Interaction of TNF with TNF receptor type 2 promotes expansion and function of mouse CD4+CD25+ T regulatory cells.

Abstract: Although TNF is a major proinflammatory cytokine, increasing evidence indicates that TNF also has immunosuppressive feedback effects.

Abstract: We have demonstrated in this study that, in both resting and activated states, mouse peripheral CD4(+)CD25(+) T regulatory cells (Tregs) expressed remarkably higher surface levels of TNFR2 than CD4(+)CD25(-) T effector cells (Teffs).

Abstract: In cocultures of Tregs and Teffs, inhibition of proliferation of Teffs by Tregs was initially transiently abrogated by exposure to TNF, but longer exposure to TNF restored suppressive effects.

Abstract: Cytokine production by Teffs remained continually suppressed by Tregs.

Abstract: The profound anergy of Tregs in response to TCR stimulation was overcome by TNF, which expanded the Treg population.

Abstract: Furthermore, in synergy with IL-2, TNF expanded Tregs even more markedly up-regulated expression of CD25 and FoxP3 and phosphorylation of STAT5, and enhanced the suppressive activity of Tregs.

Abstract: Unlike TNF, IL-1beta and IL-6 did not up-regulate FoxP3-expressing Tregs.

Abstract: Furthermore, the number of Tregs increased in wild-type mice, but not in TNFR2(-/-) mice following sublethal cecal ligation and puncture.

Abstract: Depletion of Tregs significantly decreased mortality following cecal ligation and puncture.

Abstract: Thus, the stimulatory effect of TNF on Tregs resembles the reported costimulatory effects of TNF on Teffs, but is even more pronounced because of the higher expression of TNFR2 by Tregs.

Abstract: Moreover, our study suggests that the slower response of Tregs than Teffs to TNF results in delayed immunosuppressive feedback effects.


The basics of epithelial-mesenchymal transition.

Abstract: The origins of the mesenchymal cells participating in tissue repair and pathological processes, notably tissue fibrosis, tumor invasiveness, and metastasis, are poorly understood.

Abstract: However, emerging evidence suggests that epithelial-mesenchymal transitions (EMTs) represent one important source of these cells.

Abstract: As we discuss here, processes similar to the EMTs associated with embryo implantation, embryogenesis, and organ development are appropriated and subverted by chronically inflamed tissues and neoplasias.

Abstract: The identification of the signaling pathways that lead to activation of EMT programs during these disease processes is providing new insights into the plasticity of cellular phenotypes and possible therapeutic interventions.


Child maltreatment, other trauma exposure, and posttraumatic symptomatology among children with oppositional defiant and attention deficit hyperactivity disorders.

Abstract: Consecutive child psychiatric outpatient admissions with disruptive behavior or adjustment disorders were assessed by validated instruments for trauma exposure and posttraumatic stress disorder (PTSD) symptoms and other psychopathology.

Abstract: Four reliably diagnosed groups were defined in a retrospective case-control design: Attention Deficit Hyperactivity Disorder (ADHD), Oppositional Defiant Disorder (ODD), comorbid ADHD-ODD, and adjustment disorder controls.

Abstract: ODD and (although to a lesser extent) ADHD were associated with a history of physical or sexual maltreatment.

Abstract: PTSD symptoms were most severe if (a) ADHD and maltreatment co-occurred or (b) ODD and accident/illness trauma co-occurred.

Abstract: The association between ODD and PTSD Criterion D (hyperarousal/hypervigilance) symptoms remained after controlling for overlapping symptoms, but the association of ADHD with PTSD symptoms was largely due to an overlapping symptom.

Abstract: These findings suggest that screening for maltreatment, other trauma, and PTSD symptoms may enhance prevention, treatment, and research concerning childhood disruptive behavior disorders.


Exercise-induced asthma screening of elite athletes: field versus laboratory exercise challenge.

Abstract: PURPOSE The purpose of this study was to compare a laboratory based exercise challenge (LBC) to a field based exercise challenge (FBC) for pulmonary function test (PFT) exercise-induced asthma (EIA) screening of elite athletes.

Abstract: METHODS Twenty-three elite cold weather athletes (14 men, 9 women) PFT positive for EIA (FBC screened) served as subjects.

Abstract: Twenty-three gender and sport matched controls (nonasthmatics) were randomly selected to establish PFT reference values for normal elite athletes.

Abstract: Before FBC, athletes completed a medical history questionnaire for EIA symptoms.

Abstract: FBC evaluations consisted of baseline spirometry, actual or simulated competition, and 5, 10, and 15 min postexercise spirometry.

Abstract: PFT positive athletes were evaluated in the laboratory using an exercise challenge simulating race intensity (ambient conditions: 21 degrees C, 60% relative humidity).

Abstract: PFT procedures were identical to FBC.

Abstract: RESULTS 91% of PFT positive and 48% of PFT normal athletes reported at least one symptom of EIA, with postrace cough most frequent.

Abstract: Baseline spirometry was the same for PFT positives and normal controls.

Abstract: Lower limit reference range (MN - 2 SD) of FEV1 for controls suggests that postexercise decrements of greater than approximately -7% indicate abnormal airway response in this population.

Abstract: Exercise time duration did not effect bronchial reactivity; 78% of FBC PFT positives were PFT normal post-LBC.

Abstract: CONCLUSION Self-reported symptoms by elite athletes are not reliable in identifying EIA.

Abstract: Reference range criterion for FEV1 decrement in the elite athlete postexercise contrasts current recommended guidelines.

Abstract: Moreover, a large number of false negatives may occur in this population if EIA screening is performed with inadequate exercise and environmental stress.


Use of a targeted oncolytic poxvirus, JX-594, in patients with refractory primary or metastatic liver cancer: a phase I trial.

Abstract: BACKGROUND JX-594 is a targeted oncolytic poxvirus designed to selectively replicate in and destroy cancer cells with cell-cycle abnormalities and epidermal growth factor receptor (EGFR)-ras pathway activation.

Abstract: Direct oncolysis plus granulocyte-macrophage colony-stimulating factor (GM-CSF) expression also stimulates shutdown of tumour vasculature and antitumoral immunity.

Abstract: We aimed to assess intratumoral injection of JX-594 in patients with refractory primary or metastatic liver cancer.

Abstract: METHODS Between Jan 4, 2006, and July 4, 2007, 14 patients with histologically confirmed refractory primary or metastatic liver tumours (up to 10.9 cm total diameter) that were amenable to image-guided intratumoral injections were enrolled into this non-comparative, open-label, phase I dose-escalation trial (standard 3x3 design; two to six patients for each dose with 12-18 estimated total patients).

Abstract: Patients received one of four doses of intratumoral JX-594 (10(8) plaque-forming units [pfu], 3x10(8) pfu, 10(9) pfu, or 3x10(9) pfu) every 3 weeks at Dong-A University Hospital (Busan, South Korea).

Abstract: Patients were monitored after treatment for at least 48 h in hospital and for at least 4 weeks as out-patients.

Abstract: Adverse event-monitoring according to the National Cancer Institute Common Toxicity Criteria (version 3) and standard laboratory toxicity grading for haematology, liver and renal function, coagulation studies, serum chemistry, and urinalysis were done.

Abstract: The primary aims were to ascertain the maximum-tolerated dose (MTD) and safety of JX-594 treatment.

Abstract: Data were also collected on pharmacokinetics, pharmacodynamics, and efficacy.

Abstract: Analysis was per protocol.

Abstract: This study is registered with ClinicalTrials.gov, number NCT00629759.

Abstract: FINDINGS Of 22 patients with liver tumours who were assessed for eligibility, eight patients did not meet inclusion criteria.

Abstract: Therefore, 14 patients, including those with hepatocellular, colorectal, melanoma, and lung cancer, were enrolled.

Abstract: Patients were heavily pretreated (5.6 previous treatments, SD 2.8, range 2.0-12.0) and had large tumours (7.0 cm diameter, SD 2.7, range 1.8-10.9).

Abstract: Patients received a mean of 3.4 (SD 2.2, range 1.0-8.0) cycles of JX-594.

Abstract: All patients were evaluable for toxicity.

Abstract: All patients experienced grade I-III flu-like symptoms, and four had transient grade I-III dose-related thrombocytopenia.

Abstract: Grade III hyperbilirubinaemia was dose-limiting in both patients at the highest dose; the MTD was therefore 1x10(9) pfu.

Abstract: JX-594 replication-dependent dissemination in blood was shown, with resultant infection of non-injected tumour sites.

Abstract: GM-CSF expression resulted in grade I-III increases in neutrophil counts in four of six patients at the MTD.

Abstract: Tumour responses were shown in injected and non-injected tumours.

Abstract: Ten patients were radiographically evaluable for objective responses; non-evaluable patients had contraindications to contrast medium (n=2) or no post-treatment scans (n=2).

Abstract: According to Response Evaluation Criteria in Solid Tumors (RECIST), three patients had partial response, six had stable disease, and one had progressive disease.

Abstract: INTERPRETATION Intratumoral injection of JX-594 into primary or metastatic liver tumours was generally well-tolerated.

Abstract: Direct hyperbilirubinaemia was the dose-limiting toxicity.

Abstract: Safety was acceptable in the context of JX-594 replication, GM-CSF expression, systemic dissemination, and JX-594 had anti-tumoral effects against several refractory carcinomas.

Abstract: Phase II trials are now underway.


PTH Signaling in Osteoprogenitors Is Essential for B-Lymphocyte Differentiation and Mobilization.

Abstract: Cells of the osteoblast lineage provide critical support for B lymphopoiesis in the bone marrow (BM).

Abstract: Parathyroid hormone (PTH) signaling in osteoblastic cells through its receptor (PPR) is an important regulator of hematopoietic stem cells; however, its role in regulation of B lymphopoiesis is not clear.

Abstract: Here we demonstrate that deletion of PPR in osteoprogenitors results in a significant loss of trabecular and cortical bone.

Abstract: PPR signaling in osteoprogenitors, but not in mature osteoblasts or osteocytes, is critical for B-cell precursor differentiation via IL-7 production.

Abstract: Interestingly, despite a severe reduction in B-cell progenitors in BM, mature B-lymphocytes were increased 3.5-fold in the BM of mice lacking PPR in osteoprogenitors.

Abstract: This retention of mature IgD(+) B cells in the BM was associated with increased expression of vascular cell adhesion molecule 1 (VCAM1) by PPR-deficient osteoprogenitors, and treatment with VCAM1 neutralizing antibody increased mobilization of B lymphocytes from mutant BM.

Abstract: Our results demonstrate that PPR signaling in early osteoblasts is necessary for B-cell differentiation via IL-7 secretion and for B-lymphocyte mobilization via VCAM1.


Snail coordinately regulates downstream pathways to control multiple aspects of mammalian neural precursor development.

Abstract: The Snail transcription factor plays a key role in regulating diverse developmental processes but is not thought to play a role in mammalian neural precursors.

Abstract: Here, we have examined radial glial precursor cells of the embryonic murine cortex and demonstrate that Snail regulates their survival, self-renewal, and differentiation into intermediate progenitors and neurons via two distinct and separable target pathways.

