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1.
Nat Commun ; 11(1): 5070, 2020 10 08.
Article in English | MEDLINE | ID: mdl-33033260

ABSTRACT

The evolutionary progression from primary to metastatic prostate cancer is largely uncharted, and the implications for liquid biopsy are unexplored. We infer detailed reconstructions of tumor phylogenies in ten prostate cancer patients with fatal disease, and investigate them in conjunction with histopathology and tumor DNA extracted from blood and cerebrospinal fluid. Substantial evolution occurs within the prostate, resulting in branching into multiple spatially intermixed lineages. One dominant lineage emerges that initiates and drives systemic metastasis, where polyclonal seeding between sites is common. Routes to metastasis differ between patients, and likely genetic drivers of metastasis distinguish the metastatic lineage from the lineage that remains confined to the prostate within each patient. Body fluids capture features of the dominant lineage, and subclonal expansions that occur in the metastatic phase are non-uniformly represented. Cerebrospinal fluid analysis reveals lineages not detected in blood-borne DNA, suggesting possible clinical utility.


Subject(s)
Cell Lineage , Liquid Biopsy , Prostatic Neoplasms/pathology , Body Fluids/metabolism , Chromosomes, Human, Pair 8/genetics , Clone Cells , DNA Copy Number Variations/genetics , DNA, Neoplasm/genetics , Genetic Loci , Humans , Male , Middle Aged , Neoplasm Metastasis , Phylogeny
2.
J Dent Res ; 98(9): 975-984, 2019 08.
Article in English | MEDLINE | ID: mdl-31329044

ABSTRACT

Understanding of the oral microbiome in relation to periodontal disease in older adults is limited. The composition and diversity of the subgingival microflora and their oligotypes in health and levels of periodontal disease were investigated in this study on older postmenopausal women. The 16S rRNA gene was sequenced using the Illumina MiSeq platform in 1,206 women aged 53 to 81 y. Presence and severity of periodontal disease were defined by Centers for Disease Control and Prevention/American Academy of Periodontology criteria. Composition of the microbiome was determined by 16S rRNA amplicon sequencing and the abundance of taxa described by the centered log2-ratio (CLR) transformed operational taxonomic unit (OTU) values. Differences according to periodontal disease status were determined by analysis of variance with Bonferroni correction. Bacteria oligotypes associated with periodontal disease and health were determined by minimum entropy decomposition and their functions estimated in silico using PICRUSt. Prevalence of none/mild, moderate, and severe periodontal disease was 25.1%, 58.3%, and 16.6%, respectively. Alpha diversity of the microbiome differed significantly across the 3 periodontal disease categories. ß-Diversity differed between no/mild and severe periodontal disease, although considerable overlap was noted. Of the 267 bacterial species identified at ≥0.02% abundance, 56 (20.9%) differed significantly in abundance according to periodontal disease status. Significant linear correlations for pocket depth and clinical attachment level with bacterial amounts were observed for several taxa. Of the taxa differing in abundance according to periodontal disease status, 53% had multiple oligotypes appearing to differ between none/mild and severe periodontal disease. Among older women, taxonomic differences in subgingival microbiome composition and diversity were observed in relation to clinical periodontal disease measures. Potential differences in bacterial subspecies (oligotypes) and their function were also identified in periodontal disease compared with health.


Subject(s)
Gingiva/microbiology , Microbiota , Periodontitis/microbiology , Aged , Aged, 80 and over , Bacteria , Female , Humans , Middle Aged , RNA, Ribosomal, 16S/genetics
3.
Arch Womens Ment Health ; 21(3): 365-374, 2018 06.
Article in English | MEDLINE | ID: mdl-29264646

ABSTRACT

This study aimed to investigate prospectively the contribution of maternal physical health and/or breastfeeding problems to maternal mood (depression, anxiety, fatigue, irritability, confusion, vigor) at 8-weeks postpartum. A prospective study was conducted. Participants were recruited antenatally from a public and a private maternity hospital in Melbourne, Australia. Nulliparous pregnant women (N = 229), ≥ 18 years of age, ≥ 36-week gestation, singleton pregnancy and with sufficient English were eligible. Data were collected by self-report questionnaire (pregnancy, weeks 1-4 postpartum) and telephone interview (week 8 postpartum). A high burden of physical problems was classified as ≥ 3 problems (caesarean/perineal pain; back pain; constipation; haemorrhoids; urinary and bowel incontinence) for ≥ 2 time points. A high burden of breastfeeding problems was having ≥ 2 problems (mastitis; nipple pain; frequent expressing; over- or under-supply of milk) for ≥ 2 time points. Multivariate linear regression was used to investigate the relationship between maternal mood, assessed using Profile of Mood States (8-week postpartum), and a high burden of breastfeeding and/or physical health problems. Forty-six women (20.1%) had a high burden of physical symptoms, 44 (19.2%) a high burden of breastfeeding problems only and 25 women (11.0%) had both. A high burden of breastfeeding problems alone (ß = 10.6, p = 0.01) or with co-morbid physical problems (ß = 15.35, p = 0.002) was significantly associated with poorer maternal mood at 8 weeks. Early, effective postnatal treatment of maternal health and breastfeeding problems could reduce women's risk for poor mental health.


