Superior efficacy of neoadjuvant chemotherapy and radical cystectomy in cT3-4aN0M0 compared to cT2N0M0 bladder cancer.

In this study, we compared complete pathological downstaging (pCD, ≤(y)pT1N0) and overall survival (OS) in patients with cT2 versus cT3-4aN0M0 UC of the bladder undergoing radical cystectomy (RC) with or without neoadjuvant chemo- (NAC) or radiotherapy (NAR). A population-based sample of 5,517 patients, who underwent upfront RC versus NAC + RC or NAR + RC for cT2-4aN0M0 UC between 1995-2013, was identified from the Netherlands Cancer Registry. Data were retrieved from individual patient files and pathology reports. pCD-rates were compared using Chi-square tests and OS was estimated by Kaplan-Meier analyses. Multivariable analyses were conducted to determine odds (OR) and hazard ratios (HR) for pCD-status and OS, respectively. We included 4,504 (82%) patients with cT2 and 1,013 (18%) with cT3-4a UC. Median follow-up was 9.2 years. In cT2 UC, pCD-rate was 25% after upfront RC versus 43% (p < 0.001) and 33% (p = 0.130) after NAC + RC and NAR + RC, respectively. In cT3-4a UC, pCD-rate was 8% after upfront RC versus 37% (p < 0.001) and 16% (p = 0.281) after NAC + RC and NAR + RC, respectively. In cT2 UC, 5-year OS was 57% and 51% for NAC + RC and upfront RC, respectively (p = 0.135), whereas in cT3-4a UC, 5-year OS was 55% for NAC + RC versus 36% for upfront RC (p < 0.001). In multivariable analysis for OS, NAC was beneficial in cT3-4a UC (HR: 0.67, 95%CI 0.51-0.89) but not in cT2 UC (HR: 0.91, 95%CI 0.72-1.15). NAR did not influence OS. In conclusion, NAC + RC was associated with superior pCD compared to RC alone and NAR + RC. Superior OS for NAC + RC compared to RC alone was especially evident in cT3-4a disease.

Tumor heterogeneity of fibroblast growth factor receptor 3 (FGFR3) mutations in invasive bladder cancer: implications for perioperative anti-FGFR3 treatment.

BACKGROUND: Fibroblast growth factor receptor 3 (FGFR3) is an actionable target in bladder cancer. Preclinical studies show that anti-FGFR3 treatment slows down tumor growth, suggesting that this tyrosine kinase receptor is a candidate for personalized bladder cancer treatment, particularly in patients with mutated FGFR3. We addressed tumor heterogeneity in a large multicenter, multi-laboratory study, as this may have significant impact on therapeutic response. PATIENTS AND METHODS: We evaluated possible FGFR3 heterogeneity by the PCR-SNaPshot method in the superficial and deep compartments of tumors obtained by transurethral resection (TUR, n = 61) and in radical cystectomy (RC, n = 614) specimens and corresponding cancer-positive lymph nodes (LN+, n = 201). RESULTS: We found FGFR3 mutations in 13/34 (38%) T1 and 8/27 (30%) ≥T2-TUR samples, with 100% concordance between superficial and deeper parts in T1-TUR samples. Of eight FGFR3 mutant ≥T2-TUR samples, only 4 (50%) displayed the mutation in the deeper part. We found 67/614 (11%) FGFR3 mutations in RC specimens. FGFR3 mutation was associated with pN0 (P < 0.001) at RC. In 10/201 (5%) LN+, an FGFR3 mutation was found, all concordant with the corresponding RC specimen. In the remaining 191 cases, RC and LN+ were both wild type. CONCLUSIONS: FGFR3 mutation status seems promising to guide decision-making on adjuvant anti-FGFR3 therapy as it appeared homogeneous in RC and LN+. Based on the results of TUR, the deep part of the tumor needs to be assessed if neoadjuvant anti-FGFR3 treatment is considered. We conclude that studies on the heterogeneity of actionable molecular targets should precede clinical trials with these drugs in the perioperative setting.

Psychometric properties of the Exercise Self-efficacy Scale in Dutch Primary care patients with type 2 diabetes mellitus.

