Solitary Fibrous Tumour: A Single Institution Retrospective Study and Further Validation of a Prognostic Risk Assessment System.

AIMS: Solitary fibrous tumour (SFT) is a rare mesenchymal-derived neoplasm that can arise in any anatomical location in the body. SFT rarely metastasises, but aggressive behaviour is seen in a minority of cases, and relapses can occur several years after treatment. It would be a clinical advantage if high-risk patients could be identified before treatment. MATERIALS AND METHODS: We retrospectively analysed a population-based cohort of SFT to describe treatment, outcome, prognostic factors and to further validate a previously published risk assessment tool (D-score) based on age, tumour size and mitotic index. Seventy-two patients diagnosed with SFT in the Central, North and Southern Denmark regions between 1979 and 2013 were included in the study. RESULTS: For patients with localised disease at the time of diagnosis (n = 64) the 5 and 10 year overall survival was 86% (95% confidence interval 74-92) and 65% (95% confidence interval 50-78), respectively. Seventeen of 62 patients (27%) who were in remission after radical treatment developed recurrence with either local or distant disease. The 5 year recurrence-free survival was 83% (95% confidence interval 70-90) and the 10 year was 69% (95% confidence interval 53-81). The 5 year local recurrence-free survival was 96% (95% confidence interval 86-99) and the 10 year was 92% (95% confidence interval 76-96). The median time to both overall recurrence and local recurrence was 4.3 years. Metastatic or inoperable SFT had a poor prognosis with a median overall survival of 8.4 months (range 3.6-26.4) and a 5 year overall survival of 11% (95% confidence interval 2-30). A further validation of a risk assessment tool (D-score) confirmed that patients classified as high-risk had a significantly decreased overall survival, with a hazard ratio of 3.7 (95% confidence interval 1.1-12.3). CONCLUSIONS: This study showed that our management and outcome were comparable with other published studies describing SFT and confirmed the value of the D-score as a risk assessment tool. Because of late recurrences, long-term (e.g. 10 years) follow-up for moderate- and high-risk patients is recommended.

Attenuation of transcriptional and signaling responses limits viability of ρ(0)Saccharomyces cerevisiae during periods of glucose deprivation.

BACKGROUND: The maintenance of viability during periods when a glycolytic carbon source is limited (or absent) is a major obstacle for cells whose mitochondrial DNA (mtDNA) has been damaged or lost. METHODS: We utilized genome wide transcriptional profiling and in gel mobility analyses to examine the transcriptional response and characterize defects in the phosphorylation dependent signaling events that occur during acute glucose starvation in ρ(0) cells that lack mtDNA. Genetic and pharmacological interventions were employed to clarify the contribution of nutrient responsive kinases to regulation of the transcription factors that displayed abnormal phosphoregulation in ρ(0) cells. RESULTS: The transcriptional response to glucose deprivation is dampened but not blocked in ρ(0) cells. Genes regulated by the transcription factors Mig1, Msn2, Gat1, and Ume6 were noticeably affected and phosphorylation of these factors in response to nutrient depletion is abnormal in ρ(0) cells. Regulation of the nutrient responsive kinases PKA and Snf1 remains normal in ρ(0) cells. The phosphorylation defect results from ATP depletion and loss of the activity of kinases including GSK3ß, Rim15, and Yak1. Interventions which rescue phosphoregulation of transcription factors bolster maintenance of viability in ρ(0) cells during subsequent glucose deprivation. CONCLUSIONS: A subset of nutrient responsive kinases is especially sensitive to ATP levels and their misregulation may underlie regulatory defects presented by ρ(0) cells. GENERAL SIGNIFICANCE: Abnormal regulation of mitochondrial function is implicated in numerous human disorders. This work illustrates that some signaling pathways are more sensitive than others to metabolic defects caused by mitochondrial dysfunction.

High-Performance Chemical Isotope Labeling Liquid Chromatography-Mass Spectrometry for Profiling the Metabolomic Reprogramming Elicited by Ammonium Limitation in Yeast.

