Home-based fecal calprotectin utilization in a general pediatric gastroenterology clinic.

OBJECTIVE: Remote investigation and monitoring have gained importance in ambulatory practice. A home-based fecal calprotectin (FC) test has been developed where the sample is processed and analyzed at home through a smartphone application. We aimed to assess the use of standard ELISA (sFC) versus home-based (hFC) FC testing in a general pediatric gastroenterology clinic. METHODS: Ambulatory pediatric patients with hFC or sFC performed between August 2019 and November 2020 were included. Data regarding demographics, clinical characteristics, medication use, investigations, and final diagnosis, categorized as inflammatory bowel disease (IBD), functional gastrointestinal (GI) disorders, organic non-IBD (ONI) GI disorders, non-GI disorders, and undetermined after 6 months of investigation, were recorded. RESULTS: A total of 453 FC tests from 453 unique patients were included. Of those, 249 (55%) were hFC. FC levels (median) were higher in children with IBD compared to non-IBD diagnosis (sFC 795 vs. 57 µg/g, hFC 595 vs. 47 µg/g, p < 0.001), and in ONI compared to functional GI disorders (sFC 85 vs. 54 µg/g, p = 0.003, hFC 57 vs. 40 µg/g, p < 0.001). No significant difference was observed between different ONI GI disorders or subtypes of functional disorders. Age did not significantly influence levels. CONCLUSIONS: Overall, hFC and sFC provide similar results in the general pediatric GI ambulatory setting. FC is a sensitive but not disease-specific marker to identify patients with IBD. Values appear to be higher in ONI GI disorders over functional disorders, although cut-off values have yet to be determined.

Mistimed origin licensing and activation stabilize common fragile sites under tight DNA-replication checkpoint activation.

Genome integrity requires replication to be completed before chromosome segregation. The DNA-replication checkpoint (DRC) contributes to this coordination by inhibiting CDK1, which delays mitotic onset. Under-replication of common fragile sites (CFSs), however, escapes surveillance, resulting in mitotic chromosome breaks. Here we asked whether loose DRC activation induced by modest stresses commonly used to destabilize CFSs could explain this leakage. We found that tightening DRC activation or CDK1 inhibition stabilizes CFSs in human cells. Repli-Seq and molecular combing analyses showed a burst of replication initiations implemented in mid S-phase across a subset of late-replicating sequences, including CFSs, while the bulk genome was unaffected. CFS rescue and extra-initiations required CDC6 and CDT1 availability in S-phase, implying that CDK1 inhibition permits mistimed origin licensing and firing. In addition to delaying mitotic onset, tight DRC activation therefore supports replication completion of late origin-poor domains at risk of under-replication, two complementary roles preserving genome stability.

Epidemiology of Musculoskeletal Manifestations in Pediatric Inflammatory Bowel Disease: A Systematic Review.

OBJECTIVE: Pediatric inflammatory bowel disease (p-IBD) is a chronic relapsing gastrointestinal disorder of childhood with long-term morbidity. Several extraintestinal manifestations are described, the most common being joint pain and/or inflammation. However, patient and disease characteristics, treatments, and outcomes of p-IBD-associated musculoskeletal disease are not well established. Our study aims to summarize the recent literature on the epidemiology of musculoskeletal manifestations in p-IBD in the era of biologics. METHODS: A systematic search of PubMed, Embase, Cochrane Library, Web of Science Core Collection, and Cumulative Index to Nursing and Allied Health Literature databases was performed with relevant keywords. Studies in English published from January 1, 2000, to December 21, 2020, were included. In total, 3893 articles were identified and screened. Study and population characteristics and outcomes of interest were recorded. Risk of bias assessment was performed using the Joanna Briggs Institute Critical Appraisal Tools. RESULTS: Thirteen studies were included for full review, which were primarily single-center observational studies with retrospective or cross-sectional designs. The diagnostic criteria and definitions used for musculoskeletal manifestations varied. Musculoskeletal manifestation prevalence ranged from 2% to 35%. Only one study assessed the response of musculoskeletal manifestations to biologics. Risk of bias demonstrated heterogeneity in study quality. CONCLUSION: This is the first systematic review of musculoskeletal manifestations in p-IBD. Analysis was limited because of variability in study design and data-reporting methods. Definitions varied among included studies, with a clear lack in standardization. Our study demonstrates the need for standardized assessment of musculoskeletal manifestations of p-IBD and further research to explore optimal management to advance care for this group of children.

