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The Murdoch family’s private legal battle has now spilled into the public forum
with James Murdoch making explosive comments about both his media mogul dad Rupert and older brother Lachlan
In scenes that could have been ripped straight from the hit drama series Succession, James, 52, gave a scathing interview to The Atlantic
detailing a long history of betrayal and dysfunction amid the legal battle over control of the family’s $66 billion media empire
The article recalled a moment in the lead-up to the November 2024 case
when James faced a barrage of “withering” questions from his father’s lawyer
This included referencing James and his siblings as “white
multi-billionaire trust-fund babies” – only to realise
that his father had been texting the questions to the lawyer from across the room
“How f–king twisted is that?” James told The Atlantic
James also claimed to the reporter that a recent spate of slanderous stories targeting him and his wife
could be traced back to sources associated with Rupert’s trusted inner circle
Both Rupert and his protégé Lachlan disputed James’ points in the article
describing them as a “litany of falsehoods”
The explosive comments are a huge break with convention for the normally intensely private family
But it’s also an indictment of how far relations have crumbled
James remains aligned with elder siblings Prudence
The fracture was laid bare when Rupert and Lachlan attempted to wrest control of the business by making changes to the family trust
Rupert tried to implement ‘Project Family Harmony’
which would effectively grant eldest son Lachlan full control of the Murdoch empire after his death
James and his sisters vehemently disputed this
with Elisabeth telling their father: “You are completely disenfranchising me and my siblings
You’ve blown a hole in the family.”
Yet Rupert still wanted to tear up the family trust
which had previously assured them all an equal stake in the control of Fox and News Corp after Rupert’s passing
who shares his father’s conservative values
couldn’t be outvoted 3-1 by his siblings
Rupert had become convinced that under Lachlan’s sole leadership
“These companies are my legacy,” Rupert has said
“I have put everything into them over my life.”
James and Kathryn had already publicly voiced their disdain over Murdoch-owned News Corp’s coverage on issues such as climate change and the recent US presidential election
James resigned from the News Corp board in 2020
By the time of the legal battle over the family trust that played out in a Nevada courtroom last year
“James and Rupert had barely spoken in years,” The Atlantic article claimed
a judge ruled against Rupert and Lachlan in the matter
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The Original Article was published on 26 February 2016
Correction: Breast Cancer Research (2016) 18:26 https://doi.org/10.1186/s13058-016-0686-4
the author reported that PARP blots of JIMT-1 and JIMT-1-BR3 cells in Fig
7C of the original publication were mistakenly duplicated
The correct figure below depicts the correct PARP blot for JIMT-1 cells
The error does not affect any of the interpretations or conclusions of the article
The email address of the Corresponding Author has also been updated
from klaus.podar@nct-heidelberg.de to klaus.podar@krems.lknoe.at as shown in this Correction article
Bashari MH, Fan F, Vallet S, et al. Mcl-1 confers protection of Her2-positive breast cancer cells to hypoxia: therapeutic implications. Breast Cancer Res. 2016;18:26. https://doi.org/10.1186/s13058-016-0686-4
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DOI: https://doi.org/10.1186/s13058-024-01811-y
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Deutsche Post DHL Group’s contract logistics division DHL Supply Chain is to open an automated fulfillment center in Bergkamen
The facility was commissioned to help IKEA handle its B2C online business in Germany
The multi-customer site in Bergkamen has 40,000m2 of warehouse space and is part of the neighboring DHL Supply Chain Unna Campus
20,000m2 of warehouse space will be available for IKEA alone
with some 350 employees working for the customer on-site six days a week in three shifts
The site will be fully operational at the end of June 2022
at which point an average of 8,000 stock-keeping units (SKUs) will be processed on-site
the site is capable of processing 11,000 SKUs
Packaging machines are available on-site for sensitive goods that require special packaging
complex conveyor automation is used in conjunction with robots from warehouse automation company Locus Robotics and picking machines from logistics automation company Packsize
the implementation represents another important building block in our drive to grow e-commerce
as well as the continuation of our commercial automation and digitization strategy for continuous improvement in operations
the distances our warehouse staff need to walk have been radically reduced and picking effectiveness significantly increased
the system reduces individual order process times
optimizes employee workflows and improves operational efficiency.”
country customer fulfillment manager for IKEA Deutschland
“The consumer sector around the world is undergoing change at the moment
Part of this change is the ever-increasing importance of online retail
Our focus is set on the changing needs of our customers
The commissioning of the Bergkamen fulfillment center for IKEA Germany allows us to take another step in providing an outstanding shopping experience for our customers
who has shown us new ways forward and has been able to set up a highly complex
customized fulfillment solution for us at the Bergkamen site in just a short space of time
“Having two leading companies in their respective industries partner here on-site offers a degree of security for the city of Bergkamen as well as for employees
This is an important factor given the current pace of change
the A2 Logistics Park remains the right decision at the right time
I am convinced that the site offers the potential and opportunities to continue to profit from the growth of logistics – the largest business sector in Germany after automotive and trade
Logistics is a strong industry with a powerful focus on growth and the future
William Douglas has been writing The Color of Hockey blog since 2012
Douglas joined NHL.com in 2019 and writes about people of color in the sport
as part of NHL.com's celebration of Women's History Month
a coach who was one of the first Black women to play in Germany
Miriam Thimm knew hockey was for her from the moment an uncle took her skating at a rink in Dorsten
"Figure skating is not really me," Thimm said
"And I saw a couple of guys in my neighborhood playing hockey
The first time I had a hockey stick in my hand
Thimm became one of the first Black women to play and advance in hockey in Germany
The 44-year-old retired forward played in the German Women's Ice Hockey League -- or DFEL -- and for the women's national team
She has coached men's and women's hockey in Duisburg
Thimm has become a face and a force for diversity in German hockey and beyond
an independent arbitration panel that examines cases of racism and sexism in the German Ice Hockey League
the country's highest professional men's league
Thimm is also member of the NHL Coaches' Association's Female Coaches Program
which was launched on International Women's Day in 2020 to help support the development of female hockey coaches at all levels of the sport
The program helps women coaches in several areas including skills development
networking and career advancement opportunities
"It's obvious there is an underrepresentation of women in North America
both in coaching and other roles in hockey operation," NHLCA executive director Lindsay Pennal said
Having Miriam coaching at the women's pro level in Germany is incredibly important
She's acting as a trailblazer for other women who aspire to coach at that level
and as a role model to young women and girls who are playing."
a former DEL player who is CEO and co-founder of Hockey is Diversity
a Berlin-based international nonprofit group dedicated to ending discrimination in the sport
said Thimm "means a lot to the hockey world."
"She's important because as all the Western hockey countries like Sweden
all of them are affected by demographic change," Hyun said
"That means the sport has a chance to diversify
but also having experience within the hockey world
I think that would inspire a lot of young girls
the way she brings a much different perspective to the game are important."
Thimm said it was challenging being a Black woman playing hockey in Germany in the 1990s
"I think the first time I really recognized and experienced that I was different was when I went from my hometown to the DFEL
players started yelling at me and stuff like that
you try to keep it in the back of your mind because you don't want it to change your game
But it was really tough for me because the crowds were yelling at me the whole time
we'd have to call the police to bring me safe to the bus."
But the negative receptions didn't discourage her because hockey was her sanctuary
"Hockey was my island because my mom was pretty sick at the time
and I didn't want to let people in my mind who want to interrupt or stop me."
Thimm recently stepped down as coach of Bergkamen
a position she held while she also worked as a high school physical education
It helped me out in difficult situations in my life
The move is part of the company's strategy to bid for a portion of the German Government's upcoming tender for 10GW of new capacity
German energy company Iqony is planning to construct hydrogen-ready gas-fired power plants at three of its existing sites in the country, Reuters has reported
The move is part of the company’s strategy to bid for a portion of the German Government’s upcoming tender for 10GW of new capacity
Reuters quoted the company’s spokesperson as saying: “We are preparing to build new hydrogen-ready gas plants at Bergkamen
the plants would have a capacity of 2.5GW.”
The locations of the proposed gas-fired plants were first reported by Germany’s Frankfurter Allgemeine Zeitung newspaper
Iqony’s commitment to these plants is contingent upon the specific conditions outlined in the government’s strategy
which are expected to be clarified in the tenders scheduled for 2024
Don’t let policy changes catch you off guard
Stay proactive with real-time data and expert analysis
The government has been encouraged by interested parties to provide more details regarding the operation of a capacity mechanism scheduled to take effect in 2028
the process will compensate for capacity that will only operate part-time
Power generating company Uniper recently disclosed its potential development of 1–2GW of capacity
These developments are part of a broader effort by Berlin to accelerate the phase-out of domestic brown coal plants before the 2038 deadline and to facilitate the closure of hard-coal power plants by 2030
Iqony is also planning to construct electrolysis plants for hydrogen production at Duisburg-Walsum and Voelklingen-Fenne
and large power batteries at Duisburg-Walsum and Bexbach
These projects are part of the company’s strategy to diversify into renewable energies and technologies
Iqony, which was separated from coal-fired power producer Steag in a sales process in August 2023
involving the consortium Kommunale Beteiligungsgesellschaft
Steag was divided into two divisions: Steag Power
which operates hard-coal-fired power plants
which focuses on renewable energies including solar
engineering services and gas-fired power plants
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Germany – Iqony eyes plans for three hydrogen-ready gas plants
The spokesperson said in reply to an enquiry:
We are preparing to build new hydrogen-ready gas plants at Bergkamen as well as in Bexbach and Quierschied-Weiher
the plants would have a capacity of around 2.5 gigawatts (GW).”