BACKGROUND: Excercise self-efficacy is believed to influence physical activity bahavior. PURPOSE: The purpose of this study is to assess the psychometric aspects of the Exercise Self-efficacy Scale (ESS) in a type 2 diabetes Dutch Primary care sample. METHOD: Type 2 diabetes patients (n = 322; <80 years old) filled in the ESS and the short questionnaire to assess health enhancing physical activity (SQUASH). The structural validity of the ESS was assessed by means of principal axis factor analyses and confirmatory factor analysis. In addition, reliability and concurrent validity with the SQUASH outcomes "total" and "leisure time minutes/week of moderate to vigorous intensity physical activity" were evaluated. T tests and ANOVAs were used to examine ESS scores in subgroups. In addition, a 13-item version of the ESS was developed. RESULTS: Analyses were performed on complete cases (n = 255). Exploratory factor analysis suggested one underlying factor (total explained variance 54 %), with good internal consistency (α = 0.95). Confirmatory factor analysis showed a poor fit, as did a three-factor model suggested in an earlier research. Therefore, a 13-item ESS was developed with one underlying factor (total explained variance 59 %) and good internal consistency (α = 0.95). Both the 18-item and 13-item ESS correlated significantly with total and leisure time physical activity. ESS scores differed significantly between categories of education level and physical activity level. CONCLUSION: The 13-item ESS had sound psychometric properties in a large sample of primary care type 2 diabetes patients. The 13-item ESS could be useful in (intervention) research on physical activity in type 2 diabetes patients.

Frequency selectivity in monkey auditory nerve studied with suprathreshold multicomponent stimuli.

Data from non-human primates can help extend observations from non-primate species to humans. Here we report measurements on the auditory nerve of macaque monkeys in the context of a controversial topic important to human hearing. A range of techniques have been used to examine the claim, which is not generally accepted, that human frequency tuning is sharper than traditionally thought, and sharper than in commonly used animal models. Data from single auditory-nerve fibers occupy a pivotal position to examine this claim, but are not available for humans. A previous study reported sharper tuning in auditory-nerve fibers of macaque relative to the cat. A limitation of these and other single-fiber data is that frequency selectivity was measured with tonal threshold-tuning curves, which do not directly assess spectral filtering and whose shape is sharpened by cochlear nonlinearity. Our aim was to measure spectral filtering with wideband suprathreshold stimuli in the macaque auditory nerve. We obtained responses of single nerve fibers of anesthetized macaque monkeys and cats to a suprathreshold, wideband, multicomponent stimulus designed to allow characterization of spectral filtering at any cochlear locus. Quantitatively the differences between the two species are smaller than in previous studies, but consistent with these studies the filters obtained show a trend of sharper tuning in macaque, relative to the cat, for fibers in the basal half of the cochlea. We also examined differences in group delay measured on the phase data near the characteristic frequency versus in the low-frequency tail. The phase data are consistent with the interpretation of sharper frequency tuning in monkey in the basal half of the cochlea. We conclude that use of suprathreshold, wide-band stimuli supports the interpretation of sharper frequency selectivity in macaque nerve fibers relative to the cat, although the difference is less marked than apparent from the assessment with tonal threshold-based data.

Effects of exercise training on quality of life, symptoms of depression, symptoms of anxiety and emotional well-being in type 2 diabetes mellitus: a systematic review.

AIMS/HYPOTHESIS: Psychological problems are relatively common in people with type 2 diabetes. It is unclear whether exercise training exerts an effect on quality of life, symptoms of depression, symptoms of anxiety and emotional well-being in people with type 2 diabetes. The aim of this study was to conduct a systematic review to assess the effects of exercise training on these outcomes in people with type 2 diabetes. METHODS: MEDLINE, PsycINFO, Embase and ClinicalTrials.gov databases were searched. The review included randomised controlled trials (RCTs) of at least 4 weeks' duration in people with type 2 diabetes that evaluated the effect of exercise training on quality of life, symptoms of depression, symptoms of anxiety and/or emotional well-being compared with usual care. RESULTS: Of 1,261 retrieved articles, 20 RCTs were included with a total of 1,719 participants. Quality of life was assessed in 16 studies. Between-group comparisons showed no significant results for aerobic training with the exception of one study, and mixed results for resistance and combined training. Symptoms of depression were assessed in four studies. In only one study did the intervention decrease symptoms of depression. Emotional well-being was evaluated in four studies, which also showed conflicting results. Symptoms of anxiety were evaluated in one study, which showed a significant improvement. CONCLUSIONS/INTERPRETATION: The effects of exercise training on psychological outcomes in people with type 2 diabetes are conflicting. Therefore, there is a need for further high-quality RCTs in order to gain greater insight into the role of exercise training in people with type 2 diabetes.

Data analysis supports monitoring and surveillance of goat health and welfare in the Netherlands.

