UNASSIGNED: Epithelial-mesenchymal transition (EMT) plays an important role in bladder carcinoma (BC) invasiveness and metastasis. Studies have shown that muscle-invasive BC (MIBC) and non-MIBC (NMIBC) are different at the molecular level owing to different EMT-related programming. Recent studies suggest that dysregulation of specific miRNAs is linked to EMT in BC. With this background, we aimed to study the immunoexpression of EMT-markers and its correlation with miRNA-200c expression in a series of MIBCs and NMIBCs.
UNASSIGNED: Quantitative real-time-polymerase chain reaction for the quantification of miR-200c expression was performed on 50 cases of urinary BC obtained from transurethral resection of bladder tumor (TURBT), cystectomy specimens, and ten peritumoral bladder tissue. Immunohistochemistry for ZEB1, ZEB2, TWIST, E-cadherin, and β-catenin was performed on tumor and peritumoral bladder tissue.
UNASSIGNED: Thirty-five TURBT and 15 cystectomy specimens were assessed. Among MIBC, loss of expression of E-cadherin (72.3%), β-catenin (66.7%), and ZEB1, ZEB2, and TWIST2 immunoreactivity was noted in 53.3%, 86.7%, and 73.3% of cases, respectively. Among NMIBC, loss of expression of E-cadherin (22.5%), β-catenin (17.1%) and ZEB1, ZEB2, and TWIST immunoreactivity was noted in 11.5%, 51.4%, and 91.4% of cases, respectively. Upregulation of miRNA-200c was noted in cases with retained E-cadherin and negative TWIST expression. Downregulation of miRNA-200c expression was noted in all the cases showing loss of E-cadherin, β-catenin, and in cases immunoreactive for ZEB1, ZEB2, and TWIST in MIBC. Downregulation of miRNA-200c expression was also noted in cases of MIBC with retained β-catenin and those immunonegative for ZEB1 and ZEB2. A similar trend was noted in NMIBC. Median miRNA-200c expression was low in both high-grade and low-grade NMIBC compared to peritumoral bladder tissue and was not statistically significant.
UNASSIGNED: This study for the first time explores the relation of miR200C with E-cadherin, b-catenin, and its direct transcriptional regulators, namely Zeb1, Zeb2, and Twist in the same cohort of BC. We observed that miRNA-200c is downregulated in both MIBC and NMIBC. We identified novel expression of TWIST in cases of BC showing downregulation of miR200Cs suggesting that it is one of the protein targets of altered miRNA-200c expression contributing to EMT and can serve as a promising diagnostic marker and therapeutic target. Loss of E-cadherin and ZEB1 immunoexpression in high-grade NMIBC suggests an aggressive clinical behavior. However, ZEB2 heterogeneous expression in BC limits its diagnostic and prognostic utility.

The phase 3 POLO study demonstrated a significant progression-free survival (PFS) benefit and preserved health-related quality of life (HRQOL) for active maintenance treatment with olaparib vs placebo in patients with metastatic pancreatic cancer and a germline BRCA mutation. Here, we present a post hoc analysis of the patient-centered outcomes: time without significant symptoms of disease progression or toxicity (TWiST) and quality-adjusted TWiST (Q-TWiST).
Patients were randomized 3:2 to maintenance olaparib (300 mg tablets twice daily) or placebo. Overall survival time was divided into TWiST, toxicity (TOX; time before disease progression with significant symptoms of toxicity), and relapse (REL; time after disease progression until death or censoring). Q-TWiST was the sum of TWiST, TOX, and REL, each weighted by HRQOL utility scores during the relevant health-state period. A base-case and three sensitivity analyses were performed using differing definitions of TOX.
In total, 154 patients were randomized (olaparib, n = 92; placebo, n = 62). TWiST was significantly longer for olaparib than placebo in the base-case analysis (14.6 vs 7.1 months; 95% CI, 2.9-12.0; p = .001) and all sensitivity analyses. No statistically significant benefit for Q-TWiST was observed in the base-case analysis (18.4 vs 15.9 months; 95% CI, -1.1 to 6.1; p = .171) or the sensitivity analyses.
These results support the previous findings that maintenance olaparib significantly improves PFS relative to placebo without compromising HRQOL and demonstrate that the clinically meaningful benefits of olaparib persist even when symptoms of toxicity are considered.

