In a up-front randomized study, 939 adult patients up to the age of 60 years received a double induction therapy. One course of MAV (mitoxantrone 10 mg/m2 days 4–8, cytarabine 100 mg/m2 continuous ...infusion days 1–8, etoposide 100 m g/m2 days 4–8) was followed by one cycle of MAMAC (cytarabine 1 g/m2, every 12h days 1–5; amsacrine 100 mg/m2 days 1–5). Patients with intermediate risk cytogenetic (IRCG) and a HLA matched sibling received an allogeneic transplantation, those with poor risk cytogenetic (PRCG) were intended to be transplanted from a sibling or unrelated donor. All AML patients without an available donor received the randomly assigned first postremission therapy (PRT) mitoxantrone combined with intermediate-dose cytarabine (I-MAC; total dose 12 g/m2) or high-dose cytarabine (H-MAC; total dose 36 g/m2). As second PRT, patients with t(8;21) received an additional cycle of chemotherapy. An autologous transplantation was scheduled for IRCG and PRCG without an allogeneic donor. The CR rate was 88% for patients with t(8;21), with IRCG 71%, and 50% with PRCG. The 5-year-survival was 21% (95% CI: 16–27%) in the PRCG, 40% (95% CI: 36–45%) in the IRCG and 74% (95% CI: 60–88%) in the t(8;21) group. No difference was observed between the I-MAC and the H-MAC group. In a multivariate analysis, a significant (p<.01 for each parameter) better overall survival was observed in patients under the age of 37 years, blast count <10% at day 15, high myeloperoxidase positivity, low CD34 expression, WBC <15*10^9/L, thrombocytes >50*10^9/L, and IRCG compared to PRCG. The relapse incidence was higher in patients without an allogeneic donor, a Flt3 mutant/wildtype ratio > 0.8 or PRCG. A risk score build out of the sum of the individual hazard ratios (SHR) was able to discriminate two groups for the IRCG with a marked difference in the 5-year-survival (low SHR: 55% 95% CI: 48–62%; high SHR: 33% 95% CI: 28–38%) was well as for the PRCG group (low SHR: 44% 95% CI: 32–56%; high SHR: 13% 95% CI: 7–18%). The risk score identified in this large patient cohort may allow individual tailoring of therapeutic interventions in future AML trials.
Abstract
While survival has improved for Burkitt lymphoma patients, potential differences in outcome between pediatric and adult patients remain unclear. In both age groups, survival remains poor at ...relapse. Therefore, we conducted a comparative study in a large pediatric cohort, including 191 cases and 97 samples from adults. While
TP53
and
CCND3
mutation frequencies are not age related, samples from pediatric patients showed a higher frequency of mutations in
ID3
,
DDX3X, ARID1A
and
SMARCA4
, while several genes such as
BCL2
and
YY1AP1
are almost exclusively mutated in adult patients. An unbiased analysis reveals a transition of the mutational profile between 25 and 40 years of age. Survival analysis in the pediatric cohort confirms that
TP53
mutations are significantly associated with higher incidence of relapse (25 ± 4% versus 6 ± 2%, p-value 0.0002). This identifies a promising molecular marker for relapse incidence in pediatric BL which will be used in future clinical trials.
The mechanistic target of rapamycin (mTOR) promotes pathological remodeling in the heart by activating ribosomal biogenesis and mRNA translation. Inhibition of mTOR in cardiomyocytes is protective; ...however, a detailed role of mTOR in translational regulation of specific mRNA networks in the diseased heart is unknown. We performed cardiomyocyte genome‐wide sequencing to define mTOR‐dependent gene expression control at the level of mRNA translation. We identify the muscle‐specific protein Cullin‐associated NEDD8‐dissociated protein 2 (Cand2) as a translationally upregulated gene, dependent on the activity of mTOR. Deletion of Cand2 protects the myocardium against pathological remodeling. Mechanistically, we show that Cand2 links mTOR signaling to pathological cell growth by increasing Grk5 protein expression. Our data suggest that cell‐type‐specific targeting of mTOR might have therapeutic value against pathological cardiac remodeling.
SYNOPSIS
Genome‐wide translational profiling identifies mTORC1‐dependent genes in cardiomyocytes in response to neurohumoral stimulation. Expression of the muscle‐specific gene Cand2 is controlled by mTORC1 and Cand2 regulates cardiac function and pathological hypertrophy.
Cand2 is translationally upregulated during pathological stress in cardiac myocytes.
Cand2 expression depends on the activity of mTORC1.
Cand2 promotes the expression of G‐protein coupled receptor 5 (Grk5), which in turn links to myocyte enhancer factor 2 (MEF2)‐driven transcription of cardiac hypertophy genes.
