6-Sulphated Chondroitins Have a Positive Influence on Axonal Regeneration

Abstract: Chondroitin sulphate proteoglycans (CSPGs) upregulated in the glial scar inhibit axon regeneration via their sulphated glycosaminoglycans (GAGs).

Abstract: Chondroitin 6-sulphotransferase-1 (C6ST-1) is upregulated after injury leading to an increase in 6-sulphated GAG.

Abstract: In this study, we ask if this increase in 6-sulphated GAG is responsible for the increased inhibition within the glial scar, or whether it represents a partial reversion to the permissive embryonic state dominated by 6-sulphated glycosaminoglycans (GAGs).

Abstract: Using C6ST-1 knockout mice (KO), we studied post-injury changes in chondroitin sulphotransferase (CSST) expression and the effect of chondroitin 6-sulphates on both central and peripheral axon regeneration.

Abstract: After CNS injury, wild-type animals (WT) showed an increase in mRNA for C6ST-1, C6ST-2 and C4ST-1, but KO did not upregulate any CSSTs.

Abstract: After PNS injury, while WT upregulated C6ST-1, KO showed an upregulation of C6ST-2.

Abstract: We examined regeneration of nigrostriatal axons, which demonstrate mild spontaneous axon regeneration in the WT.

Abstract: KO showed many fewer regenerating axons and more axonal retraction than WT.

Abstract: However, in the PNS, repair of the median and ulnar nerves led to similar and normal levels of axon regeneration in both WT and KO.

Abstract: Functional tests on plasticity after the repair also showed no evidence of enhanced plasticity in the KO.

Abstract: Our results suggest that the upregulation of 6-sulphated GAG after injury makes the extracellular matrix more permissive for axon regeneration, and that the balance of different CSs in the microenvironment around the lesion site is an important factor in determining the outcome of nervous system injury.


Tissue Mechanics Orchestrate Wnt-Dependent Human Embryonic Stem Cell Differentiation.

Abstract: Regenerative medicine is predicated on understanding the mechanisms regulating development and applying these conditions to direct stem cell fate.

Abstract: Embryogenesis is guided by cell-cell and cell-matrix interactions, but it is unclear how these physical cues influence stem cells in culture.

Abstract: We used human embryonic stem cells (hESCs) to examine whether mechanical features of the extracellular microenvironment could differentially modulate mesoderm specification.

Abstract: We found that, on a hydrogel-based compliant matrix, hESCs accumulate β-catenin at cell-cell adhesions and show enhanced Wnt-dependent mesoderm differentiation.

Abstract: Mechanistically, Src-driven ubiquitination of E-cadherin by Cbl-like ubiquitin ligase releases P120-catenin to facilitate transcriptional activity of β-catenin, which initiates and reinforces mesoderm differentiation.

Abstract: By contrast, on a stiff hydrogel matrix, hESCs show elevated integrin-dependent GSK3 and Src activity that promotes β-catenin degradation and inhibits differentiation.

Abstract: Thus, we found that mechanical features of the microenvironmental matrix influence tissue-specific differentiation of hESCs by altering the cellular response to morphogens.


Predictive performance of the quick Sequential Organ Failure Assessment score as a screening tool for sepsis, mortality, and intensive care unit admission in patients with febrile neutropenia

Abstract: In Sepsis-3, the quick Sequential Organ Failure Assessment (qSOFA) score was developed as criteria to use for recognizing patients who may have poor outcomes.

Abstract: This study was performed to evaluate the predictive performance of the qSOFA score as a screening tool for sepsis, mortality, and intensive care unit (ICU) admission in patients with febrile neutropenia (FN).

Abstract: We also tried to compare its performance with that of the systemic inflammatory response syndrome (SIRS) criteria and Multinational Association of Supportive Care in Cancer (MASCC) score for FN.

Abstract: We used a prospectively collected adult FN data registry.

Abstract: The qSOFA and SIRS scores were calculated retrospectively using the preexisting data.

Abstract: The primary outcome was the development of sepsis.

Abstract: The secondary outcomes were ICU admission and 28-day mortality.

Abstract: Of the 615 patients, 100 developed sepsis, 20 died, and 38 were admitted to ICUs.

Abstract: In multivariate analysis, qSOFA was an independent factor predicting sepsis and ICU admission.

Abstract: However, compared to the MASCC score, the area under the receiver operating curve of qSOFA was lower.

Abstract: qSOFA showed a low sensitivity (0.14, 0.2, and 0.23) but high specificity (0.98, 0.97, and 0.97) in predicting sepsis, 28-day mortality, and ICU admission.

Abstract: Performance of the qSOFA score was inferior to that of the MASCC score.

Abstract: The preexisting risk stratification tool is more useful for predicting outcomes in patients with FN.


LIN28 Regulates Stem Cell Metabolism and Conversion to Primed Pluripotency.

Abstract: The RNA-binding proteins LIN28A and LIN28B play critical roles in embryonic development, tumorigenesis, and pluripotency, but their exact functions are poorly understood.

Abstract: Here, we show that, like LIN28A, LIN28B can function effectively with NANOG, OCT4, and SOX2 in reprogramming to pluripotency and that reactivation of both endogenous LIN28A and LIN28B loci are required for maximal reprogramming efficiency.

Abstract: In human fibroblasts, LIN28B is activated early during reprogramming, while LIN28A is activated later during the transition to bona fide induced pluripotent stem cells (iPSCs).

Abstract: In murine cells, LIN28A and LIN28B facilitate conversion from naive to primed pluripotency.

Abstract: Proteomic and metabolomic analysis highlighted roles for LIN28 in maintaining the low mitochondrial function associated with primed pluripotency and in regulating one-carbon metabolism, nucleotide metabolism, and histone methylation.

Abstract: LIN28 binds to mRNAs of proteins important for oxidative phosphorylation and modulates protein abundance.

Abstract: Thus, LIN28A and LIN28B play cooperative roles in regulating reprogramming, naive/primed pluripotency, and stem cell metabolism.


Genetic Drift Can Compromise Mitochondrial Replacement by Nuclear Transfer in Human Oocytes.

Abstract: Replacement of mitochondria through nuclear transfer between oocytes of two different women has emerged recently as a strategy for preventing inheritance of mtDNA diseases.

Abstract: Although experiments in human oocytes have shown effective replacement, the consequences of small amounts of mtDNA carryover have not been studied sufficiently.

Abstract: Using human mitochondrial replacement stem cell lines, we show that, even though the low levels of heteroplasmy introduced into human oocytes by mitochondrial carryover during nuclear transfer often vanish, they can sometimes instead result in mtDNA genotypic drift and reversion to the original genotype.

Abstract: Comparison of cells with identical oocyte-derived nuclear DNA but different mtDNA shows that either mtDNA genotype is compatible with the nucleus and that drift is independent of mitochondrial function.

Abstract: Thus, although functional replacement of the mitochondrial genome is possible, even low levels of heteroplasmy can affect the stability of the mtDNA genotype and compromise the efficacy of mitochondrial replacement.


TAL effectors are remote controls for gene activation.

Abstract: TAL (transcription activator-like) effectors constitute a novel class of DNA-binding proteins with predictable specificity.

Abstract: They are employed by Gram-negative plant-pathogenic bacteria of the genus Xanthomonas which translocate a cocktail of different effector proteins via a type III secretion system (T3SS) into plant cells where they serve as virulence determinants.

Abstract: Inside the plant cell, TALs localize to the nucleus, bind to target promoters, and induce expression of plant genes.

Abstract: DNA-binding specificity of TALs is determined by a central domain of tandem repeats.

Abstract: Each repeat confers recognition of one base pair (bp) in the DNA.

Abstract: Rearrangement of repeat modules allows design of proteins with desired DNA-binding specificities.

Abstract: Here, we summarize how TAL specificity is encoded, first structural data and first data on site-specific TAL nucleases.


Vascular heterogeneity and specialization in development and disease

Abstract: Blood and lymphatic vessels pervade almost all body tissues and have numerous essential roles in physiology and disease.

Abstract: The inner lining of these networks is formed by a single layer of endothelial cells, which is specialized according to the needs of the tissue that it supplies.

Abstract: Whereas the general mechanisms of blood and lymphatic vessel development are being defined with increasing molecular precision, studies of the processes of endothelial specialization remain mostly descriptive.

Abstract: Recent insights from genetic animal models illuminate how endothelial cells interact with each other and with their tissue environment, providing paradigms for vessel type- and organ-specific endothelial differentiation.

Abstract: Delineating these governing principles will be crucial for understanding how tissues develop and maintain, and how their function becomes abnormal in disease.


Association of Intracerebral Hemorrhage Among Patients Taking Non–Vitamin K Antagonist vs Vitamin K Antagonist Oral Anticoagulants With In-Hospital Mortality

Abstract: Importance Although non–vitamin K antagonist oral anticoagulants (NOACs) are increasingly used to prevent thromboembolic disease, there are limited data on NOAC-related intracerebral hemorrhage (ICH).

Abstract: Objective To assess the association between preceding oral anticoagulant use (warfarin, NOACs, and no oral anticoagulants [OACs]) and in-hospital mortality among patients with ICH.

Abstract: Design, Setting, and Participants Retrospective cohort study of 141 311 patients with ICH admitted from October 2013 to December 2016 to 1662 Get With The Guidelines–Stroke hospitals.

Abstract: Exposures Anticoagulation therapy before ICH, defined as any use of OACs within 7 days prior to hospital arrival.

Abstract: Main Outcomes and Measures In-hospital mortality.

Abstract: Results Among 141 311 patients with ICH (mean [SD] age, 68.3 [15.3] years; 48.1% women), 15 036 (10.6%) were taking warfarin and 4918 (3.5%) were taking NOACs preceding ICH, and 39 585 (28.0%) and 5783 (4.1%) were taking concomitant single and dual antiplatelet agents, respectively.

Abstract: Patients with prior use of warfarin or NOACs were older and had higher prevalence of atrial fibrillation and prior stroke.

Abstract: Acute ICH stroke severity (measured by the National Institutes of Health Stroke Scale) was not significantly different across the 3 groups (median, 9 [interquartile range, 2-21] for warfarin, 8 [2-20] for NOACs, and 8 [2-19] for no OACs).

Abstract: The unadjusted in-hospital mortality rates were 32.6% for warfarin, 26.5% for NOACs, and 22.5% for no OACs.

Abstract: Compared with patients without prior use of OACs, the risk of in-hospital mortality was higher among patients with prior use of warfarin (adjusted risk difference [ARD], 9.0% [97.5% CI, 7.9% to 10.1%]; adjusted odds ratio [AOR], 1.62 [97.5% CI, 1.53 to 1.71]) and higher among patients with prior use of NOACs (ARD, 3.3% [97.5% CI, 1.7% to 4.8%]; AOR, 1.21 [97.5% CI, 1.11-1.32]).

Abstract: Compared with patients with prior use of warfarin, patients with prior use of NOACs had a lower risk of in-hospital mortality (ARD, −5.7% [97.5% CI, −7.3% to −4.2%]; AOR, 0.75 [97.5% CI, 0.69 to 0.81]).

