Η συμβολή της συνεχούς καταγραφής της γλυκόζης στην επιλογή της κατάλληλης αντιδιαβητικής αγωγής σε ασθενείς υπό χημειοθεραπεία

Είναι κοινή πρακτική στην ογκολογία η έναρξη αντικαρκινικής θεραπείας (συμπεριλαμβανομένης της χημειοθεραπείας, της στοχευμένης θεραπείας, της ανοσοθεραπείας και στεροειδών) σε άτομα με προϋπάρχοντα διαβήτη για μια σειρά καρκίνων. Αυτές οι κατευθυντήριες γραμμές απαιτούν την αναγνώριση των κινδύνων εμφάνισης υπεργλυκαιμίας και νέας εμφάνισης διαβήτη σε άτομα με καρκίνο από τη διεπιστημονική ομάδα και να παρέχει τα εργαλεία για την κατάλληλη διαχείριση αυτών των ατόμων ώστε να μειωθούν οι κίνδυνοι επιπλοκών. Η υπεργλυκαιμία είναι μια κοινή πρόκληση κατά τη διάρκεια της θεραπείας του καρκίνου. Επιπλέον, πολλοί ασθενείς με προϋπάρχοντα διαβήτη τύπου 1 ή τύπου 2 που υποβάλλονται σε θεραπεία του καρκίνου αναπτύσσουν ιατρογενή υπεργλυκαιμία με μοναδικά χαρακτηριστικά. Το πιο κοινό παράδειγμα είναι η υπεργλυκαιμία που προκαλείται από στεροειδή. Συχνά είναι απαραίτητες ειδικές εκτιμήσεις όσον αφορά τις συστάσεις τρόπου ζωής, τη βέλτιστη επιλογή των φαρμάκων και τους στόχους της θεραπείας. Σε ασθενείς με ενεργό καρκίνο, η εστίαση της διαχείρισης της υπεργλυκαιμίας μετατοπίζεται από την πρόληψη μακροπρόθεσμων επιπλοκών προς την αποφυγή οξέων και υποξέων αποτελεσμάτων, όπως αφυδάτωση από πολυουρία, λοίμωξη, καταβολή, απώλεια βάρους, υπεροσμωτικές μη κετωτικές καταστάσεις με κετωσικό υπεροσμωτικό κώμα (HNK), και διαβητική κετοξέωση (DKA). Θα πρέπει να σημειωθεί ότι καταστάσεις όπως HNK και DKA είναι σπάνιες. Για τους παραπάνω λόγους έχει βρεθεί ένα σύστημα καταγραφής της γλυκόζης που διευκολύνει αρκετά και αποτρέπει σε μεγάλο βαθμό όλα τα προβλήματα που μπορεί να προκύψουν. Λέγεται Freestyle Libre και χρησιμοποιείται υποδόρια, περιλαμβάνει τεχνολογία ανίχνευσης επιπέδων γλυκόζης στο διάμεσο υγρό. Μετριέται αυτόματα η γλυκόζη κάθε λεπτό και οι αναγνώσεις αποθηκεύονται σε διαστήματα 15 λεπτών. Ο αναγνώστης κρατιέται κοντά στον αισθητήρα όταν απαιτείται ανάγνωση γλυκόζης. Στη συνέχεια, η συσκευή θα εμφανίσει τις πληροφορίες γλυκόζης των τελευταίων 8 ωρών, συμπεριλαμβανομένης της τρέχουσας γλυκόζης, ένα γράφημα τάσης, και ένα βέλος τάσης. Το βέλος δείχνει την κατεύθυνση και την ταχύτητα του τρέχοντος επιπέδου γλυκόζης του ασθενούς. Αυτή η τεχνολογία είναι βαθμονομημένη στο εργοστάσιο και οι ασθενείς δεν χρειάζεται να βαθμονομήσουν με ενδείξεις τον μετρητή γλυκόζης δείγματος αίματος. Το Freestyle Libre για προσωπική χρήση προσθέτει μια άλλη επιλογή στους ασθενείς να παρακολουθούν τη γλυκόζη τους. Μπορεί να αντικαταστήσει έναν μετρητή γλυκόζης και να εξαλείψει την ανάγκη για πολλαπλά τρυπήματα του δακτύλου, επιτρέποντας στους ασθενείς να λαμβάνουν πληροφορίες για τη γλυκόζη κατόπιν αιτηματός τους. Αν και μπορεί να μην είναι για όλους, δίνει στους ασθενείς και τους θεράποντες ιατρούς μια καλύτερη επιλογή. Μετά από ανάλυση των μετρήσεων της συσκευής Free style Libre σε ένα δείγμα 11 ασθενών διαπιστώθηκε μείωση στη μέση τιμή της γλυκόζης, στην μεταβλητότητα των τιμών της γλυκόζης καθώς επίσης και μειωμένα επεισόδια υπογλυκαιμίας και πολύ υψηλών τιμών σακχάρου. Συμπερασματικά, διαπιστώνεται ότι η συσκευή συνεχούς καταμέτρησης γλυκόζης free style libre είναι μια πολύ χρήσιμη συσκευή για την καλύτερη αντιμετώπιση του διαβήτη στους ασθενείς υπό χημειοθεραπεία.It is common practice in oncology to start anti-cancer treatment (including chemotherapy, targeted therapy, immunotherapy and steroids) in people with pre-existing diabetes for a number of cancers. These guidelines require the interdisciplinary team to identify the risks of hyperglycemia and recurrence of diabetes in people with cancer and to provide the tools to properly manage these individuals to reduce the risk of complications. Hyperglycemia is a common challenge during cancer treatment. In addition, many patients with pre-existing type 1 or type 2 diabetes who are being treated for cancer develop iatrogenic hyperglycemia with unique characteristics. The most common example is steroid-induced hyperglycemia. Specific assessments of lifestyle recommendations, optimal drug selection, and treatment goals are often necessary. In patients with active cancer, the focus of hyperglycaemia management shifts from the prevention of long-term complications to the avoidance of acute and subacute effects, such as polyuria dehydration, infection,weakness, weight loss, hyperosmotic non-ketotic conditions with ketotic and diabetic ketoacidosis (DKA). It should be noted that conditions such as HNK and DKA are rare. For the above reasons, a glucose recording system has been found that greatly facilitates and largely prevents all problems that may arise. It is called Freestyle Libre and is used subcutaneously, it includes technology for detecting glucose levels in the interstitial fluid. Glucose is automatically measured every minute and readings are stored at 15 minute intervals. The reader is held close to the sensor when a glucose reading is required. The device will then display the glucose information for the last 8 hours, including the current glucose, a voltage graph, and a voltage arrow. The arrow indicates the direction and speed of the patient's current glucose level. This technology is factory calibrated and patients do not need to calibrate the blood sample glucose meter. Freestyle Libre for personal use adds another option for patients to monitor their glucose. It can replace a glucose meter and eliminate the need for multiple finger piercings, allowing patients to receive glucose information upon request. Although it may not be for everyone, it gives patients and treating physicians a better choice. After analyzing the measurements of the Free style Libre device in a sample of 11 patients, a decrease in the mean glucose value, in the variability of the glucose values as well as reduced episodes of hypoglycemia and very high sugar values were found. In conclusion, the free style libre continuous glucose meter is a very useful device for the better treatment of diabetes in patients undergoing chemotherapy