Genome‐wide translational profiling identifies mTORC1‐dependent genes in cardiomyocytes in response to neurohumoral stimulation. Expression of the muscle‐specific gene Cand2 is controlled by mTORC1 and Cand2 regulates cardiac function and pathological hypertrophy.
Non-invasive tests (NITs) for clinically significant portal hypertension (CSPH) require validation in patients with hepatitis D virus (HDV)-related compensated advanced chronic liver disease (cACLD). ...Therefore, we aimed to validate existing NIT algorithms for CSPH in this context.
Patients with HDV-cACLD (LSM ≥10 kPa or histological METAVIR F3/F4 fibrosis) who underwent paired HVPG and NIT assessment at Medical University of Vienna or Hannover Medical School between 2013 and 2023 were retrospectively included. Liver stiffness measurement (LSM), von Willebrand factor to platelet count ratio (VITRO), and spleen stiffness measurement (SSM) were assessed. Individual CSPH risk was calculated according to previously published models (ANTICIPATE, 3P/5P). The diagnostic performance of Baveno VII criteria and refined algorithms (Baveno VII-VITRO, Baveno VII-SSM) was evaluated. The prognostic utility of NITs was investigated in the main cohort and an independent, multicenter, validation cohort.
Fifty-one patients (HVPG ≥10 mmHg/CSPH prevalence: 62.7%, varices: 42.2%) were included. Patients with CSPH had significantly higher LSM (25.8 17.2-31.0 vs. 14.0 10.5-19.8 kPa; p <0.001), VITRO (n = 31, 3.5 2.7-4.5 vs. 1.3 0.6-2.0 %/G/L; p <0.001), and SSM (n = 20, 53.8 41.7-75.5 vs. 24.0 17.0-33.9 kPa; p <0.001). Composite CSPH risk models yielded excellent AUROCs (ANTICIPATE: 0.885, 3P: 0.903, 5P: 0.912). Baveno VII criteria ruled out CSPH with 100% sensitivity and ruled in CSPH with 84.2% specificity. The Baveno VII ‘grey zone’ (41.1%) was significantly reduced by Baveno VII-VITRO or Baveno VII-SSM algorithms, which maintained diagnostic accuracy. Hepatic decompensation within 2 years only occurred in patients who had CSPH or met Baveno VII rule-in criteria. The prognostic value of NITs was confirmed in the validation cohort comprising 92 patients.
Standalone and composite NIT/diagnostic algorithms are useful for CSPH diagnosis in patients with HDV-cACLD. Thus, NITs may be applied to identify and prioritize patients with CSPH for novel antiviral treatments against chronic hepatitis D.
Non-invasive tests (NITs) for clinically significant portal hypertension (CSPH) have been developed to identify patients with compensated advanced chronic liver disease (cACLD) at risk of decompensation, but conflicting data has been published regarding the accuracy of liver stiffness measurement (LSM) for the staging of fibrosis in patients infected with hepatitis D virus (HDV). In our study, including 51 patients with HDV-cACLD, LSM- and lab-based NITs yielded high AUROCs for CSPH. Moreover, only patients with CSPH or high non-invasively assessed CSPH risk were at risk of decompensation within 2 years, with the prognostic value of NITs confirmed in a validation cohort. Thus, NITs should be applied and updated in yearly intervals in clinical routine to identify patients with HDV-cACLD at short-term risk of clinical events; NITs may also guide prioritization for novel antiviral treatment options.
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•NITs for CSPH can be applied in HDV-cACLD with high accuracy.•NITs have similar ability to identify high-risk patients with HDV-cACLD as HVPG.•NITs may guide novel antiviral treatment prioritization and individualized prognostication.
Asymmetric dimethylarginine (ADMA;
N
G,
N
G-dimethyl-
l-arginine) is the most important endogenous inhibitor of nitric oxide synthase and a potential risk factor for cardiovascular diseases. This ...article describes a gas chromatographic–tandem mass spectrometric (GC–tandem MS) method for the accurate quantification of ADMA in human plasma or serum and urine using de novo synthesized
2
H
3
-methyl ester ADMA (d
3Me-ADMA) as the internal standard. Aliquots (100
μl) of plasma/serum ultrafiltrate or native urine and of aqueous solutions of synthetic ADMA (1
μM for plasma and serum; 20
μM for urine) are evaporated to dryness. The residue from plasma/serum ultrafiltrate or urine is treated with a 100
μl aliquot of 2
M HCl in methanol, whereas the residue of the ADMA solution is treated with a 100
μl aliquot of 2
M HCl in tetradeuterated methanol. Methyl esters are prepared by heating for 60
min at 80
°C. After cooling to room temperature, the plasma or urine sample is combined with the d
3Me-ADMA sample, the mixture is evaporated to dryness, the residue treated with a solution of pentafluoropropionic (PFP) anhydride in ethyl acetate (1:4, v/v) and the sample is incubated for 30
min at 65
°C. Solvent and reagents are evaporated under a stream of nitrogen gas, the residue is treated with a 200
μl aliquot of 0.4
M borate buffer, pH 8.5, and toluene (0.2
ml for plasma, 1
ml for urine). Reaction products are extracted by vortexing for 1
min, the toluene phase is decanted, and a 1
μl aliquot is injected into the GC–tandem MS instrument. Quantitation is performed by selected reaction monitoring (SRM) of the common product ion at
m/
z 378 which is produced by collision-induced dissociation of the ions at
m/
z 634 for endogenous ADMA and
m/
z 637 for d
3Me-ADMA. In plasma and urine of healthy humans ADMA was measured at concentrations of 0.39±0.06
μM (
n=12) and 3.4±1.1
μmol/mmol creatinine (
n=9), respectively. The limits of detection and quantitation of the method are approximately 10
amol and 320
pM of d
3Me-ADMA, respectively.