Abstract: The difference in mortality between NOAC-treated patients and warfarin-treated patients was numerically greater among patients with prior use of dual antiplatelet agents (32.7% vs 47.1%; ARD, −15.0% [95.5% CI, −26.3% to −3.8%]; AOR, 0.50 [97.5% CI, 0.29 to 0.86]) than among those taking these agents without prior antiplatelet therapy (26.4% vs 31.7%; ARD, −5.0% [97.5% CI, −6.8% to −3.2%]; AOR, 0.77 [97.5% CI, 0.70 to 0.85]), although the interaction P value (.07) was not statistically significant.

Abstract: Conclusions and Relevance Among patients with ICH, prior use of NOACs or warfarin was associated with higher in-hospital mortality compared with no OACs.

Abstract: Prior use of NOACs, compared with prior use of warfarin, was associated with lower risk of in-hospital mortality.


Elevated red blood cell distribution width contributes to poor prognosis in patients undergoing resection for nonmetastatic rectal cancer

Abstract: Several studies have reported that elevated red blood cell distribution width (RDW) was associated with the poor prognosis of different kinds of cancers.

Abstract: The aim of this study was to investigate the prognostic role of RDW in patients undergoing resection for nonmetastatic rectal cancer.

Abstract: We retrospectively reviewed a database of 625 consecutive patients who underwent curative resection for nonmetastatic rectal cancer at our institution from January 2009 to December 2014.

Abstract: The cutoff value of RDW was calculated by receiver-operating characteristic curve.

Abstract: The results demonstrated that patients in high RDW-cv group had a lower overall survival (OS) (P = .018) and disease-free survival (P = .004).

Abstract: We also observed that patients in high RDW-sd group were associated with significantly lower OS (P = .033), whereas the disease-free survival (DFS) was not significantly different (P = .179).In multivariate analysis, we found elevated RDW-cv was associated poor DFS (hazard ratio [HR] = 1.56, P = .010) and RDW-sd can predict a worse OS (HR = 1.70, P = .009).We confirmed that elevated RDW can be an independently prognostic factor in patients undergoing resection for nonmetastatic rectal cancer.

Abstract: So more intervention or surveillance might be paid to the patients with nonmetastatic rectal cancer and elevated RDW values in the future.


Health workers cohort study: methods and study design.

Abstract: Objective: To examine different health outcomes that are associated with specific lifestyle and genetic factors.

Abstract: Materials and methods: From March 2004 to April 2006, a sample of employees from three different health and academic institutions, as well as their family members, were enrolled in the study after providing informed consent.

Abstract: At baseline and follow-up (2010-2013), participants completed a self-administered questionnaire, a physical examination, and provided blood samples.

Abstract: Results: A total of 10 729 participants aged 6 to 94 years were recruited at baseline.

Abstract: Of these, 70% were females, and 50% were from the Mexican Social Security Institute.

Abstract: Nearly 42% of the adults in the sample were overweight, while 20% were obese.

Abstract: Conclusion: Our study can offer new insights into disease mechanisms and prevention through the analysis of risk factor information in a large sample of Mexicans.


Aberrant pro-atrial natriuretic peptide/corin/natriuretic peptide receptor signaling is present in maternal vascular endothelium in preeclampsia.

Abstract: OBJECTIVE Corin is a serine protease that converts pro-atrial natriuretic peptide (pro-ANP) to atrial natriuretic peptide (ANP), a cardiac hormone that regulates salt-water balance and blood pressure.

Abstract: ANP is degraded by natriuretic peptide receptor (NPR).

Abstract: This study was to determine if aberrant pro-ANP/corin/NPR signaling is present in maternal vascular system in preeclampsia.

Abstract: STUDY DESIGN Maternal venous blood was obtained from 197 pregnant women (84 normotensive, 16 complicated with chronic hypertension (CHT), 11 mild and 86 severe preeclampsia).

Abstract: Plasma corin and pro-ANP concentrations were measured by enzyme-linked immunosorbent assay.

Abstract: Maternal subcutaneous fat tissue was obtained from 12 pregnant women with cesarean section delivery (6 normotensive and 6 preeclampsia).

Abstract: Vascular ANP and its receptors NPR-A, NPR-B, and NPR-C expression were examined by immunostaining of paraffin embedded subcutaneous fat tissue sections.

Abstract: RESULTS Corin concentrations were significantly higher in mild (2.78 ± 0.67 ng/ml, p < .05) and severe (2.53 ± 0.18 ng/ml, p < .01) preeclampsia than in normotensive (1.58 ± 0.08 ng/ml) and CHT (1.55 ± 0.20 ng/ml) groups.

Abstract: Pro-ANP concentrations were significantly higher in CHT (1.59 ± 0.53 ng/ml, p < .05) and severe preeclampsia (1.42 ± 0.24 ng/ml, p < .01) than in normotensive (0.48 ± 0.06 ng/ml) and mild preeclampsia (0.52 ± 0.09 ng/ml) groups.

Abstract: ANP and NPR-B expression was undetectable in maternal vessels from normotensive and preeclamptic pregnancies, but reduced NPR-A expression and increased NPR-C expression was found in maternal vessel endothelium in preeclampsia.

Abstract: CONCLUSIONS ANP is a vasodilator and NPR-C is a clearance receptor for ANP.

Abstract: The finding of upregulation of NPR-C expression suggests that circulating ANP clearance or degradation is increased in preeclampsia.

Abstract: These results also suggest that pro-ANP/corin/NPR signaling is dominant in the vascular system in preeclampsia.


Optimising Strategies for Plasmodium falciparum Malaria Elimination in Cambodia: Primaquine, Mass Drug Administration and Artemisinin Resistance

Abstract: BACKGROUND Malaria elimination requires a variety of approaches individually optimized for different transmission settings.

Abstract: A recent field study in an area of low seasonal transmission in South West Cambodia demonstrated dramatic reductions in malaria parasite prevalence following both mass drug administration (MDA) and high treatment coverage of symptomatic patients with artemisinin-piperaquine plus primaquine.

Abstract: This study employed multiple combined strategies and it was unclear what contribution each made to the reductions in malaria.

Abstract: METHOD AND FINDINGS A mathematical model fitted to the trial results was used to assess the effects of the various components of these interventions, design optimal elimination strategies, and explore their interactions with artemisinin resistance, which has recently been discovered in Western Cambodia.

Abstract: The modelling indicated that most of the initial reduction of P. falciparum malaria resulted from MDA with artemisinin-piperaquine.

Abstract: The subsequent continued decline and near elimination resulted mainly from high coverage with artemisinin-piperaquine treatment.

Abstract: Both these strategies were more effective with the addition of primaquine.

Abstract: MDA with artemisinin combination therapy (ACT) increased the proportion of artemisinin resistant infections, although much less than treatment of symptomatic cases with ACT, and this increase was slowed by adding primaquine.

Abstract: Artemisinin resistance reduced the effectiveness of interventions using ACT when the prevalence of resistance was very high.

Abstract: The main results were robust to assumptions about primaquine action, and immunity.

Abstract: CONCLUSIONS The key messages of these modelling results for policy makers were: high coverage with ACT treatment can produce a long-term reduction in malaria whereas the impact of MDA is generally only short-term; primaquine enhances the effect of ACT in eliminating malaria and reduces the increase in proportion of artemisinin resistant infections; parasite prevalence is a better surveillance measure for elimination programmes than numbers of symptomatic cases; combinations of interventions are most effective and sustained efforts are crucial for successful elimination.


Role of the Parasite-Derived Prostaglandin D2 in the Inhibition of Epidermal Langerhans Cell Migration during Schistosomiasis Infection

Abstract: Epidermal Langerhans cells (LCs) play a key role in immune defense mechanisms and in numerous immunological disorders.

Abstract: In this report, we show that percutaneous infection of C57BL/6 mice with the helminth parasite Schistosoma mansoni leads to the activation of LCs but, surprisingly, to their retention in the epidermis.

Abstract: Moreover, using an experimental model of LC migration induced by tumor necrosis factor (TNF)-α, we show that parasites transiently impair the departure of LCs from the epidermis and their subsequent accumulation as dendritic cells in the draining lymph nodes.

Abstract: The inhibitory effect is mediated by soluble lipophilic factors released by the parasites and not by host-derived antiinflammatory cytokines, such as interleukin-10.

Abstract: We find that prostaglandin (PG)D2, but not the other major eicosanoids produced by the parasites, specifically impedes the TNF-α–triggered migration of LCs through the adenylate cyclase–coupled PGD2 receptor (DP receptor).

Abstract: Moreover, the potent DP receptor antagonist BW A868C restores LC migration in infected mice.

Abstract: Finally, in a model of contact allergen-induced LC migration, we show that activation of the DP receptor not only inhibits LC emigration but also dramatically reduces the contact hypersensitivity responses after challenge.

Abstract: Taken together, we propose that the inhibition of LC migration could represent an additional stratagem for the schistosomes to escape the host immune system and that PGD2 may play a key role in the control of cutaneous immune responses.


Axicabtagene Ciloleucel CAR T‐Cell Therapy in Refractory Large B‐Cell Lymphoma

Abstract: Background In a phase 1 trial, axicabtagene ciloleucel (axi‐cel), an autologous anti‐CD19 chimeric antigen receptor (CAR) T‐cell therapy, showed efficacy in patients with refractory large B‐cell lymphoma after the failure of conventional therapy.

Abstract: Methods In this multicenter, phase 2 trial, we enrolled 111 patients with diffuse large B‐cell lymphoma, primary mediastinal B‐cell lymphoma, or transformed follicular lymphoma who had refractory disease despite undergoing recommended prior therapy.

Abstract: Patients received a target dose of 2×106 anti‐CD19 CAR T cells per kilogram of body weight after receiving a conditioning regimen of low‐dose cyclophosphamide and fludarabine.

Abstract: The primary end point was the rate of objective response (calculated as the combined rates of complete response and partial response).

Abstract: Secondary end points included overall survival, safety, and biomarker assessments.

Abstract: Results Among the 111 patients who were enrolled, axi‐cel was successfully manufactured for 110 (99%) and administered to 101 (91%).

Abstract: The objective response rate was 82%, and the complete response rate was 54%.With a median follow‐up of 15.4 months, 42% of the patients continued to have a response, with 40% continuing to have a complete response.

Abstract: The overall rate of survival at 18 months was 52%.

Abstract: The most common adverse events of grade 3 or higher during treatment were neutropenia (in 78% of the patients), anemia (in 43%), and thrombocytopenia (in 38%).

Abstract: Grade 3 or higher cytokine release syndrome and neurologic events occurred in 13% and 28% of the patients, respectively.

Abstract: Three of the patients died during treatment.

Abstract: Higher CAR T‐cell levels in blood were associated with response.

Abstract: Conclusions In this multicenter study, patients with refractory large B‐cell lymphoma who received CAR T‐cell therapy with axi‐cel had high levels of durable response, with a safety profile that included myelosuppression, the cytokine release syndrome, and neurologic events.

