K inhibitors have started to appear, with a particular focus on the development of compounds with enhanced isoform selectivity for use as anti-cancer and anti-inflammatory therapies.

Abstract: However, challenges remain for the efforts to pharmacologically target this enzyme family in a successful manner.


Infralimbic BDNF/TrkB enhancement of GluN2B currents facilitates extinction of a cocaine-conditioned place preference.

Abstract: Brain-derived neurotrophic factor (BDNF) regulates synaptic activity and behavioral flexibility, and reduction of BDNF strongly predicts psychiatric disorders and cognitive dysfunction.

Abstract: Restoration of BDNF-dependent activity could alleviate these impairments, but BDNF has limited clinical utility due to its pharmacokinetics.

Abstract: Here we demonstrate that activation of a primary BDNF target, the tropomyosin-related kinase B (TrkB) receptor, enhances the amplitude and prolongs the decay kinetics of N-methyl-d-aspartate receptor (NMDAR) currents in male rat infralimbic prefrontal pyramidal neurons.

Abstract: Moreover, these effects were prevented and reversed by blockade of NMDARs containing the GluN2B subunit.

Abstract: Our results show that this signaling cascade bidirectionally regulates extinction of a cocaine-induced conditioned place preference (CPP), a task that requires behavioral flexibility.

Abstract: Blockade of infralimbic TrkB receptors or GluN2B-containing NMDARs disrupted consolidation of extinction of the CPP.

Abstract: In contrast, extinction was strengthened by potentiation of TrkB receptor activity with infralimbic infusions of BDNF or systemic injections of 7,8 dihydroxyflavone (7,8DHF), the newly synthesized TrkB receptor agonist.

Abstract: The 7,8DHF-induced enhancement of extinction was prevented by infralimbic infusions of a GluN2B-specific receptor antagonist, demonstrating that TrkB receptor activation enhances extinction of cocaine-CPP via GluN2B-containing NMDARs.

Abstract: Together, infralimbic TrkB receptor activation strengthens GluN2B-containing NMDAR currents to support extinction learning.

Abstract: TrkB receptor agonists would therefore be useful as pharmacological adjuncts for extinction-based therapies for treatment of psychiatric disorders associated with reduced BDNF activity.


Compaction of chromatin by diverse Polycomb group proteins requires localized regions of high charge.

Abstract: Polycomb group (PcG) proteins are required for the epigenetic maintenance of developmental genes in a silent state.

Abstract: Proteins in the Polycomb-repressive complex 1 (PRC1) class of the PcG are conserved from flies to humans and inhibit transcription.

Abstract: One hypothesis for PRC1 mechanism is that it compacts chromatin, based in part on electron microscopy experiments demonstrating that Drosophila PRC1 compacts nucleosomal arrays.

Abstract: We show that this function is conserved between Drosophila and mouse PRC1 complexes and requires a region with an overrepresentation of basic amino acids.

Abstract: While the active region is found in the Posterior Sex Combs (PSC) subunit in Drosophila, it is unexpectedly found in a different PRC1 subunit, a Polycomb homolog called M33, in mice.

Abstract: We provide experimental support for the general importance of a charged region by predicting the compacting capability of PcG proteins from species other than Drosophila and mice and by testing several of these proteins using solution assays and microscopy.

Abstract: We infer that the ability of PcG proteins to compact chromatin in vitro can be predicted by the presence of domains of high positive charge and that PRC1 components from a variety of species conserve this highly charged region.

Abstract: This supports the hypothesis that compaction is a key aspect of PcG function.


Angiopoietin-like proteins stimulate ex vivo expansion of hematopoietic stem cells

Abstract: Successful ex vivo expansion of hematopoietic stem cells (HSCs) would greatly benefit the treatment of disease and the understanding of crucial questions of stem cell biology.

Abstract: Here we show, using microarray studies, that the HSC-supportive mouse fetal liver CD3+ cells specifically express the proteins angiopoietin-like 2 (Angptl2) and angiopoietin-like 3 (Angptl3).

Abstract: We observed a 24- or 30-fold net expansion of long-term HSCs by reconstitution analysis when we cultured highly enriched HSCs for 10 days in the presence of Angptl2 or Angptl3 together with saturating levels of other growth factors.

Abstract: The coiled-coil domain of Angptl2 was capable of stimulating expansion of HSCs.

Abstract: Furthermore, angiopoietin-like 5, angiopoietin-like 7 and microfibril-associated glycoprotein 4 also supported expansion of HSCs in culture.


Balancing co-stimulation and inhibition with BTLA and HVEM

Abstract: The interaction between B- and T-lymphocyte attenuator (BTLA), an inhibitory receptor whose extracellular domain belongs to the immunoglobulin superfamily, and herpesvirus-entry mediator (HVEM), a co-stimulatory tumour-necrosis factor receptor, is unique in that it is the only receptor–ligand interaction that directly bridges these two families of receptors.

Abstract: This interaction has raised many questions about how receptors from two different families could interact and what downstream signalling events might occur as a result of receptor ligation.

Abstract: As we discuss, recent studies show that engagement of HVEM with its endogenous ligand (LIGHT) from the tumour-necrosis factor family induces a powerful immune response, whereas HVEM interactions with BTLA negatively regulate T-cell responses.


Lin28b is sufficient to drive liver cancer and necessary for its maintenance in murine models.

Abstract: Lin28a/b are RNA-binding proteins that influence stem cell maintenance, metabolism, and oncogenesis.

Abstract: Poorly differentiated, aggressive cancers often overexpress Lin28, but its role in tumor initiation or maintenance has not been definitively addressed.

