transcription reaction was first carried out with 11 mL of mixture containing 2 mL of RNA extract, 2 mL of RT primer (Ribo, China), and 7 mL of nuclease-free water. The 11-mL mixture was incubated at 70uC for 10 min and in ice for 2 min. Next, 5 mL of RT buffer, 2 mL of dNTP (2.5 mM), 0.5 mL of RNase inhibitor (40 U/ml), 0.5 mL of reverse transcriptase (200 U/mL), and 6 mL of nuclease-free water were added to the 11-mL mixture. The reverse transcription reaction continued at 42uC for 60 min, 70uC for 10 min, and 4uC for ‘. cDNA solution (3 mL) was amplified using 9 mL of SYBR Premix Ex Taq (TaKaRa, China), 2 mL of miRNA forward primer, 2 mL of miRNA backward primer, and 4 mL of nuclease-free water in a final volume of 20 mL. Quantitative PCR was run on a Biorad CFX96 2.1(Biorad Biosystems), and the reaction mixtures were incubated at 95 uC for 2 min, followed by 50 cycles of 95 uC for 5 s and 60uC for 10 s. At the end of the PCR cycles, melting curve analysis was performed to validate generation of the expected PCR product. The setting of melting curve was 65.0 to 95.0 uC at increments of 0.5uC for 0.05 min + plate read. Each sample was analyzed in triplicate. All Ct values were ,36. The expression levels of each target miRNA were normalized to that of miR-16. miR-16 is the most widely used internal control microRNA in body-fluid samples, including plasma [10,19]. And most importantly, in the discovery phase and validation phase, our study demonstrated that miR-16 could stably express in saliva and work as an internal control for salivary miRNAs. The raw data in the discovery phase can be downloaded from the website of GEO: www.ncbi.nlm.nih.gov/geo/info/ linking.html, and the accession number is GSE41268. The methodoloy, the raw data and pictures of the validation of miR- 16 as an internal control in our research were presented in the Table 2. Characteristics of EC patients and healthy controls.
Previous studies have shown substantial inconsistencies although these were based on limit- ed sample sizes or used candidate miRNA approaches. Additionally, a level of controversy ex- ists in that miRNAs identified in the blood merely indicate changes in blood cells secondary to an overall poor health condition and are not specific to the presence ofcancer in a target organ. Although the measurement of miRNA expression in plasma or serum has been postulated as a promising approach in diagnosing lung cancer, the concept requires further investigations to demonstrate its potential use as a clinical application. There is no strong evidence as to whether serum or plasma is superior for miRNA evaluation, however, recent reports suggest that plas- ma may be the preferred sample choice given that RNA released during the coagulation process may alter the composition of circulating miRNAs in serum samples .
Although circulating miRNAs have a promising potential as relevant novel non-invasive cancerbiomarkers in future as shown in the current study, several limitations need to be addressed. First, methodologies for an accurate absolute quantification of miRNAs suffer from a lack of convention, which limits the cross-comparison between studies performed by different laboratories. Standardized protocol, which should be preferably followed across all studies, needs to be established aiming to minimize protocol-based bias. In addition, some researchers have showed correlations of grade and stage of cancers with specific circulating miRNAs. Therefore, further studies addressing the relationships between miRNAs expression and clinical/pathological parameters are very impor- tant and desirable. Third, most included studies in this meta- analysis only distinguished the cancer patients from healthy controls. It is vital to identify and develop panels of miRNAs that
Prof Alberto Briganti’s presentation began with the case of a 57-year-old male diagnosed with a 4 + 3 bilateral extended prostate cancer. The patient displayed some CV risk factors, including diabetes and obesity. Following staging, the patient appeared to have no systemic disease in the bone or in the abdomen and pelvis; however, a prostate magnetic resonance imaging (MRI) scan showed a suspicious area of minimal extracapsular extension at the right apex. Prostate-speciic antigen (PSA) levels were 21.6 ng/mL, indicating that he was a high-risk patient. He consequently underwent bilateral extended pelvic lymph node dissection (PLND). The inal pathological report revealed that the patient had a Gleason score of 8, 2/21 positive lymph nodes, and a positive surgical margin, with complete recovery of urinary continence at 4 weeks after surgery. Post-surgery evaluation showed that the patient had a PSA of 0.07 ng/mL, had no spontaneous erections, and did not require a protective pad at 40 days.