Abstract: (Funded by Kite Pharma and the Leukemia and Lymphoma Society Therapy Acceleration Program; ZUMA‐1 ClinicalTrials.gov number, NCT02348216.)


Beta adrenergic blockade decreases the immunomodulatory effects of social disruption stress

Abstract: During physiological or psychological stress, catecholamines produced by the sympathetic nervous system (SNS) regulate the immune system.

Abstract: Previous studies report that the activation of β-adrenergic receptors (βARs) mediates the actions of catecholamines and increases pro-inflammatory cytokine production in a number of different cell types.

Abstract: The impact of the SNS on the immune modulation of social defeat has not been examined.

Abstract: The following studies were designed to determine whether SNS activation during social disruption stress (SDR) influences anxiety-like behavior as well as the activation, priming, and glucocorticoid resistance of splenocytes after social stress.

Abstract: CD-1 mice were exposed to one, three, or six cycles of SDR and HPLC analysis of the plasma and spleen revealed an increase in catecholamines.

Abstract: After six cycles of SDR the open field test was used to measure behaviors characteristic of anxiety and indicated that the social defeat induced increase in anxiety-like behavior was blocked by pre-treatment with the β-adrenergic antagonist propranolol.

Abstract: Pre-treatment with the β-adrenergic antagonist propranolol did not significantly alter corticosterone levels indicating no difference in activation of the hypothalamic-pituitary-adrenal axis.

Abstract: In addition to anxiety-like behavior the SDR induced splenomegaly and increase in plasma IL-6, TNFα, and MCP-1 were each reversed by pre-treatment with propranolol.

Abstract: Furthermore, flow cytometric analysis of cells from propranolol pretreated mice reduced the SDR-induced increase in the percentage of CD11b(+) splenic macrophages and significantly decreased the expression of TLR2, TLR4, and CD86 on the surface of these cells.

Abstract: In addition, supernatants from 18h LPS-stimulated ex vivo cultures of splenocytes from propranolol-treated SDR mice contained less IL-6.

Abstract: Likewise propranolol pre-treatment abrogated the glucocorticoid insensitivity of CD11b(+) cells ex vivo when compared to splenocytes from SDR vehicle-treated mice.

Abstract: Together, this study demonstrates that the immune activation and priming effects of SDR result, in part, as a consequence of SNS activation.


Stereotactic radiosurgery plus whole-brain radiation therapy vs stereotactic radiosurgery alone for treatment of brain metastases: a randomized controlled trial.

Abstract: CONTEXT In patients with brain metastases, it is unclear whether adding up-front whole-brain radiation therapy (WBRT) to stereotactic radiosurgery (SRS) has beneficial effects on mortality or neurologic function compared with SRS alone.

Abstract: OBJECTIVE To determine if WBRT combined with SRS results in improvements in survival, brain tumor control, functional preservation rate, and frequency of neurologic death.

Abstract: DESIGN, SETTING, AND PATIENTS Randomized controlled trial of 132 patients with 1 to 4 brain metastases, each less than 3 cm in diameter, enrolled at 11 hospitals in Japan between October 1999 and December 2003.

Abstract: INTERVENTIONS Patients were randomly assigned to receive WBRT plus SRS (65 patients) or SRS alone (67 patients).

Abstract: MAIN OUTCOME MEASURES The primary end point was overall survival; secondary end points were brain tumor recurrence, salvage brain treatment, functional preservation, toxic effects of radiation, and cause of death.

Abstract: RESULTS The median survival time and the 1-year actuarial survival rate were 7.5 months and 38.5% (95% confidence interval, 26.7%-50.3%) in the WBRT + SRS group and 8.0 months and 28.4% (95% confidence interval, 17.6%-39.2%) for SRS alone (P = .42).

Abstract: The 12-month brain tumor recurrence rate was 46.8% in the WBRT + SRS group and 76.4% for SRS alone group (P<.001).

Abstract: Salvage brain treatment was less frequently required in the WBRT + SRS group (n = 10) than with SRS alone (n = 29) (P<.001).

Abstract: Death was attributed to neurologic causes in 22.8% of patients in the WBRT + SRS group and in 19.3% of those treated with SRS alone (P = .64).

Abstract: There were no significant differences in systemic and neurologic functional preservation and toxic effects of radiation.

Abstract: CONCLUSIONS Compared with SRS alone, the use of WBRT plus SRS did not improve survival for patients with 1 to 4 brain metastases, but intracranial relapse occurred considerably more frequently in those who did not receive WBRT.

Abstract: Consequently, salvage treatment is frequently required when up-front WBRT is not used.

Abstract: TRIAL REGISTRATION umin.ac.jp/ctr Identifier: C000000412.


RANK signals from CD4+3− inducer cells regulate development of Aire-expressing epithelial cells in the thymic medulla

Abstract: Aire-expressing medullary thymic epithelial cells (mTECs) play a key role in preventing autoimmunity by expressing tissue-restricted antigens to help purge the emerging T cell receptor repertoire of self-reactive specificities.

Abstract: Here we demonstrate a novel role for a CD4+3− inducer cell population, previously linked to development of organized secondary lymphoid structures and maintenance of T cell memory in the functional regulation of Aire-mediated promiscuous gene expression in the thymus.

Abstract: CD4+3− cells are closely associated with mTECs in adult thymus, and in fetal thymus their appearance is temporally linked with the appearance of Aire+ mTECs.

Abstract: We show that RANKL signals from this cell promote the maturation of RANK-expressing CD80−Aire− mTEC progenitors into CD80+Aire+ mTECs, and that transplantation of RANK-deficient thymic stroma into immunodeficient hosts induces autoimmunity.

Abstract: Collectively, our data reveal cellular and molecular mechanisms leading to the generation of Aire+ mTECs and highlight a previously unrecognized role for CD4+3−RANKL+ inducer cells in intrathymic self-tolerance.


Proteomic approach to characterize biochemistry of meat quality defects.

Abstract: Proteomics can be used to characterize quality defects including pale, soft, and exudative (PSE) meat (pork and poultry), woody broiler breast meat, reddish catfish fillets, meat toughness, and beef myoglobin oxidation.

Abstract: PSE broiler meat was characterized by 15 proteins that differed in abundance in comparison to normal broiler breast meat, and eight proteins were differentially expressed in woody breast meat in comparison to normal breast meat.

Abstract: Hemoglobin was the only protein that was differentially expressed between red and normal catfish fillets.

Abstract: However, inducing low oxygen and/or heat stress conditions to catfish fillets did not lead to the production of red fillets.

Abstract: Proteomic data provided information pertaining to the protein differences that exist in meat quality defects.

Abstract: However, these data need to be evaluated in conjunction with information pertaining to genetics, nutrition, environment of the live animal, muscle to meat conversion, meat quality analyses and sensory attributes to understand causality, protein biomarkers, and ultimately how to prevent quality defects.


Phosphorylation of Janus kinase 1 (JAK1) by AMP-activated protein kinase (AMPK) links energy sensing to anti-inflammatory signaling

Abstract: Adenosine 5′-monophosphate–activated protein kinase (AMPK) is a pivotal regulator of metabolism at cellular and organismal levels.

Abstract: AMPK also suppresses inflammation.

Abstract: We found that pharmacological activation of AMPK rapidly inhibited the Janus kinase (JAK)–signal transducer and activator of transcription (STAT) pathway in various cells.

Abstract: In vitro kinase assays revealed that AMPK directly phosphorylated two residues (Ser515 and Ser518) within the Src homology 2 domain of JAK1.

Abstract: Activation of AMPK enhanced the interaction between JAK1 and 14-3-3 proteins in cultured vascular endothelial cells and fibroblasts, an effect that required the presence of Ser515 and Ser518 and was abolished in cells lacking AMPK catalytic subunits.

Abstract: Mutation of Ser515 and Ser518 abolished AMPK-mediated inhibition of JAK-STAT signaling stimulated by either the sIL-6Rα/IL-6 complex or the expression of a constitutively active V658F-mutant JAK1 in human fibrosarcoma cells.

Abstract: Clinically used AMPK activators metformin and salicylate enhanced the inhibitory phosphorylation of endogenous JAK1 and inhibited STAT3 phosphorylation in primary vascular endothelial cells.

Abstract: Therefore, our findings reveal a mechanism by which JAK1 function and inflammatory signaling may be suppressed in response to metabolic stress and provide a mechanistic rationale for the investigation of AMPK activators in a range of diseases associated with enhanced activation of the JAK-STAT pathway.


IAP antagonists induce anti-tumor immunity in multiple myeloma

Abstract: The cellular inhibitors of apoptosis (cIAP) 1 and 2 are amplified in about 3% of cancers and have been identified in multiple malignancies as being potential therapeutic targets as a result of their role in the evasion of apoptosis.

Abstract: Consequently, small-molecule IAP antagonists, such as LCL161, have entered clinical trials for their ability to induce tumor necrosis factor (TNF)-mediated apoptosis of cancer cells.

Abstract: However, cIAP1 and cIAP2 are recurrently homozygously deleted in multiple myeloma (MM), resulting in constitutive activation of the noncanonical nuclear factor (NF)-κB pathway.

Abstract: To our surprise, we observed robust in vivo anti-myeloma activity of LCL161 in a transgenic myeloma mouse model and in patients with relapsed-refractory MM, where the addition of cyclophosphamide resulted in a median progression-free-survival of 10 months.

Abstract: This effect was not a result of direct induction of tumor cell death, but rather of upregulation of tumor-cell-autonomous type I interferon (IFN) signaling and a strong inflammatory response that resulted in the activation of macrophages and dendritic cells, leading to phagocytosis of tumor cells.

Abstract: Treatment of a MM mouse model with LCL161 established long-term anti-tumor protection and induced regression in a fraction of the mice.

Abstract: Notably, combination of LCL161 with the immune-checkpoint inhibitor anti-PD1 was curative in all of the treated mice.


Psychobiological Protective Factors Modifying the Association Between Age and Sexual Health in Men: Findings From the Men’s Health 40+ Study

Abstract: Sexual health severely decreases with age.

Abstract: For males older than 40 years, erectile dysfunction (ED) is the most common sexual disorder.

Abstract: Although physical and psychological risk factors for ED have been identified, protective factors are yet to be determined.

Abstract: To date, no study has examined endocrine and psychosocial factors in parallel with regard to their modifying effect on the age-related increase in ED.

Abstract: Two hundred and seventy-one self-reporting healthy men aged between 40 and 75 years provided both psychometric data on sexual function and a set of potential psychosocial protective factors, and saliva samples for the analysis of steroid hormones and proinflammatory cytokines.

Abstract: Around 35% of the participants reported at least a mild form of ED.

Abstract: Direct associations with ED were identified for perceived general health, emotional support, relationship quality, intimacy motivation but not for steroid hormones or proinflammatory markers.

Abstract: Moderation analyses for the association between age and ED revealed positive effects for testosterone (T), dehydroepiandrosterone (DHEA), perceived general health, emotional support, intimacy motivation, and a negative effect for interleukin-6 (all p < .05; f2 > .17).

Abstract: Group differences between older men with and without ED emerged for T, DHEA, and psychometric measures such as perceived general health, emotional support, satisfaction with life, and intimacy motivation (all p < .05; d > .3).