Abstract: First, Snail promotes cell survival by antagonizing a p53-dependent death pathway because coincident p53 knockdown rescues survival deficits caused by Snail knockdown.

Abstract: Second, we show that the cell cycle phosphatase Cdc25b is regulated by Snail in radial precursors and that Cdc25b coexpression is sufficient to rescue the decreased radial precursor proliferation and differentiation observed upon Snail knockdown.

Abstract: Thus, Snail acts via p53 and Cdc25b to coordinately regulate multiple aspects of mammalian embryonic neural precursor biology.


Endothelial KLF2 links local arterial shear stress levels to the expression of vascular tone-regulating genes.

Abstract: Lung Krüppel-like factor (LKLF/KLF2) is an endothelial transcription factor that is crucially involved in murine vasculogenesis and is specifically regulated by flow in vitro.

Abstract: We now show a relation to local flow variations in the adult human vasculature: decreased LKLF expression was noted at the aorta bifurcations to the iliac and carotid arteries, coinciding with neointima formation.

Abstract: The direct involvement of shear stress in the in vivo expression of LKLF was determined independently by in situ hybridization and laser microbeam microdissection/reverse transcriptase-polymerase chain reaction in a murine carotid artery collar model, in which a 4- to 30-fold induction of LKLF occurred at the high-shear sites.

Abstract: Dissection of the biomechanics of LKLF regulation in vitro demonstrated that steady flow and pulsatile flow induced basal LKLF expression 15- and 36-fold at shear stresses greater than approximately 5 dyne/cm2, whereas cyclic stretch had no effect.

Abstract: Prolonged LKLF induction in the absence of flow changed the expression of angiotensin-converting enzyme, endothelin-1, adrenomedullin, and endothelial nitric oxide synthase to levels similar to those observed under prolonged flow.

Abstract: LKLF repression by siRNA suppressed the flow response of endothelin-1, adrenomedullin, and endothelial nitric oxide synthase (P < 0.05).

Abstract: Thus, we demonstrate that endothelial LKLF is regulated by flow in vivo and is a transcriptional regulator of several endothelial genes that control vascular tone in response to flow.


Synapsin I Cre transgene expression in male mice produces germline recombination in progeny.

Abstract: The cre/LoxP system can produce conditional loss of gene function in specific cell types such as neurons.

Abstract: A transgenic mouse line, utilized by multiple studies, used the Synapsin I promoter to drive expression of cre (SynCre) to achieve neuronal-specific cre expression.

Abstract: Herein we describe that cre expression can also be observed in SynCre mice within the testes after being bred into a floxed transgenic mouse line.

Abstract: Cre transcript was expressed in testes resulting in recombination of the floxed substrate in testes.

Abstract: In the majority of cases, progeny of male SynCre mice inherited a germline recombined floxed allele, while this was never observed in progeny from female mice carrying the SynCre allele.

Abstract: This observation should alert investigators to a potential confound using these mice and enables male germ cell "deletor" strategies.


Hospital mortality, length of stay, and preventable complications among critically ill patients before and after tele-ICU reengineering of critical care processes.

Abstract: CONTEXT The association of an adult tele-intensive care unit (ICU) intervention with hospital mortality, length of stay, best practice adherence, and preventable complications for an academic medical center has not been reported.

Abstract: OBJECTIVE To quantify the association of a tele-ICU intervention with hospital mortality, length of stay, and complications that are preventable by adherence to best practices.

Abstract: DESIGN, SETTING, AND PATIENTS Prospective stepped-wedge clinical practice study of 6290 adults admitted to any of 7 ICUs (3 medical, 3 surgical, and 1 mixed cardiovascular) on 2 campuses of an 834-bed academic medical center that was performed from April 26, 2005, through September 30, 2007.

Abstract: Electronically supported and monitored processes for best practice adherence, care plan creation, and clinician response times to alarms were evaluated.

Abstract: MAIN OUTCOME MEASURES Case-mix and severity-adjusted hospital mortality.

Abstract: Other outcomes included hospital and ICU length of stay, best practice adherence, and complication rates.

Abstract: RESULTS The hospital mortality rate was 13.6% (95% confidence interval [CI], 11.9%-15.4%) during the preintervention period compared with 11.8% (95% CI, 10.9%-12.8%) during the tele-ICU intervention period (adjusted odds ratio [OR], 0.40 [95% CI, 0.31-0.52]).

Abstract: The tele-ICU intervention period compared with the preintervention period was associated with higher rates of best clinical practice adherence for the prevention of deep vein thrombosis (99% vs 85%, respectively; OR, 15.4 [95% CI, 11.3-21.1]) and prevention of stress ulcers (96% vs 83%, respectively; OR, 4.57 [95% CI, 3.91-5.77], best practice adherence for cardiovascular protection (99% vs 80%, respectively; OR, 30.7 [95% CI, 19.3-49.2]), prevention of ventilator-associated pneumonia (52% vs 33%, respectively; OR, 2.20 [95% CI, 1.79-2.70]), lower rates of preventable complications (1.6% vs 13%, respectively, for ventilator-associated pneumonia [OR, 0.15; 95% CI, 0.09-0.23] and 0.6% vs 1.0%, respectively, for catheter-related bloodstream infection [OR, 0.50; 95% CI, 0.27-0.93]), and shorter hospital length of stay (9.8 vs 13.3 days, respectively; hazard ratio for discharge, 1.44 [95% CI, 1.33-1.56]).

Abstract: The results for medical, surgical, and cardiovascular ICUs were similar.

Abstract: CONCLUSION In a single academic medical center study, implementation of a tele-ICU intervention was associated with reduced adjusted odds of mortality and reduced hospital length of stay, as well as with changes in best practice adherence and lower rates of preventable complications.


CD11b+ monocytes abrogate Th17 CD4+ T cell-mediated experimental autoimmune myocarditis.

Abstract: Experimental autoimmune myocarditis (EAM) represents a Th17 T cell-mediated mouse model of postinflammatory heart disease.

Abstract: In BALB/c wild-type mice, EAM is a self-limiting disease, peaking 21 days after alpha-myosin H chain peptide (MyHC-alpha)/CFA immunization and largely resolving thereafter.

Abstract: In IFN-gammaR(-/-) mice, however, EAM is exacerbated and shows a chronic progressive disease course.

Abstract: We found that this progressive disease course paralleled persistently elevated IL-17 release from T cells infiltrating the hearts of IFN-gammaR(-/-) mice 30 days after immunization.

Abstract: In fact, IL-17 promoted the recruitment of CD11b(+) monocytes, the major heart-infiltrating cells in EAM.

Abstract: In turn, CD11b(+) monocytes suppressed MyHC-alpha-specific Th17 T cell responses IFN-gamma-dependently in vitro.

Abstract: In vivo, injection of IFN-gammaR(+/+)CD11b(+), but not IFN-gammaR(-/-)CD11b(+), monocytes, suppressed MyHC-alpha-specific T cells, and abrogated the progressive disease course in IFN-gammaR(-/-) mice.

Abstract: Finally, coinjection of MyHC-alpha-specific, but not OVA-transgenic, IFN-gamma-releasing CD4(+) Th1 T cell lines, together with MyHC-alpha-specific Th17 T cells protected RAG2(-/-) mice from EAM.

Abstract: In conclusion, CD11b(+) monocytes play a dual role in EAM: as a major cellular substrate of IL-17-induced inflammation and as mediators of an IFN-gamma-dependent negative feedback loop confining disease progression.


The cytokine RANKL produced by positively selected thymocytes fosters medullary thymic epithelial cells that express autoimmune regulator.

Abstract: The thymic medulla provides a microenvironment where medullary thymic epithelial cells (mTECs) express autoimmune regulator and diverse tissue-restricted genes, contributing to launching self-tolerance.

Abstract: Positive selection is essential for thymic medulla formation via a previously unknown mechanism.

Abstract: Here we show that the cytokine RANK ligand (RANKL) was produced by positively selected thymocytes and regulated the cellularity of mTEC by interacting with RANK and osteoprotegerin.

Abstract: Forced expression of RANKL restored thymic medulla in mice lacking positive selection, whereas RANKL perturbation impaired medulla formation.

Abstract: These results indicate that RANKL produced by positively selected thymocytes is responsible for fostering thymic medulla formation, thereby establishing central tolerance.


Fibrosis and adipogenesis originate from a common mesenchymal progenitor in skeletal muscle.

Abstract: Accumulation of adipocytes and collagen type-I-producing cells (fibrosis) is observed in muscular dystrophies.

Abstract: The origin of these cells had been largely unknown, but recently we identified mesenchymal progenitors positive for platelet-derived growth factor receptor alpha (PDGFRα) as the origin of adipocytes in skeletal muscle.

Abstract: However, the origin of muscle fibrosis remains largely unknown.

Abstract: In this study, clonal analyses show that PDGFRα(+) cells also differentiate into collagen type-I-producing cells.

Abstract: In fact, PDGFRα(+) cells accumulated in fibrotic areas of the diaphragm in the mdx mouse, a model of Duchenne muscular dystrophy.

Abstract: Furthermore, mRNA of fibrosis markers was expressed exclusively in the PDGFRα(+) cell fraction in the mdx diaphragm.

Abstract: Importantly, TGF-β isoforms, known as potent profibrotic cytokines, induced expression of markers of fibrosis in PDGFRα(+) cells but not in myogenic cells.

Abstract: Transplantation studies revealed that fibrogenic PDGFRα(+) cells mainly derived from pre-existing PDGFRα(+) cells and that the contribution of PDGFRα(-) cells and circulating cells was limited.

Abstract: These results indicate that mesenchymal progenitors are the main origin of not only fat accumulation but also fibrosis in skeletal muscle.


The pecking order of free radicals and antioxidants: lipid peroxidation, alpha-tocopherol, and ascorbate.

Abstract: Free radicals vary widely in their thermodynamic properties, ranging from very oxidizing to very reducing.

Abstract: These thermodynamic properties can be used to predict a pecking order, or hierarchy, for free radical reactions.

Abstract: Using one-electron reduction potentials, the predicted pecking order is in agreement with experimentally observed free radical electron (hydrogen atom) transfer reactions.

Abstract: These potentials are also in agreement with experimental data that suggest that vitamin E, the primary lipid soluble small molecule antioxidant, and vitamin C, the terminal water soluble small molecule antioxidant, cooperate to protect lipids and lipid structures against peroxidation.

Abstract: Although vitamin E is located in membranes and vitamin C is located in aqueous phases, vitamin C is able to recycle vitamin E; i.e., vitamin C repairs the tocopheroxyl (chromanoxyl) radical of vitamin E, thereby permitting vitamin E to function again as a free radical chain-breaking antioxidant.

Abstract: This review discusses: (i) the thermodynamics of free radical reactions that are of interest to the health sciences; (ii) the fundamental thermodynamic and kinetic properties that are associated with chain-breaking antioxidants; (iii) the unique interfacial nature of the apparent reaction of the tocopherol free radical (vitamin E radical) and vitamin C; and (iv) presents a hierarchy, or pecking order, for free radical electron (hydrogen atom) transfer reactions.