OBJECTIVE: By using conventional echocardiographic indices, cardiac pumping function remains unaltered during pregnancy. However, two-dimensional speckle tracking echocardiography (2D-STE) can detect subclinical changes of myocardial function even in patients with normal and stable left ventricular ejection fraction (LVEF).The aim of this study was to evaluate LV systolic performance during normal low risk pregnancy by using both conventional 2D and 3D echo indices and 2D-STE.
METHODS: One hundred and twelve pregnant women without any history of heart disease were prospectively recruited. They underwent serial echocardiographic evaluation in each pregnancy trimester and 6 months after delivery (time indicated as 1,2,3 and 4). 2D LVEF, 3D LVEF, LV global longitudinal strain (LVGLS), LV global circumferential strain (LVGCS) and LV-twist were measured and compared to the control group (c).
RESULTS: 2D-LVEF and 3D-LVEF were not significantly different among the three trimesters, postpartum and controls. LVGLS progressively decreased during pregnancy (1st :21.71 ± 2.13%, 2nd : 21.20 ± 2.30%, 3rd : 19.82 ± 2.10%, 4th : 21.81 ± 2.05%, c: 21.71 ± 2.2%, overall p < 0,001) which receded during puerperium. No significant difference was noted in LVGCS (1st : 18.08 ± 5.54%, 2nd : 18.57 ± 3.41%, 3rd :18.20 ± 3.33%, 4th : 17.95 ± 3.39%, c: 18.8 ± 2.2%, p > 0.3). LV-Twist was significantly higher in the 1st trimester compared to controls (p = 0.04) and remained constantly high during the rest of the pregnancy and puerperium (1st :13.80 ± 5.09°, 2nd :13.46 ± 5.35°, 3rd :13.58 ± 4.32°, 4th :13.37 ± 4.26°, c: 11.5 ± 4.3°).
CONCLUSIONS: In low risk individuals with normal pregnancy, a redistribution of force occurs especially in the 3rd trimester. Longitudinal strain decreases, while torsional movement of the heart increases and counterbalances the temporal change of longitudinal systolic function. These changes would probably reflect the pathophysiological alterations related to pregnancy.

BACKGROUND: Patients with wake-up ischemic stroke are frequently excluded from thrombolytic treatment due to unknown symptom onset time and limited availability of advanced imaging modalities. The Tenecteplase in Wake-up Ischaemic Stroke Trial (TWIST) is a randomized controlled trial of intravenous tenecteplase 0.25 mg/kg and standard care versus standard care alone (no thrombolysis) in patients who wake up with acute ischemic stroke and can be treated within 4.5 h of wakening based on non-contrast CT findings.
OBJECTIVE: To publish the detailed statistical analysis plan for TWIST prior to unblinding.
METHODS: The TWIST statistical analysis plan is consistent with the Consolidating Standard of Reporting Trials (CONSORT) statement and provides clear and open reporting.
CONCLUSIONS: Publication of the statistical analysis plan serves to reduce potential trial reporting bias and clearly outlines the pre-specified analyses.
BACKGROUND: ClinicalTrials.gov NCT03181360 . EudraCT Number 2014-000096-80 . WHO ICRTP registry number ISRCTN10601890 .