Germany’s Frankfurter Allgemeine Zeitung newspaper reported the locations earlier
A switch to clean hydrogen from polluting fossil fuels is part of Germany’s plans for the green energy transition
Iqony was spun off from coal-fired power producer Steag as part of a sales process to Spanish investor Asterion and includes renewable power
If and how its three planned plants take shape will depend on conditions attached to the government strategy ahead of its planned tenders this year
Interested parties have urged the government to clarify how a capacity mechanism meant to kick in in 2028 would work
The mechanism will provide remuneration for capacity that will only run part-time as renewable power gets priority
Sector peer Uniper (UN0k.DE), opens new tab on Wednesday said it might develop between 1-2 GW of capacity provided the conditions are right
The plans are part of Berlin’s bid to persuade producers of domestic brown coal to close plants earlier than the official deadline of 2038 and ensure that the closure of hard-coal power plants can happen by 2030
Iqony also said it was planning electrolysis plants for hydrogen at Duisburg-Walsum and Voelklingen-Fenne
and big power batteries at Duisburg-Walsum and Bexbach
READ the latest news shaping the hydrogen market at Hydrogen Central
Germany – Iqony eyes plans for three hydrogen-ready gas plants. source
Hydrogen on show at the TotalEnergies 6 Hours of Spa-Francorchamps A pillar of endurance racing and the Automobile Club de l’Ouest’s efforts to decarbonise motorsport
hydrogen will be highlighted during the forthcoming..
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a hydrogen hub in upheaval as natural gas producer pauses involvement while four others bail on the project
South Australia closes Hydrogen Power SA office The state government of South Australia has rolled its Office of Hydrogen Power SA (OHPSA) into the Department of Energy and Mining (DEM)
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Organic acid (OAD) and fatty acid oxidation disorders (FAOD) are inborn errors of metabolism often presenting with life-threatening metabolic decompensation followed by (irreversible) organ failure
Most of these diseases are considered as treatable
and metabolic decompensations can be avoided by early diagnosis and start of therapy
Confirmation of suspected diagnosis currently relies on enzymatic and mutation analyses and in vitro loading of palmitic acid in human skin fibroblast cultures
in some cases potentially life-threatening in vivo loading or fasting tests are still performed
we established a standardized in vitro loading test in peripheral blood mononuclear cells (PBMC) that allows reliable biochemical confirmation of a suspected diagnosis within 1 week
Patients with confirmed diagnosis of short-
and long-chain 3-hydroxyacyl-CoA dehydrogenase deficiencies
and glutaric aciduria type I were included in the study
isolated from heparinized venous blood samples of these individuals were incubated for 5 days with palmitic acid or 2-oxoadipic acid (glutaric aciduria type I)
and quantitative acylcarnitine profiling was subsequently performed in supernatants using electrospray ionization tandem mass spectrometry
including those with mild biochemical phenotypes who
are at risk to be missed under balanced metabolic conditions
the same results were also obtained using lymphoblasts
our assay allows biochemical confirmation of a number of FAOD and OAD and could easily be implemented into the confirmatory diagnostic work-up
specifically increasing the flux through deficient metabolic pathways of mitochondrial β-oxidation of fatty acids
potentially life-threatening in vivo loading or fasting tests are still performed
we present a modified method for the diagnosis of FAOD (SCADD
and the cerebral OAD GA-I using in vitro loading of peripheral blood mononuclear cells (PBMC) or lymphoblasts with palmitic acid and other substrates
This alternative method allows the reliable detection of mild and classical biochemical phenotypes of these diseases
It is more rapidly available and less cost-intensive than enzyme and mutation analysis as well as in vitro loading in human skin fibroblasts
This method is helpful for the biochemical confirmation of patients with suspected diagnosis of FAOD and OAD
in particular those with mild biochemical phenotypes
and to bridge the time period until conventional confirmatory diagnosis is performed
have not been used for biochemical confirmation of diagnosis or biochemical monitoring in these patients
heterozygous carriers for disease-causing MCAD mutations (n = 8) and healthy volunteers for the analysis in PBMC (n = 26) respectively lymphoblasts (n = 10) were included
Informed consent was obtained from the parents and healthy volunteers
The study was approved by the Institutional Review Board (#083/2003)
Blood samples were diluted 1:1 (vol/vol) with RPMI medium
The diluted samples were superposed on a Ficoll phase (twice the volume of the diluted sample
25°C) in 15 mL tubes (Falcon®; BD Biosciences)
carefully avoiding mixture of sample and Ficoll
Centrifugation was performed for 35 minutes at 2000 × g and 20°C
the PBMC interphase was carefully removed and washed twice for each 5 minutes with RPMI medium
The PBMC pellet was resolved in standard growing medium (see later) and maintained at 37°C
B lymphocytes of at least 5 mL venous EDTA blood samples from GA-I patients (n = 3) and healthy volunteers (n = 10) were immortalized using EBV virus supernatant and cultivated for 6 wk prior application
All preparations and subsequent loading experiments were performed in a blinded fashion
Standard cultivation procedures were as follows
PBMC and lymphoblasts were maintained in RPMI 1640 medium (PAA Laboratories GmbH
Austria) supplemented with 10% fetal calf serum (PAA Laboratories GmbH
1000 IU/mL penicillin and 1 mg/mL streptomycin (all purchased from PAA Laboratories)
Twenty-five U/mL IL-2 (IL-2) were added to PBMC
Cell viability was determined by trypan blue exclusion
and 3 × 106 viable cells were used immediately after isolation (PBMC) or after 6 weeks in culture (lymphoblasts) for the loading experiments with unlabeled substrates (palmitic acid
and tryptophan (all obtained from Sigma Chemical Co.
Cells were washed twice in phosphate-buffered saline and resuspended in 1 mL SMEM (Invitrogen
Germany) supplemented with 0.4% bovine serum albumin (wt/vol; Sigma Chemical Co.)
1 mg/mL streptomycin and 25 μg/mL amphotericin B
2 mM l-glutamine (Invitrogen) and 100 μM palmitic (all except for GA-I)
followed by quantitative analysis of acylcarnitines
Aliquots of PBMC or lymphoblasts were used to determine cell viability at the end of the experiments using FACS analysis (Becton-Dickinson
different parameters were stepwise modified
concentrations of l-glutamine and palmitic acid
propionic acid) were investigated in sister cultures
aliquots of 20 μL supernatant were collected after 5 days of incubation and were mixed for 20 minutes with 100 μL of isotope-labeled internal standard solution (Cambridge Isotope Laboratories
Samples were filtered and methanol was removed
followed by butylation of the dried samples
Twenty-five μL of the butylated samples were injected by a CTC-PAL autosampler and a Rheos Flux 2000 HPLC pump into the electrospray ion source of a triple quadruple MS/MS (Micromass Quattro Ultima)
Each analysis was performed at least in triplicate
Further experiments were performed in sister cultures at different time points (1–6 d of incubation) to determine the optimal time point for quantitative acylcarnitine profiling
Cutoffs for acylcarnitines and acylcarnitine ratios were defined as more than mean + 1.96 × SD
All experiments were performed at least in triplicate
Data were analyzed by t test (two groups) or one-way analysis of variance (ANOVA) followed by post hoc Scheffé's test (three or more group)
Influence of assay conditions on cell viability of PBMC and acylcarnitine concentrations
(A) Time-dependent decrease in cell viability of PBMC (n = 6) during incubation with palmitic acid (100 μM)
Cell viability was determined by FACS analysis (forward/sideward Scatter)
*p < 0.05 vs 1 day of incubation (one-way ANOVA followed by post hoc Scheffé's test)
(B) PBMC from controls (n = 8) were loaded with palmitic acid (100 μM) in the presence of l-glutamine (0–2 mM; SMEM medium)
Cell viability was determined at day 5 of incubation by FACS analysis
Cell viability in l-glutamine–treated (2 mM) cells was taken as 100%
*p < 0.001 vs glutamine-free media (one-way ANOVA followed by post hoc Scheffé's test)
(C) PBMC from controls (n = 5) were incubated with 0–800 μM palmitic acid in SMEM medium
and cell viability was determined at day 5 of incubation by FACS analysis
Cell viability in palmitic acid-free media was normalized to 100%
*p < 0.001 vs palmitic acid-free media (one-way ANOVA followed by post hoc Scheffé's test)
(D) Quantitative acylcarnitine profiling of the control group (n = 26) after 5 days of incubation with palmitic acid (100 μM) under standard conditions
Palmitic acid induced a concentration-dependent decrease in cell viability in PBMC from healthy volunteers (n = 5), reaching significance at concentrations of 400 and 800 μM palmitic acid (Fig. 1B)
A concentration of 100 μM palmitic acid was chosen for the investigation of FAOD and classical OAD
(A) The effect of different metabolic in vitro conditions (anabolic state applying RPMI medium vs catabolic state using SMEM medium) on C8 levels in MCADD patients (n = 4; squares) and healthy volunteers (n = 10; circles) was investigated at day 5 of incubation with palmitic acid (100 μM)
(B) All patients were clearly identified by elevated C8 (100% sensitivity)
whereas none of the controls showed an pathologically elevated concentration of C8 after incubation with palmitic acid (100% specificity)