Monitoring and surveillance systems have an increasingly important role in contemporary society ensuring high levels of animal health and welfare, securing export positions, and protecting public health by ensuring animal health and product safety. In the Netherlands, a voluntary monitoring and surveillance system is in place since 2003 to provide a broad overview of livestock trends in addition to disease-specific surveillance systems, including insight into the occurrence and prevalence of new and emerging non-notifiable diseases and disorders. Being a major surveillance component of this monitoring and surveillance system for small ruminant health in the Netherlands, an annual data analysis on routine census data is performed to retrospectively monitor trends and developments regarding goat health and welfare. This paper aims to describe the process of the data analysis on goat farms in the Netherlands in 2020 and subsequent results are discussed. The data analysis provides key monitoring indicators such as animal and farm density, mortality, animal movements, and numbers and origin of imported small ruminants. Trends were analysed over a five-year, period and associations between herd characteristics and herd health are evaluated. Results showed that in 2020 the Dutch goat population consisted of 670,842 goats, distributed over 14,730 unique herds and increased by 2.3 % compared to 2019. Between 2016 and 2020, although probably underestimated, recorded mortality rates showed a decline on both small-scale and professional farms, with a strongest decrease on farms with herd sizes over more than 200 animals. Seventy-five percent of all professional farms registered animal introductions, in addition to 63 % of small-scale farms, including 2439 imported goats. Performing risks analyses requires demographic knowledge of the goat industry. During and after several disease outbreaks, such as bluetongue and Schmallenberg virus disease, the data analysis proved to function as a valuable tool, however, appeared just as important for recording outbreak-free data. Since its start in 2006, the concept of the data-analysis has continuously been improved, and will in the future be further developed, especially if more complete data sets become available. Subsequently, data analysis will increasingly support monitoring and surveillance of goat health and welfare.

Induction therapy with ipilimumab and nivolumab followed by consolidative chemoradiation as organ-sparing treatment in urothelial bladder cancer: study protocol of the INDIBLADE trial.

Introduction: Studies that assessed the efficacy of pre-operative immune checkpoint blockade (ICB) in locally advanced urothelial cancer of the bladder showed encouraging pathological complete response rates, suggesting that a bladder-sparing approach may be a viable option in a subset of patients. Chemoradiation is an alternative for radical cystectomy with similar oncological outcomes, but is still mainly used in selected patients with organ-confined tumors or patients ineligible to undergo radical cystectomy. We propose to sequentially administer ICB and chemoradiation to patients with (locally advanced) muscle-invasive bladder cancer. Methods: The INDIBLADE trial is an investigator-initiated, single-arm, multicenter phase 2 trial. Fifty patients with cT2-4aN0-2M0 urothelial bladder cancer will be treated with ipilimumab 3 mg/kg on day 1, ipilimumab 3 mg/kg plus nivolumab 1 mg/kg on day 22, and nivolumab 3 mg/kg on day 43 followed by chemoradiation. The primary endpoint is the bladder-intact event-free survival (BI-EFS). Events include: local or distant recurrence, salvage cystectomy, death and switch to platinum-based chemotherapy. We will also evaluate the potential of multiparametric magnetic resonance imaging of the bladder to identify non-responders, and we will assess the clearance of circulating tumor DNA as a biomarker for ICB treatment response. Discussion: This is the first trial in which the efficacy of induction combination ICB followed by chemoradiation is being evaluated to provide bladder-preservation in patients with (locally advanced) urothelial bladder cancer. Clinical Trial Registration: The INDIBLADE trial was registered on clinicaltrials.gov on January 21, 2022 (NCT05200988).

The state of the art in beyond 5G distributed massive multiple-input multiple-output communication system solutions.

Beyond fifth generation (5G) communication systems aim towards data rates in the tera bits per second range, with improved and flexible coverage options, introducing many new technological challenges in the fields of network architecture, signal pro- cessing, and radio frequency front-ends. One option is to move towards cell-free, or distributed massive Multiple-Input Multiple-Output (MIMO) network architectures and highly integrated front-end solutions. This paper presents an outlook on be- yond 5G distributed massive MIMO communication systems, the signal processing, characterisation and simulation challenges, and an overview of the state of the art in millimetre wave antennas and electronics.

Epigenetic profiling demarcates molecular subtypes of muscle-invasive bladder cancer.

