Cocaine versus xylometazoline to prevent epistaxis after nasotracheal intubation: A randomized trial.

BACKGROUND: Nasotracheal intubation is associated with a risk of epistaxis. Several drugs, including cocaine and xylometazoline may be used as decongestants prior to nasotracheal intubation to prevent this. We hypothesized that xylometazoline would prevent epistaxis more effectively than cocaine, demonstrated by a lower proportion of patients with bleeding after nasotracheal intubation. METHODS: We conducted a single-center, outcome assessor and analyst-blinded, clinical randomized controlled trial following approval from the local research ethics committee and the national medicine agency. Written informed consent was obtained from all patients. Patients scheduled for surgery under general anesthesia with nasotracheal intubation were randomized to receive either 2 mL 4% cocaine or 2 mL 0.05% xylometazoline prior to nasotracheal intubation. Immediately following intubation, epistaxis was evaluated by the blinded intubating anesthetist on a four-point scale. We measured heart rate and blood pressure the first 5 min after drug administration. Adverse events were followed up after 24 h. RESULTS: A total of 53 patients received cocaine and 49 patients received xylometazoline. Bleeding occurred in 32 patients receiving cocaine (60.4%) and in 34 patients receiving xylometazoline (69.4%) (p = .41, Fisher's exact test) with a difference of 9.0% (95% CI: -9.4% to 27%). There was no statistically significant difference between groups regarding the heart rate or blood pressure. No adverse cardiac events were recorded in either group. CONCLUSION: We found no statistically significant difference between cocaine and xylometazoline in preventing epistaxis after nasotracheal intubation, and the choice of vasoconstrictor should be based on other considerations, such as pricing, availability and medicolegal issues.

Improved overall survival for Stage III NSCLC patients treated with curative-intended therapy from 2010 to 2018-a cohort study in Denmark.

BACKGROUND: Despite advances in treatment strategies and improved clinical outcomes, an unmet need remains for NSCLC patients. With an increased real-world knowledge of NSCLC, clinicians could offer patients optimal tailored treatment and disease management. In this retrospective cohort study, we describe patient characteristics, treatment patterns and modality, and survival in NSCLC patients diagnosed and treated at Aarhus University Hospital, Denmark. METHODS: Data on Stage III NSCLC patients aged ≥18 years diagnosed 2010-2018 were obtained from a regional cancer database and linked to national registries for information on socioeconomic and vital status. Patients were stratified by planned treatment intention at diagnosis (curative/palliative). Treatment patterns and overall survival (OS) were estimated using time-to-event methods. RESULTS: Broad patient and diseases characteristics and multiple treatment options demonstrated the heterogeneity of this patient cohort. Of 851 Stage III NSCLC patients, 599 (70%) and 252 (30%) were offered curative- and palliative-intended treatment, respectively, upon evaluation by a multidisciplinary team (MDT). The most frequent treatment modalities were CRT (n = 328; 55%) and RT (n = 97; 38%) in the curative and palliative setting, respectively. Age, disease stage, performance status and comorbidity were associated with curative-intended treatment initiation. Curative-intended treatment was associated with an improved OS of 14.6 months (median OS 24.4 months, 95% CI 21.1-27.6). Being offered curative-intended treatment and/or being diagnosed in the more contemporary study period (2016-2018) were significantly correlated with better OS (p < 0.001). CONCLUSION: Stage III NSCLC is a heterogeneous disease as regards patient and clinical characteristics, multiple treatment options, and outcomes. Age, disease staging, performance status, and comorbidity, as well as MDT evaluation and matching treatment intent, are important determinants of curative-intended treatment. Notably, an NSCLC diagnosis in the more contemporary study period was statistically significantly associated with better OS.

Improvements in cuirass ventilation for airway surgery: origins in Copenhagen on the 70th anniversary of the first intensive care unit.