Biochemical differentiation of MCADD patients (n = 6) from heterozygous carriers of disease-causing mutations (n = 8) and controls (n = 26) was reliably performed by plotting of C8 against C8/C2 and C8/C12 (C)
plotting of C8 against C8/C6 or C8/C10 did not result in a clear differentiation of these three groups (not shown)
Grey boxes represent mean ± 1.96 × SD of controls
and carriers for disease-causing MCAD mutations (n = 8) were incubated according to the previously established standard protocol for in vitro loading with palmitic acid (SMEM medium
Data represent mean of at least triplicates at day 5 of incubation
Grey boxes represent mean ± 1.96 × SD of controls (n = 26)
Data are represented mean of at least triplicates determined at day 5 of incubation
Data are presented as mean of at least triplicates determined at 5th day of incubation
In conclusion, in vitro loading in PBMC and lymphoblasts reliably detected patients with known FAOD and OAD by pathognomonic acylcarnitines and acylcarnitine ratios (Table 4)
revealing intermittently normal biochemical phenotypes (SCADD
GA-I) were clearly unraveled by this assay
reaching a diagnostic sensitivity and specificity of 100% in the patients included
heterozygous carriers of disease-causing MCAD mutations were differentiated from controls and MCADD patients
a number of patients with FAOD but not with OAD showed an intermittently normal biochemical phenotype in serial investigations
Loading tests in vivo may induce metabolic derangements in affected children
in vitro loading of PBMC has several advantages compared with conventional procedures: (1) patients are not threatened by in vitro loading tests; (2) the variability can be kept low and the reliability of this assay is high because of standardized biochemical conditions; (3) selective increase in flux through deficient metabolic pathways using in vitro loading also allows a reliable detection of patients with a mild biochemical phenotype; (4) PBMC are easily available
and results can be obtained within less than 1 week
whereas cultivation of fibroblasts requires at least 6 weeks; (5) loading tests with PBMC can be easily repeated if necessary (although this should be restricted
and venous puncture is less invasive than a skin biopsy (except for a punch biopsy); and (6) no radiochemicals are used for this assay
In addition to their diagnostic properties
PBMC or lymphocytes are an easily available model system for the simulation of metabolic derangement in vitro
by comparison of cell viability or various biochemical parameters varying metabolic conditions and treatment protocols (preliminary data not shown)
in vitro loading in PBMC is a reliable and safe tool for the biochemical confirmation of patients with suspected diagnosis of FAOD and OAD
including those with an (intermittently) normal biochemical phenotype using MS/MS in DBS or GC/MS in urine under balanced metabolic conditions
long-chain 3-hydroxyacyl-CoA dehydrogenase [EC 1.1.1.211] (deficiency)
medium-chain acyl-CoA dehydrogenase [EC 1.3.99.3] (deficiency)
electrospray ionisation-tandem mass spectrometry
short-chain acyl-CoA dehydrogenase [EC 1.3.99.2] (deficiency)
very long-chain acyl-CoA dehydrogenase [EC 1.3.99.13] (deficiency)
Roscher AA 2004 Frequencies of inherited organic acidurias and disorders of mitochondrial fatty acid transportation and oxidation in Germany
Hoffmann GF 2002 Incidence and short-term outcome of children with symptomatic presentation of organic acid and fatty acid oxidation disorders in Germany
Hoffmann GF 2003 Expanded newborn screening for inborn errors of metabolism by electrospray ionization-tandem mass spectrometry: results
Hoffmann GF 1999 Sensitivity and specificity of free and total glutaric acid and 3-hydroxyglutaric acid measurements by stable-isotope dilution assays for the diagnosis of glutaric aciduria type 1
Naylor EW 2003 Use of tandem mass spectrometry for multianalyte screening of dried blood specimens from newborns
Sweetman L 1989 Quantitative analysis for organic acids in biological samples: batch isolation followed by gas chromatographic-mass spectrometric analysis
Lehnert W 1994 Long-term results of selective screening for inborn errors of metabolism
Bennett MJ 1998 Clinical and biochemical features of fatty acid oxidation disorders
Grady GF 2001 Tandem mass spectrometry analysis for amino
and fatty acid disorders in newborn dried blood spots: a two-year summary from the New England screening program
Wijburg FA 2002 Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency: Clinical presentation and follow-up of 50 patients
and prevention of disease in glutaryl-CoA dehydrogenase deficiency
Walter JH 2003 The impact of screening for propionic and methylmalonic acidaemia
Surtees R 1998 Neurological outcome of methylmalonic acidaemia
Wendel U 2003 MS/MS-based newborn and family screening detects asymptomatic patients with very-long-chain acyl-CoA dehydrogenase deficiency
Am J Med Genet C Semin Med Genet 121: 38–52
Leonard JV 1992 Neurologic outcome of propionic academia
Wiley V 2002 Neonatal screening for medium-chain acyl-CoA dehydrogenase deficiency
Hoffmann GF 2004 Pathomechanisms of neurodegeneration in glutaryl-CoA dehydrogenase deficiency
Leonard JV 1995 The management and outcome of propionic and methylmalonic acidaemia
Saudubray JM 1994 Clinical outcome of long-term management of patients with vitamin B12-unresponsive methylmalonic acidemia
Roe C 1998 Mitochondrial very-long-chain acyl-coenzyme A dehydrogenase deficiency: clinical characteristics and diagnostic considerations in 30 patients
Gregersen N 2001 Medium-chain acyl-CoA dehydrogenase (MCAD) mutations identified by MS/MS-based prospective screening of newborns differ from those observed in patients with clinical symptoms: identification and characterization of a new
prevalent mutation that results in mild MCAD deficiency
Duran M 2003 Short-chain acyl-CoA dehydrogenase deficiency: studies in a large family adding to the complexity of the disorder
Prasad C 2002 Outcome of the first 3-years of a DNA-based neonatal screening program for glutaric acidemia type 1 in Manitoba and northwestern Ontario
Bross P 2001 Mutation analysis in mitochondrial fatty acid oxidation defects: Exemplified by acyl-CoA dehydrognease deficiencies
with special focus on genotype-phenotype relationship
Ribes A 2000 Glutaryl-CoA dehydrogenase deficiency in Spain: evidence of two groups of patients
Mayatepek E 2001 Molecular and functional characterisation of mild MCAD deficiency
Hoffmann GF 2004 Neonatal screening for glutaryl-CoA dehydrogenase deficiency
Bartlett K 2001 Neonatal screening for medium-chain acyl-CoA dehydrogenase deficiency
Christensen E 1993 A fibroblast glutaryl-CoA dehydrogenase assay using detritiation of 3H-labelled glutaryl-CoA: application in the genotyping of the glutaryl-CoA dehydrogenase locus
Ijlst L 1999 Disorders of mitochondrial fatty acyl-CoA β-oxidation
Hoffmann GF 2000 Mutation analysis in glutaric aciduria type I
Wanders RJ 1999 Quantitative acylcarnitine profiling in fibroblasts using [U-13C] palmitic acid: an improved tool for the diagnosis of fatty acid oxidation defects
Mayatepek E 2002 A method for quantitative acylcarnitine profiling in human skin fibroblasts using unlabelled palmitic acid: Diagnosis of fatty acid oxidation disorders and differentiation between biochemical phenotypes of MCAD deficiency
Jakobs C 2000 Quantitative analysis of urinary acylglycines for the diagnosis of beta-oxidation defects using GC-NCI-MS
Blau N 2000 Evaluation of urinary acylglycines by electrospray tandem mass spectrometry in mitochondrial energy metabolism defects and organic acidurias
Davies D 1971 Separation of peripheral leukocytes by Ficoll density gradient centrifugation
Cohen L 1966 The population-dependent requirement by cultured mammalian cells for metabolites
Glutamic acid and glutamine; aspartic acid and asparagine
Calder PC 1999 Dietary glutamine enhances murine T-lymphocyte responsiveness
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Andrea Schulze-Bergkamen and Jürgen G Okun: A.S.B
Andrea Schulze-Bergkamen & Henning Schulze-Bergkamen
Department of Biochemistry and Medical Genetics
This study was supported by a grant from the Medical Faculty of the University of Heidelberg to A
(# F.206611) and by a grant from the German Federal Ministry of Education and Science (BMBF # 01GM0305; project # 8) to S
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DOI: https://doi.org/10.1203/01.PDR.0000181378.98593.3E
Journal of Inherited Metabolic Disease (2017)
Analytical and Bioanalytical Chemistry (2013)
Journal of Inherited Metabolic Disease (2007)
You can read this article in 3 minutesGregor Gowans
DHL Supply Chain has announced the opening a new
highly automated fulfillment centre in the German city of Bergkamen
The facility has been commissioned to help the furniture retailer with its fast-growing ecommerce business in Germany
boasts 40,000 square meters of warehouse space and is part of the neighboring DHL Supply Chain Unna Campus
DHL said that 20,000 square meters of warehouse space will be available for IKEA alone
350 employees will work on-site six days a week in three shifts
The fulfillment centre will be fully operational at the end of this month
at which point an average of 8,000 stock keeping units (SKUs = identical products with the same stock number) will be processed on-site
DHL says the site is capable of processing 11,000 SKUs
The facility also includes a number of machines to accelerate and automate processes
Chief among them are the automated packaging machines available on-site for sensitive goods that require special packaging
Chief Executive Officer DHL Supply Chain Germany & Alps
Country Customer Fulfilment Manager for IKEA Deutschland
“Having two leading companies in their respective industries partner here on-site offers a degree of security for the city of Bergkamen as well as for employees
It is therefore a welcome partner for Bergkamen.”