Assay methods based on the Griess reaction are frequently used to measure nitrite and nitrate in urine, plasma, and other biological fluids. With minor exceptions, careful attention has not been paid ...in extending the Griess assay from aqueous solutions to biological fluids. In the present study, parallel measurements of nitrite and nitrate were performed in urine, plasma, and aqueous solutions with a published batch assay based on the Griess reaction and with gas chromatography–mass spectrometry (GC–MS). We report here further interferences by free reduced thiols, proteins, and other plasma constituents in the Griess assay but not in GC–MS. The best correlation (r2= 0.985) between the Griess assay and GC–MS was observed for aqueous solutions in the absence of thiols. Unlike GC–MS, the Griess assay was not applicable to whole human plasma and urine samples. For the measurement of nitrate in diluted human urine samples, reduction by cadmium was performed both under acidic (pH 2 or 5) and alkaline (pH 8.8) conditions. The mean recovery rate of nitrate from urine samples was quantitative in the GC–MS but amounted to only 30–80% in the Griess assay. Measurement of nitrate in human urine samples (n= 33) resulted in an excellent correlation between two GC–MS techniques (r2= 0.979) but only in a poor correlation (r2< 0.64) between the Griess assay and GC–MS. Unlike GC–MS, the batch Griess assay is associated with many problems in measuring nitrate in biological fluids.
In the year 1992,
S-nitrosoalbumin (SNALB) has been proposed to be the most abundant physiological carrier and pool of nitric oxide (NO) activity in human circulation, by which NO-dependent ...biological functions are regulated. The concentration, the metabolism and the mechanisms of the biological actions of SNALB are controversial and still incompletely understood. Moreover, the suitability of SNALB as a biomarker of diseases associated with altered NO bioactivity in human circulation has not been demonstrated convincingly so far. In the present study, we report on the development and application of a stable-isotope technique to study the pharmacokinetics of
15N-labelled SNALB (S
15NALB) in anesthetized rats. S
15NALB was synthesized from albumin isolated by affinity chromatography from freshly prepared human plasma. This technique was also applied to study and quantify the formation of S
15NALB from endogenous rat plasma albumin and intravenously applied
S-
15Nnitrosoglutathione (GS
15NO) or
S-
15Nnitrosocysteine (S
15NC) in anesthetized rats. In these investigations the mean arterial pressure (MAP) was monitored continuously. The elimination half-life (
t
1/2) of S
15NALB from rat plasma was determined to be 4.1
min (
t
1/2
α) and 9.4
min (
t
1/2
β). S
15NALB (125
nmol) produced long-lasting decreases in MAP (by 49% for 18
min). Thirty minutes after intravenous (i.v.) injection of S
15NALB (125
nmol), repeated i.v. injection of
l-cysteine or
d-cysteine (10
μmol each) produced repeatedly potent (by 44–55%) but short-lasting (about 4
min) MAP falls. Intravenously administered GS
15NO and S
15NC (each 500
nmol) could not be isolated from rat blood.
15N-Labelled nitrite and nitrate were identified as the major metabolites of all investigated
S-nitrosothiols in rat plasma. The results of this study suggest that in the rat S
15NALB is a potent
S-transnitrosylating agent and that the blood pressure-lowering effect of S
15NALB and other
S-nitrosothiols are mediated largely by
l-cysteine via
S-transnitrosylation to form S
15NC that subsequently releases
15NO. Our results also suggest that
S-transnitrosylation of the single reduced cysteine moiety of albumin by endogenous GSNO or SNC in blood is possible but does not represent an effective mechanism to produce SNALB in vivo. This stable-isotope dilution GC–MS technique is suitable to perform in vivo studies on SNALB using physiologically and pharmacologically relevant doses.
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