Abstract: We report that LIN28B overexpression is sufficient to initiate hepatoblastoma and hepatocellular carcinoma in murine models.

Abstract: We also detected Lin28b overexpression in MYC-driven hepatoblastomas, and liver-specific deletion of Lin28a/b reduced tumor burden, extended latency, and prolonged survival.

Abstract: Both intravenous siRNA against Lin28b and conditional Lin28b deletion reduced tumor burden and prolonged survival.

Abstract: Igf2bp proteins are upregulated, and Igf2bp3 is required in the context of LIN28B overexpression to promote growth.

Abstract: Therefore, multiple murine models demonstrate that Lin28b is both sufficient to initiate liver cancer and necessary for its maintenance.


Nanoenzymology of the 20S proteasome: proteasomal actions are controlled by the allosteric transition.

Abstract: The proteasome is a major cytosolic proteolytic assembly, essential for the physiology of eukaryotic cells.

Abstract: Both the architecture and enzymatic properties of the 20S proteasome are relatively well understood.

Abstract: However, despite longstanding interest, the integration of structural and functional properties of the proteasome into a coherent model explaining the mechanism of its enzymatic actions has been difficult.

Abstract: Recently, we used tapping mode atomic force microscopy (AFM) in liquid to demonstrate that the alpha-rings of the proteasome imaged in a top-view position repeatedly switched between their open and closed conformations, apparently to control access to the central channel.

Abstract: Here, we show with AFM that the molecules in a side-view position acquired two stable conformations.

Abstract: The overall shapes of the 20S particles were classified as either barrel-like or cylinder-like.

Abstract: The relative abundance of the two conformers depended on the nature of their interactions with ligands.

Abstract: Similarly to the closed molecules in top view, the barrels predominated in control or inhibited molecules.

Abstract: The cylinders and open molecules prevailed when the proteasome was observed in the presence of peptide substrates.

Abstract: Based on these data, we developed the two-state model of allosteric transitions to explain the dynamics of proteasomal structure.

Abstract: This model helps to better understand the observed properties of the 20S molecule, and sets foundations for further studies of the structural dynamics of the proteasome.


Changes in plasma copeptin, the c-terminal portion of arginine vasopressin during water deprivation and excess in healthy subjects.

Abstract: CONTEXT The measurement of arginine vasopressin (AVP) is often cumbersome because it is unstable with a short half-life time.

Abstract: AVP is derived from a larger precursor peptide along with the more stable peptide copeptin.

Abstract: Copeptin is the C-terminal part of provasopressin and has been shown to be a useful tool to indicate AVP concentration in critically ill patients.

Abstract: OBJECTIVE The objective of the study was to evaluate the clinical usefulness of copeptin as a new marker in disordered states of blood volume and plasma osmolality.

Abstract: DESIGN AND SETTING This was a prospective observational study in a university hospital.

Abstract: PARTICIPANTS AND MAIN OUTCOME MEASURES Three techniques with respective control studies were used in 24 healthy adults to produce changes in plasma osmolality and/or volume: 1) a 28-h water deprivation, 2) a 17-h hypertonic saline infusion combined with thirsting, and 3) a hypotonic saline infusion with iv desmopressin administration during free water intake.

Abstract: RESULTS Water deprivation produced a weight loss of 1.7 kg, an increase in plasma osmolality to 294.8 +/-

Abstract: 4.3 mosmol/kg, and an increase of copeptin from 4.6 +/-

Abstract: 1.7 pmol/liter to 9.2 +/-

Abstract: 5.2 pmol/liter (P < 0.0001).

Abstract: During hypertonic saline infusion and thirsting with a raise of plasma osmolality to 296.1 +/-

Abstract: 3.4 mosmol/kg, copeptin increased from 4.9 +/-

Abstract: 3.0 pmol/liter to 19.9 +/-

Abstract: 4.8 pmol/liter (P < 0.0001).

Abstract: Conversely, during hypotonic saline infusion, plasma osmolality decreased to 271.3 +/-

Abstract: 4.1 mosmol/kg, and copeptin decreased from 6.2 +/-

Abstract: 2.4 pmol/liter to 2.4 +/-

Abstract: 2.1 pmol/liter (P < 0.01).

Abstract: CONCLUSION Copeptin shows identical changes during disordered water states as previously shown for AVP.

Abstract: It might be a reliable marker of AVP secretion and substitute for the measurement of circulating AVP levels in clinical routine.


Propagation and recovery of intact, infectious Epstein-Barr virus from prokaryotic to human cells.

Abstract: With current techniques, genetic alterations of herpesviruses are difficult to perform, mostly because of the large size of their genomes.

Abstract: To solve this problem, we have designed a system that allows the cloning of any gamma-herpesvirus in Escherichia coli onto an F factor-derived plasmid.

Abstract: Immortalized B cell lines were readily established with recombinant Epstein-Barr virus (EBV), demonstrating that the F factor-cloned EBV genome has all the characteristics of wild-type EBV.

Abstract: Because any genetic modification is possible in E. coli, this experimental approach opens the way to the genetic analysis of all EBV functions.

Abstract: Moreover, it is now feasible to generate attenuated EBV strains in vitro such that vaccine strains can be designed.

Abstract: Because we incorporated the genes for hygromycin resistance and green fluorescent protein onto the E. coli cloned EBV genome, the still open question of the EBV target cells other than B lymphocytes will be addressed.


Chemosensitivity profile assay of circulating cancer cells: prognostic and predictive value in epithelial tumors.