Despite the clinical development of anti-PD-L1 therapies, the prognostic value of PD-L1 overexpression across different solid tumors is still unclear. Recently, it is reported that mela- noma patients with PD-L1-expressing cells at the invasive tumor margin and inside tumors are more sensitive to anti-PD-1 therapy . Another two studies showed that across multiple cancer types, responses were observed in patients with tumors expressing high levels of PD-L1, especially when PD-L1 expressed on tumor-infiltrating immune cells [18, 19]. The evidence suggests along with development of PD-L1/PD-1 targeted therapy, some biomarkers are needed for guiding individualized anti-PD-1 therapy option. It would be desirable to explore whether PD-L1 overexpression is associated with worse outcome. Moreover, PD-L1 overex- pression may serve as a potential biomarker for prognostic prediction and PD-L1/PD-1 tar- geted treatment option in solid tumors.
The presence of significant hypoxia in tissues can result in elevated blood levels of lactate. Lactate is produced when tissues deprived of oxygen switch from oxidative phosphorylation to anaerobic metabolism. Thus, elevated lactate in umbilical cord blood reflects significant fetal hypoxia, and indeed, higher levels have been shown epidemiologically to be associated with neonatal encephalopathy. We chose a cord lactate .6 mmol/L to represent a hypoxic fetus in our cohort, as this has been suggested as the cut- off to predict which fetuses will develop moderate-severe HIE . In our study, no single miR was able to discriminate between those fetuses with a low lactate (,4 mmol/L) or high lactate (.6 mmol/ L). However, there was a 3 fold increase in the combined expression of miR 21 and miR 20b in those fetuses with a high lactate (median expression low lactate 2.45 vs high lactate 5.94, p ,0.05). When the expression of miR21 and miR 20b were combined, there was a significant correlation (Spearman’s rank) with umbilical cord lactate levels at delivery (r = 0.79, p = 0.03). (Fig. 2).
ABSTRACT. Secretory cells of the cephalic salivary glands (CSGs) of eusocial bees produce and accumulate lipid-like secretion in the lumens of their alveoli. Correspondingly, secretory cells present typical ultrastructural features of lipid-compound producers. Previous work on bees has revealed inter-specific differences in the chemical composition of secretion, and the production mechanisms and secretory cycle of secretory cells. In this work a comparative analysis of the mechanisms of lipid storage in the CSGs of Apis mellifera (Linnaeus, 1758) and Scaptotrigona postica (Latreille, 1807) workers was carried out. The ultrastructural location of lipids was ascertained using imidazole-osmium (IO), using individuals in different stages of their life cycles. Lipid deposits were identified inside glandular cells and in the alveolar lumens in all individuals, but differences were observed between the species. The glandular cells of A. mellifera workers presented positive reactions to IO as droplets dispersed in the cytoplasm, as vesicles and in the channels formed by apical plasma membrane infolds. In S. postica, lipid compounds were detected inside the mitochondrial matrix and in smooth endoplasmic reticulum cisterns. In both species, forager workers exhibited the largest amounts of lipids stored in the alveolar lumen. The differences between the species are discussed, taking into account specific behavioral differences.
This protocol was approved by the Ethical Committee (Comité de Protection des Personnes Sud-Ouest et Outre Mer N°1, number 1-10-21). To avoid blood contamination, patients were asked not to brush their teeth within 45 minutes prior to sample collection. Saliva was collected using sterile tips and micropipettes during endoscopic examination under general anesthesia with propofol. Saliva was immediately placed in pre-chilled 1.5-ml microcentrifuge tubes con- taining and equal volume of Saliva protect reagent (Qiagen) and stored at -80°C until ready for use. In this pilot study, we included patients aged >18 years who had given their written informed consent. Other criteria for inclusion were no contraindications for general anesthesia or for endoscopic ultrasound. Fine needle aspiration material was used for histological, cytolog- ical and molecular (KRAS activating mutation analysis) diagnosis of pancreatitis or pan- creatic cancer. Twenty-one patients were included in this study; 7 were diagnosed with locally advanced, unresectable pancreatic cancer, 4 were diagnosed with pancreatitis (either acute or chronic) and 4 had unrelated digestive diseases (control group) (Table 1). Patients diagnosed with intraductal papillary mucinous neoplasia (IPMN) (n = 2) were also included. Patients were not treated before saliva collection.
In the United States, approximately 40,000 people die every year from pancreatic adenocarcinoma, making it the fourth leading cause ofcancer related death. Patients diagnosed in the early stage of pancreatic cancer have a 5-year survival rate of 24%, compared to 1.8% when diagnosed in the advanced stage (Li et al., 2004). Clinical manifestations of pancreatic cancer do not appear until after the cancer has undergone metastasis (Holly et al., 2004), emphasizing the need for early detectionbiomarkers. The etiology of pancreatic cancer remains elusive, with cigarette smoking being the most established risk factor (Vrieling et al., 2010; Nakamura et al., 2011; Fuchs, Colditz & Stampfer, 1996; Zheng et al., 1993), although links have also been made to diabetes (Haugvik et al., 2015; Liu et al., 2015), obesity (Bracci, 2012), and chronic pancreatitis (Malka et al., 2002). Recent research has also shown that men with periodontal disease have a two-fold greater risk of developing pancreatic cancer after adjusting for smoking, diabetes, and body mass index (Michaud et al., 2007).