Can gene expression profiling predict survival for patients with squamous cell carcinoma of the lung?

BACKGROUND: Lung cancer remains to be the leading cause of cancer death worldwide. Patients with similar lung cancer may experience quite different clinical outcomes. Reliable molecular prognostic markers are needed to characterize the disparity. In order to identify the genes responsible for the aggressiveness of squamous cell carcinoma of the lung, we applied DNA microarray technology to a case control study. Fifteen patients with surgically treated stage I squamous cell lung cancer were selected. Ten were one-to-one matched on tumour size and grade, age, gender, and smoking status; five died of lung cancer recurrence within 24 months (high-aggressive group), and five survived more than 54 months after surgery (low-aggressive group). Five additional tissues were included as test samples. Unsupervised and supervised approaches were used to explore the relationship among samples and identify differentially expressed genes. We also evaluated the gene markers' accuracy in segregating samples to their respective group. Functional gene networks for the significant genes were retrieved, and their association with survival was tested. RESULTS: Unsupervised clustering did not group tumours based on survival experience. At p < 0.05, 294 and 246 differentially expressed genes for matched and unmatched analysis respectively were identified between the low and high aggressive groups. Linear discriminant analysis was performed on all samples using the 27 top unique genes, and the results showed an overall accuracy rate of 80%. Tests on the association of 24 gene networks with study outcome showed that 7 were highly correlated with the survival time of the lung cancer patients. CONCLUSION: The overall gene expression pattern between the high and low aggressive squamous cell carcinomas of the lung did not differ significantly with the control of confounding factors. A small subset of genes or genes in specific pathways may be responsible for the aggressive nature of a tumour and could potentially serve as panels of prognostic markers for stage I squamous cell lung cancer

Optimization of laser capture microdissection and RNA amplification for gene expression profiling of prostate cancer

BACKGROUND: To discover prostate cancer biomarkers, we profiled gene expression in benign and malignant cells laser capture microdissected (LCM) from prostate tissues and metastatic prostatic adenocarcinomas. Here we present methods developed, optimized, and validated to obtain high quality gene expression data. RESULTS: RNase inhibitor was included in solutions used to stain frozen tissue sections for LCM, which improved RNA quality significantly. Quantitative PCR assays, requiring minimal amounts of LCM RNA, were developed to determine RNA quality and concentration. SuperScript II™ reverse transcriptase was replaced with SuperScript III™, and SpeedVac concentration was eliminated to optimize linear amplification. The GeneChip(® )IVT labeling kit was used rather than the Enzo BioArray™ HighYield™ RNA transcript labeling kit since side-by-side comparisons indicated high-end signal saturation with the latter. We obtained 72 μg of labeled complementary RNA on average after linear amplification of about 2 ng of total RNA. CONCLUSION: Unsupervised clustering placed 5/5 normal and 2/2 benign prostatic hyperplasia cases in one group, 5/7 Gleason pattern 3 cases in another group, and the remaining 2/7 pattern 3 cases in a third group with 8/8 Gleason pattern 5 cases and 3/3 metastatic prostatic adenocarcinomas. Differential expression of alpha-methylacyl coenzyme A racemase (AMACR) and hepsin was confirmed using quantitative PCR