Abstract: The prognostic value associated with the detection of circulating tumor cells (CTCs) in metastatic breast cancer by the CellSearch technology raise additional issues regarding the biological value of this information.

Abstract: We postulated that a drug-resistance profile of CTCs may predict response to chemotherapy in cancer patients and therefore could be used for patient selection.

Abstract: One hundred 5 patients with diagnosis of carcinoma were enrolled in a prospective trial.

Abstract: CTCs were isolated from peripheral blood, and positive samples were evaluated for the expression of a panel of genes involved in anticancer drugs resistance.

Abstract: The drug-resistance profile was correlated with disease-free survival (DFS; patients in adjuvant setting) and time to progression (TTP; metastatic patients) in a 24-months follow-up.

Abstract: Objective response correlation was a secondary end point.

Abstract: Fifty-one percent of patients were found positive for CTCs while all blood samples from healthy donors were negative.

Abstract: The drug-resistance profile correlates with DFS and TTP (p < 0.001 in both).

Abstract: Sensitivity of the test: able to predict treatment response in 98% of patients.

Abstract: Specificity of the test: 100%; no sample from healthy subject was positive for the presence of CTCs.

Abstract: Positive and negative predictive values were found to be 96.5 and 100%, respectively.

Abstract: We identified a drug-resistance profile of CTCs, which is predictive of response to chemotherapy, independent of tumor type and stage of disease.

Abstract: This approach may represent a first step toward the individualization of chemotherapy in cancer patients.


Trends in heart failure incidence and survival in a community-based population.

Abstract: CONTEXT The epidemic of heart failure has yet to be fully investigated, and data on incidence, survival, and sex-specific temporal trends in community-based populations are limited.

Abstract: OBJECTIVE To test the hypothesis that the incidence of heart failure has declined and survival after heart failure diagnosis has improved over time but that secular trends have diverged by sex.

Abstract: DESIGN, SETTING, AND PARTICIPANTS Population-based cohort study using the resources of the Rochester Epidemiology Project conducted in Olmsted County, Minnesota.

Abstract: Patients were 4537 Olmsted County residents (57% women; mean [SD] age, 74 [14] years) with a diagnosis of heart failure between 1979 and 2000.

Abstract: Framingham criteria and clinical criteria were used to validate the diagnosis   MAIN OUTCOME MEASURES Incidence of heart failure and survival after heart failure diagnosis.

Abstract: RESULTS The incidence of heart failure was higher among men (378/100 000 persons; 95% confidence interval [CI], 361-395 for men; 289/100 000 persons; 95% CI, 277-300 for women) and did not change over time among men or women.

Abstract: After a mean follow-up of 4.2 years (range, 0-23.8 years), 3347 deaths occurred, including 1930 among women and 1417 among men.

Abstract: Survival after heart failure diagnosis was worse among men than women (relative risk, 1.33; 95% CI, 1.24-1.43) but overall improved over time (5-year age-adjusted survival, 43% in 1979-1984 vs 52% in 1996-2000, P<.001).

Abstract: However, men and younger persons experienced larger survival gains, contrasting with less or no improvement for women and elderly persons.

Abstract: CONCLUSION In this community-based cohort, the incidence of heart failure has not declined during 2 decades, but survival after onset of heart failure has increased overall, with less improvement among women and elderly persons.


Doxycycline induces apoptosis in PANC-1 pancreatic cancer cells.

Abstract: BACKGROUND Tetracyclines such as doxycycline are reported to possess cytotoxic activity against mammalian tumor cells, but the mechanism of their effects on cell proliferation remains unclear.

Abstract: MATERIALS AND METHODS The antitumor effect of doxycycline was investigated in human pancreatic cancer cell line, PANC-1.

Abstract: We also investigated the effect of doxycycline on expression of a potent proangiogenic factor, interleukin (IL)-8.

Abstract: RESULTS In excess of 20 microg/ml, cytotoxic effects of doxycycline were accompanied by G(1)-S cell cycle arrest and DNA fragmentation in PANC-1 cells.

Abstract: Doxycycline consistently activated transcription of p53, p21 and Fas/FasL-cascade-related genes, while reducing the expression of Bcl-xL and Mcl-1.

Abstract: Doxycycline (5 microg/ml) below the cytotoxic level suppressed endogenous and paclitaxel-induced IL-8 expression.

Abstract: In the mouse xenograft model, doxycycline treatment was shown to suppress tumor growth by 80%.

Abstract: CONCLUSION These data suggest that doxycycline exerts its antitumor effect by activating proapoptotic genes, inhibiting IL-8 expression, and suppressing antiapoptotic genes.


Orientationally invariant indices of axon diameter and density from diffusion MRI.

Abstract: This paper proposes and tests a technique for imaging orientationally invariant indices of axon diameter and density in white matter using diffusion magnetic resonance imaging.

Abstract: Such indices potentially provide more specific markers of white matter microstructure than standard indices from diffusion tensor imaging.

Abstract: Orientational invariance allows for combination with tractography and presents new opportunities for mapping brain connectivity and quantifying disease processes.

Abstract: The technique uses a four-compartment tissue model combined with an optimized multishell high-angular-resolution pulsed-gradient-spin-echo acquisition.

Abstract: We test the method in simulation, on fixed monkey brains using a preclinical scanner and on live human brains using a clinical 3T scanner.

Abstract: The human data take about one hour to acquire.

Abstract: The simulation experiments show that both monkey and human protocols distinguish distributions of axon diameters that occur naturally in white matter.

Abstract: We compare the axon diameter index with the mean axon diameter weighted by axon volume.