Take your Positions and Shine: Effects of Positioning Aggregation-Induced Emission Luminophores within Sequence-Defined Macromolecules.

A luminophore with aggregation-induced emission (AIE) is employed for the conjugation onto supramolecular ligands to allow for detection of ligand binding. Supramolecular ligands are based on the combination of sequence-defined oligo(amidoamine) scaffolds and guanidiniocarbonyl-pyrrole (GCP) as binding motif. We hypothesize that AIE properties are strongly affected by positioning of the luminophore within the ligand scaffold. Therefore, we systematically investigate the effects placing the AIE luminophore at different positions within the overall construct, for example, in the main or side chain of the olig(amidoamine). Indeed, we can show that the position within the ligand structure strongly affects AIE, both for the ligand itself as well as when applying the ligand for the detection of different biological and synthetic polyanions.

Secondary Anticoagulation Prophylaxis for Catheter-Related Thrombosis in Pediatric Intestinal Failure: Comparison of Short- Vs Long-Term Treatment Protocols.

INTRODUCTION: Catheter-related thrombosis (CRT) is a devastating complication of central venous catheters in children with intestinal failure (IF), but the optimal preventive therapy of CRT is unknown. This study assessed the efficacy and safety of 2 protocols of secondary anticoagulation prophylaxis with low-molecular-weight heparin (LMWH). METHODS: This is a comparative cohort study of children from 2 IF programs who received secondary anticoagulation prophylaxis with LMWH for CRT. The short-term protocol group (N = 13) received therapeutic dosing until thrombus resolution or ≤3 months. In the long-term protocol group (N = 26), prophylactic dosing continued until line removal. Patients underwent routine annual vascular ultrasound and were followed for ≥1 year. The primary outcome was development of secondary thrombosis; post hoc analysis assessed rates of secondary thrombosis at 12 months. RESULTS: Patient demographics were similar between groups. Secondary thrombosis occurred in 8 of 13 (62%) patients in the short-term group and in 9 of 26 (35%) in the long-term protocol group (P = .019) in a median time of 144.5 and 689 days, respectively (P = .01). Secondary thrombosis within 12 months occurred in 7 of 13 (54%, short term) and 2 of 26 (8%, long term) patients (P = .001). Secondary thrombosis was associated with catheter replacements (23.5 vs 5.5 catheters per 1000 catheter days; P = .016) and longer daily parenteral nutrition (PN) infusion (24 vs 15.25 hours; P = .044). Compliance was good (>80% of doses) in 92% of patients. CONCLUSIONS: Long-term secondary anticoagulation prophylaxis with LMWH reduces the incidence of secondary thrombosis and should be considered in children with CRT that require PN for prolonged periods of time.

Selective Adhesion and Switchable Release of Breast Cancer Cells via Hyaluronic Acid Functionalized Dual Stimuli-Responsive Microgel Films.