The first modern intensive care unit was established in Copenhagen 70 yr ago. This cornerstone of anaesthesia was largely based on experience gained using positive pressure ventilation to save hundreds of patients during the polio epidemic in 1952. Ventilation approaches, monitoring techniques, and pharmacological innovations have developed to such an extent that cuirass ventilation, which proved inadequate during the polio epidemic, might now have novel applications for both anaesthesia and treatment of the critically ill.

Tube tip in pharynx-a conduit for awake oral intubation in patients with extremely restricted mouth opening.

PURPOSE: Awake flexible bronchoscope-guided intubation is challenging in patients with extremely limited mouth opening (when there is inadequate space for an oropharyngeal airway), especially when nasal access is unavailable. Alternatives include awake front of neck access, which is an invasive procedure and not suitable for elective surgery. We present a novel technique to facilitate flexible bronchoscope-guided oral intubation in these patients. CLINICAL FEATURES: Tube tip in pharynx (TTIP) is a technique for establishing a patent airway if ventilation is difficult or has failed using a face mask, supraglottic airway, or endotracheal tube. The technique involves placing the tip of the endotracheal tube in the pharynx, 10-14 cm past the teeth, filling the cuff with air, closing the mouth and nose of the patient, and then initiating ventilation. The TTIP method thus combines the function of an oropharyngeal airway and a face mask akin to a supraglottic airway device, but is more flexible with regard to insertion depth and cuff inflation and demands only minimal mouth opening. We have adapted the TTIP technique for awake flexible bronchoscope-guided oral intubation and report the technique illustrated with three cases where mouth opening was so restricted that it precluded insertion of an oropharyngeal airway. CONCLUSION: By placing an endotracheal tube with the tip in the pharynx, TTIP can establish a conduit for awake oral flexible bronchoscope-guided intubation in patients with extremely limited mouth opening and unavailable nasal access. This technique requires equipment that is readily available and may help avoid unnecessary awake tracheostomy.

RéSUMé: OBJECTIF: L'intubation éveillée guidée par bronchoscope flexible est un défi chez les patients présentant une ouverture buccale extrêmement restreinte (lorsqu'il n'y a pas suffisamment d'espace pour une canule oropharyngée), et tout particulièrement lorsqu'un accès nasal est non disponible. Les alternatives incluent l'accès antérieur du cou chez patient éveillé, une procédure invasive qui ne convient pas pour la chirurgie élective. Nous présentons une technique innovante pour faciliter l'intubation orale guidée par bronchoscope flexible chez ces patients. CARACTéRISTIQUES CLINIQUES: La technique TTIP (de l'anglais Tube Tip In Pharynx) permet d'établir un accès aux voies aériennes si la ventilation est difficile ou a échoué à l'aide d'un masque, d'un dispositif supraglottique ou d'un tube endotrachéal. La technique consiste à placer l'extrémité du tube endotrachéal dans le pharynx, à 10-14 cm au-delà des dents, à remplir le ballonnet d'air, à fermer la bouche et le nez du patient, puis à amorcer la ventilation. La méthode TTIP combine ainsi la fonction d'une canule oropharyngée et d'un masque facial, de façon similaire à un dispositif supraglottique, mais est plus flexible en ce qui concerne la profondeur d'insertion et le gonflage du ballonnet et ne nécessite qu'une ouverture minimale de la bouche. Nous avons adapté la technique TTIP pour l'intubation orale éveillée guidée par bronchoscope flexible et rapportons la technique en l'illustrant par trois cas où l'ouverture de la bouche était si restreinte qu'elle empêchait l'insertion d'une canule oropharyngée. CONCLUSION: En plaçant l'extrémité d'un tube endotrachéal dans le pharynx, la technique TTIP peut établir un passage pour l'intubation orale éveillée guidée par bronchoscope flexible chez les patients ayant une ouverture de la bouche extrêmement limitée et un accès nasal indisponible. Cette technique nécessite du matériel facilement disponible et peut contribuer à éviter une trachéotomie non nécessaire chez patient éveillé.