Subject(s)
Breast Feeding/psychology , Depression, Postpartum/psychology , Maternal Behavior/psychology , Maternal Health , Mood Disorders/psychology , Depression, Postpartum/epidemiology , Female , Health Status , Humans , Maternal Age , Mood Disorders/epidemiology , Postpartum Period , Prospective Studies , Young Adult
4.
Ann Oncol ; 29(3): 737-743, 2018 03 01.
Article in English | MEDLINE | ID: mdl-29267856

ABSTRACT

Background: Health-related quality of life (HRQoL) was a secondary end point in AGO-OVAR 16, which randomized 940 patients with EOC after first-line chemotherapy to maintenance pazopanib (PZ) or placebo (P). Additional post hoc analyses were carried out to investigate additional patient-centered end points. Patients and methods: HRQoL was measured with EORTC-QLQ-C30, QLQ-OV28 and EQ-5D-3L. Pre-specified end points included mean differences in HRQoL between treatment arms. Exploratory analyses included quality-adjusted progression-free survival (QAPFS), impact of specific symptoms and progressive disease (PD) on HRQoL and time to second-line chemotherapy. The objective was to provide clinical perspective to the significant median PFS gain of 5.6 months with PZ. Results: There were statistically significant differences between PZ and P in QLQ-C30 global health status [5.5 points; 95% confidence interval (CI), 0.7-10.4, P = 0.024] from baseline to 25 months, but not EQ-5D-3L (0.018 points; 95% CI - 0.033 to 0.069, P = 0.485). The impact of diarrhea was captured in QLQ-OV28 Abdominal/GI-Symptoms scale (8.1 points; 95% CI 3.6-12.5, P = 0.001). QAPFS was 386 days (95% CI 366-404 days) with PZ versus 359 days (95% CI 338-379 days) with placebo (P = 0.052). PD was associated with a decline in HRQoL (P < 0.0001). Median time to second-line chemotherapy was 19.7 months with PZ and 15.0 months with P [hazard ratio (HR) 0.72, 95% CI 0.69-0.86, P = 0.0001]. Conclusions: There were small to no significant mean score differences in global HRQoL and EQ5D-3L between PZ and placebo, respectively, despite the increased toxicity of PZ. Exploratory end points including QAPFS, impact of specific symptoms on HRQoL during treatment and at PD help place the PFS gain with PZ in context and interpret the results. Additional patient-centered end points should be considered in trials of maintenance therapy in EOC beyond mean differences in HRQoL scores alone, to support the benefit to patients of prolongation of PFS. Clinical Trials Registration Number: NCT00866697.


Subject(s)
Antineoplastic Agents/adverse effects , Carcinoma, Ovarian Epithelial/drug therapy , Maintenance Chemotherapy/adverse effects , Ovarian Neoplasms/drug therapy , Pyrimidines/adverse effects , Quality of Life , Sulfonamides/adverse effects , Adult , Angiogenesis Inhibitors/adverse effects , Double-Blind Method , Female , Humans , Indazoles , Maintenance Chemotherapy/methods , Middle Aged , Neoplasm Recurrence, Local/drug therapy , Patient Reported Outcome Measures , Progression-Free Survival , Time-to-Treatment
5.
Clin Exp Immunol ; 187(1): 113-123, 2017 Jan.
Article in English | MEDLINE | ID: mdl-27667787