Abstract: Both psychosocial and endocrine parameters moderated the association between age and sexual health.

Abstract: Perceived general health, emotional support, intimacy motivation, and relationship quality emerged as psychosocial protective factors against ED.

Abstract: Higher T and DHEA and lower interleukin-6 levels also buffered against an age-related increase in ED.


Patient-derived induced pluripotent stem cells in cancer research and precision oncology

Abstract: Together with recent advances in the processing and culture of human tissue, bioengineering, xenotransplantation and genome editing, Induced pluripotent stem cells (iPSCs) present a range of new opportunities for the study of human cancer.

Abstract: Here we discuss the main advantages and limitations of iPSC modeling, and how the method intersects with other patient-derived models of cancer, such as organoids, organs-on-chips and patient-derived xenografts (PDXs).

Abstract: We highlight the opportunities that iPSC models can provide beyond those offered by existing systems and animal models and present current challenges and crucial areas for future improvements toward wider adoption of this technology.


Structural basis for sequence-specific recognition of DNA by TAL effectors.

Abstract: TAL (transcription activator-like) effectors, secreted by phytopathogenic bacteria, recognize host DNA sequences through a central domain of tandem repeats.

Abstract: Each repeat comprises 33 to 35 conserved amino acids and targets a specific base pair by using two hypervariable residues [known as repeat variable diresidues (RVDs)] at positions 12 and 13.

Abstract: Here, we report the crystal structures of an 11.5-repeat TAL effector in both DNA-free and DNA-bound states.

Abstract: Each TAL repeat comprises two helices connected by a short RVD-containing loop.

Abstract: The 11.5 repeats form a right-handed, superhelical structure that tracks along the sense strand of DNA duplex, with RVDs contacting the major groove.

Abstract: The 12th residue stabilizes the RVD loop, whereas the 13th residue makes a base-specific contact.

Abstract: Understanding DNA recognition by TAL effectors may facilitate rational design of DNA-binding proteins with biotechnological applications.


Caveolin-1 deletion exacerbates cardiac interstitial fibrosis by promoting M2 macrophage activation in mice after myocardial infarction.

Abstract: Adverse remodeling following myocardial infarction (MI) leading to heart failure is driven by an imbalanced resolution of inflammation.

Abstract: The macrophage cell is an important control of post-MI inflammation, as macrophage subtypes secrete mediators to either promote inflammation and extend injury (M1 phenotype) or suppress inflammation and promote scar formation (M2 phenotype).

Abstract: We have previously shown that the absence of caveolin-1 (Cav1), a membrane scaffolding protein, is associated with adverse cardiac remodeling in mice, but the mechanisms responsible remain to be elucidated.

Abstract: We explore here the role of Cav1 in the activation of macrophages using wild type C57BL6/J (WT) and Cav1(tm1Mls/J) (Cav1(-/-)) mice.

Abstract: By echocardiography, cardiac function was comparable between WT and Cav1(-/-) mice at 3days post-MI.

Abstract: In the absence of Cav1, there were a surprisingly higher percentage of M2 macrophages (arginase-1 positive) detected in the infarcted zone.

Abstract: Conversely, restoring Cav1 function after MI in WT mice by adding back the Cav1 scaffolding domain reduced the M2 activation profile.

Abstract: Further, adoptive transfer of Cav1 null macrophages into WT mice on d3 post-MI exacerbated adverse cardiac remodeling at d14 post-MI.

Abstract: In vitro studies revealed that Cav1 null macrophages had a more pronounced M2 profile activation in response to IL-4 stimulation.

Abstract: In conclusion, Cav1 deletion promotes an array of maladaptive repair processes after MI, including increased TGF-β signaling, increased M2 macrophage infiltration and dysregulation of the M1/M2 balance.

Abstract: Our data also suggest that cardiac remodeling can be improved by therapeutic intervention regulating Cav1 function during the inflammatory response phase.


Lipid hydroperoxide measurement by oxidation of Fe2+ in the presence of xylenol orange. Comparison with the TBA assay and an iodometric method

Abstract: Study of the role of hydroperoxides and lipid peroxidation in disease requires simple and sensitive methods for direct hydroperoxide measurement.

Abstract: We report on a technique for measuring hydroperoxide which relies upon the rapid hydroperoxide-mediated oxidation of Fe2+ under acidic conditions.

Abstract: Fe3+ forms a chromophore with xylenol orange which absorbs strongly at 560 nm, yielding an apparent E560 (for H2O2, butyl hydroperoxide and cumene hydroperoxide) of 4.3×104 M−1 cm−1.

Abstract: The assay was validated in a study of liposomal lipid peroxidation and shown to give results comparable with those obtained by an iodometric method or by measuring conjugated dienes.

Abstract: The assay involving thiobarbituric acid, by comparison, underestimates lipid peroxidation and does not measure hydroperoxideper se.


Roles of exosomes in cardioprotection.

Abstract: Exosomes are extracellular vesicles of endosomal origin which have emerged as key mediators of intercellular communication.

Abstract: All major cardiac cell types-including cardiomyocytes, endothelial cells, and fibroblasts-release exosomes that modulate cellular functions.

Abstract: Exosomes released from human cardiac progenitor cells (CPCs) are cardioprotective and improve cardiac function after myocardial infarction to an extent comparable with that achieved by their parent cells.

Abstract: Cardiac progenitor cell-derived exosomes are enriched in cardioprotective microRNAs, particularly miR-146a-3p.

Abstract: Circulating exosomes mediate remote ischaemic preconditioning.

Abstract: Moreover, they currently are being investigated as diagnostic markers.

Abstract: The discovery that cell-derived extracellular signalling organelles mediate the paracrine effects of stem cells suggests that cell-free strategies could supplant cell transplantation.

Abstract: This review discusses emerging roles of exosomes in cardiovascular physiology, with a focus on cardioprotective activities of CPC-derived exosomes.


The rate of change in declining steroid hormones: a new parameter of healthy aging in men?

Abstract: Research on healthy aging in men has increasingly focused on age-related hormonal changes.

Abstract: Testosterone (T) decline is primarily investigated, while age-related changes in other sex steroids (dehydroepiandrosterone [DHEA], estradiol [E2], progesterone [P]) are mostly neglected.

Abstract: An integrated hormone parameter reflecting aging processes in men has yet to be identified.

Abstract: 271 self-reporting healthy men between 40 and 75 provided both psychometric data and saliva samples for hormone analysis.

Abstract: Correlation analysis between age and sex steroids revealed negative associations for the four sex steroids (T, DHEA, E2, and P).

Abstract: Principal component analysis including ten salivary analytes identified a principal component mainly unifying the variance of the four sex steroid hormones.

Abstract: Subsequent principal component analysis including the four sex steroids extracted the principal component of declining steroid hormones (DSH).

Abstract: Moderation analysis of the association between age and DSH revealed significant moderation effects for psychosocial factors such as depression, chronic stress and perceived general health.

Abstract: In conclusion, these results provide further evidence that sex steroids decline in aging men and that the integrated hormone parameter DSH and its rate of change can be used as biomarkers for healthy aging in men.

Abstract: Furthermore, the negative association of age and DSH is moderated by psychosocial factors.


Development of a novel helper-dependent adenovirus-Epstein-Barr virus hybrid system for the stable transformation of mammalian cells.

Abstract: Epstein-Barr virus (EBV) episomes are stably maintained in permissive proliferating cell lines due to EBV nuclear antigen 1 (EBNA-1) protein-mediated replication and segregation.

Abstract: Previous studies showed the ability of EBV episomes to confer long-term transgene expression and correct genetic defects in deficient cells.

Abstract: To achieve quantitative delivery of EBV episomes in vitro and in vivo, we developed a binary helper-dependent adenovirus (HDA)-EBV hybrid system that consists of one HDA vector for the expression of Cre recombinase and a second HDA vector that contains all of the sequences for the EBV episome flanked by loxP sites.

Abstract: Upon coinfection of cells, Cre expressed from the first vector recombined loxP sites on the second vector.

Abstract: The resulting circular EBV episomes expressed a transgene and contained the EBV-derived family of repeats, an EBNA-1 expression cassette, and 19 kb of human DNA that functions as a replication origin in mammalian cells.

Abstract: This HDA-EBV hybrid system transformed 40% of cultured cells.

Abstract: Transgene expression in proliferating cells was observed for over 20 weeks under conditions that selected for the expression of the transgene.

Abstract: In the absence of selection, EBV episomes were lost at a rate of 8 to 10% per cell division.

Abstract: Successful delivery of EBV episomes in vivo was demonstrated in the liver of transgenic mice expressing Cre from the albumin promoter.

Abstract: This novel gene transfer system has the potential to confer long-term episomal transgene expression and therefore to correct genetic defects with reduced vector-related toxicity and without insertional mutagenesis.


Spliceosome assembly pathways for different types of alternative splicing converge during commitment to splice site pairing in the A complex.

Abstract: Differential splice site pairing establishes alternative splicing patterns resulting in the generation of multiple mRNA isoforms.

Abstract: This process is carried out by the spliceosome, which is activated by a series of sequential structural rearrangements of its five core snRNPs.

Abstract: To determine when splice sites become functionally paired, we carried out a series of kinetic trap experiments using pre-mRNAs that undergo alternative 5' splice site selection or alternative exon inclusion.

Abstract: We show that commitment to splice site pairing in both cases occurs in the A complex, which is characterized by the ATP-dependent association of the U2 snRNP with the branch point.

Abstract: Interestingly, the timing of splice site pairing is independent of the intron or exon definition modes of splice site recognition.

Abstract: Using the ATP analog ATPgammaS, we showed that ATP hydrolysis is required for splice site pairing independent from U2 snRNP binding to the pre-mRNA.

Abstract: These results identify the A complex as the spliceosomal assembly step dedicated to splice site pairing and suggest that ATP hydrolysis locks splice sites into a splicing pattern after stable U2 snRNP association to the branch point.


Transcripts of the MHM region on the chicken Z chromosome accumulate as non-coding RNA in the nucleus of female cells adjacent to the DMRT1 locus

Abstract: The male hypermethylated (MHM) region, located near the middle of the short arm of the Z chromosome of chickens, consists of approximately 210 tandem repeats of a BamHI 2.2-kb sequence unit.

Abstract: Cytosines of the CpG dinucleotides of this region are extensively methylated on the two Z chromosomes in the male but much less methylated on the single Z chromosome in the female.

Abstract: The state of methylation of the MHM region is established after fertilization by about the 1-day embryonic stage.

Abstract: The MHM region is transcribed only in the female from the particular strand into heterogeneous, high molecular-mass, non-coding RNA, which is accumulated at the site of transcription, adjacent to the DMRT1 locus, in the nucleus.

Abstract: The transcriptional silence of the MHM region in the male is most likely caused by the CpG methylation, since treatment of the male embryonic fibroblasts with 5-azacytidine results in hypo-methylation and active transcription of this region.