BACKGROUND: Hemophilia is an X-linked inherited disorder primarily affecting males, its major types are type A (deficiency in factor VIII) and B (deficiency in factor IX), and is considered to be the most common severe congenital coagulation factor deficiency. The present study was designed to test whether any differences in left ventricular (LV) rotational mechanics could be demonstrated between male patients with hemophilia and healthy controls using three-dimensional speckle-tracking echocardiography (3DSTE)-derived virtual LV models.
METHODS: The present study consisted of 17 patients with hemophilia, however, 3 patients were excluded due to insufficient image quality. In the remaining patient population, 12 patients had hemophilia A and 2 patients had hemophilia B (mean age: 42.2±18.9 years, all males). The control group comprised 16 age-matched healthy subjects (46.0±5.9 years, all males).
RESULTS: None of the routine two-dimensional echocardiographic data differ between patients with hemophilia and controls. None of the patients and controls showed ≥ grade 1 valvular regurgitations and had valvular stenoses. In one subject, the near absence of LV twist called as LV \"rigid body rotation\" could be detected, data of which were managed separately. While 3DSTE-derived apical LV rotation was 3.65 degrees, basal LV rotation proved to be 3.57 degrees leading to 0.08-degree LV apico-basal gradient suggesting counterclockwise LV \"rigid body rotation\". In the remaining patients, both LV apical rotation (7.25±6.20 vs. 10.39±4.16 degrees, P<0.02) and LV twist (10.24±5.60 vs. 14.38±3.93 degrees, P<0.003) showed significant impairment in patients with hemophilia.
CONCLUSIONS: LV rotational abnormalities are present in hemophilia with reduced LV apical rotation and twist.

OBJECTIVE: The clinical significance of circulating tumor cells (CTCs) and TWIST expression in CTCs remains unelucidated in patients with gastric cancer (GC). Here, we evaluated CTCs and TWIST expression in CTCs and explored their correlation with prognosis in patients with metastatic GC.
METHODS: Peripheral blood samples were prospectively obtained from 31 patients with metastatic GC between September 2017 and December 2018, prior to treatment. CTCs were detected using a centrifugal microfluidic system and CTCs positive for TWIST immunostaining were defined as TWIST (+) CTCs.
RESULTS: CTCs and TWIST (+) CTCs were detected in 25 (80.6%) and 24 (77.4%) of the 31 patients, respectively. CTC count in patients with first diagnosis of metastatic cancer tended to be higher than that in those with recurrent metastatic cancer, but TWIST (+) CTC count was not different between the two groups. There was no difference in CTC and TWIST (+) CTC counts according to histopathologic type, peritoneal dissemination, hematogenous metastasis, serum tumor makers, or response to first-line chemotherapy. Patients with CTCs > 7.5/7.5 mL of blood showed shorter overall survival (OS) than those with CTCs ≤ 7.5/7.5 mL of blood (p = 0.049). Additionally, patients with TWIST (+) CTCs > 2.5/7.5 mL of blood tended to show shorter OS than those with TWIST (+) CTCs ≤ 2.5/7.5 mL of blood (p = 0.105).
CONCLUSIONS: Our study demonstrated that high levels of CTCs and TWIST (+) CTCs were associated with worse OS.

Left ventricular (LV) twist is calculated from the net difference of counterclockwise apical and clockwise basal rotation during systole. The current study was designed to evaluate correlations between autonomic function and LV rotational mechanics in healthy subjects. The present study comprised 18 healthy subjects (mean age: 36±12 years, 12 men). Three-dimensional speckle tracking echocardiography (3DSTE) could be used for non-invasive evaluation of LV rotation and twist. Autonomic function was assessed by means of 5 standard cardiovascular reflex tests. During 3DSTE, basal LV rotation proved to be -3.24±2.02 degree, while apical LV rotation was 9.08±3.04 degree, therefore LV twist was 11.70±6.80 degree. Valsalva test showed significant correlations with LV basal (r=0.529, P=0.019) and apical rotations (r=-0.534, P=0.022), and LV twist (r=-0.467, P=0.044). Heart rate response to deep breathing significantly correlated with LV twist, as well (r=-0.452, P=0.052). The other tests had no any relationship with rotational characteristics. Correlations exist between parasympathetic autonomic function and 3DSTE-derived LV rotation and twist in healthy subjects.