ABSTRACT

Tolerogenic dendritic cells (tolDC) are a new immunotherapeutic tool for the treatment of rheumatoid arthritis (RA) and other autoimmune disorders. We have established a method to generate stable tolDC by pharmacological modulation of human monocyte-derived DC. These tolDC exert potent pro-tolerogenic actions on CD4+ T cells. Lack of interleukin (IL)-12p70 production is a key immunoregulatory attribute of tolDC but does not explain their action fully. Here we show that tolDC express transforming growth factor (TGF)-ß1 at both mRNA and protein levels, and that expression of this immunoregulatory cytokine is significantly higher in tolDC than in mature monocyte-derived DC. By inhibiting TGF-ß1 signalling we demonstrate that tolDC regulate CD4+ T cell responses in a manner that is at least partly dependent upon this cytokine. Crucially, we also show that while there is no significant difference in expression of TGF-ßRII on CD4+ T cells from RA patients and healthy controls, RA patient CD4+ T cells are measurably less responsive to TGF-ß1 than healthy control CD4+ T cells [reduced TGF-ß-induced mothers against decapentaplegic homologue (Smad)2/3 phosphorylation, forkhead box protein 3 (FoxP3) expression and suppression of (IFN)-γ secretion]. However, CD4+ T cells from RA patients can, nonetheless, be regulated efficiently by tolDC in a TGF-ß1-dependent manner. This work is important for the design and development of future studies investigating the potential use of tolDC as a novel immunotherapy for the treatment of RA.


Subject(s)
Arthritis, Rheumatoid/therapy , CD4-Positive T-Lymphocytes/immunology , Dendritic Cells/immunology , Immune Tolerance , Immunotherapy/methods , Transforming Growth Factor beta1/metabolism , Arthritis, Rheumatoid/immunology , Cells, Cultured , Cholecalciferol/pharmacology , Dendritic Cells/drug effects , Dendritic Cells/transplantation , Dexamethasone/pharmacology , Forkhead Transcription Factors/metabolism , Humans , Immunomodulation , Interleukin-12/genetics , Interleukin-12/metabolism , Lymphocyte Activation , Smad2 Protein/metabolism
6.
J Fish Biol ; 89(3): 1866-72, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27329496

ABSTRACT

In this study, population level lateralisation and the suitability of mirror tests as a test of natural aggressive behaviour in male rainbow kribs Pelvicachromis pulcher was investigated. Aggressive behaviour in live agonistic trials correlated positively with behaviours towards a mirror image and no visual lateralisation was detected.


Subject(s)
Agonistic Behavior , Cichlids , Aggression , Animals , Male
7.
Pathologe ; 36(4): 372-84, 2015 Jul.
Article in German | MEDLINE | ID: mdl-26175197

ABSTRACT

Chronic myeloid leukemia (CML) is diagnostically defined by the reciprocal translocation t(9;22)(q34;q11). This aberration can be detected by the BCR-ABL fluorescence in situ hybridization (FISH) technique. This article presents a comparative analysis of different commercially available FISH probes and different FISH protocols in order to optimize this technique on formalin-fixed and paraffin-embedded bone marrow trephine biopsies.


Subject(s)
Bone Marrow/pathology , Fusion Proteins, bcr-abl/analysis , Fusion Proteins, bcr-abl/genetics , In Situ Hybridization, Fluorescence/methods , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology , Biopsy, Needle , Chromosomes, Human, Pair 22 , Chromosomes, Human, Pair 9 , Humans , In Situ Hybridization, Fluorescence/instrumentation , Sensitivity and Specificity , Translocation, Genetic
8.
Ann Rheum Dis ; 74(2): 341-6, 2015 Feb.
Article in English | MEDLINE | ID: mdl-24285491

ABSTRACT

AIM: To assess whether in early (rheumatoid) arthritis (RA) patients, metacarpal bone mineral density (BMD) loss after 4 months predicts radiological progression after 1 year of antirheumatic treatment. METHODS: Metacarpal BMD was measured 4 monthly during the first year by digital X-ray radiogrammetry (DXR-BMD) in patients participating in the IMPROVED study, a clinical trial in 610 patients with recent onset RA (2010 criteria) or undifferentiated arthritis, treated according to a remission (disease activity score<1.6) steered strategy. With Sharp/van der Heijde progression ≥0.5 points after 1 year (yes/no) as dependent variable, univariate and multivariate logistic regression analyses were performed. RESULTS: Of 428 patients with DXR-BMD results and progression scores available, 28 (7%) had radiological progression after 1 year. Independent predictors for radiological progression were presence of baseline erosions (OR (95% CI) 6.5 (1.7 to 25)) and early DXR-BMD loss (OR (95% CI) 1.5 (1.1 to 2.0)). In 366 (86%) patients without baseline erosions, early DXR-BMD loss was the only independent predictor of progression (OR (95% CI) 2.0 (1.4 to 2.9)). CONCLUSIONS: In early RA patients, metacarpal BMD loss after 4 months of treatment is an independent predictor of radiological progression after 1 year. In patients without baseline erosions, early metacarpal BMD loss is the main predictor of radiological progression.