Abstract: In ZZW triploid chickens, MHM regions are hypomethylated and transcribed on the two Z chromosomes, whereas MHM regions are hypermethylated and transcriptionally inactive on the three Z chromosomes in ZZZ triploid chickens, suggesting a possible role of the W chromosome on the state of the MHM region.


Macropinocytosis of protein is an amino acid supply route in Ras-transformed cells

Abstract: Macropinocytosis is a highly conserved endocytic process by which extracellular fluid and its contents are internalized into cells through large, heterogeneous vesicles known as macropinosomes.

Abstract: Oncogenic Ras proteins have been shown to stimulate macropinocytosis but the functional contribution of this uptake mechanism to the transformed phenotype remains unknown.

Abstract: Here we show that Ras-transformed cells use macropinocytosis to transport extracellular protein into the cell.

Abstract: The internalized protein undergoes proteolytic degradation, yielding amino acids including glutamine that can enter central carbon metabolism.

Abstract: Accordingly, the dependence of Ras-transformed cells on free extracellular glutamine for growth can be suppressed by the macropinocytic uptake of protein.

Abstract: Consistent with macropinocytosis representing an important route of nutrient uptake in tumours, its pharmacological inhibition compromises the growth of Ras-transformed pancreatic tumour xenografts.

Abstract: These results identify macropinocytosis as a mechanism by which cancer cells support their unique metabolic needs and point to the possible exploitation of this process in the design of anticancer therapies.


Linking Splicing to Pol II Transcription Stabilizes Pre-mRNAs and Influences Splicing Patterns

Abstract: RNA processing is carried out in close proximity to the site of transcription, suggesting a regulatory link between transcription and pre-mRNA splicing.

Abstract: Using an in vitro transcription/splicing assay, we demonstrate that an association of RNA polymerase II (Pol II) transcription and pre-mRNA splicing is required for efficient gene expression.

Abstract: Pol II-synthesized RNAs containing functional splice sites are protected from nuclear degradation, presumably because the local concentration of the splicing machinery is sufficiently high to ensure its association over interactions with nucleases.

Abstract: Furthermore, the process of transcription influences alternative splicing of newly synthesized pre-mRNAs.

Abstract: Because other RNA polymerases do not provide similar protection from nucleases, and their RNA products display altered splicing patterns, the link between transcription and RNA processing is RNA Pol II-specific.

Abstract: We propose that the connection between transcription by Pol II and pre-mRNA splicing guarantees an extended half-life and proper processing of nascent pre-mRNAs.


The emerging role of skeletal muscle extracellular matrix remodelling in obesity and exercise.

Abstract: Skeletal muscle extracellular matrix remodelling has been proposed as a new feature associated with obesity and metabolic dysfunction.

Abstract: Exercise training improves muscle function in obesity, which may be mediated by regulatory effects on the muscle extracellular matrix.

Abstract: This review examined available literature on skeletal muscle extracellular matrix remodelling during obesity and the effects of exercise.

Abstract: A non-systematic literature review was performed on PubMed of publications from 1970 to 2015.

Abstract: A total of 37 studies from humans and animals were retained.

Abstract: Studies reported overall increases in gene and protein expression of different types of collagen, growth factors and enzymatic regulators of the skeletal muscle extracellular matrix in obesity.

Abstract: Only two studies investigated the effects of exercise on skeletal muscle extracellular matrix during obesity, with both suggesting a regulatory effect of exercise.

Abstract: The effects of exercise on muscle extracellular matrix seem to be influenced by the duration and type of exercise training with variable effects from a single session compared with a longer duration of exercise.

Abstract: More studies are needed to elucidate the mechanisms behind skeletal muscle extracellular matrix remodelling during obesity and the effects of exercise.


Effect of specific exercise strategy on need for surgery in patients with subacromial impingement syndrome: randomised controlled study

Abstract: OBJECTIVE To evaluate if a specific exercise strategy, targeting the rotator cuff and scapula stabilisers, improves shoulder function and pain more than unspecific exercises in patients with subacromial impingement syndrome, thereby decreasing the need for arthroscopic subacromial decompression.

Abstract: DESIGN Randomised, participant and single assessor blinded, controlled study.

Abstract: SETTING Department of orthopaedics in a Swedish university hospital.

Abstract: PARTICIPANTS 102 patients with long standing (over six months) persistent subacromial impingement syndrome in whom earlier conservative treatment had failed, recruited through orthopaedic specialists.

Abstract: INTERVENTIONS The specific exercise strategy consisted of strengthening eccentric exercises for the rotator cuff and concentric/eccentric exercises for the scapula stabilisers in combination with manual mobilisation.

Abstract: The control exercise programme consisted of unspecific movement exercises for the neck and shoulder.

Abstract: Patients in both groups received five to six individual guided treatment sessions during 12 weeks.

Abstract: In between these supervised sessions the participants performed home exercises once or twice a day for 12 weeks.

Abstract: MAIN OUTCOME MEASURES The primary outcome was the Constant-Murley shoulder assessment score evaluating shoulder function and pain.

Abstract: Secondary outcomes were patients' global impression of change because of treatment and decision regarding surgery.

Abstract: RESULTS Most (97, 95%) participants completed the 12 week study.

Abstract: There was a significantly greater improvement in the Constant-Murley score in the specific exercise group than in the control exercise group (24 points (95% confidence interval 19 to 28.0) v 9 points (5 to 13); mean difference between group: 15 points (8.5 to 20.6)).

Abstract: Significantly more patients in the specific exercise group reported successful outcome (defined as large improvement or recovered) in the patients' global assessment of change because of treatment: 69% (35/51) v 24% (11/46); odds ratio 7.6, 3.1 to 18.9; P<0.001.

Abstract: A significantly lower proportion of patients in the specific exercise group subsequently chose to undergo surgery: 20% (10/51) v 63% (29/46); odds ratio 7.7, 3.1 to 19.4; P<0.001).

Abstract: CONCLUSION A specific exercise strategy, focusing on strengthening eccentric exercises for the rotator cuff and concentric/eccentric exercises for the scapula stabilisers, is effective in reducing pain and improving shoulder function in patients with persistent subacromial impingement syndrome.

Abstract: By extension, this exercise strategy reduces the need for arthroscopic subacromial decompression within the three month timeframe used in the study.

Abstract: TRIAL REGISTRATION Clinical trials NCT01037673.


Nucleotide sequence of an avian sarcoma virus oncogene (src) and proposed amino acid sequence for gene product

Abstract: The transforming gene (src) of avian sarcoma virus (ASV) and adjacent regions of the viral genome have been isolated by molecular cloning of viral DNA.

Abstract: Their nucleotide sequence encompasses the whole of src and the portion of the gene env that encodes gp 37, one of two glycoproteins found in the viral envelope.

Abstract: Src encodes a single, hydrophobic protein with structural features that conform to previous descriptions of the gene product (pp60src).

Abstract: It appears that a single viral protein is responsible for both the initiation and maintenance of neoplastic transformation by avian sarcoma virus.

Abstract: Neither src nor its product bear any obvious structural relationship to several other viral oncogenes and their encoded proteins.

Abstract: Src is flanked by a repeated nucleotide sequence that may facilitate frequent deletion of the gene from the viral genome.


Issues in design and interpretation of MDR-TB clinical trials: report of the first Global MDR-TB Clinical Trials Landscape Meeting

Abstract: Recognizing that the current MDR-TB regimen is suboptimal and based on low-quality evidence, the Global MDR-TB Clinical Trials Landscape Meeting was held in December, 2014 to strategize about coordination of research and development of new treatment regimens for this disease that affects millions of people worldwide every year.

Abstract: Sixty international experts on multidrug-resistant tuberculosis (MDR-TB) met in Washington D.C. and Cape Town, South Africa to consider key MDR-TB trial-related issues, including: standardization of definitions; clinical trial capacity building and; regimens optimized to foster compliance, avoid the emergence of resistance and have clinical relevance for special populations, including children and those co-infected with HIV.

Abstract: Underpinning all of this is the generation of a sufficient evidence base to facilitate regulatory approval and improved normative guidance.

Abstract: Participants discussed treatment combinations currently being studied in Phase 2B and Phase 3 trials as well as other promising new regimens and combinations that may be evaluated in the near future.

Abstract: These include regimens designed specifically to enable shorter duration and all-oral treatment as a means of maximizing treatment completion.

Abstract: It is hoped that clear definition of these challenges will facilitate the process of identifying solutions that accelerate progress towards effective, non-toxic treatments that can be programmatically implemented.


Para-aortic splanchnopleura from early mouse embryos contains B1a cell progenitors

Abstract: DEFINITIVE erythropoiesis in birds originates from stem cells that emerge in the splanchnopleural mesoderm near the embryonic aorta1–4.

Abstract: The yolk sac is still generally held to be the unique provider of haematopoietic stem cells during mammalian ontogeny5, although there may be an alternative intraembryonic source of stem cells in the mouse fetus6,7.

Abstract: Here we search for a possible non-yolk-sac source of stem cells by grafting intraembryonic splanchnopleura from 10- to 18-somite mouse embryos into adult immunodeficient SCID mice.

Abstract: We find significant amounts of donor-derived serum IgM, normal numbers of IgM-secreting plasma cells, and the Bla (IgMa brightB220dullCD5+) cell subset to be fully reconstituted by donor progenitors 3 to 6 months after engraftment.

Abstract: The haematogenic capacity revealed in our experiments is present in a previously unrecognized site, the earliest described in the embryo, 12 hours before fetal liver colonization.


Establishment in culture of pluripotential cells from mouse embryos

Abstract: Pluripotential cells are present in a mouse embryo until at least an early post-implantation stage, as shown by their ability to take part hi the formation of chimaeric animals1 and to form teratocarcinomas2.

Abstract: Until now it has not been possible to establish progressively growing cultures of these cells in vitro, and cell lines have only been obtained after teratocarcinoma formation in vivo.

Abstract: We report here the establishment in tissue culture of pluripotent cell lines which have been isolated directly from in vitro cultures of mouse blastocysts.

Abstract: These cells are able to differentiate either in vitro or after innoculation into a mouse as a tumour in vivo.

Abstract: They have a normal karyotype.


MHC class II interaction with CD4 mediated by a region analogous to the MHC class I binding site for CD8

Abstract: INTERACTIONS between major histocompatibility complex (MHC) molecules and the CD4 or CDS coreceptors have a major role in intrathymic T-cell selection1.

Abstract: On mature T cells, each of these two glycoproteins is associated with a class-specific bias in MHC molecule recognition by the T-cell receptor.

Abstract: CD4+ T cells respond to antigen in association with MHC class II molecules and CD8+ T cells respond to antigen in association with MHC class I molecules.

Abstract: Physical interaction between the CD4/MHC class II molecules and CD8/MHC class I molecules has been demonstrated by cell adhesion assay2–5, and a binding site for CDS on class I has been identified6,7.

Abstract: Here we demonstrate that a region of the MHC class IIβ-chain β2 domain, structurally analogous to the CDS-binding loop in the MHC class I α3 domain, is critical for function with both mouse and human CD4.


Instability and decay of the primary structure of DNA

Abstract: Although DNA is the carrier of genetic information, it has limited chemical stability.