Mitochondrial respiration in subcutaneous and visceral adipose tissue from patients with morbid obesity.

Abstract: Adipose tissue exerts important endocrine and metabolic functions in health and disease.

Abstract: Yet the bioenergetics of this tissue is not characterized in humans and possible regional differences are not elucidated.

Abstract: Using high resolution respirometry, mitochondrial respiration was quantified in human abdominal subcutaneous and intra-abdominal visceral (omentum majus) adipose tissue from biopsies obtained in 20 obese patients undergoing bariatric surgery.

Abstract: Mitochondrial DNA (mtDNA) and genomic DNA (gDNA) were determined by the PCR technique for estimation of mitochondrial density.

Abstract: Adipose tissue samples were permeabilized and respirometric measurements were performed in duplicate at 37 degrees C. Substrates (glutamate (G) + malate (M) + octanoyl carnitine (O) + succinate (S)) were added sequentially to provide electrons to complex I + II.

Abstract: ADP ((D)) for state 3 respiration was added after GM.

Abstract: Uncoupled respiration was measured after addition of FCCP.

Abstract: Visceral fat contained more mitochondria per milligram of tissue than subcutaneous fat, but the cells were smaller.

Abstract: Robust, stable oxygen fluxes were found in both tissues, and coupled state 3 (GMOS(D)) and uncoupled respiration were significantly (P < 0.05) higher in visceral (0.95 +/- 0.05 and 1.15 +/-

Abstract: 0.06 pmol O(2) s(1) mg(1), respectively) compared with subcutaneous (0.76 +/- 0.04 and 0.98 +/- 0.05 pmol O(2) s(1) mg(1), respectively) adipose tissue.

Abstract: Expressed per mtDNA, visceral adipose tissue had significantly (P < 0.05) lower mitochondrial respiration.

Abstract: Substrate control ratios were higher and uncoupling control ratio lower (P < 0.05) in visceral compared with subcutaneous adipose tissue.

Abstract: We conclude that visceral fat is bioenergetically more active and more sensitive to mitochondrial substrate supply than subcutaneous fat.

Abstract: Oxidative phosphorylation has a higher relative activity in visceral compared with subcutaneous adipose tissue.


Endogenous gamma-H2AX-ATM-Chk2 checkpoint activation in Bloom's syndrome helicase deficient cells is related to DNA replication arrested forks.

Abstract: The Bloom syndrome helicase (BLM) is critical for genomic stability.

Abstract: A defect in BLM activity results in the cancer-predisposing Bloom syndrome (BS).

Abstract: Here, we report that BLM-deficient cell lines and primary fibroblasts display an endogenously activated DNA double-strand break checkpoint response with prominent levels of phosphorylated histone H2AX (gamma-H2AX), Chk2 (p(T68)Chk2), and ATM (p(S1981)ATM) colocalizing in nuclear foci.

Abstract: Interestingly, the mitotic fraction of gamma-H2AX foci did not seem to be higher in BLM-deficient cells, indicating that these lesions form transiently during interphase.

Abstract: Pulse labeling with iododeoxyuridine and immunofluorescence microscopy showed the colocalization of gamma-H2AX, ATM, and Chk2 together with replication foci.

Abstract: Those foci costained for Rad51, indicating homologous recombination at these replication sites.

Abstract: We therefore analyzed replication in BS cells using a single molecule approach on combed DNA fibers.

Abstract: In addition to a higher frequency of replication fork barriers, BS cells displayed a reduced average fork velocity and global reduction of interorigin distances indicative of an elevated frequency of origin firing.

Abstract: Because BS is one of the most penetrant cancer-predisposing hereditary diseases, it is likely that the lack of BLM engages the cells in a situation similar to precancerous tissues with replication stress.

Abstract: To our knowledge, this is the first report of high ATM-Chk2 kinase activation and its linkage to replication defects in a BS model.


Impact of TNF-alpha and IL-6 levels on development of cachexia in newly diagnosed NSCLC patients.

Abstract: OBJECTIVES We investigated the role of cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in cachexia development in newly diagnosed nonsmall cell lung cancer (NSCLC) patients.

Abstract: METHODS : We evaluated 44 (M/F:41/3) NSCLC patients and 12 (M/F:10/2) age matched healthy smokers.

Abstract: NSCLC cases with a weight loss of > or =10% consisted the cachectic group (n:23, M/F:21/2) and the ones with <10% weight loss consisted the noncachectic group (n:21, M/F:19/2).

Abstract: RESULTS Body mass index (BMI) of cachectics was significantly lower than that of noncachectics (21.0 +/- 2.9 versus 24.5 +/-

Abstract: 3.6, P = 0.02) and controls (21.0 +/- 2.9 versus 25.5 +/- 2.6, P = 0.01).

Abstract: Serum TNF-alpha level did not differ between cachectic and noncachectics (37.3 +/-

Abstract: 39.1 and 51.6 +/-

Abstract: 84.2 pg/mL, respectively).

Abstract: However, it was significantly higher in NSCLC patients compared with controls (44.1 +/- 64.3 and 15.1 +/-

Abstract: 14.3 pg/mL, P = 0.03).

Abstract: Serum IL-6 level was not different between 3 groups (6.4 +/- 4.1, 8.9 +/-

Abstract: 16.3, and 4.1 +/-

Abstract: 3.5 pg/mL, respectively)

Abstract: but it correlated significantly with TNF-alpha (r = 0.4, P = 0.006) and BMI (r = -0.3, P = 0.03).

Abstract: Erythrocyte sedimentation rate (ESR) correlated significantly with TNF-alpha (r = 0.4, P = 0.003) and BMI (r = -0.3, P = 0.03).

Abstract: Among 44 cases, survival of 12 and 17 patients was recorded in cachectics and noncachectics, with no statistical difference (12.2 +/- 3.7 and 11.2 +/-

Abstract: 1.0 months, respectively).

Abstract: CONCLUSIONS TNF-alpha and IL-6 levels did not differ significantly between cachectics and noncachectics.

Abstract: However, significant correlations between IL-6, BMI, and TNF-alpha suggested that these cytokines acted as cofactors in weight loss.

Abstract: Survival was neither influenced by BMI, nor the cytokine levels in the present study.

Abstract: The significant correlation of ESR with TNF-alpha suggested that ESR could provide valuable clue for considerable weight loss in the follow-up of NSCLC patients.


Treatment of antidepressant-associated sexual dysfunction with sildenafil: a randomized controlled trial.

Abstract: CONTEXT Sexual dysfunction is a common adverse effect of antidepressants that frequently results in treatment noncompliance.

Abstract: OBJECTIVE To assess the efficacy of sildenafil citrate in men with sexual dysfunction associated with the use of selective and nonselective serotonin reuptake inhibitor (SRI) antidepressants.

Abstract: DESIGN, SETTING, AND PATIENTS Prospective, parallel-group, randomized, double-blind, placebo-controlled trial conducted between November 1, 2000, and January 1, 2001, at 3 US university medical centers among 90 male outpatients (mean [SD] age, 45 [8] years) with major depression in remission and sexual dysfunction associated with SRI antidepressant treatment.

Abstract: INTERVENTION Patients were randomly assigned to take sildenafil (n = 45) or placebo (n = 45) at a flexible dose starting at 50 mg and adjustable to 100 mg before sexual activity for 6 weeks.

Abstract: MAIN OUTCOME MEASURES The primary outcome measure was score on the Clinical Global Impression-Sexual Function (CGI-SF); secondary measures were scores on the International Index of Erectile Function, Arizona Sexual Experience Scale, Massachusetts General Hospital-Sexual Functioning Questionnaire, and Hamilton Rating Scale for Depression (HAM-D).

Abstract: RESULTS Among the 90 randomized patients, 93% (83/89) of patients treated per protocol took at least 1 dose of study drug and 85% (76/89) completed week 6 end-point assessments with last observation carried forward analyses.

Abstract: At a CGI-SF score of 2 or lower, 54.5% (24/44) of sildenafil compared with 4.4% (2/45) of placebo patients were much or very much improved (P<.001).

Abstract: Erectile function, arousal, ejaculation, orgasm, and overall satisfaction domain measures improved significantly in sildenafil compared with placebo patients.

Abstract: Mean depression scores remained consistent with remission (HAM-D score < or =10) in both groups for the study duration.

Abstract: CONCLUSION In our study, sildenafil effectively improved erectile function and other aspects of sexual function in men with sexual dysfunction associated with the use of SRI antidepressants.

Abstract: These improvements may allow patients to maintain adherence with effective antidepressant treatment.


Noncoding transcription at enhancers: general principles and functional models.

Abstract: Mammalian genomes are extensively transcribed outside the borders of protein-coding genes.

Abstract: Genome-wide studies recently demonstrated that cis-regulatory genomic elements implicated in transcriptional control, such as enhancers and locus-control regions, represent major sites of extragenic noncoding transcription.

Abstract: Enhancer-templated transcripts provide a quantitatively small contribution to the total amount of cellular nonribosomal RNA; nevertheless, the possibility that enhancer transcription and the resulting enhancer RNAs may, in some cases, have functional roles, rather than represent mere transcriptional noise at accessible genomic regions, is supported by an increasing amount of experimental data.

Abstract: In this article we review the current knowledge on enhancer transcription and its functional implications.


Microarray analysis of pneumococcal gene expression during invasive disease.

Abstract: Streptococcus pneumoniae is a leading cause of invasive bacterial disease.

Abstract: This is the first study to examine the expression of S. pneumoniae genes in vivo by using whole-genome microarrays available from The Institute for Genomic Research.

Abstract: Total RNA was collected from pneumococci isolated from infected blood, infected cerebrospinal fluid, and bacteria attached to a pharyngeal epithelial cell line in vitro.

Abstract: Microarray analysis of pneumococcal genes expressed in these models identified body site-specific patterns of expression for virulence factors, transporters, transcription factors, translation-associated proteins, metabolism, and genes with unknown function.

Abstract: Contributions to virulence predicted for several unknown genes with enhanced expression in vivo were confirmed by insertion duplication mutagenesis and challenge of mice with the mutants.

Abstract: Finally, we cross-referenced our results with previous studies that used signature-tagged mutagenesis and differential fluorescence induction to identify genes that are potentially required by a broad range of pneumococcal strains for invasive disease.


Integration of Smad and MAPK pathways: a link and a linker revisited.

Abstract: Cells develop by reading mixed signals.

Abstract: Nowhere is this clearer than in the highly dynamic processes that propel embryogenesis, when critical cell-fate decisions are made swiftly in response to well-orchestrated growthfactor combinations.

Abstract: Learning how diverse signaling pathways are integrated is therefore essential for understanding physiology.

Abstract: This requires the identification, in tangible molecular terms, of key nodes for pathway integration that operate in vivo.

Abstract: A report in this issue, on the integration of Smad and Ras/MAPK pathways during neural induction (Pera et al. 2003), provides timely insights into the relevance of one such node.