Subject(s)
Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/diagnostic imaging , Arthritis, Rheumatoid/drug therapy , Bone Density , Metacarpal Bones/diagnostic imaging , Absorptiometry, Photon , Arthritis, Rheumatoid/pathology , Disease Progression , Female , Humans , Hydroxychloroquine/therapeutic use , Male , Methotrexate/therapeutic use , Middle Aged , Prednisone/therapeutic use , Sulfasalazine/therapeutic use
9.
Chem Commun (Camb) ; 50(76): 11129-31, 2014 Oct 04.
Article in English | MEDLINE | ID: mdl-25105178

ABSTRACT

Thiol-yne click chemistry is used to covalently link a ferrocenyl derivative to the pore walls of a fully organic porous polymer coating (SURGEL). By cyclic voltammetry, it is demonstrated that the ferrocene bound to the SURGEL via a flexible alkyl linker can be reversibly reduced and oxidised. Surprisingly, when adding ferrocene as an electrolyte, a Nernstian diffusion limited process is observed. We explain this observation in terms of a high permeability of the SURGELs for ferrocene after the post synthetic modification.


Subject(s)
Electrochemical Techniques , Ferrous Compounds/chemistry , Polymers/chemistry , Diffusion , Electrolytes/chemistry , Metallocenes , Molecular Structure , Porosity , Surface Properties
10.
Science ; 345(6196): 573-7, 2014 Aug 01.
Article in English | MEDLINE | ID: mdl-24968940

ABSTRACT

Mammals are coinfected by multiple pathogens that interact through unknown mechanisms. We found that helminth infection, characterized by the induction of the cytokine interleukin-4 (IL-4) and the activation of the transcription factor Stat6, reactivated murine γ-herpesvirus infection in vivo. IL-4 promoted viral replication and blocked the antiviral effects of interferon-γ (IFNγ) by inducing Stat6 binding to the promoter for an important viral transcriptional transactivator. IL-4 also reactivated human Kaposi's sarcoma-associated herpesvirus from latency in cultured cells. Exogenous IL-4 plus blockade of IFNγ reactivated latent murine γ-herpesvirus infection in vivo, suggesting a "two-signal" model for viral reactivation. Thus, chronic herpesvirus infection, a component of the mammalian virome, is regulated by the counterpoised actions of multiple cytokines on viral promoters that have evolved to sense host immune status.


Subject(s)
Gammaherpesvirinae/physiology , Herpesvirus 8, Human/physiology , Interferon-gamma/immunology , Interleukin-4/metabolism , STAT6 Transcription Factor/metabolism , Schistosoma mansoni/immunology , Schistosomiasis mansoni/immunology , Virus Activation/physiology , Animals , Gammaherpesvirinae/genetics , Gene Expression Regulation, Viral , Herpesvirus 8, Human/genetics , Humans , Interferon-gamma/pharmacology , Interleukin-4/pharmacology , Macrophages/immunology , Mice , Mice, Inbred C57BL , Nematospiroides dubius/immunology , Ovum/immunology , Promoter Regions, Genetic , Strongylida Infections/immunology , Virus Activation/drug effects , Virus Activation/genetics , Virus Latency/physiology , Virus Replication/physiology
11.
Ann Rheum Dis ; 73(7): 1356-61, 2014 Jul.
Article in English | MEDLINE | ID: mdl-23716067

ABSTRACT

OBJECTIVES: To assess which treatment strategy is most effective in inducing remission in early (rheumatoid) arthritis. METHODS: 610 patients with early rheumatoid arthritis (RA 2010 criteria) or undifferentiated arthritis (UA) started treatment with methotrexate (MTX) and a tapered high dose of prednisone. Patients in early remission (Disease Activity Score <1.6 after 4 months) tapered prednisone to zero and those with persistent remission after 8 months, tapered and stopped MTX. Patients not in early remission were randomised to receive either MTX plus hydroxychloroquine plus sulfasalazine plus low-dose prednisone (arm 1) or to MTX plus adalimumab (ADA) (arm 2). If remission was present after 8 months both arms tapered to MTX monotherapy; if not, arm 1 changed to MTX plus ADA and arm 2 increased the dose of ADA. Remission rates and functional and radiological outcomes were compared between arms and between patients with RA and those with UA. RESULTS: 375/610 (61%) patients achieved early remission. After 1 year 68% of those were in remission and 32% in drug-free remission. Of the randomised patients, 25% in arm 1 and 41% in arm 2 achieved remission at year 1 (p<0.01). Outcomes were comparable between patients with RA and those with UA. CONCLUSIONS: Initial MTX and prednisone resulted in early remission in 61% of patients with early (rheumatoid) arthritis. Of those, 68% were in remission and 32% were in drug-free remission after 1 year. In patients not in early remission, earlier introduction of ADA resulted in more remission at year 1 than first treating with disease-modifying antirheumatic drug combination therapy plus prednisone.