The detection of tumor cells from liquid biopsy samples is of critical importance for early cancer diagnosis, malignancy assessment, and treatment. In this work, coatings of hyaluronic acid (HA)-functionalized dual-stimuli responsive poly(N-isopropylacrylamide) (PNIPAM) microgels are used to study the specificity of breast cancer cell binding and to assess cell friendly release mechanisms for further diagnostic procedures. The microgels are established by straightforward precipitation polymerization with amine bearing comonomers and postfunctionalization with a UV-labile linker that covalently binds HA to the microgel network. Well-defined microgel coatings for cell binding are established via simple physisorption and annealing. The HA-presenting PNIPAM microgel films are shown to specifically adhere CD44 expressing breast cancer cell lines (MDA-MB-231 and MCF-7), where an increase in adhesion correlates with higher CD44 expression and HA functionalization. Upon cooling below the lower critical solution temperature of PNIPAM microgels, the cells could be released; however, 10-30% of the cells still remained on the surface even after prolonged cooling and mild mechanical agitation. A complete cell release is achieved after applying the light stimulus by short UV treatment cleaving HA units from the microgels. Owing to the comparatively straightforward preparation procedures, such dual-responsive microgel films could be considered for the effective capture, release, and diagnostics of tumor cells.

Implementation of a Postpartum Hemorrhage Safety Bundle at an Urban Safety-Net Hospital.

Background Postpartum hemorrhage (PPH) is a leading cause of preventable maternal morbidity and mortality. Standardized response to obstetric hemorrhage is associated with significant improvement in maternal outcomes, yet implementation can be challenging. Objective The primary objective is to describe the methodology for program implementation of the Alliance for Innovation on Maternal Health Safety Bundle on PPH at an urban safety-net hospital. Methods Over an 18-month period, interventions geared toward (1) risk assessment and stratification, (2) hemorrhage identification and management, (3) team communication and simulation, and (4) debriefs and case review were implemented. Hemorrhage risk assessment stratification rates were tracked overtime as an early measure of bundle compliance. Results Hemorrhage risk assessment stratification rates improved to >90% during bundle implementation. Conclusion Keys to implementation included multidisciplinary stakeholder commitment, stepwise and iterative approach, and parallel systems for monitoring and evaluation Implementation of a PPH safety bundle is feasible in a resource-constrained setting.

Transcription-mediated organization of the replication initiation program across large genes sets common fragile sites genome-wide.

Common fragile sites (CFSs) are chromosome regions prone to breakage upon replication stress known to drive chromosome rearrangements during oncogenesis. Most CFSs nest in large expressed genes, suggesting that transcription could elicit their instability; however, the underlying mechanisms remain elusive. Genome-wide replication timing analyses here show that stress-induced delayed/under-replication is the hallmark of CFSs. Extensive genome-wide analyses of nascent transcripts, replication origin positioning and fork directionality reveal that 80% of CFSs nest in large transcribed domains poor in initiation events, replicated by long-travelling forks. Forks that travel long in late S phase explains CFS replication features, whereas formation of sequence-dependent fork barriers or head-on transcription-replication conflicts do not. We further show that transcription inhibition during S phase, which suppresses transcription-replication encounters and prevents origin resetting, could not rescue CFS stability. Altogether, our results show that transcription-dependent suppression of initiation events delays replication of large gene bodies, committing them to instability.

Transcription-dependent regulation of replication dynamics modulates genome stability.

Common fragile sites (CFSs) are loci that are hypersensitive to replication stress and hotspots for chromosomal rearrangements in cancers. CFSs replicate late in S phase, are cell-type specific and nest in large genes. The relative impact of transcription-replication conflicts versus a low density in initiation events on fragility is currently debated. Here we addressed the relationships between transcription, replication, and instability by manipulating the transcription of endogenous large genes in chicken and human cells. We found that inducing low transcription with a weak promoter destabilized large genes, whereas stimulating their transcription with strong promoters alleviated instability. Notably, strong promoters triggered a switch to an earlier replication timing, supporting a model in which high transcription levels give cells more time to complete replication before mitosis. Transcription could therefore contribute to maintaining genome integrity, challenging the dominant view that it is exclusively a threat.