2D speckle tracking echocardiography (2DSTE) is established to analyse left ventricular (LV) longitudinal function. The analysis of LV rotational deformation is challenging and requires standardization of image acquisition as well as postprocessing analysis. The aim of this study was to test the feasibility to analyse LV rotational deformation using 2DSTE by introducing a novel algorithm for the detection of artefacts. The study was performed in 20 healthy subjects serving as a control group and in 53 competitive sportsmen. Circumferential, radial strain (CS, RS) and LV rotation were analysed by 2DSTE in parasternal short axis views. The stepwise algorithm to exclude potential artefacts starts with the visual estimation of the image quality with respect to complete visualization of all myocardial segments during the entire cardiac cycle followed by the exclusion of data sets in participants with conduction abnormalities. The next step is the optimization of tracking areas and a cross-check of implausible strain waveforms in multiple acquired comparable cineloops. The last step is the exclusion of strain curves with persisting implausible waveforms if standardization failures and incorrect LV wall tracking are fixed. Plausible physiological strain curves were observed in 89% (n = 65/73) of all subjects. In controls all implausible waveforms could be verified as artefacts. The algorithm was applied in 53 professional athletes to test and confirm its feasibility. Abnormal CS waveforms were documented in 25 athletes, verified as artefacts due to tracking failures in 22 athletes and due to incorrect image acquisition in 3 athletes. CS artefacts were mostly located in the basal posterior and lateral LV segments. (endocardial: 6%, n = 4/70; p < 0.05) and basal posterior (endocardial: 8%, n = 5/70; p < 0.05) segments were highly susceptible to artefacts. 2DSTE of parasternal short axis views to analyse circumferential and radial deformation as well as LV rotation is feasible in athletes. The proposed algorithm helps to avoid artefacts and might contribute to standardization of this technique. 2DSTE might provide an interesting diagnostic tool for the detection of viral myocarditis, e.g. in athletes.

Patients with wake-up ischemic stroke who have evidence of salvageable tissue on advanced imaging can benefit from intravenous thrombolysis. It is not known whether patients who do not fulfil such imaging criteria might benefit from treatment, but studies indicate that treatment based on non-contrast CT criteria may be safe. Tenecteplase has shown promising results in patients with acute ischemic stroke. The aim of the Tenecteplase in Wake-up Ischemic Stroke Trial (TWIST) is to compare the effect of thrombolytic treatment with tenecteplase and standard care versus standard care alone in patients with wake-up ischemic stroke selected by non-contrast CT.
TWIST is an international, investigator-initiated, multi-centre, prospective, randomized-controlled, open-label, blinded end-point trial of tenecteplase (n = 300) versus standard care (n = 300) in patients who wake up with an acute ischemic stroke and can be treated within 4.5 h upon awakening. Seventy-seven centres in 10 countries (Denmark, Estonia, Finland, Latvia, Lithuania, New Zealand, Norway, Sweden, Switzerland, and the United Kingdom) participate. The primary outcome is the modified Rankin Scale on the ordinal scale (0-6) at three months.
TWIST aims to determine the effect and safety of thrombolytic treatment with tenecteplase in patients with wake-up ischemic stroke selected by non-contrast CT.
ClinicalTrials.gov NCT03181360. EudraCT Number 2014-000096-80.

Lipedema is a lymphedema-masquerading symmetrical, bilateral and disproportional obesity. Its conservative maintenance treatment comprises the use of flat-knitted compression pantyhoses. Lipedema is known to be associated with left ventricular morphological and functional alterations. The present study aimed to assess the effects of graduated compression stockings on left ventricular (LV) rotational mechanics measured by three-dimensional speckle-tracking echocardiography (3DSTE) in lipedema patients. The present study comprised twenty lipedema patients (mean age: 45.8 ± 11.0 years, all females) undergoing 3DSTE who were also compared to 51 age- and gender-matched healthy controls (mean age: 39.8 ± 14.1 years, all females). 3DSTE analysis was performed at rest, and subsequent to 1 hour application of compression class 2 made-to-measure flat-knitted pantyhose. Six lipedema patients showed significant LV rotational abnormalities. Of the remaining fourteen lipedema patients LV basal rotation rotation showed significant reduction, while LV apical rotation showed significant increase with unchanged LV twist after a 60-minute use of compression garment. Significant changes in LV rotational mechanics could be detected among 14 women with lipedema after the use of compression garment however six probands have special LV rotational abnormalities at baseline and/or after compression.