Pölös Zsófia Journalist Trans.info | 5.05.2025
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GXO to manage Northern Italy transport operations for PRG retail groupPölös Zsófia Journalist Trans.info | 5.05.2025
Metrics details
Colorectal cancer (CRC) is one of the most common cancers in the Western world
5-Fluorouracil (5FU)-based chemotherapy (CT) remains the mainstay treatment of CRC in the advanced setting
and activates executioner caspases in target cells
Executioner caspases are key proteins involved in cell disassembly during apoptosis
Activation of executioner caspases also has a role in tissue regeneration and repopulation by stimulating signal transduction and cell proliferation in neighbouring
Tissue microarrays (TMAs) consisting of tumour tissue from 93 stage II and III colon cancer patients were analysed by immunohistochemistry
patients with low levels of active Caspase-3 had an increased disease-free survival time
This was particularly pronounced in patients who received 5FU-based adjuvant CT
lower serum levels of active Caspase-3 were found in patients with metastasised CRC who revealed stable disease or tumour regression compared with those with disease progression
The role of Caspase-3 in treatment responses was explored further in primary human tumour explant cultures from fresh patient tumour tissue
Exposure of explant cultures to 5FU-based CT increased the percentage of cells positive for active Caspase-3 and Terminal Deoxynucleotidyl Transferase dUTP Nick end Labelling (TUNEL)
but also the expression of regeneration and proliferation markers β-Catenin and Ki-67
selective inhibition of Caspase-3 with Ac-DNLD-CHO
significantly reduced the expression of proliferation markers as well as COX-2
Inhibition of COX-2 with aspirin or celecoxib did not affect Caspase-3 levels but also reduced Ki-67 and β-Catenin levels
suggesting that Caspase-3 acted via COX-2 to stimulate cell proliferation and tissue regeneration
This indicates that low levels of active Caspase-3 may represent a new predictor of CT responsiveness
or antagonising downstream effectors of Caspase-3 paracrine signalling
such as COX-2 may improve patient outcomes following CT in advanced CRC
Colorectal cancer (CRC) has the third highest incidence rate and the fourth highest mortality rate of any cancer in the Western world
or combined with 5-Fluorouracil (5FU)-based chemotherapies
is the current treatment option for stage II and advanced
The response rate to palliative 5FU-based treatments for metastatic stage IV CRC is c
but median overall survival (OS) remains low at c.16–19 months
we therefore examined Caspase-3 as a prognostic marker in stage 2/3 as well as in metastatic CRC patients
and further explored the role of caspase-3 as a potential predictive biomarker for 5FU-based CT and therapeutic target in CRC using primary human tumour explant culture
Low active Caspase-3 levels in patient tumours results in a significantly better disease-free survival time compared with high active Caspase-3 levels in stage 2/3 CRC patients receiving 5FU-based CT
were stained for Pro- and active Caspase-3 and scored based on extent and intensity of staining (scale bar=100 μm)
(b) Protein levels were correlated to patient outcome
and patients with low ProCaspase-3 (blue) did similarly to those with high ProCaspase-3 (green)
40 patients expressed low protein levels and 53 patients expressed high levels)
(c and d) This survival pattern was observed whether patients received CT (c; N=35
with 5 experiencing recurrence) or not (d; N=58
(e) Patients with low levels of active Caspase-3 were found to have a significantly better disease-free survival than patients with high Caspase-3 (P=0.045; N=93
64 patients expressed low protein levels and 29 patients expressed high levels)
this effect was further emphasised and disease-free survival was again increased in patients with low active Caspase-3 (P=0.013; N=35
(g) In the patients who received no chemotherapy this effect on disease-free survival was lost (N=58
(h) Western blot analysis on HCT116 cell line
confirmed that the antibodies (cleaved Caspase-3 antibody
Atlas Antibodies) only detected the active (cleaved) form of Caspase-3 and the proform
MCF7 cells were included as a negative control for specificity determination of the ProCaspase-3 antibody
These data suggested active Caspase-3 in tumour tissue as a predictive marker for CRC patients receiving 5FU-based CT
allowing for the identification of patients who are likely to respond to therapy
Western blot analysis performed on staurosporine-treated human colon cancer HCT116 cells confirmed that the ProCaspase-3 and the active Caspase-3 antibodies used only detected the pro-enzyme and cleaved form, respectively (Figure 1h)
High levels of Caspase-3/-7 activity in the serum are indicative of PD in metastasised colorectal carcinoma patients (a)
Caspase-3/-7 activity levels in the serum were evaluated at baseline (before therapy) according to their treatment response following the first cycle of CT
PD (N=7) was associated with higher Caspase-3/-7 activity compared with SD (N=14) and partial response (PR; N=9)
(b) Patients who displayed PD revealed significantly higher levels of Caspase-3/-7 activity in their serum when compared with patients who displayed SD or partial tumour regression at day 14 following commencement of therapy
(c) Caspase-3/-7 activity levels in these patients could predict PD at day 14 with a sensitivity of 86% and a specificity of 78%
Caspase-3 is activated and apoptosis is induced following CT
and COX-2 inhibitors do not affect this process
(a) Fresh patient colon tumour tissue (N=19 patients) was treated with 5FU-based CT alone and in combination with specific Caspase-3 inhibitor (DNLD) and COX-2 inhibitors (Aspirin and Celecoxib)
Tissue was sectioned and stained for ProCaspase-3 and active Caspase-3 (a) and scored based on extent and intensity of staining (scale bar=100 μm)
(b and c) Levels of ProCaspase-3 (b) and active Caspase-3 (c) were quantified
ProCaspase-3 levels were unaffected by treatment regimes
while active Caspase-3 was significantly reduced (5FU/Oxali versus 5FU/Oxali+DNLD
P=0.021) by specific Caspase-3 inhibition alongside 5FU-based CT but unaffected by other treatments
Chemotherapy-induced apoptosis (Untreated versus 5FU/Oxali
which was reduced with Caspase-3 inhibition (5FU/Oxali versus 5FU/Oxali+DNLD
COX-2 inhibition did not reduce apoptosis induction
*Denotes statistically significant difference
Active Caspase-3 and Ki-67 do not colocalise
(a) Tumour explant cultures remained untreated or were treated with 5FU/Oxali alone or in combination with the specific Caspase-3 inhibitor
Tissues were sectioned and double stained for immunofluorescence for active Caspase-3 (green
Represented images from n=x–y explant cultures per treatment condition are shown (scale bar=50 μm)
(b) Detail of active Caspase-3 and Ki-67 staining in a 5FU/Oxaliplatin-treated culture
showing that Ki-67 nuclear staining corresponded to the zone of proliferating epithelial progenitors of the colonic crypts
and differentiating compartments are localised (scale bar=50 μm)
Aspirin co-treatment did not significantly reduce expression of COX-2
Explant samples were examined for ZEB1 expression
Expression was not altered across treatment groups
suggesting that the EMT does not accompany Caspase-3-driven proliferation
active Caspase-3 levels gave an even more notable survival advantage
suggesting that while CT is activating Caspase-3
high levels of the active protein may contribute to the eventual relapse
levels of low ProCaspase-3 were suggestive of improved DFS for patients
implying that ProCaspase-3 levels may be determinant of active Caspase-3 levels
Caspase-3 levels were not associated with DFS
indicating that this phenomenon is present only when CT is administered
This indicates that active Caspase-3 levels represent a potential predictive biomarker of patient outcome and response to 5FU-based CT
helping to identify individuals likely to respond to CT
these data present evidence pointing to Caspase-3 as a potential predictive biomarker of response to 5FU-based CT in advanced CRC
Measurement of Caspase-3 in the tumour or of Caspase-3/-7 activity in the serum may enable reliable stratification of patients based on response and allow for the identification of patients likely to respond poorly to 5FU-based CT
analysis in larger cohorts and long-term studies are required to further validate Caspase-3 or Caspase-3/-7 activity as a biomarker for early prediction of treatment outcome or disease progression in CRC patients
these results suggest that COX-2 inhibitors may have potential as novel co-treatments to compliment traditional chemotherapies in advanced CRC
Patients with hereditary forms of colorectal cancer were excluded
Informed consent was obtained from all patients; and ethical approval was obtained by the Beaumont Hospital Ethics Committee
Three 1-mm cores were taken from every case where available
using the Beecher Instruments arrayer (Sun Prairie
Arrays consisted of 90–100 cores per array
floated onto adhesive slides and baked overnight at 65 °C
Antibodies were chosen based on their suitability for use on FFPE tissue and optimised
All staining was performed on a Leica Bond III automated immunostainer (Leica Microsystems
The following primary antibodies were used: ProCaspase-3 (1 : 200; Rabbit polyclonal
Active Caspase-3 (1:750; Rabbit polyclonal
COX-2 (1 : 400; Mouse monoclonal; CAY160112-1
Negative controls for both rabbit and mouse antibodies (Negative Control Mouse Cocktail from mouse IgG1
Dako; Negative Control Rabbit Immunoglobulin fraction of serum from non-immunized rabbits
Dako) were included; and no staining was observed in these controls
Visualisation was performed using the enhanced diaminobenzidine tetrachloride with Harris′ haematoxylin as counterstain
cores were scored based on the extent and intensity of the protein of interest
As multiple cores for each patient were arrayed
mean intensity scores were used for all subsequent analysis
Patients were allocated as those with either low or high protein levels
based on whether the score was above or below the median
The specificity of ProCaspase-3 and active Caspase-3 antibodies was confirmed by western blotting using the HCT116 cell line+STS treatment
MCF7 cells were included as a negative control
The study was conducted according to the Ethics Committees of the National Centre for Tumour Diseases and Hannover Medical School
Collection media consists of Dulbecco’s modified Eagle’s medium (Gibco
USA) supplemented with penicillin (200 IU/l)
all tissues were cultured at 37 °C and 5% CO2 until harvest
Under sterile conditions and 2 h before tissue dissection
1 cm3 haemostatic gelatine dental sponges (Humanus Dental AB) were hydrated in 1 ml collection media at 37 °C in individual wells of a 12-well cell-culture dish