Abstract: Hydrolysis, oxidation and nonenzymatic methylation of DNA occur at significant ratesin vivo, and are counteracted by specific DNA repair processes.

Abstract: The spontaneous decay of DNA is likely to be a major factor in mutagenesis, carcinogenesis and ageing, and also sets limits for the recovery of DNA fragments from fossils.


Single-cell proteomic analysis of S. cerevisiae reveals the architecture of biological noise

Abstract: A major goal of biology is to provide a quantitative description of cellular behaviour.

Abstract: This task, however, has been hampered by the difficulty in measuring protein abundances and their variation.

Abstract: Here we present a strategy that pairs high-throughput flow cytometry and a library of GFP-tagged yeast strains to monitor rapidly and precisely protein levels at single-cell resolution.

Abstract: Bulk protein abundance measurements of >2,500 proteins in rich and minimal media provide a detailed view of the cellular response to these conditions, and capture many changes not observed by DNA microarray analyses.

Abstract: Our single-cell data argue that noise in protein expression is dominated by the stochastic production/destruction of messenger RNAs.

Abstract: Beyond this global trend, there are dramatic protein-specific differences in noise that are strongly correlated with a protein's mode of transcription and its function.

Abstract: For example, proteins that respond to environmental changes are noisy whereas those involved in protein synthesis are quiet.

Abstract: Thus, these studies reveal a remarkable structure to biological noise and suggest that protein noise levels have been selected to reflect the costs and potential benefits of this variation.


Direct conversion of fibroblasts to functional neurons by defined factors

Abstract: Cellular differentiation and lineage commitment are considered to be robust and irreversible processes during development.

Abstract: Recent work has shown that mouse and human fibroblasts can be reprogrammed to a pluripotent state with a combination of four transcription factors.

Abstract: This raised the question of whether transcription factors could directly induce other defined somatic cell fates, and not only an undifferentiated state.

Abstract: We hypothesized that combinatorial expression of neural-lineage-specific transcription factors could directly convert fibroblasts into neurons.

Abstract: Starting from a pool of nineteen candidate genes, we identified a combination of only three factors, Ascl1, Brn2 (also called Pou3f2) and Myt1l, that suffice to rapidly and efficiently convert mouse embryonic and postnatal fibroblasts into functional neurons in vitro.

Abstract: These induced neuronal (iN) cells express multiple neuron-specific proteins, generate action potentials and form functional synapses.

Abstract: Generation of iN cells from non-neural lineages could have important implications for studies of neural development, neurological disease modelling and regenerative medicine.


Inflammasome-mediated dysbiosis regulates progression of NAFLD and obesity

Abstract: Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of metabolic syndrome and the leading cause of chronic liver disease in the Western world.

Abstract: Twenty per cent of NAFLD individuals develop chronic hepatic inflammation (non-alcoholic steatohepatitis, NASH) associated with cirrhosis, portal hypertension and hepatocellular carcinoma, yet the causes of progression from NAFLD to NASH remain obscure.

Abstract: Here, we show that the NLRP6 and NLRP3 inflammasomes and the effector protein IL-18 negatively regulate NAFLD/NASH progression, as well as multiple aspects of metabolic syndrome via modulation of the gut microbiota.

Abstract: Different mouse models reveal that inflammasome-deficiency-associated changes in the configuration of the gut microbiota are associated with exacerbated hepatic steatosis and inflammation through influx of TLR4 and TLR9 agonists into the portal circulation, leading to enhanced hepatic tumour-necrosis factor (TNF)-α expression that drives NASH progression.

Abstract: Furthermore, co-housing of inflammasome-deficient mice with wild-type mice results in exacerbation of hepatic steatosis and obesity.

Abstract: Thus, altered interactions between the gut microbiota and the host, produced by defective NLRP3 and NLRP6 inflammasome sensing, may govern the rate of progression of multiple metabolic syndrome-associated abnormalities, highlighting the central role of the microbiota in the pathogenesis of heretofore seemingly unrelated systemic auto-inflammatory and metabolic disorders.


Chromatin remodelling at promoters suppresses antisense transcription

Abstract: Chromatin allows the eukaryotic cell to package its DNA efficiently.

Abstract: To understand how chromatin structure is controlled across the Saccharomyces cerevisiae genome, we have investigated the role of the ATP-dependent chromatin remodelling complex Isw2 in positioning nucleosomes.

Abstract: We find that Isw2 functions adjacent to promoter regions where it repositions nucleosomes at the interface between genic and intergenic sequences.

Abstract: Nucleosome repositioning by Isw2 is directional and results in increased nucleosome occupancy of the intergenic region.

Abstract: Loss of Isw2 activity leads to inappropriate transcription, resulting in the generation of both coding and noncoding transcripts.

Abstract: Here we show that Isw2 repositions nucleosomes to enforce directionality on transcription by preventing transcription initiation from cryptic sites.

Abstract: Our analyses reveal how chromatin is organized on a global scale and advance our understanding of how transcription is regulated.


The Human Mitochondrial DEAD-Box Protein DDX28 Resides in RNA Granules and Functions in Mitoribosome Assembly.

Abstract: Human mitochondrial ribosomes are specialized in the synthesis of 13 proteins, which are fundamental components of the oxidative phosphorylation system.

Abstract: The pathway of mitoribosome biogenesis, the compartmentalization of the process, and factors involved remain largely unknown.

Abstract: Here, we have identified the DEAD-box protein DDX28 as an RNA granule component essential for the biogenesis of the mitoribosome large subunit (mt-LSU).

Abstract: DDX28 interacts with the 16S rRNA and the mt-LSU.

Abstract: RNAi-mediated DDX28 silencing in HEK293T cells does not affect mitochondrial mRNA stability or 16S rRNA processing or modification.

Abstract: However, it leads to reduced levels of 16S rRNA and mt-LSU proteins, impaired mt-LSU assembly, deeply attenuated mitochondrial protein synthesis, and consequent failure to assemble oxidative phosphorylation complexes.

Abstract: Our findings identify DDX28 as essential during the early stages of mitoribosome mt-LSU biogenesis, a process that takes place mainly near the mitochondrial nucleoids, in the compartment defined by the RNA granules.


Conversion of Adult Pancreatic α-cells to β-cells After Extreme β-cell Loss

Abstract: Pancreatic insulin-producing beta-cells have a long lifespan, such that in healthy conditions they replicate little during a lifetime.

Abstract: Nevertheless, they show increased self-duplication after increased metabolic demand or after injury (that is, beta-cell loss).

Abstract: It is not known whether adult mammals can differentiate (regenerate) new beta-cells after extreme, total beta-cell loss, as in diabetes.

Abstract: This would indicate differentiation from precursors or another heterologous (non-beta-cell) source.

Abstract: Here we show beta-cell regeneration in a transgenic model of diphtheria-toxin-induced acute selective near-total beta-cell ablation.

Abstract: If given insulin, the mice survived and showed beta-cell mass augmentation with time.

Abstract: Lineage-tracing to label the glucagon-producing alpha-cells before beta-cell ablation tracked large fractions of regenerated beta-cells as deriving from alpha-cells, revealing a previously disregarded degree of pancreatic cell plasticity.

Abstract: Such inter-endocrine spontaneous adult cell conversion could be harnessed towards methods of producing beta-cells for diabetes therapies, either in differentiation settings in vitro or in induced regeneration.


Driver mutations in histone H3.3 and chromatin remodelling genes in paediatric glioblastoma

Abstract: Glioblastoma multiforme (GBM) is a lethal brain tumour in adults and children.

Abstract: However, DNA copy number and gene expression signatures indicate differences between adult and paediatric cases.

Abstract: To explore the genetic events underlying this distinction, we sequenced the exomes of 48 paediatric GBM samples.

Abstract: Somatic mutations in the H3.3-ATRX-DAXX chromatin remodelling pathway were identified in 44% of tumours (21/48).

Abstract: Recurrent mutations in H3F3A, which encodes the replication-independent histone 3 variant H3.3, were observed in 31% of tumours, and led to amino acid substitutions at two critical positions within the histone tail (K27M, G34R/G34V) involved in key regulatory post-translational modifications.

Abstract: Mutations in ATRX (α-thalassaemia/mental retardation syndrome X-linked) and DAXX (death-domain associated protein), encoding two subunits of a chromatin remodelling complex required for H3.3 incorporation at pericentric heterochromatin and telomeres, were identified in 31% of samples overall, and in 100% of tumours harbouring a G34R or G34V H3.3 mutation.

Abstract: Somatic TP53 mutations were identified in 54% of all cases, and in 86% of samples with H3F3A and/or ATRX mutations.

Abstract: Screening of a large cohort of gliomas of various grades and histologies (n = 784) showed H3F3A mutations to be specific to GBM and highly prevalent in children and young adults.

Abstract: Furthermore, the presence of H3F3A/ATRX-DAXX/TP53 mutations was strongly associated with alternative lengthening of telomeres and specific gene expression profiles.

Abstract: This is, to our knowledge, the first report to highlight recurrent mutations in a regulatory histone in humans, and our data suggest that defects of the chromatin architecture underlie paediatric and young adult GBM pathogenesis.


Translational control of intron splicing in eukaryotes

Abstract: Most eukaryotic genes are interrupted by non-coding introns that must be accurately removed from pre-messenger RNAs to produce translatable mRNAs.

Abstract: Splicing is guided locally by short conserved sequences, but genes typically contain many potential splice sites, and the mechanisms specifying the correct sites remain poorly understood.

Abstract: In most organisms, short introns recognized by the intron definition mechanism cannot be efficiently predicted solely on the basis of sequence motifs.

Abstract: In multicellular eukaryotes, long introns are recognized through exon definition and most genes produce multiple mRNA variants through alternative splicing.

Abstract: The nonsense-mediated mRNA decay (NMD) pathway may further shape the observed sets of variants by selectively degrading those containing premature termination codons, which are frequently produced in mammals.

Abstract: Here we show that the tiny introns of the ciliate Paramecium tetraurelia are under strong selective pressure to cause premature termination of mRNA translation in the event of intron retention, and that the same bias is observed among the short introns of plants, fungi and animals.

Abstract: By knocking down the two P. tetraurelia genes encoding UPF1, a protein that is crucial in NMD, we show that the intrinsic efficiency of splicing varies widely among introns and that NMD activity can significantly reduce the fraction of unspliced mRNAs.

Abstract: The results suggest that, independently of alternative splicing, species with large intron numbers universally rely on NMD to compensate for suboptimal splicing efficiency and accuracy.


Alternatively activated macrophages produce catecholamines to sustain adaptive thermogenesis

Abstract: All homeotherms use thermogenesis to maintain their core body temperature, ensuring that cellular functions and physiological processes can continue in cold environments.

Abstract: In the prevailing model of thermogenesis, when the hypothalamus senses cold temperatures it triggers sympathetic discharge, resulting in the release of noradrenaline in brown adipose tissue and white adipose tissue.

Abstract: Acting via the β(3)-adrenergic receptors, noradrenaline induces lipolysis in white adipocytes, whereas it stimulates the expression of thermogenic genes, such as PPAR-γ coactivator 1a (Ppargc1a), uncoupling protein 1 (Ucp1) and acyl-CoA synthetase long-chain family member 1 (Acsl1), in brown adipocytes.