Abstract: Pera et al. (2003) report that FGF8 and IGF2—two growth factors that activate the Ras/MAPK pathway— favor neural differentiation and mesoderm dorsalization in Xenopus by inhibiting BMP (Bone Morphogenetic Protein) signaling.

Abstract: Mesoderm is formed from ectoderm in response to Nodal-related signals from the endoderm at the blastula stage and beyond (Fig. 1; for review, see De Robertis et al. 2000).

Abstract: BMP induces differentiation of ectoderm into epidermal cell fates at the expense of neural fates, and it ventralizes the mesoderm at the expense of dorsal fates (for review, see Weinstein and HemmatiBrivanlou 1999; De Robertis et al. 2000).

Abstract: Accordingly, neural differentiation and dorsal mesoderm formation are favored when BMP signaling is attenuated.

Abstract: Noggin, Chordin, Cerberus, and Follistatin, secreted by the Spemann organizer on the dorsal side at the gastrula stage, facilitate the formation of neural tissue by sequestering BMP (Weinstein and Hemmati-Brivanlou 1999; De Robertis et al. 2000).

Abstract: Experimentally blocking BMP signaling with a dominant-negative BMP receptor has a similar effect of promoting ectoderm neuralization (Weinstein and Hemmati-Brivanlou 1999).

Abstract: As it turns out, neural induction can also be achieved with FGF (fibroblast growth factor; Kengaku and Okamoto 1993; Lamb and Harland 1995; Hongo et al. 1999; Hardcastle et al. 2000; Streit et al. 2000; Wilson et al. 2000) and IGF (insulin-like growth factor; Pera et al. 2001; Richard-Parpaillon et al. 2002).

Abstract: Injection of transcripts encoding FGF8 or IFG2 into one animal-pole blastomere of a fourto eight-cell embryo results in an expanded neural plate at the injected side (Pera et al. 2003).

Abstract: Surprisingly, expression of a dominant-negative FGF receptor prevents neuralization of ectoderm explants by the BMP blocker Noggin (Launay et al. 1996).

Abstract: Likewise, the potent neuralizing effect of Chordin can be blocked by a dominant-negative FGF receptor or a morpholino oligonucleotide targeting the IGF receptor (Pera et al. 2003).

Abstract: Thus, the neuralizing effect of BMP inhibitors is somehow tied to FGF and IFG signaling.

Abstract: The question is, how?

Abstract: Because FGF8 and IFG2 activate MAPK, Pera et al. (2003) took heed from previous work showing that MAPK inhibits the BMP signal-transduction factor Smad1 (Kretzschmar et al. 1997a).

Abstract: Smad1 is directly phosphorylated by the BMP receptor, resulting in Smad1 activation (Kretzschmar et al. 1997b), and by MAPK in response to EGF, resulting in Smad1 inhibition (Kretzschmar et al. 1997a; Fig. 2).

Abstract: Smad transcription factors mediate gene responses to the entire TGF (Transforming Growth Factor) family, to which the BMPs belong (for review, see Massague 2000; Derynck and Zhang 2003).

Abstract: Smads 1, 5, and 8 act primarily downstream of BMP receptors and Smads 2 and 3 downstream of TGF , Activin and Nodal receptors.

Abstract: Smad proteins have two conserved globular domains—the MH1 and MH2 domains (Fig. 2).

Abstract: The MH1 domain is involved in DNA binding and the MH2 domain in binding to cytoplasmic retention factors, activated receptors, nucleoporins in the nuclear pore, and DNA-binding cofactors, coactivators, and corepressors in the nucleus (for review, see Shi and Massague 2003).

Abstract: Receptor-mediated phosphorylation occurs at the carboxy-terminal sequence SXS.

Abstract: This enables the nuclear accumulation of Smads and their association with the shared partner Smad4 to form transcriptional complexes that are interpreted by the cell as a function of the context (Massague 2000).

Abstract: Between the MH1 and MH2 domains lies a linker region of variable sequence and length.

Abstract: Attention was drawn to this region when it was found that EGF (epidermal growth factor), a classical activator of the Ras/ MAPK pathway, causes phosphorylation of the Smad1 linker at four MAPK sites (PXSP sequences; Kretzschmar et al. 1997a).

Abstract: This prevents the nuclear localization of Smad1 and inhibits BMP signaling.

Abstract: Mutation of these E-MAIL j-massague@ski.mskcc.org; FAX (212) 717-3298.

Abstract: Article and publication are at http://www.genesdev.org/cgi/doi/10.1101/ gad.1167003.


A brief analysis of clinical pharmacy interventions undertaken in an Australian teaching hospital.

Abstract: Selected clinical pharmacy interventions undertaken during a 30-day data capture period were analysed, seeking to gain a greater understanding of the nature of the drug-related problems involved.

Abstract: Pharmacists were asked to record only interventions that were of potentially major significance.

Abstract: A total of 67 interventions were submitted for analysis.

Abstract: In 28 cases (41.7% of the initial total) the intervention reports were excluded from further analysis after initial review.

Abstract: For the remaining 39 interventions, 20 patients (51%) were under the care of a medical unit, and cardiovascular/antithrombotic agents accounted for 17 reports (43.5%).

Abstract: The majority of interventions were implemented at the time of inpatient medication order review by the clinical pharmacist (n=25, 64%).

Abstract: The most common category of drug-related problem addressed in the interventions related to the prescription of inappropriately high doses of the correct drug for the patient (n=17, 43.6%).

Abstract: Deficiencies in technical knowledge accounted for less than 25% of all cases.


Lethal complications of typhoid-cholera-vaccination. (Case report and review of the literature).

Abstract: Simultaneous parenteral vaccination against typhoid and cholera lead to death through either anaphylactic shock or endotoxic shock in a 36-year-old male.

Abstract: At autopsy the charactertic features of shock as well as chronic interstitial myocarditis were noted.

Abstract: Moreover, fresh histiocytic and lymphocytic nodules were found in the liver, heart and meninges.

Abstract: A review of the literature dealing with lethal complications following parenteral tyhoid vaccinations shows an increased risk in debilitated persons (emaciation, stress, cold).

Abstract: Most of the fatalities occurred in persons who had previous disturbances of the cardiovascular system, as in the case reviewed here.

Abstract: Cardiac failure, Landry's paralysis, renal failure and disturbances of skin, joints and intestines may also follow typhoid vaccinations.

Abstract: However, these latter complications are usually not lethal.

Abstract: The patients presented here had many of the conditions which are known to aggravate the situation and to lead to a lethal culmination.

Abstract: The review of this case and the disucussion following it shows that only healthy persons should receive the parenteral typhoid vaccination.

Abstract: Hopefully, the presentation of this material will help prevent fatalities of this type in the future.


Functional genomics and proteomics approaches to study the ERBB network in cancer.

Abstract: Substantial progress in functional genomic and proteomic technologies has opened new perspectives in biomedical research.

Abstract: The sequence of the human genome has been mostly determined and opened new visions on its complexity and regulation.

Abstract: New technologies, like RNAi and protein arrays, allow gathering knowledge beyond single gene analysis.

Abstract: Increasingly, biological processes are studied with systems biological approaches, where qualitative and quantitative data of the components are utilized to model the respective processes, to predict effects of perturbations, and to then refine these models after experimental testing.

Abstract: Here, we describe the potential of applying functional genomics and proteomics, taking the ERBB family of growth-factor receptors as an example to study the signaling network and its impact on cancer.


Evaluation of spatial filters to create smoothed maps of health data.

Abstract: Spatial filters have been used as an easy and intuitive way to create smoothed disease maps.

Abstract: Birth weight data from New York State for 1994 and 1995 are used to compare the traditional filter type of fixed geographical size with a filter size of constant or nearly constant population size.

Abstract: The latter are more appropriate for mapping disease in geographic areas with widely varying population density, such as New York State.

Abstract: Issues such as the choice of population size for the filter, the scale of smoothing, the ability to detect true spatial variation and the ability to smooth over random spatial noise are evaluated and discussed.


Glucose tolerance and blood pressure: long term follow up in middle aged men.

Abstract: OBJECTIVE to investigate the role of glucose tolerance in the development of hypertension.

Abstract: DESIGN Retrospective analysis of the results of a health check up in a group of clinically healthy middle aged men in the late 1960s (median year 1968).

Abstract: The subjects were invited to enter into a primary prevention trial for cardiovascular disease in 1974, when they underwent clinical examination for risk factors.

Abstract: The trial was completed in 1979, when the men were re-examined.

Abstract: Follow up was in 1986.

Abstract: SETTING Institute of Occupational Health, Helsinki, Finland and second department of medicine, University of Helsinki.

Abstract: SUBJECTS In all, 3490 men born during 1919-34 participated in a health check up in the late 1960s.

Abstract: In 1974, 1815 of these men who were clinically healthy were entered into a primary prevention trial for cardiovascular disease.

Abstract: On clinical examination 1222 of the men were considered at high risk of cardiovascular disease.

Abstract: Of these, 612 received an intervention and were excluded from the study.

Abstract: A total of 593 men were without risk factors.

Abstract: The study comprised all of the men who did not have an intervention (n = 1203).

Abstract: In 1979, 1120 men were re-examined, and in 1986 945 men attended follow up.

Abstract: There were two groups for analysis: one comprising all subjects and the other comprising only men who were normotensive in 1968 and for whom complete information was available.

Abstract: INTERVENTIONS By 1979, 103 men were taking antihypertensive drugs, and by 1986, 131 were taking antihypertensive drugs and 12 were taking drugs for hyperglycaemia.

Abstract: MAIN OUTCOME MEASURES Blood glucose concentration one hour after a glucose load, blood pressure, and body weight were measured in 1968, 1974, and 1979.

Abstract: In 1986 blood pressure and body weight were recorded.

Abstract: RESULTS Men who were hypertensive in 1986 had significantly higher blood pressures (p less than 0.0001) and (after adjustment for body mass index and alcohol intake) significantly higher blood glucose concentrations one hour after a glucose load at all examinations than those who were normotensive in 1986.

Abstract: Regression analysis showed that the higher the blood glucose concentration after a glucose load in 1968 the higher the blood pressure during the following years.

Abstract: Those men between the second and third tertiles of blood glucose concentration in 1968 had a significantly higher risk of developing hypertension (odds ratio 1.71, 95% confidence interval 1.05 to 2.77) compared with those below the first tertile.

Abstract: CONCLUSION In this study men who developed hypertension tended to have shown an increased intolerance to glucose up to 18 years before the clinical manifestation of their disorder.

Abstract: Blood glucose concentration one hour after a glucose load was an independent predictor of future hypertension.


At the gates of death.

Abstract: Apoptosis that proceeds via the mitochondrial pathway involves mitochondrial outer membrane permeabilization (MOMP), responsible for the release of cytochrome c and other proteins of the mitochondrial intermembrane space.

Abstract: This essential step is controlled and mediated by proteins of the Bcl-2 family.

Abstract: The proapoptotic proteins Bax and Bak are required for MOMP, while the antiapoptotic Bcl-2 proteins, including Bcl-2, Bcl-xL, Mcl-1, and others, prevent MOMP.

Abstract: Different proapoptotic BH3-only proteins act to interfere with the function of the antiapoptotic Bcl-2 members and/or activate Bax and Bak.