Managing and securing the bleeding upper airway: a narrative review.

Failure to manage bleeding in the airway is an important cause of airway-related death. The purpose of this narrative review is to identify techniques and strategies that can be employed when severe bleeding in the upper airway may render traditional airway management (e.g., facemask ventilation, intubation via direct/video laryngoscopy, flexible bronchoscopy) impossible because of impeded vision. An extensive literature search was conducted of bibliographic databases, guidelines, and textbooks using search terms related to airway management and bleeding. We identified techniques that establish a definitive airway, even in cases of impeded visibility resulting from severe bleeding in the airway. These include flexible video-/optical- scope-guided intubation via a supraglottic airway device; cricothyroidotomy or tracheotomy; and retrograde-, blind nasal-, oral-digital-, light-, and ultrasound-guided intubation. We provide a structured approach to managing bleeding in the airway that accounts for the source of bleeding and the estimated risk of failure to intubate using direct laryngoscopy or to achieve a front-of-neck access for surgical airway rescue. In situations where these techniques are predicted to be successful, the recommended approach is to identify the cricothyroid membrane (in preparation for rescue cricothyroidotomy), followed by rapid sequence induction. In situations where traditional management of the airway is likely to fail, we recommend an awake approach with one of the aforementioned techniques.

Depth to the airway lumen at the level of the cricothyroid membrane measured by ultrasound.

BACKGROUND: Knowing the likely depth to the airway before emergency cricothyroidotomy may improve success in cases where it cannot be measured. Our aim was to measure the depth to the airway at the cricothyroid membrane by ultrasound in a large group of adult patients. METHOD: Prospective, observational study in two centres, Oxford and Gloucester. Patients presenting for a large variety of surgical operations were studied. Patients under 18 years; pregnant; critically ill; had a history of neck surgery were not included. Ultrasound examination was performed pre-operatively while participants lay supine with their head and neck extended, with light transducer pressure. We measured depth to the airway lumen in mm; age; weight; height and sex. RESULTS: In total 352 patients were studied. We found that depth to the airway lumen strongly correlated with weight (r = 0.855, P < 0.001) and to a lesser extent body mass index (r = 0.781, P < 0.001). Statistical analysis produced an equation to predict upper 95% CI of depth to the airway from the patient's weight: Depth to the airway lumen in mm = (0.13 × weight in kg) + 0.86. CONCLUSIONS: If ultrasound measurement is not possible before emergency cricothyroidotomy, the clinician could use our results to predict the depth to the airway by using the patient's weight. If the upper 95% CI were used as the depth of incision, it would enter the airway in 39 out of 40 patients of that weight, without damage to posterior structures in those with a shallower airway.

Sun behaviour and physical activity associated with autumn vitamin D status in 4-8-year-old Danish children.

OBJECTIVE: To explore determinants of serum 25-hydroxyvitamin D (s-25(OH)D) during autumn in young, Caucasian children not consuming vitamin D-fortified foods or supplements, and explore differences in sun behaviours between pre-school and school children. DESIGN: In September-October, s-25(OH)D was measured by LC-MS/MS; physical activity, sun behaviours and vitamin D intake were assessed with questionnaires. SETTING: Baseline data from the ODIN Junior trial at 55°N. SUBJECTS: Children aged 4-8 years (n 130), of whom 96% gave blood samples. RESULTS: Mean s-25(OH)D was 56·8 (sd 12·5) nmol/l and positively associated with fat-free mass index (P=0·014). Children being active 6-7 h/week had 5·6 (95% CI 1·1, 10·0) nmol/l higher s-25(OH)D than less active children (P=0·014). Children seeking shade sometimes or rarely/never had 7·0 (95% CI 1·2, 12·9; P=0·018) and 7·2 (95% CI 0·8, 13·6; P=0·028) nmol/l higher s-25(OH)D, respectively, than children always/often seeking shade. Pre-school children had more sun-safe behaviour than school children in terms of use of a hat, sunscreen and sunscreen sun protection factor (P<0·05). In school but not pre-school children, using a hat rarely/never was associated with 12·1 (95% CI 2·5, 21·7; P=0·014) nmol/l higher s-25(OH)D v. always/often (P interaction=0·019). Vitamin D intake was not associated with s-25(OH)D (P=0·241). CONCLUSIONS: Physical activity and sun behaviours are associated with s-25(OH)D in young children. Identifying factors influencing autumn s-25(OH)D is relevant to optimize levels before sun exposure diminishes. Strategies to reduce risk of inadequacy should consider risk of skin cancer and sunburn, and could include fortification and/or vitamin D supplementation.