Abstract: However, the precise nature of all the cell types involved in this efferent loop is not well established.

Abstract: Here we report in mice an unexpected requirement for the interleukin-4 (IL-4)-stimulated program of alternative macrophage activation in adaptive thermogenesis.

Abstract: Exposure to cold temperature rapidly promoted alternative activation of adipose tissue macrophages, which secrete catecholamines to induce thermogenic gene expression in brown adipose tissue and lipolysis in white adipose tissue.

Abstract: Absence of alternatively activated macrophages impaired metabolic adaptations to cold, whereas administration of IL-4 increased thermogenic gene expression, fatty acid mobilization and energy expenditure, all in a macrophage-dependent manner.

Abstract: Thus, we have discovered a role for alternatively activated macrophages in the orchestration of an important mammalian stress response, the response to cold.


Alternative Lengthening of Telomeres Mediated by Mitotic DNA Synthesis Engages Break-Induced Replication Processes.

Abstract: Alternative lengthening of telomeres (ALT) is a telomerase-independent telomere maintenance mechanism that occurs in a subset of cancers.

Abstract: By analyzing telomerase-positive cells and their human TERC knockout-derived ALT human cell lines, we show that ALT cells harbor more fragile telomeres representing telomere replication problems.

Abstract: ALT-associated replication defects trigger mitotic DNA synthesis (MiDAS) at telomeres in a RAD52-dependent, but RAD51-independent, manner.

Abstract: Telomeric MiDAS is a conservative DNA synthesis process, potentially mediated by break-induced replication, similar to type II ALT survivors in Saccharomyces cerevisiae Replication stresses induced by ectopic oncogenic expression of cyclin E, G-quadruplexes, or R-loop formation facilitate the ALT pathway and lead to telomere clustering, a hallmark of ALT cancers.

Abstract: The TIMELESS/TIPIN complex suppresses telomere clustering and telomeric MiDAS, whereas the SMC5/6 complex promotes them.

Abstract: In summary, ALT cells exhibit more telomere replication defects that result in persistent DNA damage responses at telomeres, leading to the engagement of telomeric MiDAS (spontaneous mitotic telomere synthesis) that is triggered by DNA replication stress, a potential driver of genomic duplications in cancer.


Light acts directly on organs and cells in culture to set the vertebrate circadian clock.

Abstract: The expression of clock genes in vertebrates is widespread and not restricted to classical clock structures.

Abstract: The expression of the Clock gene in zebrafish shows a strong circadian oscillation in many tissues in vivo and in culture, showing that endogenous oscillators exist in peripheral organs.

Abstract: A defining feature of circadian clocks is that they can be set or entrained to local time, usually by the environmental light-dark cycle.

Abstract: An important question is whether peripheral oscillators are entrained to local time by signals from central pacemakers such as the eyes or are themselves directly light-responsive.

Abstract: Here we show that the peripheral organ clocks of zebrafish are set by light-dark cycles in culture.

Abstract: We also show that a zebrafish-derived cell line contains a circadian oscillator, which is also directly light entrained.


Small molecule inhibition of the KRAS–PDEδ interaction impairs oncogenic KRAS signalling

Abstract: The KRAS oncogene product is considered a major target in anticancer drug discovery.

Abstract: However, direct interference with KRAS signalling has not yet led to clinically useful drugs.

Abstract: Correct localization and signalling by farnesylated KRAS is regulated by the prenyl-binding protein PDEδ, which sustains the spatial organization of KRAS by facilitating its diffusion in the cytoplasm.

Abstract: Here we report that interfering with binding of mammalian PDEδ to KRAS by means of small molecules provides a novel opportunity to suppress oncogenic RAS signalling by altering its localization to endomembranes.

Abstract: Biochemical screening and subsequent structure-based hit optimization yielded inhibitors of the KRAS–PDEδ interaction that selectively bind to the prenyl-binding pocket of PDEδ with nanomolar affinity, inhibit oncogenic RAS signalling and suppress in vitro and in vivo proliferation of human pancreatic ductal adenocarcinoma cells that are dependent on oncogenic KRAS.

Abstract: Our findings may inspire novel drug discovery efforts aimed at the development of drugs targeting oncogenic RAS.


Evolution of novel cooperative swarming in the bacterium Myxococcus xanthus

Abstract: Cooperation among individuals is necessary for evolutionary transitions to higher levels of biological organization.

Abstract: In such transitions, groups of individuals at one level (such as single cells) cooperate to form selective units at a higher level (such as multicellular organisms).

Abstract: Though the evolution of cooperation is difficult to observe directly in higher eukaryotes, microorganisms do offer such an opportunity.

Abstract: Here we report the evolution of novel cooperative behaviour in experimental lineages of the bacterium Myxococcus xanthus.

Abstract: Wild-type strains of M. xanthus exhibit socially dependent swarming across soft surfaces by a mechanism known as ‘S-motility’ that requires the presence of extracellular type IV pili.

Abstract: In lineages of M. xanthus unable to make pili, a new mechanistic basis for cooperative swarming evolved.

Abstract: Evolved swarming is mediated, at least in part, by enhanced production of an extracellular fibril matrix that binds cells—and their evolutionary interests—together.

Abstract: Though costly to individuals, fibril production greatly enhanced population expansion in groups of interconnected cells.

Abstract: These results show that fundamental transitions to primitive cooperation can readily occur in bacteria.


Exon duplication and divergence in the human preproglucagon gene

Abstract: Glucagon is a 29-amino acid pancreatic hormone which counteracts the blood glucose-lowering action of insulin by stimulating hepatic glycogenolysis and gluconeogenesis1.

Abstract: The structure of the hamster pancreatic glucagon precursor has recently been determined from the sequence of a cloned cDNA2.

Abstract: Hamster preproglucagon is a 180-amino acid protein which contains five functional regions; a signal or pre-peptide, an NH2-terminal peptide (also called glicentin-related pancreatic peptide, GRPP), glucagon, and two carboxy-terminal glucagon-like peptides (GLP-1 and GLP-2).

Abstract: The sequences of two non-allelic anglerfish pancreatic glucagon precursors3–5 have also been determined and their organization is similar but not identical to the hamster protein; they lack the polypeptide segment corresponding to hamster GLP-2.

Abstract: The presence of three regions possessing internal homology, that is, glucagon, GLP-1 and GLP-2, within proglucagon, and the absence of GLP-2 in the anglerfish precursors suggests that the structure of the preproglucagon gene might provide insight into the evolution of this polyprotein.

Abstract: We have isolated and sequenced the human preproglucagon gene and report here that the organization of the human precursor deduced from this sequence is identical to the hamster protein.

Abstract: The gene contains at least three intervening sequences which divide the protein-coding portion of the gene into four regions corresponding to the signal peptide and part of the NH2-terminal peptide, the remainder of the NH2-terminal peptide and glucagon, GLP-1, and GLP-2.

Abstract: The data suggest that triplication and subsequent sequence divergence of an exon encoding glucagon or a glucagon-like peptide produced this polyprotein precursor.


BH1 and BH2 domains of Bcl-2 are required for inhibition of apoptosis and heterodimerization with Bax

Abstract: BCL-2 was isolated from the t(14;18) chromosomal breakpoint in follicular B-cell lymphoma1–3.

Abstract: Bcl-2 has the unique oncogenic role of extending cell survival by inhibiting a variety of apoptotic deaths4–13.

Abstract: An emerging family of Bcl-2 -related proteins share two highly conserved regions14–20 referred to here as Bcl-2 homology 1 and 2 (BH1 and BH2) domains (Fig. 1).

Abstract: This includes Bax which heterodimerizes with Bcl-2 and when overexpressed counteracts Bcl-214.

Abstract: We report here that site-specific mutagenesis of Bcl-2 establishes the two domains as novel dimerization motifs.

Abstract: Substitu-tion of Gly 145 in BHl domain or Trp 188 in BH2 domain completely abrogated Bcl-2's death-repressor activity in inter-leukin-3 deprivation, γ-irradiation and glucocorticoid-induced apoptosis.

Abstract: Mutations that affected Bcl-2's function also disrupted its heterodimerization with Bax, yet still permitted Bcl-2 homo-dimerization.

Abstract: These results establish a functional role for the BH1 and BH2 domains and suggest Bcl-2 exerts its action through heterodimerization with Bax.


KAP1 controls endogenous retroviruses in embryonic stem cells

Abstract: More than forty per cent of the mammalian genome is derived from retroelements, of which about one-quarter are endogenous retroviruses (ERVs).

Abstract: Some are still active, notably in mice the highly polymorphic early transposon (ETn)/MusD and intracisternal A-type particles (IAP).

Abstract: ERVs are transcriptionally silenced during early embryogenesis by histone and DNA methylation (and reviewed in ref.

Abstract: 7), although the initiators of this process, which is essential to protect genome integrity, remain largely unknown.

Abstract: KAP1 (KRAB-associated protein 1, also known as tripartite motif-containing protein 28, TRIM28) represses genes by recruiting the histone methyltransferase SETDB1, heterochromatin protein 1 (HP1) and the NuRD histone deacetylase complex, but few of its physiological targets are known.

Abstract: Two lines of evidence suggest that KAP1-mediated repression could contribute to the control of ERVs: first, KAP1 can trigger permanent gene silencing during early embryogenesis, and second, a KAP1 complex silences the retrovirus murine leukaemia virus in embryonic cells.

Abstract: Consistent with this hypothesis, here we show that KAP1 deletion leads to a marked upregulation of a range of ERVs, in particular IAP elements, in mouse embryonic stem (ES) cells and in early embryos.

Abstract: We further demonstrate that KAP1 acts synergistically with DNA methylation to silence IAP elements, and that it is enriched at the 5′ untranslated region (5′UTR) of IAP genomes, where KAP1 deletion leads to the loss of histone 3 lysine 9 trimethylation (H3K9me3), a hallmark of KAP1-mediated repression.

Abstract: Correspondingly, IAP 5′UTR sequences can impose in cis KAP1-dependent repression on a heterologous promoter in ES cells.

Abstract: Our results establish that KAP1 controls endogenous retroelements during early embryonic development.


The TOR signalling pathway controls nuclear localization of nutrient-regulated transcription factors.

Abstract: The rapamycin-sensitive TOR signalling pathway in Saccharomyces cerevisiae activates a cell-growth program in response to nutrients such as nitrogen and carbon.

Abstract: The TOR1 and TOR2 kinases (TOR) control cytoplasmic protein synthesis and degradation through the conserved TAP42 protein.

Abstract: Upon phosphorylation by TOR, TAP42 binds and possibly inhibits type 2A and type-2A-related phosphatases; however, the mechanism by which TOR controls nuclear events such as global repression of starvation-specific transcription is unknown.

Abstract: Here we show that TOR prevents transcription of genes expressed upon nitrogen limitation by promoting the association of the GATA transcription factor GLN3 with the cytoplasmic protein URE2.