tissues were transferred to a 10-cm cell-culture dish with 10 ml collection media for dissection
Samples were then cut into 1 mm3 sections using a sterile scalpel blade
Sample sections were washed in fresh collection media
Upon complete hydration of gelatine sponges 1 mm3 dissected specimen sections were chosen at random and placed on top of each sponge using sterile forceps
Explant cultures were incubated at 37 °C and 5% CO2
All explants were maintained in collection media for 24 h
media was aspirated carefully to prevent disturbance of plated tissues
and 1 ml of treatment media was then added to the corresponding treatment wells
media was aspirated and stored at −20 °C for prostaglandin E2 assay
tissues were processed and FFPE for immunohistochemical analysis
Sections were floated onto slides and left to air-dry
Sections were then baked overnight at 65 °C
Samples were deparaffinised by passing through xylene and 100 and 70% ethanol washes
All staining was performed using the DAKO EnVision+ System-HRP (DAB) kit for rabbit (K4010; DAKO) or mouse (K4006; DAKO) primary antibodies
The following primary antibodies were used: ProCaspase-3 (1 : 100; Rabbit polyclonal
Active Caspase-3 (1 : 200; Rabbit polyclonal
β-Catenin (1 : 300; Neomarkers RB-9035 (Fisher Scientific
Positive and negative controls were included according to the suppliers′ recommendations (Negative Control Rabbit Immunoglobulin Fraction (Normal)
on a scale of 0–3 based on the extent and intensity of staining
Specimens were washed using sodium citrate as antigen retrieval
Cells were permeabilised for 30 min using 0.3% Triton X-100 in 1 × PBS
Sections were then incubated overnight at 4 °C with Rabbit polyclonal Cleaved Caspase-3 Antibody (1 : 200; 9661
Cell Signalling Technology) and Mouse Monoclonal Ki-67 Antibody (MIB1
diluted in 1 × PBS with 1% BSA and 0.3% Triton X-100
Alexa Fluor 488 Donkey anti-Rabbit IgG (H+L) (A-21206
USA) and Alexa Fluor 568 Donkey anti-Mouse IgG (A-10037
Life Technologies) secondary antibodies were used at 1:400 dilutions and incubated for 1 h at room temperature
Nuclei were counterstained with Hoechst stain solution
Slides were mounted using DPX Slide mounting medium (06522
As a negative control to ensure specificity of staining
the omission of the primary antibodies from the staining procedure was carried out on some samples
Cells were imaged using a LSM 510 Meta (Carl Zeiss
Germany) equipped with × 40 oil immersion objective using appropriate excitation
To analyse apoptosis in each sample sections were processed using the TdT-DAB in situ apoptosis detection kit (Trevigen
Staining was carried out as per manufacturer’s instructions
and stainings were scored once again based on the extent and intensity of DAB-positive cells
To measure PGE2 levels according to the treatment used; media was collected from the corresponding samples at 96 h of treatment and was processed using the Prostaglandin E2 Parameter Assay Kit (KGE004B
Optical density was determined using a microplate reader set to 450 nm
Evaluation was carried out as per manufacturer’s instructions
Standards and controls were included as described in the kit
Statistical analyses were performed SPSS (IBM
ANOVA and subsequent Tukey test were used for normally distributed data
Relations between levels of active Caspase-3 and Ki-67 on CRC TMAs were compared by Spearman correlation test
Serum levels of activated Caspase-3 were compared by using Mann–Whitney’s U test for non-normally distributed data
P-values≤0.05 were considered statistically significant
ROC analyses were performed to calculate the cutoff value of activated Caspase-3 in the serum that correctly predicts treatment failure
Response Evaluation Criteria in solid tumours
terminal deoxynucleotidyl transferase dUTP nick end labelling
A caspase-activated DNase that degrades DNA during apoptosis
Single-cell fluorescence resonance energy transfer analysis demonstrates that caspase activation during apoptosis is a rapid process: role of CASPASE-3
Executioner caspase-3 and caspase-7 are functionally distinct proteases
Immunosuppressive effects of apoptotic cells
A perspective on mammalian caspases as positive and negative regulators of inflammation
Apoptotic cells activate the "Phoenix Rising" pathway to promote wound healing and tissue regeneration
Apoptotic cells provide an unexpected source of Wnt3 signaling to drive hydra head regeneration
The prognostic value of Ki-67 antigen in non-Hodgkin lymphoma of Waldeyer ring and the nasal cavity
Proliferation markers and survival in early breast cancer: a systematic review and meta-analysis of 85 studies in 32,825 patients
Genetic interaction of PGE2 and Wnt signaling regulates developmental specification of stem cells and regeneration
Caspase-3-dependent activation of calcium-independent phospholipase A2 enhances cell migration in non-apoptotic ovarian cancer cells
Caspase 3-mediated stimulation of tumour cell repopulation during cancer radiotherapy
American Society of Clinical Oncology/College of American Pathologists Guideline Recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer
Detection of apoptotic caspase activation in sera from patients with chronic HCV infection is associated with fibrotic liver injury
Therapeutic response to CDK4/6 inhibition in breast cancer defined by ex vivo analyses of human tumors
Evidence for efficacy of new Hsp90 inhibitors revealed by ex vivo culture of human prostate tumors
Structural and functional definition of the specificity of a novel caspase-3 inhibitor
Ki67 in breast cancer: prognostic and predictive potential
Measuring proliferation in breast cancer: practicalities and applications
Epithelial–mesenchymal transitions in development and pathologies
The role of the ZEB family of transcription factors in development and disease
Cyclooxygenase-2–dependent regulation of E-cadherin: prostaglandin E2 induces transcriptional repressors ZEB1 and Snail in non–small cell lung cancer
Paracrine control of tissue regeneration and cell proliferation by Caspase-3
A genetic model for colorectal tumorigenesis
Baseline caspase activity predicts progression free survival of temsirolimus-treated head neck cancer patients
The extent of liver steatosis in chronic hepatitis C virus infection is mirrored by caspase activity in serum
a novel selective inhibitor of caspase-3 enhances cell death and extends tumor growth delay in irradiated lung cancer models
p53-mediated induction of Cox-2 counteracts p53- or genotoxic stress-induced apoptosis
Inhibition of cyclooxygenase-2 gene expression by p53
Positive feedback regulation of COX-2 expression by prostaglandin metabolites
Positive feedback between PGE2 and COX2 redirects the differentiation of human dendritic cells toward stable myeloid-derived suppressor cells
Aspirin and COX-2 inhibitor use in patients with stage III colon cancer
Aspirin for the prevention of colorectal cancer
Best Pract Res Clin Gastroenterol 2011; 25: 461–472
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This research was supported by a grant from the Health Research Board
Department of Physiology and Medical Physics
Department of Gastroenterology and Oncology
The authors declare no conflict of interest
Supplementary Information accompanies this paper on Cell Death and Disease website
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2022 at 10:34 AM EDTBookmarkSaveLock This article is for subscribers only.Germany is deploying about 3 gigawatts of coal-fired generation to ensure there are enough electricity supplies to make it through the winter amid curtailed natural gas supplies from Russia
The Bergkamen-based tuner went to work recently on a Panamera Turbo S
The latter looks like the steering wheel of a Pagani
The exterior of the car was also modified, as per the owner’s request, and now features a matte full-body wrap in dark grey, with neon green highlights that match the interior
Watch: Just How Good Is The 2021 Porsche Panamera Turbo S?
that ‘PS-Sattlerei Evo’ steering wheel really stands out thanks to its carbon fiber upper and lower sections and their rhombic structure
while the grips on either side are upholstered in non-slip perforated black Nappa leather
Other features include the 12 o’clock stripe and the pronounced contours
Moving on to the seats and door panels, they too boast Nappa leather, to go with a Lamborghini-sourced neon-green leather
the seats and backrests have been adorned with diamond quilting
whereas Alcantara can be found on the side panels of the seats and on the dashboard
Last but certainly not least is the starlit sky headliner composed of many tiny LEDs that even allow for lighting effects such as catching a glimpse of a shooting star every now and then
the tuner created a custom duffel bag to match the interior
German specialty chemicals company Lanxess has agreed to sell its organometallics business to US chemicals firm PMC Group
PMC’s European subsidiary will acquire the business
which includes Lanxess’s global organotin catalyst
organotin specialties and intermediates product lines
The deal also includes a toll manufacturing agreement under which Lanxess will continue to manufacture certain products for PMC at its plant in Bergkamen
The transaction is scheduled to close by the end of the year
The acquisition is the second for PMC this year and follows the purchase of Solvay’s hydrocolloid product line in January through wholly owned French subsidiary PMC Ouvrie. The products are sold to the home and personal care
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The German Women's Ice Hockey League (DFEL) will start the upcoming 2024/2025 season with six teams
a team from Hungary will also take part in the competition
The five-time champion of the European Women's Hockey League (EWHL)
will strengthen the DFEL for the coming season
The other matches for the coming season have also already been confirmed
The season will be played in a double round as usual
5./ 6.10.2024 I ECDC Memmingen Indians - Mad Dogs Mannheim
5./ 6.10.2024 I Eisbären Berlin - ERC Ingolstadt
5./ 6.10.2024 I EC Berkamener Bären – Hokiklub Budapest
The last matchday of the DFEL main round is scheduled for the weekend of February 15/16
The champions of the 2024/2025 season will then be determined by March 29
The entire schedule of the German Women’s Ice Hockey League can be viewed here: https://deb-online.live/liga/damen/bundesliga/
Development Coach Women Ronja Jenike: "We are delighted to welcome a strong new team to the DFEL with the Kandai Magyar Hokiklub Budapest
Women's ice hockey in Hungary has taken a big step forward in recent years
both in the club sector and in the national team
The competition from Budapest will be a good benchmark for the DFEL teams and will contribute positively to the sporting development of the German women's ice hockey league."