Subject(s)
Anti-Inflammatory Agents/therapeutic use , Antibodies, Monoclonal, Humanized/therapeutic use , Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/drug therapy , Hydroxychloroquine/therapeutic use , Methotrexate/therapeutic use , Prednisone/therapeutic use , Sulfasalazine/therapeutic use , Adalimumab , Adult , Aged , Arthritis/diagnostic imaging , Arthritis/drug therapy , Arthritis, Rheumatoid/diagnostic imaging , Disease Progression , Drug Therapy, Combination/methods , Early Medical Intervention/methods , Female , Humans , Male , Middle Aged , Radiography , Remission Induction/methods , Single-Blind Method , Treatment Outcome
12.
Oncogene ; 33(41): 4961-5, 2014 Oct 09.
Article in English | MEDLINE | ID: mdl-24186201

ABSTRACT

Recent studies have demonstrated that in clear cell renal cell carcinoma (ccRCC) several chromatin remodeling enzymes are genetically inactivated. Although, growing evidence in cancer models has demonstrated the importance of epigenetic changes, currently only changes in DNA methylation can be accurately determined from clinical samples. To address this limitation, we have applied formaldehyde-assisted isolation of regulatory elements (FAIREs) combined with next-generation sequencing (FAIRE-seq) to identify specific changes in chromatin accessibility in clinical samples of ccRCC. We modified the FAIRE procedure to allow us to examine chromatin accessibility for small samples of solid tumors. Our FAIRE results were compared with DNA-methylation analysis and show how chromatin accessibility decreases at many sites where DNA-methylation remains unchanged. In addition, our FAIRE-seq analysis allowed us to identify regulatory elements associated with both normal and tumor tissue. We have identified decreases in chromatin accessibility at key ccRCC-linked genes, including PBRM1, SETD2 and MLL2. Overall, our results demonstrate the power of examining multiple aspects of the epigenome.


Subject(s)
Carcinoma, Renal Cell/genetics , Chromatin/metabolism , DNA Methylation , Epigenomics/methods , Kidney Neoplasms/genetics , Carcinoma, Renal Cell/pathology , Chromatin Assembly and Disassembly , DNA-Binding Proteins/genetics , Genome, Human , High-Throughput Nucleotide Sequencing , Histone-Lysine N-Methyltransferase/genetics , Humans , Kidney Neoplasms/pathology , Neoplasm Proteins/genetics , Nuclear Proteins/genetics , Regulatory Elements, Transcriptional , Reproducibility of Results , Sequence Analysis, DNA , Transcription Factors/genetics
13.
Anal Chem ; 85(15): 7139-45, 2013 Aug 06.
Article in English | MEDLINE | ID: mdl-23799655

ABSTRACT

Expression microdissection (xMD) is a high-throughput, operator-independent technology that enables the procurement of specific cell populations from tissue specimens. In this method, histological sections are first stained for cellular markers via either chemical or immuno-guided methods, placed in close contact with an ethylene vinyl acetate (EVA) film, and exposed to a light source. The focal, transient heating of the stained cells or subcellular structures melts the EVA film selectively to the targets for procurement. In this report, we introduce a custom-designed flashcube system that permits consistent and reproducible microdissection of nuclei across an FFPE rat brain tissue section in milliseconds. In addition, we present a method to efficiently recover and combine captured proteins from multiple xMD films. Both light and scanning electron microscopy demonstrated captured nuclear structures. Shotgun proteomic analysis of the samples showed a significant enrichment in nuclear localized proteins, with an average 25% of recovered proteins localized to the nucleus, versus 15% for whole tissue controls (p < 0.001). Targeted mass spectrometry using multiple reaction monitoring (MRM) showed more impressive data, with a 3-fold enrichment in histones, and a concurrent depletion of proteins localized to the cytoplasm, cytoskeleton, and mitochondria. These data demonstrate that the flashcube-xMD technology is applicable to the proteomic study of a broad range of targets in molecular pathology.