Challenges and technologies of care developed by caregivers of patients with Alzheimer's disease / Desafios e tecnologias de cuidado desenvolvidos por cuidadores de pacientes com doença de Alzheimer

Objective: to identify the challenges and technologies of care developed by caregivers of patients with Alzheimer's disease. Method: an exploratory study with a qualitative approach was carried out with nine caregivers of elderly people with Alzheimer's disease from the mutual help group of a university hospital in the south of Brazil. Data collection took place between May and August 2017 through a semi-structured interview. Content analysis was used to analyze the data. Results: two categories emerged from the analysis of the data: the challenges faced by caregivers of elderly people with Alzheimer's Disease and the care technologies developed by caregivers of elderly people with Alzheimer's disease. Conclusion: the study showed that the care strategies elaborated by the caregiver can enhance understanding, reflection and discussion among health professionals, caregivers and family members about quality care for the elderly and minimize the difficulties of care in order to provide greater quality of care for the elderly.

Objetivo: conhecer os desafios e tecnologias de cuidado desenvolvidas por cuidadores de pacientes com doença de Alzheimer. Método: estudo exploratório qualitativo realizado com nove cuidadores de idosos com Doença de Alzheimer participantes do grupo de ajuda mútua de um hospital universitário do sul do Brasil. A coleta de dados ocorreu entre maio e agosto de 2017 através de entrevistas semiestruturada. Para a análise dos dados utilizou-se a Análise de Conteúdo. Resultados: emergiram duas categorias: Desafios enfrentados por cuidadores de idosos com a Doença de Alzheimer e Tecnologias de cuidado desenvolvidas por cuidadores de idosos com Doença de Alzheimer. Conclusão: o estudo mostrou que as estratégias de cuidado elaboradas pelo cuidador podem potencializar compreensão, reflexão e discussão entre os profissionais da saúde, cuidadores e familiares acerca do cuidado de qualidade ao idoso, além de minimizar as dificuldades de cuidado a fim de proporcionar maior qualidade de cuidado ao idoso.

An in vitro model of hemogenic endothelium commitment and hematopoietic production.

Adult-type hematopoietic stem and progenitor cells are formed during ontogeny from a specialized subset of endothelium, termed the hemogenic endothelium, via an endothelial-to-hematopoietic transition (EHT) that occurs in the embryonic aorta and the associated arteries. Despite efforts to generate models, little is known about the mechanisms that drive endothelial cells to the hemogenic fate and about the subsequent molecular control of the EHT. Here, we have designed a stromal line-free controlled culture system utilizing the embryonic pre-somitic mesoderm to obtain large numbers of endothelial cells that subsequently commit into hemogenic endothelium before undergoing EHT. Monitoring the culture for up to 12 days using key molecular markers reveals stepwise commitment into the blood-forming system that is reminiscent of the cellular and molecular changes occurring during hematopoietic development at the level of the aorta. Long-term single-cell imaging allows tracking of the EHT of newly formed blood cells from the layer of hemogenic endothelial cells. By modifying the culture conditions, it is also possible to modulate the endothelial cell commitment or the EHT or to produce smooth muscle cells at the expense of endothelial cells, demonstrating the versatility of the cell culture system. This method will improve our understanding of the precise cellular changes associated with hemogenic endothelium commitment and EHT and, by unfolding these earliest steps of the hematopoietic program, will pave the way for future ex vivo production of blood cells.

Stretch Out or Fold Back? Conformations of Dinuclear Gold(I) N-Heterocyclic Carbene Macrocycles.

We report the synthesis and characterization of 12 new dinuclear gold(I) N-heterocyclic carbene (NHC) complexes and the corresponding imidazolium precursors. The focus lies in a systematic study of conformational changes and intra- and intermolecular gold-gold and π-π interactions of dinuclear gold(I) carbene complexes. Common to all members of the series of gold macrocycles are NHC ligands on the basis of imidazole with ethyl side chains and bromide as well as hexafluorophosphate counterions, respectively. The compounds vary in the length of a flexible alkyl linker between the NHC units. For the methylene and ethylene bridged macrocycles, a ring inversion movement can be observed by VT-NMR. In total, 11 molecular structures have been characterized by X-ray diffraction. Open ring conformations with intermolecular π-π and Au-Au interactions prevail, but a backfolded conformation with a short intramolecular Au-Au distance has been found for the ethylene-bridged species. The presence of Au-Au interactions could be proven by quantum chemical calculations.