Abstract: Here, we discuss an emerging view, proposed by Certo et al. in this issue of Cancer Cell, on how these interactions result in MOMP and apoptosis.


Structure of human RNase H1 complexed with an RNA/DNA hybrid: insight into HIV reverse transcription.

Abstract: We report here crystal structures of human RNase H1 complexed with an RNA/DNA substrate.

Abstract: Unlike B. halodurans RNase H1, human RNase H1 has a basic protrusion, which forms a DNA-binding channel and together with the conserved phosphate-binding pocket confers specificity for the B form and 2'-deoxy DNA.

Abstract: The RNA strand is recognized by four consecutive 2'-OH groups and cleaved by a two-metal ion mechanism.

Abstract: Although RNase H1 is overall positively charged, the substrate interface is neutral to acidic in character, which likely contributes to the catalytic specificity.

Abstract: Positions of the scissile phosphate and two catalytic metal ions are interdependent and highly coupled.

Abstract: Modeling of HIV reverse transcriptase (RT) with RNA/DNA in its RNase H active site suggests that the substrate cannot simultaneously occupy the polymerase active site and must undergo a conformational change to toggle between the two catalytic centers.

Abstract: The region that accommodates this conformational change offers a target to develop HIV-specific inhibitors.


A diffusion tensor magnetic resonance imaging study of brain tissue from patients with migraine.

Abstract: OBJECTIVE To measure in vivo, using diffusion tensor magnetic resonance imaging (DT-MRI) the extent of pathological damage of normal appearing brain tissue (NABT) from patients with migraine.

Abstract: METHODS Dual echo and DT-MRI scans of the brain were acquired from 34 patients with migraine and 17 sex and age matched healthy volunteers.

Abstract: Mean diffusivity (MD) and fractional anisotropy (FA) histograms of the NABT were obtained from all subjects and the histograms' peak heights and average NABT MD and FA measured.

Abstract: When present, average MD and FA values of T2 visible lesions were also measured.

Abstract: RESULTS In comparison with healthy volunteers, patients with migraine had lower MD histogram peak height (p=0.02) of the NABT.

Abstract: No differences were found in FA histogram derived metrics between migraine patients and healthy subjects.

Abstract: No difference was found for any MD and FA histogram derived metrics between migraine patients with and without brain MRI lesions, and between patients with and without aura.

Abstract: CONCLUSIONS This study shows that, although brain damage may extend beyond T2 weighted abnormalities in patients with migraine, the severity of these "occult" changes is mild compared with that found in other diseases associated with white matter abnormality.


Determination of chloramphenicol residues in milk by enzyme-linked immunosorbent assay: improvement by biotin-streptavidin-amplified system.

Abstract: A sensitive biotin-streptavidin amplified enzyme-linked immunosorbent assay (BA-ELISA) method was developed for the determination of chloramphenicol residues in milk.

Abstract: The biotin-streptavidin system was applied to enhance the sensitivity.

Abstract: After optimization, the detection limit of the method was found to be 0.042 +/-

Abstract: 0.006 ng mL(-1), which is 8-fold more sensitive than the traditional competitive ELISA using the same antibody and coating antigen.

Abstract: The amplification mechanism of the biotin-streptavidin system and the major factors affecting the sensitivity of detection are discussed.

Abstract: This method was successfully applied to determine the chloramphenicol residues in milk samples with a simple and rapid extraction procedure, and good recoveries (85.66-109.67%) were obtained.

Abstract: The result indicated that the biotin-streptavidin system may be a valuable tool to improve the specific detection of trace veterinary drug residues and could be widely used for routine monitoring of food samples.


Wedelolactone exhibits anti-fibrotic effects on human hepatic stellate cell line LX-2.

Abstract: Wedelolactone is a major coumarin of Eclipta prostrata, which is used for preventing liver damage.

Abstract: However the effects of wedelolactone on hepatic fibrosis remained unexplored.

Abstract: The purpose of this study was to demonstrate the anti-fibrotic effects of wedelolactone on activated human hepatic stellate cell (HSC) line LX-2 and the possible underlying mechanisms by means of MTT assay, Hoechst staining, as well as real-time quantitative PCR and western blot.

Abstract: The results showed that wedelolactone reduced the cellular viability of LX-2 in a time and dose-dependent manner.

Abstract: After treatment of wedelolactone, the expressions of collagen I and α-smooth muscle actin, two biomarkers of LX-2 activation, were remarkably decreased.

Abstract: The apoptosis of LX-2 cells was induced by wedelolactone accompanied with the decreasing expression of anti-apoptotic Bcl-2 and increasing expression of pro-apoptotic Bax.

Abstract: In addition, phosphorylated status of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) was up-regulated, but not in p38.

Abstract: Moreover, wedelolactone significantly repressed the level of phosphorylated inhibitor of nuclear factor κB (IκB) and p65 in nucleus in spite of tumor necrosis factor-α stimulation.

Abstract: In conclusion, wedelolactone could significantly inhibit the activation of LX-2 cells, the underlying mechanisms of which included inducing Bcl-2 family involved apoptosis, up-regulating phosphorylated status of ERK and JNK expressions, and inhibiting nuclear factor-κB (NF-κB) mediated activity.

Abstract: Wedelolactone might present as a useful tool for the prevention and treatment of hepatic fibrosis.


Adipose stem cells originate from perivascular cells.

Abstract: Recent research has shown that adipose tissues contain abundant MSCs (mesenchymal stem cells).

Abstract: The origin and location of the adipose stem cells, however, remain unknown, presenting an obstacle to the further purification and study of these cells.

Abstract: In the present study, we aimed at investigating the origins of adipose stem cells.

Abstract: α-SMA (α-smooth muscle actin) is one of the markers of pericytes.

Abstract: We harvested ASCs (adipose stromal cells) from α-SMA-GFP (green fluorescent protein) transgenic mice and sorted them into GFP-positive and GFP-negative cells by FACS.

Abstract: Multilineage differentiation tests were applied to examine the pluripotent ability of the α-SMA-GFP-positive and -negative cells.

Abstract: Immunofluorescent staining for α-SMA and PDGF-Rβ (platelet-derived growth factor receptor β) were applied to identify the α-SMA-GFP-positive cells.

Abstract: Then α-SMA-GFP-positive cells were loaded on a collagen-fibronectin gel with endothelial cells to test their vascularization ability both in vitro and in vivo.

Abstract: Results show that, in adipose tissue, all of the α-SMA-GFP-positive cells congregate around the blood vessels.

Abstract: Only the α-SMA-GFP-positive cells have multilineage differentiation ability, while the α-SMA-GFP-negative cells can only differentiate in an adipogenic direction.

Abstract: The α-SMA-GFP-positive cells maintained expression of α-SMA during multilineage differentiation.

Abstract: The α-SMA-GFP-positive cells can promote the vascularization of endothelial cells in three-dimensional culture both in vitro and in vivo.

Abstract: We conclude that the adipose stem cells originate from perivascular cells and congregate around blood vessels.


Interferon alpha and zidovudine therapy in adult T-cell leukaemia lymphoma: response and outcome in 15 patients.

Abstract: Adult T-cell leukaemia lymphoma (ATLL) is an aggressive disease caused by the human T-lymphotropic virus 1 (HTLV-I) with a short survival.

Abstract: Responses to interferon alpha (IFN-alpha) and zidovudine (AZT) have been documented but not with long-term follow-up.

Abstract: We treated 15 ATLL patients with IFN and AZT.

Abstract: Eleven patients had acute ATLL, two had lymphoma and two smouldering ATLL, with progression.

Abstract: The main features were: organomegaly (14), skin lesions (10), high white blood cell (WBC) count (11) and hypercalcaemia (9).

Abstract: Eleven patients had previously received chemotherapy and one had received an autograft.

Abstract: At the time of the study, seven patients had progressive disease and eight were in partial or complete clinical remission.

Abstract: Responses (PR) lasting 2+ to 44+ months were seen in 67%; 26% did not respond (NR) and one patient was not evaluable.

Abstract: Hypercalcaemia predicted a poor outcome but differences were not significant.

Abstract: Eight of the 15 patients have died 3-41 months from diagnosis.

Abstract: Median survival for the 15 patients was 18 months.

Abstract: Survival of the NR ranged from 4 to 20 months; six PR patients are alive 8-82 months from diagnosis.

Abstract: The differences in survival between NR (median: 6 months) and PR (55% of patients alive at 4 years) were statistically significant (P = 0.002).

Abstract: In conclusion, IFN and AZT improves the outcome of ATLL patients and helps maintain responses.


Replisome stalling and stabilization at CGG repeats, which are responsible for chromosomal fragility

Abstract: Expanded CGG repeats cause chromosomal fragility and hereditary neurological disorders in humans.

Abstract: Replication forks stall at CGG repeats in a length-dependent manner in primate cells and in yeast.

Abstract: Saccharomyces cerevisiae proteins Tof1 and Mrc1 facilitate replication fork progression through CGG repeats.

Abstract: Remarkably, the fork-stabilizing role of Mrc1 does not involve its checkpoint function.

Abstract: Thus, chromosomal fragility might occur when forks stalled at expanded CGG repeats escape the S-phase checkpoint.


Direct reprogramming of fibroblasts into neural stem cells by defined factors.

Abstract: Recent studies have shown that defined sets of transcription factors can directly reprogram differentiated somatic cells to a different differentiated cell type without passing through a pluripotent state, but the restricted proliferative and lineage potential of the resulting cells limits the scope of their potential applications.

Abstract: Here we show that a combination of transcription factors (Brn4/Pou3f4, Sox2, Klf4, c-Myc, plus E47/Tcf3) induces mouse fibroblasts to directly acquire a neural stem cell identity-which we term as induced neural stem cells (iNSCs).

Abstract: Direct reprogramming of fibroblasts into iNSCs is a gradual process in which the donor transcriptional program is silenced over time.

Abstract: iNSCs exhibit cell morphology, gene expression, epigenetic features, differentiation potential, and self-renewing capacity, as well as in vitro and in vivo functionality similar to those of wild-type NSCs.

Abstract: We conclude that differentiated cells can be reprogrammed directly into specific somatic stem cell types by defined sets of specific transcription factors.


Epigenetics in lung fibrosis: from pathobiology to treatment perspective.

Abstract: PURPOSE OF REVIEW Idiopathic pulmonary fibrosis (IPF) is a fatal disease with limited treatment options and extensive gene expression changes identified in the lung parenchyma.

Abstract: Multiple lines of evidence suggest that epigenetic factors contribute to dysregulation of gene expression in IPF lung.

Abstract: Most importantly, risk factors that predispose to IPF - age, sex, cigarette smoke, and genetic variants - all influence epigenetic marks.

Abstract: This review summarizes recent findings of association of DNA methylation and histone modifications with the presence of disease and fibroproliferation.

Abstract: RECENT FINDINGS In addition to targeted studies focused on specific gene loci, genome-wide profiles of DNA methylation demonstrate widespread DNA methylation changes in IPF lung tissue and a substantial effect of these methylation changes on gene expression.