Abstract: The index differs from this mean and is protocol dependent, but correlation is good for the monkey protocol and weaker, but discernible, for the human protocol where greater diffusivity and lower gradient strength limit sensitivity to only the largest axons.

Abstract: Maps of axon diameter and density indices from the monkey and human data in the corpus callosum and corticospinal tract reflect known trends from histology.

Abstract: The results show orientationally invariant sensitivity to natural axon diameter distributions for the first time with both specialist and clinical hardware.

Abstract: This demonstration motivates further refinement, validation, and evaluation of the precise nature of the indices and the influence of potential confounds.


IFI16 and cGAS cooperate in the activation of STING during DNA sensing in human keratinocytes

Abstract: Many human cells can sense the presence of exogenous DNA during infection though the cytosolic DNA receptor cyclic GMP-AMP synthase (cGAS), which produces the second messenger cyclic GMP-AMP (cGAMP).

Abstract: Other putative DNA receptors have been described, but whether their functions are redundant, tissue-specific or integrated in the cGAS-cGAMP pathway is unclear.

Abstract: Here we show that interferon-γ inducible protein 16 (IFI16) cooperates with cGAS during DNA sensing in human keratinocytes, as both cGAS and IFI16 are required for the full activation of an innate immune response to exogenous DNA and DNA viruses.

Abstract: IFI16 is also required for the cGAMP-induced activation of STING, and interacts with STING to promote STING phosphorylation and translocation.

Abstract: We propose that the two DNA sensors IFI16 and cGAS cooperate to prevent the spurious activation of the type I interferon response.


Oncostatic effects of the indole melatonin and expression of its cytosolic and nuclear receptors in cultured human melanoma cell lines.

Abstract: Melatonin has been shown to have oncostatic effects on malignant melanoma in vitro and in vivo.

Abstract: We studied the growth suppressive effects of melatonin over a wide range of concentrations in four melanoma cell lines (SBCE2, WM-98, WM-164 and SKMEL-188) representative for different growth stages and phenotype.

Abstract: Melanoma cells were incubated with melatonin 10(-12)-10(-3) M, and proliferation and clonogenicity was assessed at 12 h and 14 days, respectively.

Abstract: We also determined the expression of cytosolic quinone oxidoreductases NQO1, NQO2 (known as MT3 receptor) and nuclear receptor RORalpha by RT-PCR.

Abstract: Melatonin at pharmacological concentrations (10(-3)-10(-7) M) suppressed proliferation in all melanoma cell lines.

Abstract: In SKMEL-188 cells cultured in serum-free media, melatonin at low concentrations (10(-12)-10(-10) M) also slightly attenuated the proliferation.

Abstract: The effects of pharmacological doses of melatonin were confirmed in the clonogenic assay.

Abstract: Expression of NQO1 was detected in all cell lines, whereas NQO2 and nuclear receptor RORalpha including its isoform RORalpha4 were present only in SBCE2, WM-164 and WM-98.

Abstract: Thus, melatonin differentially suppressed proliferation in melanoma cell lines of different behaviour.

Abstract: The intensity of the oncostatic response to melatonin could be related to the cell-line specific pattern of melatonin cellular receptors and cytosolic binding protein expression.


Lysosomal localization of TRPML3 depends on TRPML2 and the mucolipidosis-associated protein TRPML1.

Abstract: Mucolipidosis type IV is an autosomal recessive lysosomal storage disorder characterized by severe neurodegeneration, achlorhydria, and visual impairments such as corneal opacity and strabismus.

Abstract: The disease arises due to mutations in a group 2 transient receptor potential (TRP)-related cation channel, TRPML1.

Abstract: Mammals encode two additional TRPML proteins named TRPML2 and TRPML3.

Abstract: Information regarding the propensity of these proteins to multimerize, their subcellular distribution and mechanisms that regulate their trafficking are limited.

Abstract: Here we demonstrate that TRPMLs interact to form homo- and heteromultimers.

Abstract: Moreover, the presence of either TRPML1 or TRPML2 specifically influences the spatial distribution of TRPML3.

Abstract: TRPML1 and TRPML2 homomultimers are lysosomal proteins, whereas TRPML3 homomultimers are in the endoplasmic reticulum.

Abstract: However, TRPML3 localizes to lysosomes when coexpressed with either TRPML1 or TRPML2 and is comparably mislocalized when lysosomal targeting of TRPML1 and TRPML2 is disrupted.

Abstract: Conversely, TRPML3 does not cause retention of TRPML1 or TRPML2 in the endoplasmic reticulum.

Abstract: These data demonstrate that there is a hierarchy controlling the subcellular distributions of the TRPMLs such that TRPML1 and TRPML2 dictate the localization of TRPML3 and not vice versa.


Evolution of metastasis revealed by mutational landscapes of chemically induced skin cancers

Abstract: Human tumors show a high level of genetic heterogeneity, but the processes that influence the timing and route of metastatic dissemination of the subclones are unknown.

Abstract: Here we have used whole-exome sequencing of 103 matched benign, malignant and metastatic skin tumors from genetically heterogeneous mice to demonstrate that most metastases disseminate synchronously from the primary tumor, supporting parallel rather than linear evolution as the predominant model of metastasis.

Abstract: Shared mutations between primary carcinomas and their matched metastases have the distinct A-to-T signature of the initiating carcinogen dimethylbenzanthracene, but non-shared mutations are primarily G-to-T, a signature associated with oxidative stress.

Abstract: The existence of carcinomas that either did or did not metastasize in the same host animal suggests that there are tumor-intrinsic factors that influence metastatic seeding.