DNA methyltransferases expression in normal tissues and various human cancer cell lines, xenografts and tumors.

DNA methylation plays an important role in carcinogenesis and aberrant methylation patterns have been found in many tumors. Methylation is regulated by DNA methyltransferases (DNMT), catalyzing DNA methylation. Therefore inhibition of DNMT is an interesting target for anticancer treatment. RX-3117 (fluorocyclopentenylcytosine) is a novel demethylating antimetabolite that is currently being studied in clinical trials in metastatic bladder and pancreatic cancers. The active nucleotide of RX-3117 is incorporated into DNA leading to downregulation of DNMT1, the maintenance DNA methylation enzyme. Since DNMT1 is a major target for the activity of RX-3117, DNMT1 may be a potential predictive biomarker. Therefore, DNMT1 protein and mRNA expression was investigated in 19 cancer cell lines, 26 human xenografts (hematological, lung, pancreatic, colon, bladder cancer) and 10 colorectal cancer patients. The DNMT1 mRNA expression showed large variation between cell lines (100-fold) and the 26 xenografts (1100-fold) investigated. The DNMT1 protein was overexpressed in colon tumours from patients compared to non-malignant mucosa from the same patients (P = 0.02). The DNA methylation in these patients was significantly higher in tumour tissues compared to normal mucosa (P = 0.001). DNMT1 expression in normal white blood cells also showed a large variation. In conclusion, the large variation in DNMT1 expression may serve as a potential biomarker for demethylating therapy such as with RX-3117.

Sensitive liquid chromatography mass spectrometry (LC-MS) assay reveals novel insights on DNA methylation and incorporation of gemcitabine, its metabolite difluorodeoxyuridine, deoxyuridine, and RX-3117 into DNA.

Antimetabolites are incorporated into DNA and RNA, affecting their function. Liquid-chromatography-mass-spectrometry (LC-MS-MS) permits the sensitive, selective analysis of normal nucleosides. The method was adapted to measure the incorporation of deoxyuridine, gemcitabine (difluorodeoxycytidine), its metabolite difluorodeoxyuridine (dFdU), and the novel compound fluorocyclopentenylcytosine (RX3117). DNA was degraded to its deoxynucleotides for quantification by LC-MS-MS, gradient chromatography on a Phenomenex prodigy-3-ODS with positive ionization. The range of deoxyuridine DNA-mis-incorporation varied nine-fold in 27 cell lines (leukemia, colon, ovarian, lung cancer). At low-folate conditions a 2.1-fold increase in deoxyuridine was observed. Global methylation (given as % 5-methyl-deoxycytidine) was comparable between the cell lines (4.6-6.5%). Exposure of A2780 cells to 1 µM gemcitabine (4 hours) resulted in 3.6 pmol gemcitabine/µg DNA, but in AG6000 cells (deoxycytidine-kinase-deficient) no incorporation was found. However, when A2780, AG6000, or CCRF-CEM cells were exposed to 100 µM dFdU we found it as gemcitabine, 20.5, 19.6, and 0.51 pmol gemcitabine/µg DNA, respectively. Preincubation of CCRF-CEM cells with cyclopentenyl-cytosine (a CTP-synthetase inhibitor) increased dFdU incorporation four-fold. Apparently dFdU is activated independently of deoxycytidine-kinase and possibly converted in-situ to dFdCMP. RX3117 was incorporated into both DNA and RNA (0.0037 and 0.00515 pmol/µg, respectively). In summary, a sensitive method to quantify the incorporation of gemcitabine, deoxyuridine, and RX-3117 was developed, which revealed that dFdU was incorporated into DNA as the parent compound gemcitabine.