Health governance by collaboration: a case study on an area-based programme to tackle health inequalities in the Dutch city of the Hague.

BACKGROUND: Area-based programmes are seen as a promising strategy for tackling health inequalities. In these programmes, local authorities and other local actors collaborate to employ health promoting interventions and policies. Little is known about the underlying processes of collaborative governance. To unravel this black box, we explored how the authority of The Hague, The Netherlands, developed a programme tackling health inequalities drawing on a collaborative mode of governance. METHODS: Case study drawing on qualitative semi-structured interviews and document review. Data were inductively analysed against the concept of collaborative governance. RESULTS: The authority's ambition was to co-produce a programme on tackling health inequalities with local actors. Three stages could be distinguished in the governing process: (i) formulating policy objectives, (ii) translating policy objectives into interventions and (iii) executing health interventions. In the stage of formulating policy objectives, the collaboration led to a reframing of the initial objectives. Furthermore, the translation of the policy objectives into health interventions was rather pragmatic and loosely based on health needs and/or evidence. As a result, the concrete actions that ensued from the programme did not necessarily reflect the initial objectives. CONCLUSION: In a local system of health governance by collaboration, factors other than the stated policy objectives played a role, eventually undermining the effectiveness of the programme in reducing health inequalities. To be effective, the processes of collaborative governance underlying area-based programmes require the attention of the local authority, including the building and governing of networks, a competent public health workforce and supportive infrastructures.

Effects of a ketogenic diet on the quality of life in 16 patients with advanced cancer: A pilot trial.

BACKGROUND: Tumor patients exhibit an increased peripheral demand of fatty acids and protein. Contrarily, tumors utilize glucose as their main source of energy supply. Thus, a diet supplying the cancer patient with sufficient fat and protein for his demands while restricting the carbohydrates (CHO) tumors thrive on, could be a helpful strategy in improving the patients' situation. A ketogenic diet (KD) fulfills these requirements. Therefore, we performed a pilot study to investigate the feasibility of a KD and its influence on the quality of life of patients with advanced metastatic tumors. METHODS: Sixteen patients with advanced metastatic tumors and no conventional therapeutic options participated in the study. The patients were instructed to follow a KD (less than 70 g CHO per day) with normal groceries and were provided with a supply of food additives to mix a protein/fat shake to simplify the 3-month intervention period. Quality of life [assessed by EORTC QLQ-C30 (version 2)], serum and general health parameters were determined at baseline, after every two weeks of follow-up, or after drop out. The effect of dietary change on metabolism was monitored daily by measuring urinary ketone bodies. RESULTS: One patient did not tolerate the diet and dropped out within 3 days. Among those who tolerated the diet, two patients died early, one stopped after 2 weeks due to personal reasons, one felt unable to stick to the diet after 4 weeks, one stopped after 6 and two stopped after 7 and 8 weeks due to progress of the disease, one had to discontinue after 6 weeks to resume chemotherapy and five completed the 3 month intervention period. These five and the one who resumed chemotherapy after 6 weeks report an improved emotional functioning and less insomnia, while several other parameters of quality of life remained stable or worsened, reflecting their very advanced disease. Except for temporary constipation and fatigue, we found no severe adverse side effects, especially no changes in cholesterol or blood lipids. CONCLUSIONS: These pilot data suggest that a KD is suitable for even advanced cancer patients. It has no severe side effects and might improve aspects of quality of life and blood parameters in some patients with advanced metastatic tumors.

Induction of programmed cell death by inhibition of AKT with the alkylphosphocholine perifosine in in vitro models of platinum sensitive and resistant ovarian cancers.