Subject(s)
Brain/cytology , Cell Nucleus/metabolism , Microdissection/methods , Proteomics/methods , Amino Acid Sequence , Animals , Chemical Precipitation , Formaldehyde/metabolism , Mass Spectrometry , Molecular Sequence Data , Paraffin Embedding , Proteolysis , Rats , Tissue Fixation
14.
Pathologe ; 34(4): 329-34, 2013 Jul.
Article in German | MEDLINE | ID: mdl-23179209

ABSTRACT

BACKGROUND: The detection of characteristic genomic aberrations by fluorescence in situ hybridization (FISH) has a high diagnostic impact on lymphomas according to the World Health Organization (WHO). To investigate the reproducibility of non-isotopic ISH results a multicenter trial was carried out involving eight institutes for hematopathology. MATERIAL AND METHODS: Analyses were performed on two diffuse large B-cell lymphomas (DLBCL) without known aberrations, on one follicular lymphoma with a IGH/BCL2 translocation and BCL6 split and on two B-cell lymphomas intermediate between DLBCL and Burkitt's lymphoma with c-MYC and BCL2 rearrangements, one with an additional BCL6 split. Break-apart probes for BCL6 and c-MYC, as well as fusion probes for the c-MYC/IGH and the IGH/BCL2 translocations were used. RESULTS: All aberrations were correctly detected by all centres and no false positive or false negative results were obtained. The numbers of positive cells varied from 25% to 94%. Pearson's correlation coefficient between the centres was always > 0.8. CONCLUSIONS: The ISH analysis of recurrent genomic aberrations in formalin-fixed paraffin-embedded (FFPE) tissue is a highly reproducible technique which yields substantial additive help for lymphoma diagnostics.


Subject(s)
Chromosome Aberrations , In Situ Hybridization/methods , Lymphoma, Non-Hodgkin/genetics , Biomarkers, Tumor/genetics , Burkitt Lymphoma/diagnosis , Burkitt Lymphoma/genetics , Burkitt Lymphoma/pathology , DNA-Binding Proteins/genetics , Diagnosis, Differential , Genes, myc/genetics , Humans , Immunoglobulin Heavy Chains/genetics , In Situ Hybridization, Fluorescence/methods , Lymphoma, Follicular/diagnosis , Lymphoma, Follicular/genetics , Lymphoma, Follicular/pathology , Lymphoma, Large B-Cell, Diffuse/diagnosis , Lymphoma, Large B-Cell, Diffuse/genetics , Lymphoma, Large B-Cell, Diffuse/pathology , Lymphoma, Non-Hodgkin/diagnosis , Lymphoma, Non-Hodgkin/pathology , Proto-Oncogene Proteins c-bcl-6 , Quality Assurance, Health Care , Reproducibility of Results , Translocation, Genetic/genetics
15.
Braz. j. med. biol. res ; 45(12): 1127-1134, Dec. 2012. ilus
Article in English | LILACS | ID: lil-659650

ABSTRACT

DNA repair is crucial to the survival of all organisms. The bacterial RecA protein is a central component in the SOS response and in recombinational and SOS DNA repairs. The RecX protein has been characterized as a negative modulator of RecA activity in many bacteria. The recA and recX genes of Herbaspirillum seropedicae constitute a single operon, and evidence suggests that RecX participates in SOS repair. In the present study, we show that the H. seropedicae RecX protein (RecX Hs) can interact with the H. seropedicaeRecA protein (RecA Hs) and that RecA Hs possesses ATP binding, ATP hydrolyzing and DNA strand exchange activities. RecX Hs inhibited 90% of the RecA Hs DNA strand exchange activity even when present in a 50-fold lower molar concentration than RecA Hs. RecA Hs ATP binding was not affected by the addition of RecX, but the ATPase activity was reduced. When RecX Hs was present before the formation of RecA filaments (RecA-ssDNA), inhibition of ATPase activity was substantially reduced and excess ssDNA also partially suppressed this inhibition. The results suggest that the RecX Hs protein negatively modulates the RecA Hs activities by protein-protein interactions and also by DNA-protein interactions.


Subject(s)
Bacterial Proteins/metabolism , Herbaspirillum/chemistry , Rec A Recombinases/metabolism , DNA, Bacterial , Escherichia coli/metabolism , Protein Binding
16.
Braz J Med Biol Res ; 45(12): 1127-34, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23044625

ABSTRACT

DNA repair is crucial to the survival of all organisms. The bacterial RecA protein is a central component in the SOS response and in recombinational and SOS DNA repairs. The RecX protein has been characterized as a negative modulator of RecA activity in many bacteria. The recA and recX genes of Herbaspirillum seropedicae constitute a single operon, and evidence suggests that RecX participates in SOS repair. In the present study, we show that the H. seropedicae RecX protein (RecX Hs) can interact with the H. seropedicaeRecA protein (RecA Hs) and that RecA Hs possesses ATP binding, ATP hydrolyzing and DNA strand exchange activities. RecX Hs inhibited 90% of the RecA Hs DNA strand exchange activity even when present in a 50-fold lower molar concentration than RecA Hs. RecA Hs ATP binding was not affected by the addition of RecX, but the ATPase activity was reduced. When RecX Hs was present before the formation of RecA filaments (RecA-ssDNA), inhibition of ATPase activity was substantially reduced and excess ssDNA also partially suppressed this inhibition. The results suggest that the RecX Hs protein negatively modulates the RecA Hs activities by protein-protein interactions and also by DNA-protein interactions.