Information about how yeast metabolism is rewired in response to internal and external cues can inform the development of metabolic engineering strategies for food, fuel, and chemical production in this organism. We report a new metabolomics workflow for the characterization of such metabolic rewiring. The workflow combines efficient cell lysis without using chemicals that may interfere with downstream sample analysis and differential chemical isotope labeling liquid chromatography mass spectrometry (CIL LC-MS) for in-depth yeast metabolome profiling. Using (12)C- and (13)C-dansylation (Dns) labeling to analyze the amine/phenol submetabolome, we detected and quantified a total of 5719 peak pairs or metabolites. Among them, 120 metabolites were positively identified using a library of 275 Dns-metabolite standards, and 2980 metabolites were putatively identified based on accurate mass matches to metabolome databases. We also applied (12)C- and (13)C-dimethylaminophenacyl (DmPA) labeling to profile the carboxylic acid submetabolome and detected over 2286 peak pairs, from which 33 metabolites were positively identified using a library of 188 DmPA-metabolite standards, and 1595 metabolites were putatively identified. Using this workflow for metabolomic profiling of cells challenged by ammonium limitation revealed unexpected links between ammonium assimilation and pantothenate accumulation that might be amenable to engineering for better acetyl-CoA production in yeast. We anticipate that efforts to improve other schemes of metabolic engineering will benefit from application of this workflow to multiple cell types.

Rewiring AMPK and mitochondrial retrograde signaling for metabolic control of aging and histone acetylation in respiratory-defective cells.

Abnormal respiratory metabolism plays a role in numerous human disorders. We find that regulation of overall histone acetylation is perturbed in respiratory-incompetent (ρ(0)) yeast. Because histone acetylation is highly sensitive to acetyl-coenzyme A (acetyl-CoA) availability, we sought interventions that suppress this ρ(0) phenotype through reprogramming metabolism. Nutritional intervention studies led to the discovery that genetic coactivation of the mitochondrion-to-nucleus retrograde (RTG) response and the AMPK (Snf1) pathway prevents abnormal histone deacetylation in ρ(0) cells. Metabolic profiling of signaling mutants uncovered links between chromatin-dependent phenotypes of ρ(0) cells and metabolism of ATP, acetyl-CoA, glutathione, branched-chain amino acids, and the storage carbohydrate trehalose. Importantly, RTG/AMPK activation reprograms energy metabolism to increase the supply of acetyl-CoA to lysine acetyltransferases and extend the chronological lifespan of ρ(0) cells. Our results strengthen the framework for rational design of nutrient supplementation schemes and drug-discovery initiatives aimed at mimicking the therapeutic benefits of dietary interventions.

Expression of secreted frizzled-related protein 4 (SFRP4) in primary serous ovarian tumours.

OBJECTIVE: Serous ovarian cancer is the most prevalent type of ovarian cancer. The majority of women present at an advanced stage and patient survival is poor. Resistance to chemotherapy is thought to relate to failure of tumours to undergo apoptosis. Secreted frizzled-related protein 4 (SFRP4) has been demonstrated to be involved in apoptosis in the ovary but not in ovarian tumours as yet. This study examined SFRP4 expression in ovarian cancers and correlated this with expression of beta-catenin, a main component of the wNT-signalling pathway it inhibits. METHODS: We examined 153 primary serous ovarian carcinomas for SFRP4 and B-catenin expression using immunohistochemistry on tissue microarrays and correlated this with clinical information. RESULTS: SFRP4 expression was inversely associated with beta-catenin expression in 84% of samples. However, high-level SFRP4 expression was not significantly associated with patient survival (p = 0.08). CONCLUSION: Elevated SFRP4 expression in serous ovarian tumours appears to correlate with reduced beta-catenin expression but long-term survival appears unaffected by this.

A glycolytic burst drives glucose induction of global histone acetylation by picNuA4 and SAGA.

Little is known about what enzyme complexes or mechanisms control global lysine acetylation in the amino-terminal tails of the histones. Here, we show that glucose induces overall acetylation of H3 K9, 18, 27 and H4 K5, 8, 12 in quiescent yeast cells mainly by stimulating two KATs, Gcn5 and Esa1. Genetic and pharmacological perturbation of carbon metabolism, combined with (1)H-NMR metabolic profiling, revealed that glucose induction of KAT activity directly depends on increased glucose catabolism. Glucose-inducible Esa1 and Gcn5 activities predominantly reside in the picNuA4 and SAGA complexes, respectively, and act on chromatin by an untargeted mechanism. We conclude that direct metabolic regulation of globally acting KATs can be a potent driving force for reconfiguration of overall histone acetylation in response to a physiological cue.

Untargeted tail acetylation of histones in chromatin: lessons from yeast.