Abstract: We also demonstrate the importance of germline polymorphisms in determining allele-specific mutations, and we identify somatic genetic alterations that are specifically related to initiation of carcinogenesis by Hras or Kras mutations.

Abstract: Mouse tumors that mimic the genetic heterogeneity of human cancers can aid our understanding of the clonal evolution of metastasis and provide a realistic model for the testing of novel therapies.


A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002).

Abstract: Phosphatidylinositol (PtdIns) 3-kinase is an enzyme implicated in growth factor signal transduction by associating with receptor and nonreceptor tyrosine kinases, including the platelet-derived growth factor receptor.

Abstract: Inhibitors of PtdIns 3-kinase could potentially give a better understanding of the function and regulatory mechanisms of the enzyme.

Abstract: Quercetin, a naturally occurring bioflavinoid, was previously shown to inhibit PtdIns 3-kinase with an IC50 of 1.3 microgram/ml (3.8 microM); inhibition appeared to be directed at the ATP-binding site of the kinase.

Abstract: Analogs of quercetin were investigated as PtdIns 3-kinase inhibitors, with the most potent ones exhibiting IC50 values in the range of 1.7-8.4 micrograms/ml.

Abstract: In contrast, genistein, a potent tyrosine kinase inhibitor of the isoflavone class, did not inhibit PtdIns 3-kinase significantly (IC50 > 30 micrograms/ml).

Abstract: Since quercetin has also been shown to inhibit other PtdIns and protein kinases, other chromones were evaluated as inhibitors of PtdIns 3-kinase without affecting PtdIns 4-kinase or selected protein kinases.

Abstract: One such compound, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (also known as 2-(4-morpholinyl)-8-phenylchromone, LY294002), completely and specifically abolished PtdIns 3-kinase activity (IC50 = 0.43 microgram/ml; 1.40 microM) but did not inhibit PtdIns 4-kinase or tested protein and lipid kinases.

Abstract: Analogs of LY294002 demonstrated a very selective structure-activity relationship, with slight changes in structure causing marked decreases in inhibition.

Abstract: LY294002 was shown to completely abolish PtdIns 3-kinase activity in fMet-Leu-Phe-stimulated human neutrophils, as well as inhibit proliferation of smooth muscle cells in cultured rabbit aortic segments.

Abstract: Since PtdIns 3-kinase appears to be centrally involved with growth factor signal transduction, the development of specific inhibitors against the kinase may be beneficial in the treatment of proliferative diseases as well as in elucidating the biological role of the kinase in cellular proliferation and growth factor response.


A simple two-step protocol for the purification of human pancreatic beta cells

Abstract: Isolated pure human beta cells would be helpful for a number of research purposes.

Abstract: However, lack of beta cell-specific surface antigens has been a major problem.

Abstract: We aimed to develop a simple method for human beta cell isolation based on the initial elimination of ductal cells by their expression of carbohydrate antigen 19-9 (CA19-9), followed by positive selection of beta cells by their expression of polysialic acid–neural cell adhesion molecule (PSA-NCAM).

Abstract: Cell type-specific expression of CA19-9, NCAM and PSA-NCAM was studied in sections of adult human pancreas and in cultured primary endocrine and exocrine cells.

Abstract: Dispersed human islet cells were purified in two steps, after 4 days of suspension culture, by binding to magnetic microbeads coupled to antibodies against CA19-9 and PSA-NCAM.

Abstract: NCAM expression was detected in ducts and islets in the human pancreas.

Abstract: In contrast, PSA-NCAM immunoreactivity was detected only in islets.

Abstract: PSA-NCAM staining in dispersed cells revealed that the marker is expressed in all endocrine cell types, but not in duct cells.

Abstract: Purification of dispersed islet cells using PSA-NCAM microbeads alone did not completely eliminate contaminating duct cells.

Abstract: However, elimination of the duct cells by CA19-9 microbeads followed by positive sorting of the PSA-NCAM-positive cells in five consecutive islet preparations resulted in 90 to 98% pure endocrine cells, of which 89 to 97% were beta cells.

Abstract: We describe a simple and reproducible method for purification of viable human pancreatic beta cells devoid of exocrine acini and ducts.


Prev | Table of Contents Reports Embryonic Stem Cell Lines Derived from Human

Abstract: Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages.

Abstract: After undifferentiated proliferation in vitro for 4 to 5 months, these cells still maintained the developmental potential to form trophoblast and derivatives of all three embryonic germ layers, including gut epithelium (endoderm); cartilage, bone, smooth muscle, and striated muscle (mesoderm); and neural epithelium, embryonic ganglia, and stratified squamous epithelium (ectoderm).

Abstract: These cell lines should be useful in human developmental biology, drug discovery, and transplantation medicine.


Ascorbic-acid transporter Slc23a1 is essential for vitamin C transport into the brain and for perinatal survival

Abstract: The only proven requirement for ascorbic acid (vitamin C) is in preventing scurvy, presumably because it is a cofactor for hydroxylases required for post-translational modifications that stabilize collagen.

Abstract: We have created mice deficient in the mouse ortholog (solute carrier family 23 member 1 or Slc23a1) of a rat ascorbic-acid transporter, Svct2 (ref.

Abstract: Cultured embryonic fibroblasts from homozygous Slc23a1−/− mice had less than 5% of normal ascorbic-acid uptake.

Abstract: Ascorbic-acid levels were undetectable or markedly reduced in the blood and tissues of Slc23a1−/− mice.