Abstract: Genetic loci that have been recently associated with IPF also contain differentially methylated regions, suggesting that genetic and epigenetic factors act in concert to dysregulate gene expression in IPF lung.

Abstract: SUMMARY Although we are in very early stages of understanding the role of epigenetics in IPF, the potential for the use of epigenetic marks as biomarkers and therapeutic targets is high and discoveries made in this field will likely bring us closer to better prognosticating and treating this fatal disease.


Novel roles for KLF1 in erythropoiesis revealed by mRNA-seq.

Abstract: KLF1 (formerly known as EKLF) regulates the development of erythroid cells from bi-potent progenitor cells via the transcriptional activation of a diverse set of genes.

Abstract: Mice lacking Klf1 die in utero prior to E15 from severe anemia due to the inadequate expression of genes controlling hemoglobin production, cell membrane and cytoskeletal integrity, and the cell cycle.

Abstract: We have recently described the full repertoire of KLF1 binding sites in vivo by performing KLF1 ChIP-seq in primary erythroid tissue (E14.5 fetal liver).

Abstract: Here we describe the KLF1-dependent erythroid transcriptome by comparing mRNA-seq from Klf1(+/+) and Klf1(-/-) erythroid tissue.

Abstract: This has revealed novel target genes not previously obtainable by traditional microarray technology, and provided novel insights into the function of KLF1 as a transcriptional activator.

Abstract: We define a cis-regulatory module bound by KLF1, GATA1, TAL1, and EP300 that coordinates a core set of erythroid genes.

Abstract: We also describe a novel set of erythroid-specific promoters that drive high-level expression of otherwise ubiquitously expressed genes in erythroid cells.

Abstract: Our study has identified two novel lncRNAs that are dynamically expressed during erythroid differentiation, and discovered a role for KLF1 in directing apoptotic gene expression to drive the terminal stages of erythroid maturation.


Self-assembling peptides optimize the post-traumatic milieu and synergistically enhance the effects of neural stem cell therapy after cervical spinal cord injury.

Abstract: INTRODUCTION The hostile environment after spinal cord injury (SCI) can compromise effects of regenerative therapies.

Abstract: We hypothesized that optimizing the post-traumatic environment with QL6 self-assembling peptides (SAPs) before neural precursor cell (NPC) transplantation would improve cell survival, differentiation and functional recovery.

Abstract: METHODS A total of 90 Wistar rats received a clip-compression SCI at C7.

Abstract: Within each of two study arms, animals were randomized into 5 groups (NPC, SAP, NPC+SAP, vehicle, and sham).

Abstract: SAPs and NPCs were injected into the spinal cord 1day and 14days post-injury, respectively.

Abstract: Animals received growth factors over 7days and were immunosuppressed.

Abstract: Rats were sacrificed at 4weeks and sections of the cervical spinal cord prepared for immunohistochemistry (first study arm).

Abstract: Neurological function was assessed weekly for 8weeks using a battery of behavioral tests.

Abstract: Nine weeks post-SCI, the corticospinal tract was assessed using fiber-tracking (second arm).

Abstract: RESULTS SAP-treated animals had significantly more surviving NPCs which showed increased differentiation to neurons and oligodendrocytes compared to controls.

Abstract: SAPs alone or in combination with NPCs resulted in smaller intramedullary cysts and larger volume of preserved tissue compared to other groups.

Abstract: The combined treatment group showed reduced astrogliosis and chondroitin sulfate proteoglycan deposition.

Abstract: Synaptic connectivity was increased in the NPC and combined treatment groups.

Abstract: Corticospinal tract preservation and behavioral outcomes improved with combinatorial treatment.

Abstract: CONCLUSION Injecting SAPs after SCI enhances subsequent NPC survival, integration and differentiation and improves functional recovery.

Abstract: STATEMENT OF SIGNIFICANCE The hostile environment after spinal cord injury (SCI) can compromise effects of regenerative therapies.

Abstract: We hypothesized that improving this environment with self-assembling peptides (SAPs) before neural precursor cell (NPC) transplantation would support their beneficial effects.

Abstract: SAPs assemble once injected, providing a supportive scaffold for repair and regeneration.

Abstract: We investigated this in a rat model of spinal cord injury.

Abstract: More NPCs survived in SAP-treated animals and these showed increased differentiation compared to controls.

Abstract: SAPS alone or in combination with NPCs resulted in smaller cysts and larger volume of preserved tissue with the combined treatment also reducing scarring and improving behavioral outcomes.

Abstract: Overall, injection of SAPs was shown to improve the efficacy of NPC treatment, a promising finding for those with SCIs.


A direct role for IFN-gamma in regulation of Th1 cell development.

Abstract: IL-12 has been identified as a major cytokine influencing the differentiation of CD4 cells to a Th1 phenotype, whereas a role for IFN-gamma is controversial.

Abstract: We investigated the interrelationship between IL-12 and IFN-gamma in promoting Th1 responses using naive CD4 cells reactive with pigeon cytochrome c from TCR transgenics and memory CD4 cells derived by in vivo priming with KLH.

Abstract: Without exogenous rIL-12 or rIFN-gamma, primary and memory effectors induced by Ag or anti-CD3 and anti-CD28 secreted variable levels of IL-2 and IFN-gamma.

Abstract: The level of IFN-gamma secreted by effectors correlated with endogenous IFN-gamma produced in primary cultures, and anti-IFN-gamma largely inhibited the development of effectors producing IFN-gamma.

Abstract: With optimal TCR stimulation and costimulation, endogenous IFN-gamma, without IL-12, was sufficient to elicit Th1 cells via an autocrine mechanism, whereas with suboptimal stimulation, exogenous rIFN-gamma or rIL-12 was required for Th1 development.

Abstract: However, rIL-12 was more effective than rIFN-gamma, partially because rIL-12 greatly enhanced autocrine production of IFN-gamma, and optimal development of the Th1 phenotype was mediated by the synergistic actions of both cytokines.

Abstract: Thus, both IFN-gamma and IL-12 can independently regulate Th1 development, but because of IFN-gamma-mediated feedback, their relative contributions are determined by the conditions of T cell stimulation.

Abstract: The extent of differentiation to a Th1 phenotype may, therefore, depend on the availability of both APC-derived IL-12 and autocrine IFN-gamma consequent to the overall strength of T cell stimulation.


Neuregulin 1 promotes excitatory synapse development and function in GABAergic interneurons.

Abstract: Neuregulin 1 (NRG1) and its receptor ErbB4 are both susceptibility genes of schizophrenia.

Abstract: However, little is known about the underlying mechanisms of their malfunction.

Abstract: Although ErbB4 is enriched in GABAergic interneurons, the role of NRG1 in excitatory synapse formation in these neurons remains poorly understood.

Abstract: We showed that NRG1 increased both the number and size of PSD-95 puncta and the frequency and amplitude of miniature EPSCs (mEPSCs) in GABAergic interneurons, indicating that NRG1 stimulates the formation of new synapses and strengthens existing synapses.

Abstract: In contrast, NRG1 treatment had no effect on either the number or size of excitatory synapses in glutamatergic neurons, suggesting its synaptogenic effect is specific to GABAergic interneurons.

Abstract: Ecto-ErbB4 treatment diminished both the number and size of excitatory synapses, suggesting that endogenous NRG1 may be critical for basal synapse formation.

Abstract: NRG1 could stimulate the stability of PSD-95 in the manner that requires tyrosine kinase activity of ErbB4.

Abstract: Finally, deletion of ErbB4 in parvalbumin-positive interneurons led to reduced frequency and amplitude of mEPSCs, providing in vivo evidence that ErbB4 is important in excitatory synaptogenesis in interneurons.

Abstract: Together, our findings suggested a novel synaptogenic role of NRG1 in excitatory synapse development, possibly via stabilizing PSD-95, and this effect is specific to GABAergic interneurons.

Abstract: In light of the association of the genes of both NRG1 and ErbB4 with schizophrenia and dysfunction of GABAergic system in this disorder, these results provide insight into its potential pathological mechanism.


Tolerance rather than immunity protects from Helicobacter pylori-induced gastric preneoplasia.

Abstract: BACKGROUND & AIMS Chronic infection with the bacterial pathogen Helicobacter pylori causes gastric disorders, ranging from chronic gastritis to gastric adenocarcinoma.

Abstract: Only a subset of infected persons will develop overt disease; most remains asymptomatic despite lifelong colonization.

Abstract: This study aims to elucidate the differential susceptibility to H pylori that is found both across and within populations.

Abstract: METHODS We have established a C57BL/6 mouse model of H pylori infection with a strain that is capable of delivering the virulence factor cytotoxin-associated gene A (CagA) into host cells through the activity of a Cag-pathogenicity island-encoded type IV secretion system.

Abstract: RESULTS Mice infected at 5-6 weeks of age with CagA(+)H pylori rapidly develop gastritis, gastric atrophy, epithelial hyperplasia, and metaplasia in a type IV secretion system-dependent manner.

Abstract: In contrast, mice infected during the neonatal period with the same strain are protected from preneoplastic lesions.

Abstract: Their protection results from the development of H pylori-specific peripheral immunologic tolerance, which requires transforming growth factor-β signaling and is mediated by long-lived, inducible regulatory T cells, and which controls the local CD4(+) T-cell responses that trigger premalignant transformation.

Abstract: Tolerance to H pylori develops in the neonatal period because of a biased ratio of T-regulatory to T-effector cells and is favored by prolonged low-dose exposure to antigen.

Abstract: CONCLUSIONS Using a novel CagA(+)H pylori infection model, we report here that the development of tolerance to H pylori protects from gastric cancer precursor lesions.

Abstract: The age at initial infection may thus account for the differential susceptibility of infected persons to H pylori-associated disease manifestations.


Impaired mitochondrial fatty acid oxidation and insulin resistance in aging: novel protective role of glutathione.

Abstract: Aging is associated with impaired fasted oxidation of nonesterified fatty acids (NEFA) suggesting a mitochondrial defect.

Abstract: Aging is also associated with deficiency of glutathione (GSH), an important mitochondrial antioxidant, and with insulin resistance.

Abstract: This study tested whether GSH deficiency in aging contributes to impaired mitochondrial NEFA oxidation and insulin resistance, and whether GSH restoration reverses these defects.

Abstract: Three studies were conducted: (i) in 82-week-old C57BL/6 mice, the effect of naturally occurring GSH deficiency and its restoration on mitochondrial (13) C1 -palmitate oxidation and glucose metabolism was compared with 22-week-old C57BL/6 mice; (ii) in 20-week C57BL/6 mice, the effect of GSH depletion on mitochondrial oxidation of (13) C1 -palmitate and glucose metabolism was studied; (iii) the effect of GSH deficiency and its restoration on fasted NEFA oxidation and insulin resistance was studied in GSH-deficient elderly humans, and compared with GSH-replete young humans.

Abstract: Chronic GSH deficiency in old mice and elderly humans was associated with decreased fasted mitochondrial NEFA oxidation and insulin resistance, and these defects were reversed with GSH restoration.