The formation and design of the TRIAGE study--baseline data on 6005 consecutive patients admitted to hospital from the emergency department.

BACKGROUND: Patient crowding in emergency departments (ED) is a common challenge and associated with worsened outcome for the patients. Previous studies on biomarkers in the ED setting has focused on identification of high risk patients, and and the ability to use biomarkers to identify low-risk patients has only been sparsely examined. The broader aims of the TRIAGE study are to develop methods to identify low-risk patients appropriate for early ED discharge by combining information from a wide range of new inflammatory biomarkers and vital signs, the present baseline article aims to describe the formation of the TRIAGE database and characteristize the included patients. METHODS: We included consecutive patients ≥ 17 years admitted to hospital after triage staging in the ED. Blood samples for a biobank were collected and plasma stored in a freezer (-80 °C). Triage was done by a trained nurse using the Danish Emergency Proces Triage (DEPT) which categorizes patients as green (not urgent), yellow (urgent), orange (emergent) or red (rescusitation). Presenting complaints, admission diagnoses, comorbidities, length of stay, and 'events' during admission (any of 20 predefined definitive treatments that necessitates in-hospital care), vital signs and routine laboratory tests taken in the ED were aslo included in the database. RESULTS: Between September 5(th) 2013 and December 6(th) 2013, 6005 patients were included in the database and the biobank (94.1 % of all admissions). Of these, 1978 (32.9 %) were categorized as green, 2386 (39.7 %) yellow, 1616 (26.9 %) orange and 25 (0.4 %) red. Median age was 62 years (IQR 46-76), 49.8 % were male and median length of stay was 1 day (IQR 0-4). No events were found in 2658 (44.2 %) and 158 (2.6 %) were admitted to intensive or intermediate-intensive care unit and 219 (3.6 %) died within 30 days. A higher triage acuity level was associated with numerous events, including acute surgery, endovascular intervention, i.v. treatment, cardiac arrest, stroke, admission to intensive care, hospital transfer, and mortality within 30 days (p < 0.001). CONCLUSION: The TRIAGE database has been completed and includes data and blood samples from 6005 unselected consecutive hospitalized patients. More than 40 % experienced no events and were therefore potentially unnecessary hospital admissions.

Positron emission tomography/computed tomography and biomarkers for early treatment response evaluation in metastatic colon cancer.

BACKGROUND: Treatment options for metastatic colon cancer (mCC) are widening. We prospectively evaluated serial 2-deoxy-2-[18F]fluoro-d-glucose positron-emission tomography/computed tomography (PET/CT) and measurements of tissue inhibitor of metalloproteinases-1 (TIMP-1), carcinoembryonic antigen (CEA), and liberated domain I of urokinase plasminogen activator receptor (uPAR(I)) for early assessment of treatment response in mCC patients. METHODS: Thirty-three mCC patients scheduled for first-line chemotherapy with capecitabine and oxaliplatin (CAPOX) and bevacizumab participated; 27 were evaluated by PET/CT before treatment, after one and four treatment series. Morphological and metabolic response was independently assessed according to Response Evaluation Criteria in Solid Tumors and European Organization for Research and Treatment of Cancer PET criteria. Plasma TIMP-1, plasma uPAR(I), and serum CEA were determined. RESULTS: Metabolic response after one treatment course predicted the ability of CAPOX and bevacizumab to induce morphological response after four treatment series with a sensitivity of 80%, specificity of 69%, and odds ratio of 13.9 (95% confidence interval [CI] 1.9; 182). Early metabolically stable or progressive disease was associated with shorter progression-free survival (hazard ratio [HR] = 3.2 [CI 1.3; 7.8]). Biomarker levels at early evaluation were associated with shorter OS (TIMP-1 per unit increase on a log-2-transformed ng/mL scale: HR = 2.6 [CI 1.4; 4.9]; uPAR(I) per 25 fmol/mL increase: HR = 1.5 [CI 1.1; 2.1]). CONCLUSION: This monocentric study demonstrated predictive value of early metabolic PET response and prognostic value of TIMP-1 and uPAR(I) levels in mCC treated with CAPOX and bevacizumab. Results support investigation of PET/CT, TIMP-1, and uPAR(I) guided early treatment adaptation in mCC.