Abstract: Prenatal supplementation of pregnant females did not elevate blood ascorbic acid in Slc23a1−/− fetuses, suggesting Slc23a1 is important in placental ascorbic-acid transport.

Abstract: Slc23a1−/− mice died within a few minutes of birth with respiratory failure and intraparenchymal brain hemorrhage.

Abstract: Lungs showed no postnatal expansion but had normal surfactant protein B levels.

Abstract: Brain hemorrhage was unlikely to be simply a form of scurvy since Slc23a1−/− mice showed no hemorrhage in any other tissues and their skin had normal skin 4-hydroxyproline levels despite low ascorbic-acid content.

Abstract: We conclude that Slc23a1 is required for transport of ascorbic acid into many tissues and across the placenta.

Abstract: Deficiency of the transporter is lethal in newborn mice, thereby revealing a previously unrecognized requirement for ascorbic acid in the perinatal period.


MVP-mediated exosomal sorting of miR-193a promotes colon cancer progression

Abstract: Exosomes are emerging mediators of intercellular communication; whether the release of exosomes has an effect on the exosome donor cells in addition to the recipient cells has not been investigated to any extent.

Abstract: Here, we examine different exosomal miRNA expression profiles in primary mouse colon tumour, liver metastasis of colon cancer and naive colon tissues.

Abstract: In more advanced disease, higher levels of tumour suppressor miRNAs are encapsulated in the exosomes.

Abstract: miR-193a interacts with major vault protein (MVP).

Abstract: Knockout of MVP leads to miR-193a accumulation in the exosomal donor cells instead of exosomes, inhibiting tumour progression.

Abstract: Furthermore, miR-193a causes cell cycle G1 arrest and cell proliferation repression through targeting of Caprin1, which upregulates Ccnd2 and c-Myc.

Abstract: Human colon cancer patients with more advanced disease show higher levels of circulating exosomal miR-193a.

Abstract: In summary, our data demonstrate that MVP-mediated selective sorting of tumour suppressor miRNA into exosomes promotes tumour progression.


Casual blood pressure and neurocognitive function in children with chronic kidney disease: a report of the children with chronic kidney disease cohort study.

Abstract: BACKGROUND AND OBJECTIVES Children with chronic kidney disease (CKD) are at risk for cognitive dysfunction, and over half have hypertension.

Abstract: Data on the potential contribution of hypertension to CKD-associated neurocognitive deficits in children are limited.

Abstract: Our objective was to determine whether children with CKD and elevated BP (EBP) had decreased performance on neurocognitive testing compared with children with CKD and normal BP.

Abstract: DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS This was a cross-sectional analysis of the relation between auscultatory BP and neurocognitive test performance in children 6 to 17 years enrolled in the Chronic Kidney Disease in Children (CKiD) project.

Abstract: RESULTS Of 383 subjects, 132 (34%) had EBP (systolic BP and/or diastolic BP ≥90(th) percentile).

Abstract: Subjects with EBP had lower mean (SD) scores on Wechsler Abbreviated Scales of Intelligence (WASI) Performance IQ than those with normal BP (normal BP versus EBP, 96.1 (16.7) versus 92.4 (14.9), P = 0.03) and WASI Full Scale IQ (97.0 (16.2) versus 93.4 (16.5), P = 0.04).

Abstract: BP index (subject's BP/95(th) percentile BP) correlated inversely with Performance IQ score (systolic, r = -0.13, P = 0.01; diastolic, r = -0.19, P < 0.001).

Abstract: On multivariate analysis, the association between lower Performance IQ score and increased BP remained significant after controlling for demographic and disease-related variables (EBP, β = -3.7, 95% confidence interval [CI]: -7.3 to -0.06; systolic BP index, β = -1.16 to 95% CI: -2.1, -0.21; diastolic BP index, β = -1.17, 95% CI: -1.8 to -0.55).

Abstract: CONCLUSIONS Higher BP was independently associated with decreased WASI Performance IQ scores in children with mild-to-moderate CKD.


Bone marrow failure and developmental delay caused by mutations in poly(A)-specific ribonuclease (PARN).

Abstract: BACKGROUND Deadenylation regulates RNA function and fate.

Abstract: Poly(A)-specific ribonuclease (PARN) is a deadenylase that processes mRNAs and non-coding RNA.

Abstract: Little is known about the biological significance of germline mutations in PARN.

Abstract: METHODS We identified mutations in PARN in patients with haematological and neurological manifestations.

Abstract: Genomic, biochemical and knockdown experiments in human marrow cells and in zebrafish have been performed to clarify the role of PARN in the human disease.

Abstract: RESULTS We identified large monoallelic deletions in PARN in four patients with developmental delay or mental illness.

Abstract: One patient in particular had a severe neurological phenotype, central hypomyelination and bone marrow failure.

Abstract: This patient had an additional missense mutation on the non-deleted allele and severely reduced PARN protein and deadenylation activity.

Abstract: Cells from this patient had impaired oligoadenylation of specific H/ACA box small nucleolar RNAs.

Abstract: Importantly, PARN-deficient patient cells manifested short telomeres and an aberrant ribosome profile similar to those described in some variants of dyskeratosis congenita.

Abstract: Knocking down PARN in human marrow cells and zebrafish impaired haematopoiesis, providing further evidence for a causal link with the human disease.

Abstract: CONCLUSIONS Large monoallelic mutations of PARN can cause developmental/mental illness.

Abstract: Biallelic PARN mutations cause severe bone marrow failure and central hypomyelination.


The changing epidemiology of malaria in Ifakara Town, southern Tanzania.

Abstract: Between 1995 and 2000 there were marked changes in the epidemiology of malaria in Ifakara, southern Tanzania.