Abstract: Acute depletion of GSH in young mice resulted in lower mitochondrial NEFA oxidation, but did not alter glucose metabolism.

Abstract: These data suggest that GSH is a novel regulator of mitochondrial NEFA oxidation and insulin resistance in aging.

Abstract: Chronic GSH deficiency promotes impaired NEFA oxidation and insulin resistance, and GSH restoration reverses these defects.

Abstract: Supplementing diets of elderly humans with cysteine and glycine to correct GSH deficiency could provide significant metabolic benefits.


Control of central and peripheral tolerance by Aire.

Abstract: The negative selection of self-reactive thymocytes depends on the expression of tissue-specific antigens by medullary thymic epithelial cells.

Abstract: The autoimmune regulator (Aire) protein plays an important role in turning on these antigens, and the absence of even one Aire-induced tissue-specific antigen in the thymus can lead to autoimmunity in the antigen-expressing target organ.

Abstract: Recently, Aire protein has been detected in peripheral lymphoid organs, suggesting that peripheral Aire plays a complementary role here.

Abstract: In these peripheral sites, Aire was found to regulate the expression of a group of tissue-specific antigens that is distinct from those expressed in the thymus.

Abstract: Furthermore, transgenic antigen expression in extrathymic Aire-expressing cells (eTACs) can mediate deletional tolerance, but the immunological relevance of Aire-dependent, endogenous tissue-specific antigens remains to be determined.


Congenital Hypogonadotropic Hypogonadism: A Trait Shared by Several Complex Neurodevelopmental Disorders.

Abstract: Reproductive function depends on the activity of the gonadotropic axis, which is controlled by a hypothalamic neural network whose main function is to regulate the secretion of gonadotropin-releasing hormone (GnRH).

Abstract: This endocrine network is not mature at birth, and several phases of activation-inactivation of the gonadotropic axis are necessary for its normal development.

Abstract: The postnatal maturation of the GnRH network lies under the control of a neurodevelopmental program that starts in fetal life and ends at puberty.

Abstract: There are many clinical situations in which this program is interrupted, leading to congenital hypogonadotropic hypogonadism (CHH) and an absence of puberty.

Abstract: For many years, attention has mainly been focused on the genetics of isolated CHH.

Abstract: More recently, the emergence of new genomics techniques has led to the description of genetic defects in very rare syndromes in which CHH is associated with complex neurological dysfunctions.

Abstract: Here, we review the clinical phenotype and genetic defects linked to such syndromic CHH.

Abstract: This analysis highlights the close link between the ubiquitin pathway, synaptic proteins and CHH, as well as unexpected mutations in genes encoding nucleolar proteins.


Elevated expression of miR-24-3p is a potentially adverse prognostic factor in colorectal adenocarcinoma.

Abstract: OBJECTIVES Several miRNAs are aberrantly expressed in cancer.

Abstract: miR-24-3p is involved in cancer-related cellular processes, including cell cycle control, cell growth, proliferation, and apoptosis.

Abstract: In this study, we examined the potential diagnostic and prognostic significance of miR-24-3p expression in colorectal adenocarcinoma.

Abstract: DESIGN AND METHODS Total RNA was isolated from 182 colorectal adenocarcinoma specimens and 86 paired non-cancerous colorectal mucosae.

Abstract: After polyadenylation of 2μg total RNA and reverse transcription into first-strand cDNA using an oligo-dT-adapter primer, miR-24-3p expression was quantified using an in-house-developed reverse-transcription real-time quantitative PCR method, based on the SYBR Green chemistry.

Abstract: SNORD43 (RNU43) was used as a reference gene.

Abstract: RESULTS miR-24-3p levels do not significantly differ between colorectal adenocarcinoma and non-cancerous colorectal mucosae.

Abstract: Thus, miR-24-3p expression cannot be used for diagnostic purposes.

Abstract: However, high miR-24-3p expression predicts poor disease-free survival (DFS) and overall survival (OS) of colorectal adenocarcinoma patients.

Abstract: Multivariate Cox regression analysis confirmed that miR-24-3p overexpression is a significant predictor of relapse in colorectal adenocarcinoma and that its prognostic significance is independent of other established prognostic factors and treatment of patients.

Abstract: Of note, miR-24-3p overexpression retains its rather unfavorable prognostic value in the subgroup of patients with advanced yet locally restricted colorectal adenocarcinoma (T3) and in those without distant metastasis (M0).

Abstract: Moreover, miR-24-3p overexpression is a potentially unfavorable prognosticator for patients who were not treated with radiotherapy.

Abstract: CONCLUSIONS Strong expression of miR-24-3p predicts poor DFS and OS of colorectal adenocarcinoma patients, independently of clinicopathological parameters that are currently used for prognosis in this human malignancy.


Reforming the health sector in developing countries: the central role of policy analysis.

Abstract: Policy analysis is an established discipline in the industrialized world, yet its application to developing countries has been limited.

Abstract: The health sector in particular appears to have been neglected.

Abstract: This is surprising because there is a well recognized crisis in health systems, and prescriptions abound of what health policy reforms countries should introduce.

Abstract: However, little attention has been paid to how countries should carry out reforms, much less who is likely to favour or resist such policies.

Abstract: This paper argues that much health policy wrongly focuses attention on the content of reform, and neglects the actors involved in policy reform (at the international, national sub-national levels), the processes contingent on developing and implementing change and the context within which policy is developed.

Abstract: Focus on policy content diverts attention from understanding the processes which explain why desired policy outcomes fail to emerge.

Abstract: The paper is organized in 4 sections.

Abstract: The first sets the scene, demonstrating how the shift from consensus to conflict in health policy established the need for a greater emphasis on policy analysis.

Abstract: The second section explores what is meant by policy analysis.

Abstract: The third investigates what other disciplines have written that help to develop a framework of analysis.

Abstract: And the final section suggests how policy analysis can be used not only to analyze the policy process, but also to plan.


The processing of Holliday junctions by BLM and WRN helicases is regulated by p53.

Abstract: BLM, WRN, and p53 are involved in the homologous DNA recombination pathway.

Abstract: The DNA structure-specific helicases, BLM and WRN, unwind Holliday junctions (HJ), an activity that could suppress inappropriate homologous recombination during DNA replication.

Abstract: Here, we show that purified, recombinant p53 binds to BLM and WRN helicases and attenuates their ability to unwind synthetic HJ in vitro.

Abstract: The p53 248W mutant reduces abilities of both to bind HJ and inhibit helicase activities, whereas the p53 273H mutant loses these abilities.

Abstract: Moreover, full-length p53 and a C-terminal polypeptide (residues 373-383) inhibit the BLM and WRN helicase activities, but phosphorylation at Ser(376) or Ser(378) completely abolishes this inhibition.

Abstract: Following blockage of DNA replication, Ser(15) phospho-p53, BLM, and RAD51 colocalize in nuclear foci at sites likely to contain DNA replication intermediates in cells.

Abstract: Our results are consistent with a novel mechanism for p53-mediated regulation of DNA recombinational repair that involves p53 post-translational modifications and functional protein-protein interactions with BLM and WRN DNA helicases.


Trim and fill: A simple funnel-plot-based method of testing and adjusting for publication bias in meta-analysis.

Abstract: We study recently developed nonparametric methods for estimating the number of missing studies that might exist in a meta-analysis and the effect that these studies might have had on its outcome.

Abstract: These are simple rank-based data augmentation techniques, which formalize the use of funnel plots.

Abstract: We show that they provide effective and relatively powerful tests for evaluating the existence of such publication bias.

Abstract: After adjusting for missing studies, we find that the point estimate of the overall effect size is approximately correct and coverage of the effect size confidence intervals is substantially improved, in many cases recovering the nominal confidence levels entirely.

Abstract: We illustrate the trim and fill method on existing meta-analyses of studies in clinical trials and psychometrics.


Familial Barrett's esophagus associated with adenocarcinoma.

Abstract: A family with six cases of Barrett's esophagus over three generations is described.

Abstract: The Barrett's esophagus affected only males, and there were three associated adenocarcinomas.

Abstract: The mechanism of inheritance is compatible with an autosomal dominant pattern with nearly complete penetrance.

Abstract: Other case reports in the literature are compared with this familial case of Barrett's esophagus with associated adenocarcinomas.


Development of an insect model for the in vivo pathogenicity testing of yeasts.

Abstract: Conventional in vivo assays to determine the relative pathogenicity of yeast isolates rely upon the use of a range of mammalian species.

Abstract: The purpose of the work presented here was to investigate the possibility of using an insect (Galleria mellonella) as a model system for in vivo pathogenicity testing.

Abstract: The haemolymph of G. mellonella larvae was inoculated with PBS containing different concentrations of stationary phase yeasts of the genus Candida by injection at the last pro-leg.

Abstract: Larvae were incubated at 30 degrees C and monitored over 72 hours.

Abstract: Results indicate that G. mellonella can be killed by the pathogenic yeast Candida albicans and by a range of other Candida species but not to a significant extent by the yeast Saccharomyces cerevisiae.

Abstract: The kill kinetics for larvae inoculated with clinical and laboratory isolates of C. albicans indicate the former class of isolates to be more pathogenic.

Abstract: Differences in the relative pathogenicity of a range of Candida species may be distinguished using G. mellonella as a model.

Abstract: This work indicates that G. mellonella may be employed to give results consistent with data previously obtained using mammals in conventional in vivo pathogenicity testing.

Abstract: Larvae of G. mellonella are inexpensive to culture, easy to manipulate and their use may reduce the need to employ mammals for routine in vivo pathogenicity testing with a concomitant reduction in mammalian suffering.


Use of TLS parameters to model anisotropic displacements in macromolecular refinement.

Abstract: An essential step in macromolecular refinement is the selection of model parameters which give as good a description of the experimental data as possible while retaining a realistic data-to-parameter ratio.

Abstract: This is particularly true of the choice of atomic displacement parameters, where the move from individual isotropic to individual anisotropic refinement involves a sixfold increase in the number of required displacement parameters.

Abstract: The number of refinement parameters can be reduced by using collective variables rather than independent atomic variables and one of the simplest examples of this is the TLS parameterization for describing the translation, libration and screw-rotation displacements of a pseudo-rigid body.

Abstract: This article describes the implementation of the TLS parameterization in the macromolecular refinement program REFMAC.

Abstract: Derivatives of the residual with respect to the TLS parameters are expanded in terms of the derivatives with respect to individual anisotropic U values, which in turn are calculated using a fast Fourier transform technique.

Abstract: TLS refinement is therefore fast and can be used routinely.

Abstract: Examples of TLS refinement are given for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and a transcription activator GerE, for both of which there is data to only 2.0 A, so that individual anisotropic refinement is not feasible.

Abstract: GAPDH has been refined with between one and four TLS groups in the asymmetric unit and GerE with six TLS groups.

Abstract: In both cases, inclusion of TLS parameters gives improved refinement statistics and in particular an improvement in R and free R values of several percent.

Abstract: Furthermore, GAPDH and GerE have two and six molecules in the asymmetric unit, respectively, and in each case the displacement parameters differ significantly between molecules.