Positron emission tomography/computed tomography for optimized colon cancer staging and follow up.

OBJECTIVES: Optimal management of colon cancer (CC) requires detailed assessment of extent of disease. This study prospectively investigates the diagnostic accuracy of 2-deoxy-2-[18F]fluoro-D-glucose positron emission tomography/computed tomography (PET/CT) for staging and detection of recurrence in primary CC. MATERIAL AND METHODS: PET/CT for preoperative staging was performed in 66 prospectively included patients with primary CC. Diagnostic accuracy for PET/CT and CT was analyzed. In addition to routine follow up, 42 stages I-III CC patients had postoperative PET/CT examinations every 6 months for 2 years. Serological levels of tissue inhibitor of metalloproteinase-1 (TIMP-1), carcinoembryonic antigen, and liberated domain I of urokinase plasminogen activator receptor were analyzed. RESULTS: Accuracy for tumor, nodal, and metastases staging by PET/CT were 82% (95% confidence interval [CI]: 70; 91), 66% (CI: 51; 78), and 89% (CI: 79; 96); for CT the accuracy was 77% (CI: 64; 87), 60% (CI: 46; 73), and 69% (CI: 57; 80). Cumulative relapse incidences for stages I-III CC at 6, 12, 18, and 24 months were 7.1% (CI: 0; 15); 14.3% (CI: 4; 25); 19% (CI: 7; 31), and 21.4% (CI: 9; 34). PET/CT diagnosed all relapses detected during the first 2 years. High preoperative TIMP-1 levels were associated with significant hazards toward risk of recurrence and shorter overall survival. CONCLUSIONS: This study indicates PET/CT as a valuable tool for staging and follow up in CC. TIMP-1 provided prognostic information potentially useful in selection of patients for intensive follow up.

Airway management in a bronchoscopic simulator based setting: an observational study.

BACKGROUND: Several simulation-based possibilities for training flexible optical intubation have been developed, ranging from non-anatomical phantoms to high-fidelity virtual reality simulators. These teaching devices might also be used to assess the competence of trainees before allowing them to practice on patients. OBJECTIVES: To evaluate the validity of airway simulation as an assessment tool for the acquisition of the preclinical basic skills in flexible optical intubation and to investigate anaesthetists' opinion on airway simulation. DESIGN: Observational study. SETTING: International airway course. PARTICIPANTS: Thirty-six consultants and residents in anaesthesiology. MAIN OUTCOME MEASURES: All participants performed one single procedure on each of the three different simulators. Their video-filmed performances were assessed by two independent, blinded experts and their opinions of simulation were surveyed. RESULTS: The mean score increased 0.33 points after each attempt (P = 0.021). The attitude towards simulation-based training was always more than 4 on a scale from 1 to 5. Only 25% of the procedures were performed to satisfaction with a learning-by-testing effect (P = 0.021). Generalisability coefficient was 0.55, and there was no correlation between the number of clinical procedures performed beforehand and test scores (P = 0.93). CONCLUSION: The increase in mean score is a learning effect indicating that simulator training allows for entry of the learning curve at a higher level. The anaesthetists in our study agreed completely that simulation-based training was useful regardless of the fidelity of the simulator. Local, practical issues such as cost and portability should decide available simulation modalities in each teaching hospital.