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We investigated the mechanisms by which TAp73β and dominant-negative p73 (ΔNp73) regulate apoptosis
TAp73β transactivated the CD95 gene via the p53-binding site in the first intron
TAp73β induced expression of proapoptotic Bcl-2 family members and led to apoptosis via the mitochondrial pathway
Endogenous TAp73 was upregulated in response to DNA damage by chemotherapeutic drugs
ΔNp73 conferred resistance to chemotherapy
Inhibition of CD95 gene transactivation was one mechanism by which ΔNp73 functionally inactivated the tumor suppressor action of p53 and TAp73β
ΔNp73 inhibited apoptosis emanating from mitochondria
ΔNp73 expression in tumors selects against both the death receptor and the mitochondrial apoptosis activity of TAp73β
The importance of these data is evidenced by our finding that upregulation of ΔNp73 in hepatocellular carcinoma patients correlates with reduced survival
Our data indicate that ΔNp73 is an important gene in hepatocarcinogenesis and a relevant prognostic factor
The discovery of a ΔNp73 (dominant-negative p73)-based p53–p73 interference network suggests that the p53 status of a tumor should no longer be regarded as the sole predictor of clinical outcome and therapeutic responsiveness
the TP73 locus encodes both a tumor suppressor (TAp73) and a putative oncogene (ΔNp73)
Given the possible role of TAp73 and ΔNp73 isoforms in cancer
it is of great interest to determine how they regulate apoptosis
as the critical process which links tumor development
treatment sensitivity and clinical outcome
We have analyzed the mechanisms of TAp73/ΔNp73-regulated apoptosis and their relevance for chemosensitivity and prognosis in hepatocellular carcinoma (HCC)
Our data suggest possible signaling pathways through which the balance between TAp73 and ΔNp73 regulates the apoptosis response of cancer cells
thereby playing a decisive role in the choice between elimination of cancer-prone cells versus hepatocarcinogenesis as well as between treatment sensitivity versus drug resistance
we show that ΔNp73 is a predictor of adverse outcome and a new prognostic factor in HCC
Data obtained in five separate experiments were averaged
(c) The caspase inhibitors DEVD-FMK (C3Inh)
Z-IETD-FMK (C8Inh) and Z-LEHD-FMK (C9Inh) significantly reduced TAp73β-dependent apoptosis
Three independent experiments were performed
(d) Determination of the increased enzymatic activity of the caspase-3
-8 and -9 class of proteases was performed by fluorometric assays 48 h following rAd-TAp73β transfer (10 MOI) into Hep3B cells
Fold activation represents the DEVDase- (consistent with caspase 3-)
IETDase- (consistent with caspase 8-) and LEHDase- (consistent with caspase 9-) activity of Hep3B cells transduced by rAd-TAp73β compared to cells transduced by rAd-GFP
Additional evidence for the involvement of receptor-mediated signaling in TAp73β-induced apoptosis was provided by the fact that the pro-caspase-8-binding adapter protein Fas-associated death domain (FADD) was found to be upregulated
Activation of caspases is clearly involved in mediating TAp73β-induced downstream apoptosis signaling. Caspase-1, -2, -3, -4, -6, -8, -9 and -10 were found to be upregulated in response to TAp73β (Table 1)
Furthermore, we identified the genes encoding the proapoptotic Bcl-2 family members BAD and BIK and the genes encoding BNIP3, HRK and RAD9 as targets for transcriptional upregulation in cells overexpressing TAp73β (Table 1)
microarray analyses provide evidence that TAp73β stimulates both
genes that regulate the extrinsic apoptosis pathway initiated by ligation of death receptors and genes that regulate the intrinsic/mitochondrial apoptosis pathway
TAp73β induces upregulation of the CD95 receptor and sensitizes towards CD95-mediated apoptosis
Semiquantitative PCR-analysis of CD95 mRNA expression following adenoviral transfer (24 h) of either rAd-GFP
rAd-p53 or rAd-TAp73β (10 MOI each) into Hep3B cells (p53−/−)
(b) FACScan® analysis of CD95 receptor expression in Hep3B cells following transduction by either rAd-GFP
Transfer of rAd-TAp73β (72 h) as well as restitution of a wt p53 status restores the ability of the p53−/− cell line Hep3B to increase the CD95 receptor
Presented are representative diagrams of FL-2 (CD95) for rAd-p53 and rAd-TAp73β
TAp73β induces upregulation of the CD95 receptor in Hep3B (dose dependent) and in Saos2 cells (50 MOI)
FACScan® analyses were performed in triplicates
six independent experiments were performed
a representative result is shown (mean±S.D.
(e) TAp73β sensitizes Hep3B cells towards CD95-mediated apoptosis
Only Hep3B cells transduced by rAd-TAp73β exhibited a significantly increased responsiveness towards induction of apoptosis by anti-APO-1
Data are expressed as % increase in specific apoptosis due to addition of anti-APO-1
FACScan® analysis following Nicoletti staining
A representative result of three independent experiments is shown
Importantly, FACS analysis revealed that overexpression of TAp73β led to an increase in the amount of CD95 displayed on the cell surface in Hep3B (Figure 2b and c). This increase in CD95 expression was also observed in a second cellular model, in Saos2 cells (Figure 2d)
Saos2 cells are p53 negative and show no detectable levels of p63 and p73 at either mRNA or protein level
The increased number of death receptors on the cell surface is likely to increase sensitivity of cancer cells towards apoptosis. We tested this hypothesis by treating Hep3B cells with agonistic anti-APO-1 antibody. Figure 2e shows that the antibody is in fact able to trigger cell death in TAp73β-overexpressing Hep3B cells
but not in cells infected with a control green fluorescent protein (GFP)-adenovirus
This indicates that TAp73β induces the expression of a functional CD95 death receptor on the cell surface
essential nucleotides for the binding of wt p53 protein have been mutated in the intronic p53-responsive element
γ and Δ transactivated the CD95 gene via binding to the p53-binding site in its first intron
Mutation of this binding site (mt p1142CD95-luc) completely abrogated binding and transactivation by TAp73-isoforms
Cells were transfected with 1 μg of plasmid p1142CD95-luc together with 100 ng of either a wt p53-plasmid
Presented is the fold p53- or TAp73-dependent activation of the p1142CD95-luc reporter plasmid
calculated relative to the value obtained with the same reporter in the absence of p53 or the TAp73-isoforms
Assays were performed in triplicates in five independent experiments
One representative experiment is shown (mean±S.D.
This argues strongly in favor of the conclusion that the CD95 gene is a direct transcriptional target of TAp73
Dominant-negative (ΔN) p73β and delta (Δ) 84p73β inhibit TAp73- and p53-mediated transactivation of the CD95 gene and counteract apoptosis induced by TAp73β
analysis of TAp73-dependent luciferase activity
Hep3B cells were transfected with 1 μg of plasmid p1142CD95-luc together with 100 ng of either a TAp73α
or an equivalent amount of empty vector alone or together with the respective ΔNp73- or Δ84-isoform
Presented is the fold TAp73- or p53-dependent activation of each reporter plasmid
calculated relative to the value obtained with the same reporter in the absence of the respective TAp73-isoform or of p53
ΔNp73 (a) and Δ84p73β (a) were not able to transactivate the CD95 gene
they act as dominant-negative being able to repress the ability of TAp73 to drive the CD95 gene
In addition to their ability to inhibit TAp73-mediated transactivation of the CD95 gene
the ΔN- and Δ84 isoforms of p73 inhibit wt p53-induced transactivation of the CD95 gene (c)
(d) ΔN- and Δ84 isoforms of p73 inhibit specific apoptosis induced by TAp73β
Hep3B cells were transduced for 72 h either with rAd TAp73β (10 MOI) or rAd Δ84p73β (10 MOI)
Data obtained in three separate experiments were averaged
In order to further characterize the molecular mechanisms of TAp73-mediated apoptosis, we investigated the influence of TAp73β on mitochondrial apoptosis signaling. FACS analysis following JC-1 staining revealed an alteration of the mitochondrial membrane potential following adenoviral TAp73β transfer in Hep3B cells (Figure 5a).
Hep3B cells were transfected with 1 μg of the reporter plasmid presented in (b) together with 10 MOI rAd-TAp73β
The fold TAp73β-dependent activation of the Bax-Pr/luc reporter plasmid
calculated relative to the value obtained with the same reporter in the absence of TAp73β is shown
TAp73β induced a significant time-dependent increase of the transactivation of the Bax gene (ANOVA)
A representative result of five independent experiments is shown
(d) Immunoblot of endogenous Bax expression following rAd-TAp73β transfer
(e) ΔNp73 and Δ84p73β act as dominant-negative being able to repress the ability of p53 and TAp73β to transactivate the Bax gene
Presented is the fold TAp73β- or p53-dependent (10 MOI) activation of the Bax-Pr/luc reporter plasmid (1 μg)
calculated relative to the value obtained with the same reporter in the absence of TAp73β or p53
Assays were performed in triplicates in three independent experiments
TAp73β contributes to apoptosis by inducing the expression of several proapoptotic genes involved in mitochondrial signaling
Blocking TAp73β by rAd-Δ84p73β significantly decreased Bax-Pr/luc activity. Furthermore, Δ84p73β did also block p53-mediated transactivation of the Bax promoter (Figure 5e)
in addition to interference with the death receptor-apoptosis activity of TAp73β
N terminus-deleted p73-isoforms exert their antiapoptotic action also on mitochondrial apoptosis signaling pathways
This has been implicated in resistance towards chemotherapy
We therefore next investigated if TAp73β restores chemosensitivity of cancer cells
One representative out of three experiments performed is shown
(g) Western blot analysis confirms the cooperative induction of endogenous Bax protein by rAd-TAp73β and bleomycin in Hep3B cells
Since TAp73 and ΔNp73 show opposite effects on apoptosis induction
we investigated if ΔNp73 interferes with chemotherapy-induced apoptosis
This supports the hypothesis that the ratio of the two isoforms TAp73/ΔNp73 is an important determinant of clinical response to chemotherapy
This prompted us to further analyze the clinical significance of ΔNp73 expression in patients with HCC
Western blot analysis revealed that TAp73 and ΔNp73 are specifically expressed in tumor tissue, but not in non-neoplastic liver tissue. Hereby, TAp73 and ΔNp73 seemed to be inversely regulated, that is, high expression of TAp73 corresponded to low expression of ΔNp73 and vice versa (Figure 8).