PURPOSE: We analyzed the anti-tumor effect and the mechanism of action of perifosine, an orally active alkylphospholipid AKT inhibitor using in vitro models of human ovarian cancer. METHODS: Ovarian cancer cells OAW42, PA-1, SKOV3, and A2780 as well as platinum resistant A2780cis cells were incubated with increasing concentrations of perifosine, with and without multi-caspase inhibitor zVAD-FMK. The effect of a combined treatment with cisplatin and perifosine was investigated in OAW42, SKOV3, A2780 and A2780cis cells. Cytotoxic effects of perifosine were analyzed using crystal violet staining, FACS analysis of DNA content as well as Annexin V/propidium iodide-double staining. The effect of perifosine on AKT phosphorylation was determined by Western blotting. RESULTS: Perifosine displayed anti-tumor activity in all five cell lines, which increased time-dependently. While IC(50) values at 24 h were >40 µM, IC(50) values after 72 h decreased to 10 µM in OAW42 and 25 µM in PA-1 and 30 µm in SKOV3 cells. In platinum resistant A2780cis cells perifosine showed good antiproliferative activity (IC(50) = 3 µm). At adequate doses, perifosine increased cytotoxic effects of cisplatin in OAW42, A2780 and A2780cis cell. Anti-tumor activity of perifosine was not confined to a specific phase of the cell cycle and could not be decreased by the pan-caspase inhibitor zVAD-FMK. AnnexinV/propidium iodide-double staining after treatment with perifosine was not indicative of classical apoptosis. AKT phosphorylation was dose-dependently inhibited by perifosine. CONCLUSIONS: Perifosine showed substantial cytotoxic effects in various in vitro models of ovarian cancer. Since anti-tumor effects were not confined to platinum-sensitive cells perifosine seems to be a good candidate for clinical studies in patients especially with platinum resistant ovarian cancer.

The clinicopathological and prognostic relevance of pyruvate kinase M2 and pAkt expression in breast cancer.

BACKGROUND: Cancer cell growth has been described to depend on glucose utilization. Activation of Akt and up-regulation of pyruvate kinase M2 (M2-PK) are attributes of tumour glycolysis. PATIENTS AND METHODS: In order to evaluate the prognostic relevance of glycolytic markers in breast cancer, the expression of pAkt and M2PK was analysed in 160 tissue samples. The staining results were compared with clinicopathological characteristics and survival data. RESULTS: Overexpression of pAkt was detected in 58% and of M2PK in 70% of breast cancer samples. Increased pAkt-expression was accompanied with shorter survival time. In contrast, M2PK expression was significantly higher in patients surviving breast cancer for more than 13 years. CONCLUSION: Strong M2PK expression seems to be a favourable prognostic factor and its role in breast cancer progression should be further explored. Our data confirm previous observations of pAkt as a negative prognostic marker.

Generating political priority to tackle health disparities: a case study in the Dutch city of The Hague.

OBJECTIVES: We sought to study the factors that determined the success of a recent initiative to generate political priority for the problem of health disparities in the city of The Hague, the Netherlands. METHODS: Our study had a prospective design. The qualitative data collection included interviews, document analyses, and observations. RESULTS: Crucial for the success of this initiative was the presence of powerful and credible actors. These actors effectively presented scientific evidence on health disparities and framed the issue in the light of shared values, priorities, and policy principles. Finally, their actions were supported by the national context, including the availability of national scientific research on health disparities. CONCLUSIONS: The project in The Hague shows that political priority for tackling health disparities can be generated at a local level. Key factors included framing the issue in the light of shared values and framing the problem and the solution as in line with existing policy principles.

Glycolytic phenotype in breast cancer: activation of Akt, up-regulation of GLUT1, TKTL1 and down-regulation of M2PK.