Regulation of APC and AXIN2 expression by intestinal tumor suppressor CDX2 in colon cancer cells.

Wnt signaling is often constitutively active in colorectal cancer cells. The expression of the intestinal specific transcription factor CDX2 is found to be transiently decreased in invasive cells at the tumor/stroma interface. A recent ChIP-Seq study has indicated that several Wnt signaling-related genes are regulated by CDX2. The aim was to investigate the role of decreased CDX2 level on the expression of APC, AXIN2 and GSK3ß in migrating colon cancer cells at the invasive front. CDX2-bound promoter and enhancer regions from APC, AXIN2 and GSK3ß were analyzed for gene regulatory activity and the expression pattern of APC and GSK3ß at the invasive front was evaluated by immunohistochemical procedures. Transfection of intestinal and non-intestinal cell lines demonstrated that CDX2 activated APC and AXIN2 promoter activities via intestinal cell-specific enhancer elements. Suppressed CDX2 expression was associated with endogenous downregulation of APC and AXIN2 expression in Caco-2 cells but did not affect GSK3ß expression. Furthermore, elevated levels of nuclear ß-catenin and reduced levels of cytoplasmic APC were correlated to a low CDX2 expression in migrating colon cancer cells in vivo. These results suggest that a low CDX2 level has influence on the Wnt signaling in invasive colon cancer cells possibly promoting cellular migration.

Induction of Anti-Tumor Immune Responses by Peptide Receptor Radionuclide Therapy with (177)Lu-DOTATATE in a Murine Model of a Human Neuroendocrine Tumor.

Peptide receptor radionuclide therapy (PRRT) is a relatively new mode of internally targeted radiotherapy currently in clinical trials. In PRRT, ionizing radioisotopes conjugated to somatostatin analogues are targeted to neuroendocrine tumors (NETs) via somatostatin receptors. Despite promising clinical results, very little is known about the mechanism of tumor control. By using NCI-H727 cells in an in vivo murine xenograft model of human NETs, we showed that (177)Lu-DOTATATE PRRT led to increased infiltration of CD86+ antigen presenting cells into tumor tissue. We also found that following treatment with PRRT, there was significantly increased tumor infiltration by CD49b+/FasL+ NK cells potentially capable of tumor killing. Further investigation into the immunomodulatory effects of PRRT will be essential in improving treatment efficacy.

Qualities of sessile serrated adenoma/polyp/lesion and its borderline variant in the context of synchronous colorectal carcinoma.

AIMS: Although much data have accumulated on sessile serrated adenoma/polyp/lesion (SSA/P/L) in general, its characteristics in specified contexts are less well elucidated. This lack of knowledge is even more conspicuous concerning its borderline counterpart, referred to as BSSA/P/L. The previous histological observations of the authors on SSA/P/L and BSSA/P/L in general are here extended to encompass attributes of these polyps in the context of synchronous colorectal carcinoma (SCRC), with a focus on the place of BSSA/P/L in the spectrum of non-dysplastic serrated polyps. METHODS: 219 SSA/P/Ls, 206 BSSA/P/Ls and 170 hyperplastic polyps (HPs) were examined for SCRC. Demographics, polyp details (size, site, BRAF((V600E))) and advanced synchronous conventional adenomas were recorded. RESULTS: SCRC was present in 12.3% of SSA/P/Ls, 7.1% of HPs (p=0.09) and 8.3% of BSSA/P/Ls. Patients' ages were comparable. Gender distribution of SSA/P/L and BSSA/P/L was equal, which differed, albeit insignificantly, from a male predominance of HPs. More SSA/P/Ls and BSSA/P/Ls than HPs exceeded 4 mm (p≤0.0001). A proximal site characterised SSA/P/L compared with BBSA/P/L and HP (p<0.0001). BRAF mutation was more prevalent in SSA/P/Ls and BSSA/P/Ls, which further coexisted with advanced synchronous conventional adenomas less commonly than HPs. CONCLUSIONS: BSSA/P/L was like SSA/P/L in most respects. The lower SCRC prevalence of BSSA/P/L could fit the idea of BSSA/P/L as a precursor to SSA/P/L, a notion that deserves attention when formulating guidelines for CRC screening.