Subject(s)
Bacterial Proteins/metabolism , Herbaspirillum/chemistry , Rec A Recombinases/metabolism , DNA, Bacterial , Escherichia coli/metabolism , Protein Binding
17.
Ann Oncol ; 22(11): 2417-2423, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21402619

ABSTRACT

BACKGROUND: CALYPSO (CAeLYx in Platinum Sensitive Ovarian) patients compared carboplatin-pegylated liposomal doxorubicin (C-PLD) with carboplatin-paclitaxel (C-P) in patients with late-relapsing recurrent ovarian cancer (ROC). We analyzed outcomes in patients ≥70 years. PATIENTS AND METHODS: Nine hundred and seventy-six patients with taxane-pretreated ROC relapsing >6 months after first- or second-line platinum-based therapy were randomly assigned to 4-weekly C area under the curve (AUC) 5 plus PLD 30 mg/m(2) or 3-weekly C AUC 5 plus P 175 mg/m(2) for six or more cycles. RESULTS: One hundred and fifty-seven (16%) patients ≥70 years (median: 74 years, C-PLD; 73 years, C-P; range 70-82 years) were included (n = 71, C-PLD; n = 86, C-P). In comparing elderly and younger, elderly patients experienced fewer grade ≥2 allergic reactions (P = 0.005) but more grade ≥2 sensory neuropathy (P = 0.007). Myelosuppression did not differ with age. Elderly patients completed planned treatment as frequently as younger (79%, C-PLD; 82%, C-P). In comparing arms within elderly patients, C-P was associated with more grade ≥2 alopecia, sensory neuropathy, arthralgia/myalgia (P < 0.001 for all), severe leukopenia plus febrile neutropenia; C-PLD was associated with more grade ≥2 hand-foot syndrome (P = 0.005). Median progression-free survival was 11.6 months (C-PLD) and 10.3 months (C-P; P = 0.44). CONCLUSIONS: Patients ≥70 years experienced more neuropathy, with a higher incidence in the C-P arm. Similar to all study patients, C-PLD provided a better therapeutic index with less toxicity than C-P in elderly women with platinum-sensitive ROC.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Neoplasm Recurrence, Local/drug therapy , Ovarian Neoplasms/drug therapy , Age Factors , Aged , Aged, 80 and over , Antineoplastic Combined Chemotherapy Protocols/adverse effects , Carboplatin/administration & dosage , Carboplatin/adverse effects , Doxorubicin/administration & dosage , Doxorubicin/adverse effects , Doxorubicin/analogs & derivatives , Female , Humans , Paclitaxel/administration & dosage , Paclitaxel/adverse effects , Polyethylene Glycols/administration & dosage , Polyethylene Glycols/adverse effects , Quality of Life , Treatment Outcome
18.
J Perinatol ; 31(6): 377-86, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21183927

ABSTRACT

With mounting evidence that hypothermia is neuroprotective in newborns with hypoxic-ischemic encephalopathy (HIE), an increasing number of centers are offering this therapy. Hypothermia is associated with a wide range of physiologic changes affecting every organ system, and awareness of these effects is essential for optimum patient management. Lowering the core temperature also alters pharmacokinetic and pharmacodynamic properties of medications commonly used in asphyxiated neonates, necessitating close attention to drug efficacy and side effects. Rewarming introduces additional risks and challenges as the hypothermia-associated physiologic and pharmacologic changes are reversed. In this review we provide an organ system-based assessment of physiologic changes associated with hypothermia. We also summarize evidence from randomized controlled trials showing lack of serious adverse effects of moderate hypothermia therapy in term and near-term newborns with moderate-to-severe HIE. Finally, we review the effects of hypothermia on drug metabolism and clearance based on studies in animal models and human adults, and limited data from neonates.