Muscle-invasive bladder cancer (MIBC) is a heterogeneous disease that often recurs despite aggressive treatment with neoadjuvant chemotherapy and (radical) cystectomy. Basal and luminal molecular subtypes have been identified that are linked to clinical characteristics and have differential sensitivities to chemotherapy. While it has been suggested that epigenetic mechanisms play a role in defining these subtypes, a thorough understanding of the biological mechanisms is lacking. This report details the first genome-wide analysis of histone methylation patterns of human primary bladder tumours by chromatin immunoprecipitations and next-generation sequencing (ChIP-seq). We profiled multiple histone marks: H3K27me3, a marker for repressed genes, and H3K4me1 and H3K4me3, which are indicators of active enhancers and active promoters. Integrated analysis of ChIP-seq data and RNA sequencing revealed that H3K4 mono-methylation demarcates MIBC subtypes, while no association was found for the other two histone modifications in relation to basal and luminal subtypes. Additionally, we identified differentially methylated H3K4me1 peaks in basal and luminal tumour samples, suggesting that active enhancers play a role in defining subtypes. Our study is the first analysis of histone modifications in primary bladder cancer tissue and provides an important resource for the bladder cancer community.

Angiopoietin-2, permeability oedema, occurrence and severity of ALI/ARDS in septic and non-septic critically ill patients.

BACKGROUND: Angiopoietin-2 and vascular endothelial growth factor (VEGF) may impair vascular barrier function while angiopoietin-1 may protect it. It was hypothesised that circulating angiopoietin-2 is associated with pulmonary permeability oedema and severity of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) during septic or non-septic critical illness. METHODS: Plasma levels of angiopoietin-1 and angiopoietin-2 were measured in mechanically ventilated patients (24 with sepsis, 88 without sepsis), together with the pulmonary leak index (PLI) for 67-gallium-labelled transferrin and extravascular lung water (EVLW) by transpulmonary thermal-dye dilution as measures of pulmonary permeability and oedema, respectively. ALI/ARDS was characterised by consensus criteria and the lung injury score (LIS). Plasma VEGF and von Willebrand factor (VWF) levels were assayed. RESULTS: Angiopoietin-2, VWF, PLI, EVLW and LIS were higher in patients with sepsis than in those without sepsis and higher in patients with ALI/ARDS (n = 10/12 in sepsis, n = 19/8 in non-sepsis) than in those without. VEGF was also higher in patients with sepsis than in those without. Patients with high PLI, regardless of EVLW, had higher angiopoietin-2 levels than patients with normal PLI and EVLW. Angiopoietin-2 correlated with the PLI, LIS and VWF levels (minimum r = 0.34, p<0.001) but not with EVLW. Angiopoietin-2 and VWF were predictive for ARDS in receiver operating characteristic curves (minimum area under the curve = 0.69, p = 0.006). Angiopoietin-1 and VEGF did not relate to the permeability oedema of ALI/ARDS. CONCLUSION: Circulating angiopoietin-2 is associated with pulmonary permeability oedema, occurrence and severity of ALI/ARDS in patients with and without sepsis. The correlation of angiopoietin-2 with VWF suggests activated endothelium as a common source.

FDG-PET/CT for response evaluation of invasive bladder cancer following neoadjuvant chemotherapy.

PURPOSE: We investigated the accuracy of FDG-PET/CT response identification following neoadjuvant or induction chemotherapy (NAIC) for invasive bladder cancer (BC) as to better select patients for radical cystectomy (RC). METHODS: Between 2010 and 2014, 37 cT1-4N1-3 BC patients received a FDG-PET/CT before and after NAIC followed by RC. Metabolic lymph node (LN) response was evaluated according to EORTC recommendations. Additionally, primary tumor response was evaluated for 23 patients by means of delayed pelvic imaging after forced diuresis. Gold standard was response on pathologic analysis of RC specimens. Response was defined as partial response (pPR, any pathologic downstaging) or complete response (pCR, 

Association Of Plasma And Urinary Mutant DNA With Clinical Outcomes In Muscle Invasive Bladder Cancer.

Muscle Invasive Bladder Cancer (MIBC) has a poor prognosis. Whilst patients can achieve a 6% improvement in overall survival with Neo-Adjuvant Chemotherapy (NAC), many do not respond. Body fluid mutant DNA (mutDNA) may allow non-invasive identification of treatment failure. We collected 248 liquid biopsy samples including plasma, cell pellet (UCP) and supernatant (USN) from spun urine, from 17 patients undergoing NAC. We assessed single nucleotide variants and copy number alterations in mutDNA using Tagged-Amplicon- and shallow Whole Genome- Sequencing. MutDNA was detected in 35.3%, 47.1% and 52.9% of pre-NAC plasma, UCP and USN samples respectively, and urine samples contained higher levels of mutDNA (p = <0.001). Longitudinal mutDNA demonstrated tumour evolution under the selective pressure of NAC e.g. in one case, urine analysis tracked two distinct clones with contrasting treatment sensitivity. Of note, persistence of mutDNA detection during NAC predicted disease recurrence (p = 0.003), emphasising its potential as an early biomarker for chemotherapy response.