Abstract: We documented these changes using parasitological and clinical data from a series of community- and hospital-based studies involving children up to the age of 5 years.

Abstract: There was a right shift and lowering in the age-specific parasite prevalence in the community-based cohort studies.

Abstract: The incidence of clinical malaria in placebo-receiving infants in additional study cohorts dropped from 0.8 in 1995 to 0.43 episodes per infant per year in 2000, an incidence rate ratio of 0.53 (95% confidence interval: 0.404, 0.70, P<0.0001).

Abstract: At the same time, there was an increase in the total number of malaria admissions and a marked right shift in the age pattern of these admissions (median age in 1995 1.55 years vs. 2.33 in 2000, P<0.0001).

Abstract: However, the burden of malaria deaths remained in infants.

Abstract: We discuss how these dramatic changes in the epidemiology of malaria may have arisen from the use of currently available malaria control tools.

Abstract: Caution is required in the interpretation of hospital-based data as it is likely to underestimate the impact of anaemia on mortality in the community, where most paediatric deaths occur.

Abstract: Even in low/moderate malaria transmission settings, where older children suffer most malaria episodes, targeting effective malaria control at infants may produce important reductions in infant mortality caused by malaria.


MicroRNA-192 in diabetic kidney glomeruli and its function in TGF-beta-induced collagen expression via inhibition of E-box repressors.

Abstract: Key features of diabetic nephropathy (DN) include the accumulation of extracellular matrix proteins such as collagen 1-alpha 1 and -2 (Col1a1 and -2).

Abstract: Transforming growth factor beta1 (TGF-beta), a key regulator of these extracellular matrix genes, is increased in mesangial cells (MC) in DN.

Abstract: By microarray profiling, we noted that TGF-beta increased Col1a2 mRNA in mouse MC (MMC) but also decreased mRNA levels of an E-box repressor, deltaEF1.

Abstract: TGF-beta treatment or short hairpin RNAs targeting deltaEF1 increased enhancer activity of upstream E-box elements in the Col1a2 gene.

Abstract: TGF-beta also decreased the expression of Smad-interacting protein 1 (SIP1), another E-box repressor similar to deltaEF1.

Abstract: Interestingly, we noted that SIP1 is a target of microRNA-192 (miR-192), a key miR highly expressed in the kidney.

Abstract: miR-192 levels also were increased by TGF-beta in MMC.

Abstract: TGF-beta treatment or transfection with miR-192 decreased endogenous SIP1 expression as well as reporter activity of a SIP1 3' UTR-containing luciferase construct in MMC.

Abstract: Conversely, a miR-192 inhibitor enhanced the luciferase activity, confirming SIP1 to be a miR-192 target.

Abstract: Furthermore, miR-192 synergized with deltaEF1 short hairpin RNAs to increase Col1a2 E-box-luc activity.

Abstract: Importantly, the in vivo relevance was noted by the observation that miR-192 levels were enhanced significantly in glomeruli isolated from streptozotocin-injected diabetic mice as well as diabetic db/db mice relative to corresponding nondiabetic controls, in parallel with increased TGF-beta and Col1a2 levels.

Abstract: These results uncover a role for miRs in the kidney and DN in controlling TGF-beta-induced Col1a2 expression by down-regulating E-box repressors.


Activation of PAR(2) receptors sensitizes primary afferents and causes leukocyte rolling and adherence in the rat knee joint.

Abstract: BACKGROUND AND PURPOSE The PAR(2) receptors are involved in chronic arthritis by mechanisms that are as yet unclear.

Abstract: Here, we examined PAR(2) activation in the rat knee joint.

Abstract: EXPERIMENTAL APPROACH PAR(2) in rat knee joint dorsal root ganglia (DRG) cells at L3-L5, retrogradely labelled with Fluoro-gold (FG) were demonstrated immunohistochemically.

Abstract: Electrophysiological recordings from knee joint nerve fibres in urethane anaesthetized Wistar rats assessed the effects of stimulating joint PAR(2) with its activating peptide, 2-furoyl-LIGRLO-NH(2) (1-100 nmol·100 μL(-1) , via close intra-arterial injection).

Abstract: Fibre firing rate was recorded during joint rotations before and 15 min after administration of PAR(2) activating peptide or control peptide.

Abstract: Leukocyte kinetics in the synovial vasculature upon PAR(2) activation were followed by intravital microscopy for 60 min after perfusion of 2-furoyl-LIGRLO-NH(2) or control peptide.

Abstract: Roles for transient receptor potential vanilloid-1 (TRPV1) or neurokinin-1 (NK(1) ) receptors in the PAR(2) responses were assessed using the selective antagonists, SB366791 and RP67580 respectively.

Abstract: KEY RESULTS PAR(2) were expressed in 59 ± 5% of FG-positive DRG cells; 100 nmol 2-furoyl-LIGRLO-NH(2) increased joint fibre firing rate during normal and noxious rotation, maximal at 3 min (normal; 110 ± 43%, noxious; 90 ± 31%).

Abstract: 2-Furoyl-LIGRLO-NH(2) also significantly increased leukocyte rolling and adhesion over 60 min.

Abstract: All these effects were blocked by pre-treatment with SB366791 and RP67580 (P < 0.05 compared with 2-furoyl-LIGRLO-NH(2) alone).

Abstract: CONCLUSIONS AND IMPLICATIONS PAR(2) receptors play an acute inflammatory role in the knee joint via TRPV1- and NK(1) -dependent mechanisms involving both PAR(2) -mediated neuronal sensitization and leukocyte trafficking.