Western Blot analysis of ΔNp73 and TAp73 expression in six HCCs (‘T’) and corresponding non-neoplastic liver tissue (‘N’) using our specific antibodies
TAp73 and ΔNp73 are distinctively expressed in tumor tissue
(b) Positive tumor cell nuclei with a slight perinuclear staining during mitosis (arrowhead): ΔNp73 immunoreactivity occurs also in the cytoplasm
Sinus endothelial cells are ΔNp73-negative (asterisk) (original magnification × 100; oil immersion)
(c) Surrounding liver tissue is negative for ΔNp73 (original magnification × 20)
p53 mutational analysis revealed that 46% (39/84) of these HCC patients carried p53 mutations. There was no correlation between ΔNp73 overexpression and the p53 mutational status (Table 3)
Model of TAp73/ΔNp73-regulated apoptosis
the death receptor- (pathway 1) and the mitochondrial- (pathway 2) apoptosis pathway
ΔNp73 is a strong inhibitor of both pathways
There may exist additional proapoptotic genes (harboring p53 binding elements) whose expression is downregulated by ΔNp73
contributing to tumor progression and a worse patient outcome
We identified the Bax promoter to be repressed by ΔNp73
resistance towards apoptosis by ΔNp73 is also imposed by inhibition of mitochondrial apoptosis pathways
ΔNp73 strongly inhibited chemotherapy-induced activation of caspase-3
our data couple the disruption of apoptosis at the death receptor- and mitochondrial level by ΔNp73 with intrinsic drug resistance
We show that the functional status of TAp73/ΔNp73 is an important determinant of cellular response to chemotherapeutic drugs
While the expression of TAp73 synergizes with chemotherapeutic drugs
the protein isoform ΔNp73 confers drug resistance
relative balance of these two isoforms may influence the clinical outcome
Our antibody raised specifically against ΔNp73
identified 31 out of 84 (37%) of patients with HCC to overexpress ΔNp73 in their tumor tissue
we provide the first evidence that ΔNp73 upregulation in HCC is correlated with a poor prognosis
Patients with tumors overexpressing ΔNp73 exhibited a significantly shorter survival time compared with patients whose tumors were ΔNp73 negative (P<0.005 log-rank test)
This is an important and clinically relevant finding
which suggests the use of ΔNp73 status as a prognostic marker for patients with HCC
Our finding that a significant percentage of HCCs select for dominant-negative p73 isoforms strongly argues for their oncogenic role during hepatocarcinogenesis
Preferential upregulation of ΔNp73 in HCCs might impose oncogenic activity that specifically interferes with the tumor suppressor function of wild-type p53 and TAp73 disabling major apoptosis pathways (death receptor- and mitochondrial pathways)
One might predict that interfering with the expression or function of ΔNp73 and/or mutant p53 and/or ΔNp63 in tumor cells may render such tumors more responsive to therapy and reduce their aggressiveness and metastatic capacity
Hep3B cells were maintained in MEM (Invitrogen
Germany) and Saos2 in a 1:1 mixture of Ham's F-12/DMEM (Biochrom
Medium was supplemented with 10% FCS (Biochrom)
2 mM L-Glutamine (Invitrogen) and 100 μg/ml Gentamycin (Invitrogen)
Cells were treated with bleomycin at a dose range of 0.3–300 μg/ml or with doxorubicin at a dose range of 0.005–0.5 μg/ml or with mitoxantrone at a dose range of 0.1–1 μg/ml for 6 h up to 72 h
The concentrations relevant for therapy are 1.5–3 μg/ml for bleomycin
and 0.03–0.5 μg/ml for mitoxantrone in patients' sera
TAp73β cDNA (rAd-TAp73β) or Δ84p73β cDNA (rAd-Δ84p73β)
each under the control of the cytomegalovirus immediate/early gene (CMV) promoter
an infection rate of 80–90% of the cells was obtained
Cells were seeded in six-well plates 24 h before transduction
rAd-TAp73β or rAd-Δ84p73β) were added to the culture medium and cells were incubated with the virus for 4 h
Cells were transfected by the use of calcium-phosphate
cells were infected with the adenovirus at a MOI of 10
cells were harvested and assayed for luciferase activity as described by the manufacturer (Promega
the broad spectrum caspase inhibitor ZVAD-FMK (z-Val-Ala-DL-Asp-fluoromethylketone
DEVD-FMK (Z-Asp(OCH3)-Glu(OCH3)-Val-Asp(OCH3)-FMK; inhibitor of caspase-3 as well as -6
Z-IETD-FMK (z-Ile-Glu(OMe)-Thr-Asp(OMe)-CH2F; inhibitor of caspase-8)
Z-LEHD-FMK (z-Leu-Glu(OMe)-His-Asp(OMe)-CH2F; inhibitor of caspase-9
Germany) and the antagonistic anti-human CD95L mAb (clone NOK-1
Hep3B cells were harvested 36 and 48 h following rAd-TAp73β or rAd-Δ84p73β or rAd-GFP transfer with or without concomitant bleomycin treatment (caspase-3
Cell surface expression of the CD95 receptor was assessed by FACScan®
Hep3B or Saos2 cells were incubated for 30 min with anti- CD95 receptor antibody (anti-APO-1 IgG3
PE-labeled affinity purified F(ab)’2 fragment goat anti-mouse IgG Fc antibody (Dianova
Germany) was used as secondary detecting reagent
To examine staining specificity of the second antibody cells were incubated with isotype control (IgG3) alone
The slides were evaluated by two observers who were blinded to clinical and pathologic information
To examine whether synergy between rAd-TAp73β transfer and concurrent chemotherapeutic treatment is observed
a balanced two-way ANOVA (model with fixed effects) was performed
we applied MANOVA (multivariate analysis of variance) and Wilcoxon analysis to test for statistical significance
The log-rank test was used to detect differences between survival curves for stratified variables
Statistical analysis was carried out using the SAS software system (SAS Institute Inc.