Subject(s)
Hypothermia, Induced/methods , Hypoxia-Ischemia, Brain/physiopathology , Hypoxia-Ischemia, Brain/therapy , Inactivation, Metabolic/physiology , Metabolic Clearance Rate/physiology , Animals , Brain/physiopathology , Humans , Hypothermia, Induced/adverse effects , Infant, Newborn , Pharmacokinetics , Randomized Controlled Trials as Topic , Rewarming/methods , Temperature
19.
Mol Biol Evol ; 28(3): 1141-55, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21059793

ABSTRACT

Sensing the environment and responding appropriately to it are key capabilities for the survival of an organism. All extant organisms must have evolved suitable sensors, signaling systems, and response mechanisms allowing them to survive under the conditions they are likely to encounter. Here, we investigate in detail the evolutionary history of one such system: The phage shock protein (Psp) stress response system is an important part of the stress response machinery in many bacteria, including Escherichia coli K12. Here, we use a systematic analysis of the genes that make up and regulate the Psp system in E. coli in order to elucidate the evolutionary history of the system. We compare gene sharing, sequence evolution, and conservation of protein-coding as well as noncoding DNA sequences and link these to comparative analyses of genome/operon organization across 698 bacterial genomes. Finally, we evaluate experimentally the biological advantage/disadvantage of a simplified version of the Psp system under different oxygen-related environments. Our results suggest that the Psp system evolved around a core response mechanism by gradually co-opting genes into the system to provide more nuanced sensory, signaling, and effector functionalities. We find that recruitment of new genes into the response machinery is closely linked to incorporation of these genes into a psp operon as is seen in E. coli, which contains the bulk of genes involved in the response. The organization of this operon allows for surprising levels of additional transcriptional control and flexibility. The results discussed here suggest that the components of such signaling systems will only be evolutionarily conserved if the overall functionality of the system can be maintained.


Subject(s)
Escherichia coli K12/genetics , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Evolution, Molecular , Trans-Activators/genetics , Trans-Activators/metabolism , Base Sequence , Escherichia coli K12/metabolism , Escherichia coli Proteins/classification , Gene Expression Profiling , Gene Expression Regulation, Bacterial , Genetic Association Studies , Genome, Bacterial , Genomic Instability/physiology , Genomics , Operon , Phylogeny , Stress, Physiological/physiology , Trans-Activators/classification , Transcription, Genetic
20.
Anaesthesist ; 60(2): 118-24, 2011 Feb.
Article in German | MEDLINE | ID: mdl-21060981

ABSTRACT

BACKGROUND: The goal of the present study was to evaluate the publication rate of abstracts presented during the German Anesthesia Congress (Deutscher Anästhesiecongress, DAC) and the meeting of the European Society of Anesthesiologists (ESA) in the years 2000 and 2005 in Medline listed journals (http://www.ncbi.nlm.nih.gov/pubmed). In addition, the respective impact factors of the journals in which the articles were published were evaluated (http://www.isiknowledge.com). METHODS: All abstracts of free papers and posters presented at the DAC and ESA from the years 2000 and 2005 were included into the study. The presence of authors and the topics of abstracts in the literature were analyzed by a Medline based inquiry over a time period of 5 years. The search was based on the last name and initials of authors and when these could not be identified in Medline the search was extended by keywords of relevant topics of the abstract. Umlauts "ä/ö/ü" were replaced by "ae/oe/ue" and "ß" was replaced by "ss". Only original papers were included in this analysis. Once an original paper was found the impact factor of the journal in that year was identified. RESULTS: A total of 465 abstracts from the DAC 2000, 378 abstracts from the DAC 2005, 644 abstracts from the ESA 2000 and 720 abstracts from the ESA 2005 were included. Of the abstracts from the DAC 2000, 183 (39%) were published in Medline listed journals, 179 (47%) from DAC 2005, 218 (34%) from ESA 2000 and 233 (32%) from ESA 2005. The ESA abstracts were published in English more often than the DAC abstracts (ESA 2000: 95%; ESA 2005: 95%; DAC 2000: 78%; DAC 2005: 86%). While the publication rate after the ESA remained nearly unchanged between 2000 and 2005, the publication rate after the DAC increased by about 7%. The average impact factors of the publications were 1.777 (DAC 2000), 2.836 (DAC 2005), 1.825 (ESA 2000) and 2.36 (ESA 2005). Independent of the congress (DAC or ESA) where the abstract was presented, most articles were published in the journal Anesthesia & Analgesia. CONCLUSION: In the year 2005 more abstracts of the DAC were published in Medline listed papers than in 2000. When comparing the number of abstracts published in Medline listed journals, more abstracts of the DAC were published compared to abstracts of the ESA. The increase in papers written in English after abstract presentation on the DAC is mostly due to the wider readership which can be reached with manuscripts in the English language. Besides a larger readership, English journals often also have a higher ranked impact factor. This analysis does not claim to be a complete registration of all published abstracts due to the limitation on Medline listed journals and publications in other journals were not rated. Medline was selected because of the widespread and international use of this database.


Subject(s)
Anesthesiology/statistics & numerical data , Congresses as Topic , Periodicals as Topic , Publications/statistics & numerical data , Europe , Germany , Journal Impact Factor , MEDLINE , Research Design
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