Dynamic acetylation of lysine residues in the amino-terminal tails of the core histones is functionally important for the regulation of diverse DNA-dependent processes in the nucleus, including replication, transcription, and DNA repair. The targeted and untargeted activities of histone lysine acetylases (KATs) and deacetylases (HDACs) both contribute to the dynamics of chromatin acetylation. While the mechanisms and functional consequences of targeted on histone acetylation are well understood, relatively little is known about untargeted histone acetylation. Here, we review the current understanding of the mechanisms by which untargeted KAT and HDAC activities modulate the acetylation state of nucleosomal histones, focusing on results obtained for H3 and H4 in budding yeast. We also highlight unresolved problems in this area, including the question of how a particular steady-state level of untargeted acetylation is set in the absence of cis-dependent mechanisms that instruct the activity of KATs and HDACs.

Expression of secreted frizzled-related protein 4 in the primate placenta.

Secreted frizzled-related protein 4 (sFRP4) blocks the Wnt signalling pathway by competitively binding Wnt ligands (frizzled receptors). This pathway is important during development and oncogenesis. It is, however, complex with a large number of interacting proteins, isoforms and receptors. The Wnt signalling pathway has a role in human placental development and implantation, particularly in the trophoblast. Humans and macaque monkeys exhibit a similar remodelling of the decidual spiral arteries. The expression of sFRP4 in human and macaque placentas at different gestational ages have been examined with immunohistochemistry, in-situ hybridization, real-time polymerase chain reaction, and western blotting. This study demonstrates that sFRP4 is expressed predominantly in the villous syncytiotrophoblast and the invasive intermediate cytotrophoblast, and in the amnion. These observational studies suggest that sFRP4 has a role in placental development and implantation, and may be an important factor in the development of the decidual fibrinoid zone, and in trophoblast apoptosis and a band of apoptosis in the underlying decidua deep into the trophoblast.

Analysis of responses of Long Beach, California residents to the Smoke-free Bars Law.

OBJECTIVES: We investigated survey responses to the Smoke-free Bars Law by residents of Long Beach, California (population 460,000), a city that reflects the state's diverse population. The research specifically aimed to determine: (1) residents' approval for the 1998 California Smoke-free Bars Law when it was implemented; and (2) changes in approval between baseline and 2-year follow-up. Data were also assessed for the demographic characteristics of the respondents and whether the respondents were self-acknowledged smokers or non-smokers. STUDY DESIGN: A random telephone survey was conducted in 1998 and 2000 in Long Beach to determine the degree of community support for the 1998 state law that prohibited smoking in all workplaces including alcohol-serving establishments. The numbers analysed were 784 in 1998 and 1237 in 2000. METHODS: Statistical analyses used in this research included univariate frequency distributions and logistic regression for 1998 and 2000. RESULTS: The major findings were as follows. Overall community approval for the 1998 state law increased from 65.2% in 1998 to 72.6% in 2000. Over this period, the rate of approval by smokers increased from 20.6% to 37.1%, and the rate of approval by non-smokers increased from 74.5% to 80.3%. CONCLUSIONS: The general public in a large city in California strongly approve of the prohibition of smoking in all indoor public places. This strong endorsement has major public health implications.

Public opinion analysis regarding the uses of master settlement agreement (MSA) funds.

OBJECTIVES: In the present research, we assessed the relationship between characteristics of the residents of Long Beach, California, a typical city in America, and their opinions regarding the uses of master settlement agreement (MSA) funds. METHODS: The statistical analyses used in the present research included univariate frequency distributions, cross-tabulations, and classification and regression trees. RESULTS: The results indicate that the majority of Long Beach residents share the opinion that the MSA funds should be allocated to health programmes. They do not, however, feel that these funds need to be earmarked solely for smoking prevention or cessation. CONCLUSIONS: Due to state budget deficits, legislators may strongly advocate for the MSA funds to be used for non-health purposes. Our findings provide support for community advocates who wish to bring the current uses of MSA funds and tobacco taxes to the forefront of national and international public debate.

The role of sFRP4, a secreted frizzled-related protein, in ovulation.

Ovulation is a complex, multi-factorial event that involves the degeneration of a specific area of the follicular and ovarian surface via apoptosis. Many apoptosis related genes have been identified in the ovary. Secreted Frizzled Related Protein 4 (sFRP4) is a protein that appears to antagonize a molecular pathway for cell survival. sFRP4 gene expression is known to be upregulated with apoptosis in the ovarian corpus luteum. In this study, ovulation was hormonally induced in immature Wistar rats and their ovaries collected for analysis of apoptosis and sFRP4. TUNEL staining identified a greater amount of dying cells in the thecal layer of treated rat ovaries compared to controls. The results of 3'-end labelling revealed a significant increase (p < 0.01) in apoptosis at 12 hours following treatment compared to other time points and control. In situ hybridization exhibited a visible increase in amounts of sFRP4 mRNA expression in the thecal layers of follicles from treated rats compared to controls. Quantitative RT-PCR revealed no significant difference in sFRP4 expression levels between treated and control tissues although a clear trend towards an increase was observed in the treated group. This study demonstrates an association between sFRP4 and apoptosis in rat ovulation.