Phosphorylation of pyruvate kinase type K in human gliomas by a cyclic adenosine 5'-monophosphate-independent protein kinase.

In recent years, we reported the isozyme shift of pyruvate kinase from the M- toward the K-type in human neuroectodermal tumors. To investigate whether this shift enables phosphorylation of pyruvate kinase in these tumors, we studied 29 different specimens of human brain tumors for endogenous pyruvate kinase phosphorylation. While in normal human brain no phosphorylation of pyruvate kinase was detected, in all brain tumors pyruvate kinase became phosphorylated. There was no correlation between the extent of the pyruvate kinase phosphorylation and the histological classification and grading or the pyruvate kinase isozyme composition of the tumors. Only pyruvate kinase type K, and not type M, served as a substrate in the phosphorylation reaction. The phosphorylation of pyruvate kinase could be completely inhibited by addition of fructose 1,6-bisphosphate, a positive effector of pyruvate kinase type K; alanine, however, a negative effector, and phospho-enol-pyruvate, a substrate in the pyruvate kinase reaction, had no effect. While pyruvate kinase type L in liver is phosphorylated by a cyclic AMP-dependent protein kinase, the incorporation of phosphate into pyruvate kinase in human brain tumors appeared to be cyclic AMP independent and occurred exclusively on serine residues.

Subunit-specific phosphorylation of pyruvate kinase in medullary thyroid carcinomas of the rat.

Pyruvate kinase from anaplastic medullary thyroid carcinomas contains predominantly K-type subunits, whereas pyruvate kinase from differentiated medullary thyroid carcinomas consist of M- and K-type subunits in about equal proportion. In order to analyse the incorporation of phosphate in the respective isozymes after endogenous phosphorylation of cytosolic extracts with [32P]ATP, homotetrameric isozymes as well as heterotetrameric hybrids of differentiated tumors were resolved by affinity chromatography on Blue-Sepharose CL-6B and, if necessary, further purified by immunoprecipitation. SDS-polyacrylamide gel electrophoresis of purified isozymes and subsequent autoradiography showed the incorporation of phosphate in the K4-type isozymes, but not in the other isozymes. The phosphorylation appeared to be cAMP-independent and occurred on a serine residue.

Partial characterization of protein tyrosine kinase activity in normal and leukemic human myeloid cells.

We have examined the expression of the protein tyrosine kinase (PTK) encoding oncogenes fes and abl in normal and malignant human myeloid cells in immunoblotting experiments. fes was markedly present in all cytosolic and most membrane fractions of normal and malignant cells. abl was only visible in normal cells, and occurred mostly in the cytosolic fractions. Molecular weights of identified proteins were different from the known products of fes and abl, possibly by alternative splicing at the mRNA level or by proteolysis. PTKs in myeloid cells were further purified by fast liquid protein chromatography (FPLC). PTK-activities of column fractions were assayed using a solid-phase non-radioactive dot-blot assay. Cytosolic and membrane fractions showed a FPLC pattern with a constant as well as a variable part in both normal and malignant cells, possibly indicative for PTKs with specialized functions in normal cell growth and transformation. Partial characterization of PTKs from different eluted peaks of AML-M4 blast cells demonstrated that PTKs from these peaks are kinetically distinct from each other.

A randomized, placebo-controlled study of loop diuretics in patients with essential hypertension: the bumetanide and furosemide on lipid profile (BUFUL) clinical study report.

This study was conducted to determine whether loop diuretics are more effective than placebo in reducing blood pressure without raising serum lipid levels, and whether bumetanide is more effective than furosemide in this respect. In a double-blind, 24-week placebo-controlled crossover study, 27 patients with essential hypertension were treated in four periods of 6 weeks each, including placebo twice, furosemide 40 mg daily, and bumetanide 1 mg daily. Several metabolic parameters, including serum lipid levels, and blood pressure were assessed. Overall levels of total cholesterol, triglycerides, and low-density lipoprotein (LDL) cholesterol were 5%, 12.4%, and 4.8% higher, respectively, during loop diuretic therapy than during placebo treatment. Overall systolic and diastolic blood pressure measurements were 12 mmHg and 4 mmHg lower, respectively, during loop diuretic therapy than during placebo treatment. Any added effect of bumetanide on serum lipid levels and blood pressure compared with furosemide, however, could not be confirmed. Our results indicate that the loop diuretics bumetanide and furosemide are effective in reducing blood pressure, and influence serum lipid levels markedly less than do thiazide diuretics or chlorthalidone. In addition, these results indicate that differences in blood pressure reduction and serum lipid levels between the two compounds were small and nonsignificant.