Polyphosphate kinase 1, a central node in the stress response network of Mycobacterium tuberculosis, connects the two-component systems MprAB and SenX3-RegX3 and the extracytoplasmic function sigma factor, sigma E.

Abstract: Polyphosphate (poly P) metabolism regulates the stress response in mycobacteria.

Abstract: Here we describe the regulatory architecture of a signal transduction system involving the two-component system (TCS) SenX3-RegX3, the extracytoplasmic function sigma factor sigma E (SigE) and the poly P-synthesizing enzyme polyphosphate kinase 1 (PPK1).

Abstract: The ppk1 promoter of Mycobacterium tuberculosis is activated under phosphate starvation.

Abstract: This is attenuated upon deletion of an imperfect palindrome likely representing a binding site for the response regulator RegX3, a component of the two-component system SenX3-RegX3 that responds to phosphate starvation.

Abstract: Binding of phosphorylated RegX3 to this site was confirmed by electrophoretic mobility shift assay.

Abstract: The activity of the ppk1 promoter was abrogated upon deletion of a putative SigE binding site.

Abstract: Pull-down of SigE from M. tuberculosis lysates of phosphate-starved cells with a biotinylated DNA harbouring the SigE binding site confirmed the likely binding of SigE to the ppk1 promoter.

Abstract: In vitro transcription corroborated the involvement of SigE in ppk1 transcription.

Abstract: Finally, the overexpression of RseA (anti-SigE) attenuated ppk1 expression under phosphate starvation, supporting the role of SigE in ppk1 transcription.

Abstract: The regulatory elements identified in ppk1 transcription in this study, combined with our earlier observation that PPK1 is itself capable of regulating sigE expression via the MprAB TCS, suggest the presence of multiple positive-feedback loops in this signalling circuit.

Abstract: In combination with the sequestering effect of RseA, we hypothesize that this architecture could be linked to bistability in the system that, in turn, could be a key element of persistence in M. tuberculosis.


Natural history of asymptomatic unruptured cerebral aneurysms evaluated at CT angiography: growth and rupture incidence and correlation with epidemiologic risk factors.

Abstract: PURPOSE To characterize the relationship between aneurysm size and epidemiologic risk factors with growth and rupture by using computed tomographic (CT) angiography.

Abstract: MATERIALS AND METHODS In this HIPAA-compliant, institutional review board approved study, patients with known asymptomatic unruptured intracerebral aneurysms were followed up longitudinally with CT angiographic examinations.

Abstract: Growth was defined as an increase in one or more dimensions above the measurement error, and at least 5% volume by using the ABC/2 method.

Abstract: Associations of epidemiologic factors with aneurysm growth and rupture were analyzed by using logistic regression analysis.

Abstract: Intra- and interobserver agreement coefficients for dimension, volume, and growth were evaluated by using the Pearson correlation coefficient and difference of means with 95% confidence intervals, the agreement statistic, and the McNemar χ(2).

Abstract: RESULTS Patients (n = 165) with aneurysms (n = 258) had a mean follow-up time of 2.24 years from time of diagnosis.

Abstract: Forty-six of 258 (18%) aneurysms in 38 patients grew larger.

Abstract: Spontaneous rupture occurred in four of 228 (1.8%) intradural aneurysms of average size (6.2 mm).

Abstract: Risk of aneurysm rupture per patient-year was 2.4% (95% CI: 0.5%, 7.12%) with growth and 0.2% (95% CI: 0.006%, 1.22%) without growth (P = .034).

Abstract: There was a 12-fold higher risk of rupture for growing aneurysms (P < .002), with high intra- and interobserver correlation coefficients for size, volume, and growth.

Abstract: Tobacco smoking (3.806, one degree of freedom; P < .015,) and initial size (5.895, two degrees of freedom; P < .051) were independent covariates, predicting 78.4% of growing aneurysms.

Abstract: CONCLUSION These results support imaging follow-up of all patients with aneurysms, including those whose aneurysms are smaller than the current 7-mm treatment threshold.

Abstract: Aneurysm growth, size, and smoking were associated with increased rupture risk.


A search for the presence of the enteroviral capsid protein VP1 in pancreases of patients with Type 1 (insulin-dependent) diabetes and pancreases and hearts of infants who died of coxsackieviral myocarditis

Abstract: Using an antiserum raised to a recombinant coxsackie virus B3 capsid protein, VP1, an immunocytochemical technique was developed which was capable of detecting the presence of all coxsackie B viruses in formalin fixed paraffin embedded infected tissue culture cells.

Abstract: This technique was tested on autopsy heart and pancreas from 21 patients who were thought to have died of acute coxsackievirus B myocarditis.

Abstract: Cardiac myocytes were positive for the VP1 protein in 12 of 20 cases where the heart was available for study.

Abstract: Insulitis was present in the pancreas in seven of these cases and in all seven islet endocrine cells containing VP1 were found.

Abstract: VP1 was only rarely found in exocrine pancreas.

Abstract: In heart and pancreas, cells shown to contain VP1 usually showed signs of necrosis.

Abstract: Autopsy pancreases from 88 patients who had died at clinical presentation of Type 1 (insulin-dependent) diabetes mellitus showed no evidence of the presence of VP1.

Abstract: The continuing destruction of insulin-secreting B cells seen at the time of death in the diabetic pancreas is unlikely to be due to a direct cytopathic effect of a coxsackie B virus.

Abstract: However, this study does not exclude the possibility that a persistent infection of B cells by a defective enterovirus may result in their destruction by an autoimmune mechanism.