The JAK2V617F allele burden and STAT3- and STAT5 phosphorylation in myeloproliferative neoplasms: early prefibrotic myelofibrosis compared with essential thrombocythemia, polycythemia vera and myelofibrosis.

Early prefibrotic myelofibrosis (early PMF) is a diagnosis that clinically and histologically mimic essential thrombocythemia (ET), but is important to distinguish from ET, polycythemia vera (PV) and primary myelofibrosis (PMF) due to its different prognosis and clinical evolution. In this study, we assessed the allele burden of JAK2V617F in bone marrow biopsies from patients with these chronic myeloproliferative neoplasms. We correlated our findings with the amount of phosphorylated STAT3 (P-STAT3) and STAT5 (P-STAT5) in megakaryocyte nuclei in the bone marrow. The JAK2V617F allele burden was significantly higher in patients with PV (median: 50.99, range: 23.08-97.29, p < 0.01 and p < 0.01) and PMF (median: 44.13, range: 33.61-92.17, p < 0.05 and p < 0.01) compared with a low allele burden in ET (median: 23.465, range: 8.67-47.92) and early PMF (median: 25.68, range: 0.61-49.13) respectively. In addition, we found a significantly higher phosphorylation of STAT5 and STAT3 in the JAK2V617F positive group than in the negative group. There was no positive correlation between increasing JAK2V617F allele burden and the amount of P-STAT3 and P-STAT5. However, we found low values of P-STAT5 in bone marrow biopsies from patients with ETJAK2V617F+ as compared with patients with early PMFJAK2V617F+. Although this difference was statistically significant, larger studies are needed to firmly support this conclusion.

Na,K-ATPase activity in mouse muscle is regulated by AMPK and PGC-1α.

Na,K-ATPase activity, which is crucial for skeletal muscle function, undergoes acute and long-term regulation in response to muscle activity. The aim of the present study was to test the hypothesis that AMP kinase (AMPK) and the transcriptional coactivator PGC-1α are underlying factors in long-term regulation of Na,K-ATPase isoform (α,ß and PLM) abundance and Na(+) affinity. Repeated treatment of mice with the AMPK activator AICAR decreased total PLM protein content but increased PLM phosphorylation, whereas the number of α- and ß-subunits remained unchanged. The K(m) for Na(+) stimulation of Na,K-ATPase was reduced (higher affinity) after AICAR treatment. PLM abundance was increased in AMPK kinase-dead mice compared with control mice, but PLM phosphorylation and Na,K-ATPase Na(+) affinity remained unchanged. Na,K-ATPase activity and subunit distribution were also measured in mice with different degrees of PGC-1α expression. Protein abundances of α1 and α2 were reduced in PGC-1α +/- and -/- mice, and the ß(1)/ß(2) ratio was increased with PGC-1α overexpression (TG mice). PLM protein abundance was decreased in TG mice, but phosphorylation status was unchanged. Na,K-ATPase V (max) was decreased in PCG-1α TG and KO mice. Experimentally in vitro induced phosphorylation of PLM increased Na,K-ATPase Na(+) affinity, confirming that PLM phosphorylation is important for Na,K-ATPase function. In conclusion, both AMPK and PGC-1α regulate PLM abundance, AMPK regulates PLM phosphorylation and PGC-1α expression influences Na,K-ATPase α(1) and α(2) content and ß(1)/ß(2) isoform ratio. Phosphorylation of the Na,K-ATPase subunit PLM is an important regulatory mechanism.