Vousden KH and Lu X (2002) Live or let die: the cell's response to p53
De Laurenzi V and Vousden KH (2002) p73: friend or foe in tumorigenesis
Moll UM and Slade N (2004) p63 and p73: roles in development and tumor formation
Melino G and Candi E (2005) p73 induces apoptosis by different mechanisms
McKeon F and Caput D (2000) p73-deficient mice have neurological
pheromonal and inflammatory defects but lack spontaneous tumours
McKeon F and Caput D (1997) Monoallelically expressed gene related to p53 at 1p36
a region frequently deleted in neuroblastoma and other human cancers
Terrinoni A and Falco M (1998) Two new p73 splice variants
a dominant-negative inhibitor of wild-type p53 and TAp73
Tonini GP and Romani M (2002) Expression of DeltaNp73 is a molecular marker for adverse outcome in neuroblastoma patients
Zaika A and Moll UM (2003) DNp73 facilitates cell immortalization and cooperates with oncogenic Ras in cellular transformation in vivo
McKeon F and Jacks T (2005) Tumor predisposition in mice mutant for p63 and p73: evidence for broader tumor suppressor functions for the p53 family
Oren M and Shaul Y (1999) Interaction of c-Abl and p73alpha and their collaboration to induce apoptosis
Levrero M and Wang JY (1999) The tyrosine kinase c-Abl regulates p73 in apoptotic response to cisplatin-induced DNA damage
Stremmel W and Müller M (2005) TAp63alpha induces apoptosis by activating signaling via death receptors and mitochondria
McKeon F and Jacks T (2002) p63 and p73 are required for p53-dependent apoptosis in response to DNA damage
Alt FW and McKeon F (2004) p63 and p73 are not required for the development and p53-dependent apoptosis of T cells
Krammer PH and Galle PR (1997) Drug-induced apoptosis in hepatoma cells is mediated by the CD95 (APO-1/Fas) receptor/ligand system and involves activation of wild-type p53
Oren M and Krammer PH (1998) p53 activates the CD95 (APO-1/Fas) gene in response to DNA damage by anticancer drugs
Hahn WC and Kaelin WG (2003) Chemosensitivity linked to p73 function
Grignani F and Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry
Angel P and Krammer PH (2000) A novel AP-1 element in the CD95 ligand promoter is required for induction of apoptosis in hepatocellular carcinoma cells upon treatment with anticancer drugs
Wu G and El-Deiry WS (1997) KILLER/DR5 is a DNA damage-inducible p53-regulated death receptor gene [letter]
Khuri FR and Sun SY (2004) p53 upregulates death receptor 4 expression through an intronic p53 binding site
DeGregori J and Wang JYJ (2004) Tumor necrosis factor alpha-induced apoptosis requires p73 and c-ABL activation downstream of RB degradation
Hammann A and Dimanche-Boitrel MT (1999) Fas ligand-independent
FADD-mediated activation of the Fas death pathway by anticancer drugs
Camonis JH and Wallach D (1995) Self-association of the ‘death domains’ of the p55 tumor necrosis factor (TNF) receptor and Fas/APO1 prompts signaling for TNF and Fas/APO1 effects
Thompson C and Vousden KH (2003) p73 induces apoptosis via PUMA transactivation and Bax mitochondrial translocation
Caput D and Davrinche C (2005) p73-dependent apoptosis through death receptor: impairment by human cytomegalovirus infection
Engeland K and Wittekind C (1999) Expression of p73 and its relation to histopathology and prognosis in hepatocellular carcinoma
Melino G and Candi E (2005) New antibodies recognizing p73: comparison with commercial antibodies
Kinzler KW and Vogelstein B (1998) A simplified system for generating recombinant adenoviruses
Miyashita T and Reed JC (1995) Tumor suppressor p53 is a direct transcriptional activator of the human bax gene
Levrero M and Melino G (2000) Induction of neuronal differentiation by p73 in a neuroblastoma cell line
Tibshirani R and Chu G (2001) Significance analysis of microarrays applied to the ionizing radiation response
Hamilton SR and Aaltonen LA (2000) WHO: Pathology and Genetics (Lyon: IARC Press)
Sobin LH and Wittekind CH (2002) UICC: TNM Classification of Malignant Tumours 6th edn (New York: Wiley-Liss)
Hauss J and Wittekind CH (2003) Identification of novel proteins associated with hepatocellular carcinomas using protein microarrays
Takizawa T and Nakanishi Y (1995) Transcription stimulation of the Fas-encoding gene by nuclear factor for interleukin-6 expression upon influenza virus infection
Kinzler KW and Vogelstein B (1992) Definition of a consensus binding site for p53
Vilo J and Vingron M (2001) Minimum information about a microarray experiment (MIAME)-toward standards for microarray data
Download references
We thank Petra Hill for expert technical assistance
This work was supported by grants of the Medizinische Forschungsförderung Heidelberg
of the Sonderforschungsbereich 601 and of the Tumorzentrum Heidelberg/Mannheim to MM and PHK
The work was in part performed thanks to grants from AIRC
EU (QLG1-1999-00739 and YLK-CT-2002-01956)
EU (QLK3-CT-2002-01956) to GM and MO and EU grant QLG1-1999-00739 to PHK
c/o Department of Experimental Medicine and Biochemical Sciences
These authors contributed equally to this work
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DOI: https://doi.org/10.1038/sj.cdd.4401774
Brad Pitt has found love again in German model Nicole Poturalski and the two are enjoying a private vacation recently while relishing in each other's enriching company.
The duo's romance can be traced back to November, 2019 when they were snapped for the first time together during Kanye West's performance at the Hollywood Bowl.
Nicole Poturalski, who goes by the name of Nico Mary, was born in Polish capital Warsaw and lives in Bergkamen, Germany.
She has advocated for important causes, like tighter gun laws, and is a staunch supporter of Black Lives Matter movement.
Pitt's new girlfriend has command over as many as five languages. She is very cultured and well-travelled. Poturalski divides her time between Berlin and Los Angeles - where Pitt lives.
View this post on Instagram Missing my Sunset Views @drewblackwellphoto
A post shared by Nico (@nico.potur) on Feb 9, 2020 at 7:51am PST
Poturalski is 27 years' old, making her 29 years younger than Pitt, who is 56 years' old . It has been reported that after Poturalski was first spotted with the Ad Astra star she hinted at a relationship with him in an Instagram post in February, referring to him as her 'love' at the time.
"Missing my strolls with my Love," she captioned a photo of herself walking down Rodeo Drive in LA.
In March, she posted about her 'better half,' again dropping hints about being in love.
View this post on Instagram Saturdays these days for me is self reflection, reading books spending time with my better half.
A post shared by Nico (@nico.potur) on Mar 28, 2020 at 6:50am PDT
Poturalski is a doting single mom, who has a son named Emil. Although she keeps things about Emil strictly under the wraps, the model called him 'the love of her life' in a now-deleted Instagram post.
“When you are back together with the love of your life! #nothing #likemomandson #motherson #love #biggestlove #weekends #familyfirst," she captioned the photo.
She is passionate about raising her voice against sexual exploitation of children and wants to see the end to child sex trafficking and child pornography.
View this post on Instagram Protect Kids not Guns @chnge
A post shared by Nico (@nico.potur) on May 6, 2020 at 5:51am PDT
A professional model, Poturalski was first dicovered by a modelling scout at Disneyland Paris when she was just 13. She reportedly wanted to be a marine biologist after growing up.
She has graced the cover of famous magazines like Elle, Harper's Bazaar and Grazia, and even Marie Claire and Cosmopolitan.
A post shared by Nico (@nico.potur) on Aug 5
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Though it's unclear how long the actor and 27-year-old German model have been dating (or if they even are an item), the two may have gotten together as early as November 2019
who obtained photos of the maybe-couple looking cozy in the VIP box during Kanye West's performance at the Hollywood Bowl
but here's everything we do know about the model
Though Nicole currently divides her time between Berlin and LA, she is originally from Bergkamen
Nicole reportedly moved to Berlin at 19 with her mother
in order to pursue modeling professionally
The model has seemingly spent a lot of time in France and Italy
so French and Italian may be the other two languages she's mastered
When Nicole was 13, she was spotted by a talent scout while visiting Disneyland Paris
In March 2020, Nicole showed off a tiny neck tattoo on Instagram
"MOON." It's unclear what the tattoo means
but she did use a moon emoji in a February 2020 IG post (which has since been deleted) that seemingly alluded to Brad
"Missing my strolls with my Love 🌙💥," she captioned the photo
The women's national team will meet in Füssen from August 18th to 21.08.2016st
almost four weeks before the league officially starts
Women's national coach Benjamin Hinterstocker has nominated five goalkeepers
Women's national coach Benjamin Hinterstocker: "As with our first ice hockey course three weeks ago in Füssen
we will use the joint training sessions to improve our ice hockey
we will try to promote the three parts of the team
we will of course try intensively to speed up our game."
Der HC TuRa Bergkamen trauert um Lukas Bommer
Der Torhüter starb am Dienstag völlig unerwartet
Am Sonntag stand er noch zwischen den Pfosten
Wie zufrieden sind Sie heute mit Ihrem Besuch auf hellwegeranzeiger.de
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As part of the second edition of the Sliema International Arts Festival
the string orchestra Backreis Bergkamen from Dusseldorf
will give a concert at the culture performance hall of the Sliema Band Club
The orchestra is formed of around 70 musicians with a varied repertoire
It will perform excerpts from the 6th Symphony by Peter Tcshaikowsy
introduction to Act III of Lohengrin by Richard Wagner
theme from the 2nd Symphony by Ludwig Van Beethoven
Night On A Bare Mountain by Modest Mussorgsky
Grand March from Aida by Giuseppe Verdi and excerpts from Carmen Suite by Georges Bizet
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it appears that the Oscar winner has finally moved on with 27-year-old model Nicole Poturalski
Here’s everything we know about the German beauty
According to The Daily Mail, Poturalski is a Capricorn who was born on January 2,1993, in Bergkamen, Germany. She is of Polish heritage, and on her Facebook page she lists her former city as Warsaw.
During her 10-year modeling career, Poturalski has graced the covers of Elle magazine, Cosmopolitan, and Marie Claire. She has also walked the catwalk at various fashion shows in New York, Paris, and Milan. Poturalski is also one of British designer’s Vivienne Westwood’s favorite models.
View this post on Instagram A post shared by Nico (@nico.potur)
Her international modeling work has taken her around the globe
and the 5’10” beauty is reportedly fluent in five languages
and her son Emil makes regular appearances on her Instagram account
Poturalski had dreams of becoming a marine biologist when she was a kid
after a talent scout spotted her at Disneyland Paris when she was a teenager
Poturalski began her modeling career while completing her Abitur
Poturalski – who goes by the first name Nico – has been vocal on social media about her political views
Before the global protests against racial injustice began in May
the brunette beauty posted a pic of herself wearing a top that read “Protect Kids
In an Instagram Story post this spring, Poturalski said, “I truly believe we need change. Black Lives Matter
Children’s lives matter…Abused children can’t protest
RELATED: Brad Pitt Was First Spotted Getting Cozy With Nicole Poturalski 9 Months Ago
She added that it is on “us young people” to educate and watch out for younger fellow humans
Poturalski declared that change is needed and asked her fans to be kind and do their best
Her politics appear to be very similar to Pitts
who voiced his dissent against President Trump and the Republican Party during his Oscar acceptance speech earlier this year
“They told me I only have 45 seconds up here,” said Pitt
“… which is 45 seconds more than the senate gave John Bolton this week.”
Poturalski was thrown into the limelight after cameras spotted her with Pitt at the South of France’s Le Castellet Airport
It is believed that they are spending time at the estate he purchased in 2011 with Jolie – Château Miraval
View this post on Instagram A post shared by Nico (@nico.potur)
“They were kissing and Brad was being super attentive to her,” claimed the source
“He was in a semi public place but didn’t seem to mind people seeing him
She’s a real beauty and obviously a lot younger than him.”
The outlet’s PDA report hasn’t been corroborated, but Entertainment Tonight reports that Pitt and Poturalski were seen “driving in a chauffeured vehicle that took them to the airport for their next flight.”