Abstract: The binding of GLN3 to URE2 requires TOR-dependent phosphorylation of GLN3.

Abstract: Phosphorylation and cytoplasmic retention of GLN3 are also dependent on the TOR effector TAP42, and are antagonized by the type-2A-related phosphatase SIT4.

Abstract: TOR inhibits expression of carbon-source-regulated genes by stimulating the binding of the transcriptional activators MSN2 and MSN4 to the cytoplasmic 14-3-3 protein BMH2.

Abstract: Thus, the TOR signalling pathway broadly controls nutrient metabolism by sequestering several transcription factors in the cytoplasm.


Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides

Abstract: Murine embryonic stem (ES) cells are pluripotent cell lines established directly from the early embryo1,2 which can contribute differentiated progeny to all adult tissues, including the germ-cell lineage3, after re-incorporation into the normal embryo.

Abstract: They provide both a cellular vector for the generation of transgenic animals4 and a useful system for the identification of polypeptide factors controlling differentiation processes in early development5.

Abstract: In particular, medium conditioned by Buffalo rat liver cells contains a polypeptide factor, ES cell differentiation inhibitory activity (DIA), which specifically suppresses the spontaneous differentiation of ES cells in vitro, thereby permitting their growth as homogeneous stem cell populations in the absence of heterologous feeder cells6.

Abstract: ES cell pluripotentiality, including the ability to give rise to functional gametes, is preserved after prolonged culture in Buffalo rat liver media as a source of DIA7.

Abstract: Here, we report that purified DIA is related in structure and function to the recently identified haemopoetic regulatory factors human interleukin for DA cells8,9 and leukaemia inhibitory factor10.

Abstract: DIA and human interleukin DA/leukaemia inhibitory factor have thus been identified as related multifunctional regulatory factors with distinct biological activities in both early embryonic and haemopoetic stem cell systems.


Pancreatic cancer genomes reveal aberrations in axon guidance pathway genes

Abstract: Pancreatic cancer is a highly lethal malignancy with few effective therapies.

Abstract: We performed exome sequencing and copy number analysis to define genomic aberrations in a prospectively accrued clinical cohort (n = 142) of early (stage I and II) sporadic pancreatic ductal adenocarcinoma.

Abstract: Detailed analysis of 99 informative tumours identified substantial heterogeneity with 2,016 non-silent mutations and 1,628 copy-number variations.

Abstract: We define 16 significantly mutated genes, reaffirming known mutations (KRAS, TP53, CDKN2A, SMAD4, MLL3, TGFBR2, ARID1A and SF3B1), and uncover novel mutated genes including additional genes involved in chromatin modification (EPC1 and ARID2), DNA damage repair (ATM) and other mechanisms (ZIM2, MAP2K4, NALCN, SLC16A4 and MAGEA6).

Abstract: Integrative analysis with in vitro functional data and animal models provided supportive evidence for potential roles for these genetic aberrations in carcinogenesis.

Abstract: Pathway-based analysis of recurrently mutated genes recapitulated clustering in core signalling pathways in pancreatic ductal adenocarcinoma, and identified new mutated genes in each pathway.

Abstract: We also identified frequent and diverse somatic aberrations in genes described traditionally as embryonic regulators of axon guidance, particularly SLIT/ROBO signalling, which was also evident in murine Sleeping Beauty transposon-mediated somatic mutagenesis models of pancreatic cancer, providing further supportive evidence for the potential involvement of axon guidance genes in pancreatic carcinogenesis.


Rejuvenation of aged progenitor cells by exposure to a young systemic environment

Abstract: The decline of tissue regenerative potential is a hallmark of ageing and may be due to age-related changes in tissue-specific stem cells.

Abstract: A decline in skeletal muscle stem cell (satellite cell) activity due to a loss of Notch signalling results in impaired regeneration of aged muscle.

Abstract: The decline in hepatic progenitor cell proliferation owing to the formation of a complex involving cEBP-α and the chromatin remodelling factor brahma (Brm) inhibits the regenerative capacity of aged liver.

Abstract: To examine the influence of systemic factors on aged progenitor cells from these tissues, we established parabiotic pairings (that is, a shared circulatory system) between young and old mice (heterochronic parabioses), exposing old mice to factors present in young serum.

Abstract: Notably, heterochronic parabiosis restored the activation of Notch signalling as well as the proliferation and regenerative capacity of aged satellite cells.

Abstract: The exposure of satellite cells from old mice to young serum enhanced the expression of the Notch ligand (Delta), increased Notch activation, and enhanced proliferation in vitro.

Abstract: Furthermore, heterochronic parabiosis increased aged hepatocyte proliferation and restored the cEBP-α complex to levels seen in young animals.

Abstract: These results suggest that the age-related decline of progenitor cell activity can be modulated by systemic factors that change with age.


Continuous single-cell imaging of blood generation from haemogenic endothelium

Abstract: Despite decades of research, the identity of the cells generating the first haematopoietic cells in mammalian embryos is unknown.

Abstract: Indeed, whether blood cells arise from mesodermal cells, mesenchymal progenitors, bipotent endothelial–haematopoietic precursors or haemogenic endothelial cells remains controversial.

Abstract: Proximity of endothelial and blood cells at sites of embryonic haematopoiesis, as well as their similar gene expression, led to the hypothesis of the endothelium generating blood.

Abstract: However, owing to lacking technology it has been impossible to observe blood cell emergence continuously at the single-cell level, and the postulated existence of haemogenic endothelial cells remains disputed.

Abstract: Here, using new imaging and cell-tracking methods, we show that embryonic endothelial cells can be haemogenic.

Abstract: By continuous long-term single-cell observation of mouse mesodermal cells generating endothelial cell and blood colonies, it was possible to detect haemogenic endothelial cells giving rise to blood cells.

Abstract: Living endothelial and haematopoietic cells were identified by simultaneous detection of morphology and multiple molecular and functional markers.

Abstract: Detachment of nascent blood cells from endothelium is not directly linked to asymmetric cell division, and haemogenic endothelial cells are specified from cells already expressing endothelial markers.

Abstract: These results improve our understanding of the developmental origin of mammalian blood and the potential generation of haematopoietic stem cells from embryonic stem cells.


Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines

Abstract: The recent availability in culture of embryo-derived pluripotential cells which exhibit both a normal karyotype and a high differentiative ability1–3 has encouraged us to assess the potential of these cells to form functional germ cells following their incorporation into chimaeric mice.

Abstract: We report here the results of blastocyst injection studies using three independently isolated XY embryo-derived cell lines (EK.CP1, EK.CC1.1 and EKCC1.2) which produce a very high proportion (>50%) of live-born animals that are overtly chimaeric.

Abstract: Seven chimaeric male mice, derived from these three lines, have, so far, proved to be functional germ-line chimaeras.


Chloroquine resistance not linked to mdr-like genes in a Plasmodium falciparum cross

Abstract: CHLOROQUINE is thought to act against falciparum malaria by accumulating in the acid vesicles of the parasite and interfering with their function14.

Abstract: Parasites resistant to chloroquine expel the drug rapidly in an unaltered form, thereby reducing levels of accumulation in the vesicles5.

Abstract: The discovery that verapamil partially reverses chloroquine resistance in vitro 6 led to the proposal that efflux may involve an ATP-driven P-glycoprotein pump similar to that in mammalian multidrug-resistant (mdr) tumor cell lines.

Abstract: Indeed, Plasmodium falciparum contains at least two mdr-like genes7,8, one of which has been suggested to confer the chloroquine resistant (CQR) phenotype7,9,10.

Abstract: To determine if either of these genes is linked to chloroquine resistance, we performed a genetic cross between CQR and chloroquine-susceptible (CQS) clones of P. falciparum.

Abstract: Examination of 16 independent recombinant progeny indicated that the rapid efflux phenotype is controlled by a single gene or a closely linked group of genes.

Abstract: But, there was no linkage between the rapid efflux, CQR phenotype and either of the mdr-like P. falciparum genes or amplification of those genes.

Abstract: These data indicate that the genetic locus governing chloroquine efflux and resistance is independent of the known mdr-like genes.


Identification of a cold receptor reveals a general role for TRP channels in thermosensation

Abstract: The cellular and molecular mechanisms that enable us to sense cold are not well understood.

Abstract: Insights into this process have come from the use of pharmacological agents, such as menthol, that elicit a cooling sensation.

Abstract: Here we have characterized and cloned a menthol receptor from trigeminal sensory neurons that is also activated by thermal stimuli in the cool to cold range.

Abstract: This cold- and menthol-sensitive receptor, CMR1, is a member of the TRP family of excitatory ion channels, and we propose that it functions as a transducer of cold stimuli in the somatosensory system.

Abstract: These findings, together with our previous identification of the heat-sensitive channels VR1 and VRL-1, demonstrate that TRP channels detect temperatures over a wide range and are the principal sensors of thermal stimuli in the mammalian peripheral nervous system.


Surface mechanics mediate pattern formation in the developing retina

Abstract: Pattern formation of biological structures involves organizing different types of cells into a spatial configuration.

Abstract: In this study, we investigate the physical basis of biological patterning of the Drosophila retina in vivo.

Abstract: We demonstrate that E- and N-cadherins mediate apical adhesion between retina epithelial cells.

Abstract: Differential expression of N-cadherin within a sub-group of retinal cells (cone cells) causes them to form an overall shape that minimizes their surface contact with surrounding cells.

Abstract: The cells within this group, in both normal and experimentally manipulated conditions, pack together in the same way as soap bubbles do.

Abstract: The shaping of the cone cell group and packing of its components precisely imitate the physical tendency for surfaces to be minimized.

Abstract: Thus, simple patterned expression of N-cadherin results in a complex spatial pattern of cells owing to cellular surface mechanics.


Role of inscuteable in orienting asymmetric cell divisions in Drosophila

Abstract: Drosophila neuroblasts and epithelial cells in the procephalic neurogenic region divide perpendicular to the surface, and segregate the proteins Numb and Prospero into the basal daughter cell.

Abstract: We demonstrate here that orientation of the mitotic spindle and correct localization of Numb and Prospero in these cells require the inscuteable gene.

Abstract: Moreover, ectopic expression of inscuteable in other epithelial cells leads to spindle reorientation.

Abstract: The Inscuteable protein localizes to the apical cell cortex before mitosis, suggesting that Inscuteable functions in establishing polarity for asymmetric cell division.


Specific interference by ingested dsRNA.

Abstract: A genetic interference phenomenon in the nematode Caenorhabditis elegans has been described in which expression of an individual gene can be specifically reduced by microinjecting a corresponding fragment of double-stranded (ds) RNA.

Abstract: One striking feature of this process is a spreading effect: interference in a broad region of the animal is observed following the injection of dsRNA into the extracellular body cavity.

Abstract: Here we show that C. elegans can respond in a gene-specific manner to dsRNA encountered in the environment.

Abstract: C. elegans normally feed on bacteria, ingesting and grinding them in the pharynx and subsequently absorbing bacterial contents in the gut.

Abstract: We find that Escherichia coli bacteria expressing dsRNAs can confer specific interference effects on the nematode larvae that feed on them.