A sensitive assay to identify volatile organic metabolites (VOMs) as biomarkers that can accurately diagnose the onset of breast cancer using non-invasively collected clinical specimens is ideal for early detection. Therefore the aim of this study was to establish the urinary metabolomic proﬁle of breast cancer patients and healthy individuals (control group) and to explore the VOMs as potential biomarkers in breast cancer diagnosis at early stage. Solid-phase microextraction (SPME) using CAR/PDMS sorbent combined with gas chromatography–mass spectrometry was applied to obtain metabolomic informa- tion patterns of 26 breast cancer patients and 21 healthy individuals (controls). A total of seventy-nine VOMs, belonging to distinct chemical classes, were detected and identiﬁed in control and breast cancer groups. Ketones and sulfur compounds were the chemical classes with highest contribution for both groups. Results showed that excretion values of 6 VOMs among the total of 79 detected were found to be statistically different (p < 0.05). A signiﬁcant increase in the peak area of (−)-4-carene, 3-heptanone, 1,2,4- trimethylbenzene, 2-methoxythiophene and phenol, in VOMs ofcancer patients relatively to controls was observed. Statiscally signiﬁcant lower abundances of dimethyl disulﬁde were found in cancer patients. Bioanalytical data were submitted to multivariate statistics [principal component analysis (PCA)], in order to visualize clusters of cases and to detect the VOMs that are able to differentiate cancer patients from healthy individuals. Very good discrimination within breast cancer and control groups was achieved. Nevertheless, a deep study using a larger number of patients must be carried out to conﬁrm the results.
Davies SM, Robison LL, Buckley JD, Radloff GA, Ross JA, Perentesis JP. Glutathione S-transferase polymorphisms in children with myeloid leukemia: a children´s cancer group study. Cancer Epidemiol Biomarkers Prev. 2000;9:563-6. Depeille P, Cuq P, Mary S, Passagne I, Evrard A, Cupissol D, et al. Glutathione S-transferase M1 and mulidrug resistance protein act in synergy to protect melanoma cells from vincristine effects. Mol Pharmacol. 2004;65:897-905. De Rijke B, Van Horssen-Zoetbrood A, Beekman JM, Otterud B, Maas F, Woestenenk R, et al. A frameshift polymorphism in P2X5 elicits an allogenic cytotoxic T lymphocyte response associated with remission of chronic myeloid leukemia. J Clin Invest. 2005;15(12):3506-16.
In the postoperative the patient showed acute respiratory dysfunction syndrome and was admitted at Intensive Care Unit for a period of ten days. Before he started oral intake, a barium esophagram was performed (Fig. 5a and 5b). After his stay at Intensive Care Unit he recovered very well and was discharged 20 days after surgery.
ABSTRACT: The use ofbiomarkers is an important recent development in veterinary medicine. Biomarkers allow non-invasive quantification of substances with diagnostic and prognostic potential in several diseases. The microRNAs are small, non-coding RNAs that regulate gene expression and are expressed in different forms in many diseases. Reduced or over-expression ofmicroRNAs showed to be part of the pathogenesis of some heart diseases in humans and animals. Diagnostic and therapeutic value of measuring microRNAs in veterinary cardiology is increased because abnormal expression can be managed by the use of antagonists (in the case of overexpression) and mimicking (in the case of underexpression). Thus, this literature review aimed to compile scientific evidence of dysregulation ofmicroRNAs expression in different cardiac diseases being one of the promises in the therapeutic field and diagnosis of veterinary cardiology. MicroRNAs not only have potential as a biomarker but may also help in elucidation of aspects of the pathogenesis of a variety of diseases.
Structural health monitoring of civil infrastructures has great practical importance for engineers, owners and stakeholders. Numerous researches have been carried out using long- term monitoring, such as the Rio–Niterói Bridge in Brazil, the former Z24 Bridge in Switzerland and the Millau Bridge in France. In fact, some structures are continuously monitored to supply dynamic measurements that can be used for the identification of structural problems such as the presence of cracks, excessive vibration or even to perform a quite extensive structural evaluation concerning its reliability and life cycle. The outputs of such an analysis, commonly entitled modal identification, are the so-called modal parameters, that is, natural frequencies, damping rations and mode shapes. Therefore, the development and validation of tools for the automatic modal identification during normal operation is fundamental, as the success of subsequent damage detection algorithms depends on the accuracy of the modal parameters’ estimates. This work proposes a novel methodology to perform, automatically, the modal identification based on the modes’ estimates data generated by any parametric system identification method. To assess the proposed methodology, several tests are conducted using numerically generated signals, as well as experimental data obtained from a simply supported beam and from a motorway bridge.