Role of DDC-4/sFRP-4, a secreted frizzled-related protein, at the onset of apoptosis in mammary involution.

Using differential display, we isolated DDC-4, a secreted frizzled-related protein (sFRP), which is induced in the physiological apoptosis of hormonally regulated, reproductive tissues such as mammary gland, prostate, corpus luteum and uterus. The role of this gene in apoptosis was studied in animals overexpressing ectopic DDC-4/sFRP-4. Transgenic mice bearing the DDC-4/sFRP-4 cDNA under the control of the MMTV-LTR promoter showed lactational insufficiency and many apoptotic cells in the alveoli between day 19 of pregnancy and day 4 of lactation as demonstrated by TUNEL reaction and the presence of activated caspase-3. We performed a PKB/Akt kinase assay and studied several of its substrates using phosphorylation-specific antibodies to show reduced phosphorylation in PKB/Akt itself, as well as in glycogen synthetase kinase-3beta (GSK-3beta), BAD, and Forkhead. Taken together, our results show a role for DDC-4/sFRP-4 in abrogating an epithelial cell survival pathway at the onset of mammary gland involution.

Apoptosis-linked in vivo regulation of the tissue transglutaminase gene promoter.

Tissue transglutaminase (tTG) is upregulated in various cells undergoing apoptosis. To investigate the transcriptional regulation of tTG a mouse strain carrying a beta-galactosidase reporter gene under the control of a 3.8 kilobase fragment of the tTG promoter was characterised. The transgene construct was shown to be expressed in the apoptotic regions of the mouse embryo. Here we report that the regulation of the transgene is also apoptosis-linked in adult animals. The transgene is induced in endocrine apoptosis involving mammary gland involution and corpus luteum regression. Induction of the reporter gene is detectable during in vivo but not in vitro apoptosis of thymocytes induced by the glucocorticoid receptor, the nur77, p53 and the retinoid receptor gamma mediated pathways. Additionally, the lacZ expression mimics the activation of the endogenous promoter in tissues characterised by high apoptotic turnover. These results suggest that the apoptosis-specific transcriptional regulation of tTG is mediated through elements of a 3.8 kb promoter and may require cosignals available only in tissue environment. Cell Death and Differentiation (2000) 7, 1225 - 1233.

Physiological apoptosis in hormone-dependent tissues: involvement of caspases.

Physiological apoptosis in mammals is a type of programmed cell death, an important element in the developmental repertoire ensuring tissue homeostasis and proper disposal of cells that are no longer needed, such as milk-producing epithelial cells in the mammary gland after lactation, luteal cells in the post partum Corpus luteum or secretory cells in the prostate after castration. Although incompletely described, apoptosis in hormone-dependent tissues is apparently initiated and executed using common biochemical strategies. These include survival pathways governed by local and systemic factors and hormones, diverse regulatory pathways and caspase-dependent execution pathways. Using an antibody that recognizes processed effector caspases or a fluorogenic caspase substrate, we present for the first time evidence that caspases are activated in the mammary gland, in the prostate and in the ovary at the time when apoptosis occurs. Most likely phagocytosis of apoptotic cells by neighboring cells may represent an important step, since only a modest involvement of professional phagocytes is apparent. Here, we will summarize and discuss recent data and will attempt to draw a generalized picture of how physiological apoptosis may occur in these organs.

Energy metabolism in the involuting mammary gland.

A strong and coordinated upregulation of the glycolytic, glutaminolytic and pentose phosphate pathway enzymes occurs during the onset of lactation in the normal mouse mammary gland. Induction of apoptosis by removing the pups led to an inactivation of the same enzymes with different time courses. While the ATP-consuming glycolytic 6-phosphofructo 1-kinase and mitochondrial bound hexokinase still remained high on days one and two of involution, the ATP-regenerating pyruvate kinase was immediately reduced. The enzymes of the pentose phosphate and glutaminolytic pathway were inactivated on the first two days of involution. In accordance with such an inactivation of the enzymes ATP, GTP, UTP, ADP, NAD NADH and lactate concentrations decreased. The synthetic product of UTP, UDP-N-acetylglucosamine, increased. AMP was found in the milk, not in the epithelial cells. The inactivation of the enzymes was caused by partial proteolysis or by a loss of the intact proteins from the cytosol without signs of proteolysis.