Colorectal Cancer with Residual Polyp of Origin: A Model of Malignant Transformation

AbstractThe majority of colorectal cancers (CRCs) arise from adenomatous polyps. In this study, we sought to present the underrecognized CRC with the residual polyp of origin (CRC RPO+) as an entity to be utilized as a model to study colorectal carcinogenesis. We identified all subjects with biopsy-proven CRC RPO+ that were evaluated over 10 years at Mayo Clinic, Rochester, MN, and compared their clinical and pathologic characteristics to CRC without remnant polyps (CRC RPO−). Overall survival and disease-free survival overlap with an equivalent hazard ratio between CRC RPO+ and RPO− cases when age, stage, and grade are adjusted. The somatic genomic profile obtained by whole genome sequencing and the gene expression profiles by RNA-seq for CRC RPO+ tumors were compared with that of age -and gender-matched CRC RPO− evaluated by The Cancer Genome Atlas. CRC RPO+ cases were more commonly found with lower-grade, earlier-stage disease than CRC RPO−. However, within the same disease stage and grade, their clinical course is very similar to that of CRC RPO−. The mutation frequencies of commonly mutated genes in CRC are similar between CRC RPO+ and RPO− cases. Likewise, gene expression patterns are indistinguishable between the RPO+ and RPO− cases. We have confirmed that CRC RPO+ is clinically and biologically similar to CRC RPO− and may be utilized as a model of the adenoma to carcinoma transition

Impact of sample acquisition and linear amplification on gene expression profiling of lung adenocarcinoma: laser capture micro-dissection cell-sampling versus bulk tissue-sampling

<p>Abstract</p> <p>Background</p> <p>The methods used for sample selection and processing can have a strong influence on the expression values obtained through microarray profiling. Laser capture microdissection (LCM) provides higher specificity in the selection of target cells compared to traditional bulk tissue selection methods, but at an increased processing cost. The benefit gained from the higher tissue specificity realized through LCM sampling is evaluated in this study through a comparison of microarray expression profiles obtained from same-samples using bulk and LCM processing.</p> <p>Methods</p> <p>Expression data from ten lung adenocarcinoma samples and six adjacent normal samples were acquired using LCM and bulk sampling methods. Expression values were evaluated for correlation between sample processing methods, as well as for bias introduced by the additional linear amplification required for LCM sample profiling.</p> <p>Results</p> <p>The direct comparison of expression values obtained from the bulk and LCM sampled datasets reveals a large number of probesets with significantly varied expression. Many of these variations were shown to be related to bias arising from the process of linear amplification, which is required for LCM sample preparation. A comparison of differentially expressed genes (cancer vs. normal) selected in the bulk and LCM datasets also showed substantial differences. There were more than twice as many down-regulated probesets identified in the LCM data than identified in the bulk data. Controlling for the previously identified amplification bias did not have a substantial impact on the differences identified in the differentially expressed probesets found in the bulk and LCM samples.</p> <p>Conclusion</p> <p>LCM-coupled microarray expression profiling was shown to uniquely identify a large number of differentially expressed probesets not otherwise found using bulk tissue sampling. The information gain realized from the LCM sampling was limited to differential analysis, as the absolute expression values obtained for some probesets using this study's protocol were biased during the second round of amplification. Consequently, LCM may enable investigators to obtain additional information in microarray studies not easily found using bulk tissue samples, but it is of critical importance that potential amplification biases are controlled for.</p

Detection and quantification of methylation in DNA using solid-state nanopores.

Epigenetic modifications in eukaryotic genomes occur primarily in the form of 5-methylcytosine (5 mC). These modifications are heavily involved in transcriptional repression, gene regulation, development and the progression of diseases including cancer. We report a new single-molecule assay for the detection of DNA methylation using solid-state nanopores. Methylation is detected by selectively labeling methylation sites with MBD1 (MBD-1x) proteins, the complex inducing a 3 fold increase in ionic blockage current relative to unmethylated DNA. Furthermore, the discrimination of methylated and unmethylated DNA is demonstrated in the presence of only a single bound protein, thereby giving a resolution of a single methylated CpG dinucleotide. The extent of methylation of a target molecule could also be coarsely quantified using this novel approach. This nanopore-based methylation sensitive assay circumvents the need for bisulfite conversion, fluorescent labeling, and PCR and could therefore prove very useful in studying the role of epigenetics in human disease