Abstract: These differences are well accounted for by the TLS parameterization, leaving residual local displacements which are very similar between molecules and to which NCS restraints can be applied.


Sequence features surrounding the translation initiation sites assigned on the genome sequence of Synechocystis sp. strain PCC6803 by amino-terminal protein sequencing.

Abstract: To characterize the sequence features surrounding the translation initiation sites on the genome of Synechocystis sp. strain 6803, the total proteins extracted from the cell were resolved by two-dimensional electrophoresis, and the amino-terminal sequences of the relatively abundant protein spots were determined.

Abstract: By comparison of the determined amino-terminal sequences with the nucleotide sequence of the entire genome, the translation initiation sites of a total of 72 proteins were successfully assigned on the genome.

Abstract: The sequence features emerged from the nucleotide sequences at and surrounding the translation initiation sites were as follows: (1) In addition to the three initiation codons, ATG, GTG, and TTG, evidence was obtained that ATT was also used as a rare initiation codon; (2) the core sequences (GAGG, GGAG and AGGA) of the Shine-Dalgarno sequence were identified in the appropriate position preceding the 35 initiation sites (48.6%); and (3) the preferential sequence surrounding the initiation codons was formulated as 5'-YY[...]R-3' where Y and R denote pyrimidine and purine nucleotides, respectively, and three dots represent the initiation codons.

Abstract: The result obtained would provide valuable information for improvement of the gene-finding software, and the approach used in this study should be applicable for comprehensive analysis of the expression profiles of cellular proteins.


Deregulated DNA polymerase beta induces chromosome instability and tumorigenesis.

Abstract: To reach the biological alterations that characterize cancer, the genome of tumor cells must acquire increased mutability resulting from a malfunction of a network of genome stability systems, e.g., cell cycle arrest, DNA repair, and high accuracy of DNA synthesis during DNA replication.

Abstract: Numeric chromosomal imbalance, referred to as aneuploidy, is the most prevalent genetic changes recorded among many types of solid tumors.

Abstract: We report here that ectopic expression in cells of DNA polymerase beta, an error-prone enzyme frequently over-regulated in human tumors, induces aneuploidy, an abnormal localization of the centrosome-associated gamma-tubulin protein during mitosis, a deficient mitotic checkpoint, and promotes tumorigenesis in nude immunodeficient mice.

Abstract: Thus, we find that alteration of polymerase beta expression appears to induce major genetic changes associated with a malignant phenotype.


Combinatorial signaling by Sonic hedgehog and Wnt family members induces myogenic bHLH gene expression in the somite.

Abstract: We have demonstrated previously that a combination of signals from the neural tube and the floor plate/notochord complex synergistically induce the expression of myogenic bHLH genes and myogenic differentiation markers in unspecified somites.

Abstract: In this study we demonstrate that Sonic hedgehog (Shh), which is expressed in the floor plate/notochord, and a subset of Wnt family members (Wnt-1, Wnt-3, and Wnt-4), which are expressed in dorsal regions of the neural tube, mimic the muscle inducing activity of these tissues.

Abstract: In combination, Shh and either Wnt-1 or Wnt-3 are sufficient to induce myogenesis in somitic tissue in vitro.

Abstract: Therefore, we propose that myotome formation in vivo may be directed by the combinatorial activity of Shh secreted by ventral midline tissues (floor plate and notochord) and Wnt ligands secreted by the dorsal neural tube.


Advances in osteoclast biology: old findings and new insights from mouse models

Abstract: The maintenance of adequate bone mass is dependent upon the controlled and timely removal of old, damaged bone.

Abstract: This complex process is performed by the highly specialized, multinucleated osteoclast.

Abstract: Over the past 15 years, a detailed picture has emerged describing the origins, differentiation pathways and activation stages that contribute to normal osteoclast function.

Abstract: This information has primarily been obtained by the development and skeletal analysis of genetically modified mouse models.

Abstract: Mice harboring mutations in specific genetic loci exhibit bone defects as a direct result of aberrations in normal osteoclast recruitment, formation or function.

Abstract: These findings include the identification of the RANK–RANKL–OPG system as a primary mediator of osteoclastogenesis, the characterization of ion transport and cellular attachment mechanisms and the recognition that matrix-degrading enzymes are essential components of resorptive activity.

Abstract: This Review focuses on the principal observations in osteoclast biology derived from genetic mouse models, and highlights emerging concepts that describe how the osteoclast is thought to contribute to the maintenance of adequate bone mass and integrity throughout life.


DNA methylation in embryonic stem cells.

Abstract: Embryonic stem cells (ESCs) are pluripotent, self-renewing cells.

Abstract: These cells can be used in applications such as cell therapy, drug development, disease modeling, and the study of cellular differentiation.

Abstract: Investigating the interplay of epigenetics, genetics, and gene expression in control of pluripotence and differentiation could give important insights on how these cells function.

Abstract: One of the best known epigenetic factors is DNA methylation, which is a major mechanism for regulation of gene expression.

Abstract: This phenomenon is mostly seen in imprinted genes and X-chromosome inactivation where DNA methylation of promoter regions leads to repression of gene expression.

Abstract: Differential DNA methylation of pluripotence-associated genes such as Nanog and Oct4/Pou5f1 has been observed between pluripotent and differentiated cells.

Abstract: It is clear that tight regulation of DNA methylation is necessary for normal development.

Abstract: As more associations between aberrant DNA methylation and disease are reported, the demand for high-throughput approaches for DNA methylation analysis has increased.

Abstract: In this article, we highlight these methods and discuss recent DNA methylation studies on ESCs.


Reserves, functional, immunoregulatory, and cytogenetic properties of bone marrow mesenchymal stem cells in patients with myelodysplastic syndromes.

Abstract: Defective hematopoiesis supporting capacity of bone marrow (BM) stroma has been implicated in the pathophysiology of myelodysplastic syndromes (MDS).

Abstract: The aim of this study is to explore whether the BM stroma progenitors, namely the mesenchymal stem cells (MSCs), are primarily affected in MDS by evaluating the reserves, the functional properties, as well as the cytogenetic characteristics, in comparison to BM hematopoietic cells, in patients with de novo MDS (n = 13).

Abstract: The number, differentiation potential toward adipocytes/chondrocytes/osteoblasts and immunosuppressive function in terms of inhibition of mitogen-induced T-cell proliferation did not differ significantly between patient and normal (n = 20) MSCs.

Abstract: Patient MSCs did not show any aberrations in the production of proinflammatory or growth-promoting cytokines and did not harbor the cytogenetic abnormalities present in hematopoietic cells.

Abstract: Occasional patient and normal MSC cultures, however, developed irrelevant chromosomal alterations (trisomies 5 and 7) with uncertain pathophysiologic significance.

Abstract: Compared to controls, patient MSCs displayed impaired proliferative and clonogenic potential through passages that might represent a nonspecific abnormality associated with the chronic inflammatory process present in patients' BM.

Abstract: These data suggest that BM MSCs from MDS patients do not belong to the abnormal clone and do not represent the main cellular source contributing to the inflammatory marrow microenvironment.


Symmetric signalling within asymmetric dimers of the Staphylococcus aureus receptor histidine kinase AgrC.

Abstract: Virulence in Staphylococcus aureus is largely under control of the accessory gene regulator (agr) quorum-sensing system.

Abstract: The AgrC receptor histidine kinase detects its autoinducing peptide (AIP) ligand and generates an intracellular signal resulting in secretion of virulence factors.

Abstract: Although agr is a well-studied quorum-sensing system, little is known about the mechanism of AgrC activation.

Abstract: By co-immunoprecipitation analysis and intermolecular complementation of receptor mutants, we showed that AgrC forms ligand-independent dimers that undergo trans-autophosphorylation upon interaction with AIP.

Abstract: Remarkably, addition of specific AIPs to AgrC mutant dimers with only one functional sensor domain caused symmetric activation of either kinase domain despite the sensor asymmetry.

Abstract: Furthermore, mutant dimers involving one constitutive protomer demonstrated ligand-independent activity, irrespective of which protomer was kinase deficient.

Abstract: These results demonstrate that signalling through either individual AgrC protomer causes symmetric activation of both kinase domains.

Abstract: We suggest that such signalling across the dimer interface may be an important mechanism for dimeric quorum-sensing receptors to rapidly elicit a response upon signal detection.


Attachment of the ubiquitin-related protein Urm1p to the antioxidant protein Ahp1p.

Abstract: Urm1p is a ubiquitin-related protein that serves as a posttranslational modification of other proteins.

Abstract: Urm1p conjugation has been implicated in the budding process and in nutrient sensing.

Abstract: Here, we have identified the first in vivo target for the urmylation pathway as the antioxidant protein Ahp1p.

Abstract: The attachment of Urm1p to Ahp1p requires the E1 for the urmylation pathway, Uba4p.

Abstract: Loss of the urmylation pathway components results in sensitivity to a thiol-specific oxidant, as does loss of Ahp1p, implying that urmylation has a role in an oxidative-stress response.

Abstract: Moreover, treatment of cells with thiol-specific oxidants affects the abundance of Ahp1p-Urm1p conjugates.

Abstract: These results suggest that the conjugation of Urm1p to Ahp1p could regulate the function of Ahp1p in antioxidant stress response in Saccharomyces cerevisiae.


Sulfasalazine in the treatment of spondylarthropathy. A randomized, multicenter, double-blind, placebo-controlled study.

Abstract: OBJECTIVE To assess the efficacy and tolerability of sulfasalazine (SSZ) in the treatment of spondylarthropathy.

Abstract: METHODS We conducted a 6-month randomized, placebo-controlled, double-blind, multicenter study of patients with spondylarthropathy whose disease had remained active despite treatment with nonsteroidal antiinflammatory drugs.

Abstract: Patients were treated with SSZ (3 gm/day) or placebo.

Abstract: The primary efficacy variables were the physician's and patient's overall assessments, pain, and morning stiffness.

Abstract: End points were analyzed in the intent-to-treat and completer patient populations; the time course of effect was analyzed in the completer patient population.

Abstract: RESULTS Of the 351 patients enrolled, 263 (75%) completed the 6-month treatment period.

Abstract: The withdrawal rates were 35 (20%) and 53 (30%) in the placebo and SSZ groups, respectively.

Abstract: In the intent-to-treat analysis of end point efficacy, the between-treatment difference reached statistical significance only for 1 of the 4 primary outcome variables, the patient's overall assessment of disease activity, for which 60% of the patients taking SSZ improved by at least 1 point on a 5-point scale, in contrast to 44% of the patients taking placebo.

Abstract: Laboratory markers of inflammation also showed statistically significant change in favor of SSZ.

Abstract: In subgroup analysis, the most impressive effects were seen in patients with psoriatic arthritis, both for the 4 primary efficacy variables and for secondary efficacy variables such as the number of inflamed joints.

Abstract: Adverse events were more frequent in the SSZ group than the placebo group, but all were transient or reversible after cessation of treatment.

Abstract: CONCLUSION The results of this study show that SSZ had greater efficacy than placebo in the treatment of active spondylarthropathy, notably in patients with psoriatic arthritis.