our PAF was much lower than the estimate from a previous population based case-control study in the US (64.4%)  and the estimate from a previous European population based pro- spective cohort study (25% for cancers of the upper aerodigestive tract) . In China, however, an increase in the prevalence of alcohol drinking among women between 1991(2.6%) and 2002 (4.5%) was observed. Thus it is expected that the contribution of alcohol drinking to the cancer burden will increase in the future. Most previous studies about the cancer burden in global or national studies have evaluated the combined effect of total fruit and vegetable intake [30,31,33]. Our report separately estimates the esophagealcancer burden attributable to low intake of fruit and low intake of vegetables. Fruit and vegetables contain different nutrients and have different nutritional values . For example, the contents of vitamins, minerals, fiber, and phytochemicals are higher in most vegetables, especially in dark vegetables, while carbohydrates and organic acids are higher in fruits. Danaei and his colleagues  calculated that 19% ofesophagealcancer deaths in low and middle-income countries were attributable to low vegetable and fruit intake. Our estimates indicated a little higher proportion ofesophagealcancer deaths were attributable to low vegetable and fruit intake, reflecting the different sources of RRs and exposure rates in the two studies.
complications . Previous radiotherapy was found to increase the risk ofesophageal stent complications [28–30]. To our opinion, these complication rates may be underestimated, as airway examination was not reported. In recent reviews, some authors drew attention to the risk of airway compromise when the neoplasic mass narrows the trachea (figure 3) [30,31]. They advised to consider airway evaluation and tracheo-bronchial stenting in these high risk situations (malignant infiltration in the proximal third part of the esophagus) [22,30,31]. Perforation and fistula are major and life-threatening complications, and should be avoided in these palliative situations where quality of life is the major end point. We think that preventive double stenting may be considered. Global improvement of respiratory symptoms and, good tolerance of airway stents were major points to promote this procedure. Moreover, scheduled stenting procedure allowed to administer planned EGC therapy for 96% patients, compared to the 14% in the emergency group. This approach may be beneficial to EGC patients. By closing fistulas and decreasing airway complications related to esophageal stenting, chronic aspiration, chronic sepsis, and recurrent atelectasis were uncommon in patients with scheduled airway stenting. Chemotherapy and/or radiotherapy were administered with no delay. Covered metallic stents (ultraflexH) were mostly used because of their high expansion properties to counteract the larger and wider esophageal metallic stent. In case of carina involvement, only Y silicone stents (novatechH) were used. However, all recent marketed stents can probably be considered for this indication depending on physician’s experience .
This study was designed to investigate the pathogen- esis of ESCC and EAC by 1) screening existing EAC- and ESCC-related miRNA expression microarray data to identify differentially expressed miRNAs and analyze the correlations between miRNA expression and the risk factors, treatment methods and survival rates of patients; 2) screening the EAC- and ESCC-related gene expres- sion microarray data for differentially expressed genes; 3) predicting the target genes of differentially expressed miRNAs and constructing regulatory networks depending on the differentially expressed target genes. Their effects on biological processes of the target genes were also investigated with pathway and gene ontology (GO) enrichment analysis.
Considering cg01847754- located in the first exon of CXorf1 gene is hypomethylated in stage IV. This is in accordance with survival analysis performed, which showed that low methylation levels are associated with a poor prognosis. Interestingly, one more time, the canonical pathway is not observed, meaning that hypomethylation of cg01847754 is associated with down-regulation of CXorf1 gene. Indeed, this gene may be an interesting candidate biomarker for cancer prognosis although its role in cancer is not clear yet. Then, were identified genes that could predict patient recurrence. CNTD2, SOX1, and HTR2C are some of the genes found as predictors of recurrence free survival in stages II, III, and IV, respectively. Specifically, CNTD2 was found over-expressed in stage II CRC tissue which is in agreement with a poor prognostic associated with high gene expression levels. CNTD2 is a member of the cyclin family which can control the cell cycle. When there are alterations on genes that regulate the cell cycle, the consequence may be uncontrolled cell growth. Recently, this gene was reported as a new oncogenic driver in lung cancer 200 .
Markers of glial damage were also tested in the CSF of MS patients. Glial fibrillary acidic protein (GFAP) is a component of astrocytes filaments. Increased GFAP levels in CSF reflect astrocyte damage. Multiple sclerosis patients have increased CSF GFAP levels when compared to controls. Multiple sclerosis patients with worse ambulation or more severe disability have higher CSF GFAP levels than less disabled patients and controls, suggesting that CSF GFAP may be a biomarker of clinical progression and prognosis. (32-35)