PURPOSE: Metabolic dependence on glucose utilisation has been described for different tumours characterised by activation of Akt, upregulation of GLUT1, M2PK and TKTL1. To date, however, little is known about glucose metabolism in breast cancer tissue. METHODS: We analysed 55 breast cancer specimens, 26 adjacent ductal carcinomas in situ (DCIS) and 23 adjacent normal breast tissues for expression of glycolytic markers by immunohistochemistry. RESULTS: We found expression of pAkt in 49%, GLUT1 in 25%, M2PK in 68% and TKTL1 in 31% of the tumours investigated. Expression of pAkt and Her2neu are positively correlated with borderline significance (P = 0.055). Expression of pAkt, GLUT1 and TKTL1 were higher in breast cancer and DCIS than in normal tissue. Surprisingly, M2PK expression was highest in normal breast tissue. CONCLUSIONS: We found a glycolytic phenotype in a high percentage of breast cancer samples. Inhibition of glycolysis might evolve as a future option for breast cancer therapy.

Expression of p-AKT characterizes adenoid cystic carcinomas of head and neck with a higher risk for tumor relapses.

BACKGROUND: Adenoid cystic carcinomas are rare tumors with an indolent clinical course, but frequent local relapses. The identification of tumors with a higher relapse risk seems to be interesting. Hence we investigated parameters of glucose metabolism, which were found associated with poor prognosis in other malignancies. METHODS: Specimen of 29 patients were investigated immunohistochemically with antibodies against p-AKT, TKTL-1 (transketolase-like 1), M2PK (M2 pyruvate kinase), and GLUT-1. Proliferation was investigated by staining with Ki67. The tumors were located at the major or minor salivary glands. Only the typical cribriform subtype was investigated. The initial tumor stage was pT1 or pT2. RESULTS: Expression of p-AKT was significantly (P = 0.036) associated with a higher relapse risk in multivariate analysis. Low expression of M2PK was non-significantly (P = 0.065) predictive for a higher risk. TKTL-1 and GLUT-1 were expressed in the majority of cases, albeit not associated with relapse risk. CONCLUSION: Adenoid cystic carcinomas positive for p-AKT show a higher relapse risk. However, other parameters of glucose metabolism investigated here or proliferation (Ki67) were not predictive in this entity. Our findings demonstrate a possible background for therapeutic approaches targeting the inhibition of PI3K/AKT pathway.

Glucose transporter GLUT1 expression is an stage-independent predictor of clinical outcome in adrenocortical carcinoma.

Owing to the rarity of adrenocortical carcinoma (ACC) no prognostic markers have been established beyond stage and resection status. Accelerated glycolysis is a characteristic feature of cancer cells and in a variety of tumour entities key factors in glucose metabolism like glucose transporter 1 and 3 (GLUT1 and -3), transketolase like-1 enzyme (TKTL1) and pyruvate kinase type M2 (M2-PK) are overexpressed and of prognostic value. Therefore, we investigated the role of these factors in ACC. Immunohistochemical analysis was performed on tissue microarrays of paraffin-embedded tissue samples from 167 ACCs, 15 adrenal adenomas and 4 normal adrenal glands. Expression was correlated with baseline parameters and clinical outcome. GLUT1 and -3 were expressed in 33 and 17% of ACC samples respectively, but in none of the benign tumours or normal adrenals glands. By contrast, TKTL1 and M2-PK were detectable in all benign tissues and the vast majority of ACCs. GLUT1 expression was strongly associated with prognosis in univariate and multivariate analysis (P<0.01), whereas GLUT3, TKTL1 and M2-PK did not correlate with clinical outcome. Patients with strong GLUT1 staining showed a considerably higher overall mortality (hazard ratio (HR) 6.34 (95% confidence interval 3.10-12.90) compared with patients with no GLUT1 staining. When analysing patients in their early stages and advanced disease separately, similar results were obtained. HR for survival was 5.31 (1.80-15.62) in patients with metastatic ACC and in patients after radical resection the HR for disease-free survival was 6.10 (2.16-16.94). In conclusion, GLUT1 is a highly promising stage-independent, prognostic marker in ACC.