Development and evaluation of a novel fast broad-range PCR and sequencing assay (FBR-PCR/S) using dual priming oligonucleotides targeting the ITS/LSU gene regions for rapid diagnosis of invasive fungal diseases: multi-year experience in a large Canadian healthcare zone.

BACKGROUND: This study evaluated the performance of a novel fast broad range PCR and sequencing (FBR-PCR/S) assay for the improved diagnosis of invasive fungal disease (IFD) in high-risk patients in a large Canadian healthcare region. METHODS: A total of 114 clinical specimens (CS) including bronchoalveolar lavages (BALs) were prospectively tested from 107 patients over a 2-year period. Contrived BALs (n = 33) inoculated with known fungi pathogens were also tested to increase diversity. Patient characteristics, fungal stain and culture results were collected from the laboratory information system. Dual-priming oligonucleotide (DPO) primers targeted to the internal transcribed spacer (ITS) (~ 350 bp) and large subunit (LSU) (~ 550 bp) gene regions were used to perform FBR-PCR/S assays on extracted BALs/CS. The performance of the molecular test was evaluated against standard microbiological methods and clinical review for the presence of IFD. RESULTS: The 107 patients were predominantly male (67, 62.6%) with a mean age of 59 years (range = 0-85 years): 74 (69.2%) patients had at least one underlying comorbidity: 19 (34.5%) had confirmed and 12 (21.8%) had probable IFD. Culture recovered 66 fungal isolates from 55 BALs/CS with Candida spp. and Aspergillus spp. being most common. For BALs, the molecular assay vs. standard methods had sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV), and efficiency of 88.5% vs.100%, 100% vs. 61.1%, 100% vs. 88.5%, 61.1% vs. 100%, and 90.2% for both. For other CS, the molecular assay had similar performance to standard methods with sensitivity, specificity, PPV, NPV and efficiency of 66.7%, 87.0%, 66.7%, 87.0% and 81.3% for both methods. Both methods also performed similarly, regardless of whether CS stain/microscopy showed yeast/fungal elements. FBR-PCR/S assays results were reported in ~ 8 h compared to fungal cultures that took between 4 and 6 weeks. CONCLUSIONS: Rapid molecular testing compared to standard methods have equivalent diagnostic efficiency but improves clinical utility by reporting a rapid species-level identification the same dayshift (~ 8 h).

[Partially French Canadians are susceptible to increased cardiovascular risk factors: A population-based retrospective cohort study]. / Les Canadiens partiellement Français sont susceptibles à une augmentation des facteurs de risque cardiovasculaire : une étude de cohorte rétrospective basée sur la population.

BACKGROUND: Through various research lead in the past, it has been made evident that Quebec is home to higher rates of acute myocardial infarction (AMI) and higher prevalence of cardiovascular risk factors than other Canadian provinces. This proposed study will perform a retrospective analysis on Caucasian populations in order to analyze the cardiovascular risk factors in partially francophone populations in comparison to French and Non-French Canadians. Furthermore, we will closely analyze both genders of aforementioned populations. METHODS: This population-based retrospective cohort study was achieved using the University of Ottawa Heart Institute CCTA registry. Included are Caucasian patients of all ages who came to UOHI for a CCTA between 2006 and 2018 and provided written informed consent. SPSS was used to compare the different populations (French Canadian, partially French Canadian and non-French Canadian) and sex. RESULTS: The PFC population more closely resembles FC, having higher incidence of cardiovascular risk factors such as smoking, dyslipidemia and type 2 diabetes. INTERPRETATION: Our results suggest that PFC, like FC, may benefit from more intensive education and lifestyle modification techniques.

Pharmacological Actions of Glucagon-Like Peptide-1, Gastric Inhibitory Polypeptide, and Glucagon.

Glucagon family of peptide hormones is a group of structurally related brain-gut peptides that exert their pleiotropic actions through interactions with unique members of class B1 G protein-coupled receptors (GPCRs). They are key regulators of hormonal homeostasis and are important drug targets for metabolic disorders such as type-2 diabetes mellitus (T2DM), obesity, and dysregulations of the nervous systems such as migraine, anxiety, depression, neurodegeneration, psychiatric disorders, and cardiovascular diseases. The current review aims to provide a detailed overview of the current understanding of the pharmacological actions and therapeutic advances of three members within this family including glucagon-like peptide-1 (GLP-1), gastric inhibitory polypeptide (GIP), and glucagon.

Conversion of atrial fibrillation after levothyroxine in a dog with hypothyroidism and arterial thromboembolism.

A six-year-old female spayed mixed-breed dog was referred following a 3-week history of lameness and progressive neurological deficits in both hindlimbs, and a 1-week history of a cardiac arrhythmia. The dog was diagnosed with hypothyroidism, atrial fibrillation, myocardial dysfunction and arterial thromboembolism. Cardioversion occurred after 2 weeks of levothyroxine supplementation, with improved systolic function over time. To the authors' knowledge, this is the first case reported in the veterinary or human literature documenting hypothyroidism with persistent atrial fibrillation being converted to sinus rhythm with levothyroxine as sole therapy.

Expressed Breast Milk as a Predictor of Neonatal Yeast Colonization in an Intensive Care Setting.

BACKGROUND: Yeast colonization is a predictor for invasive infection in neonates. Candida albicans and Candida parapsilosis are leading causes of invasive fungal infection (IFI) in this population. This study examines maternal breast milk as a predictor of colonization of infants with yeast. METHODS: Inclusion criteria were admission longer than 72 hours to the neonatal intensive care unit and parental consent. Cultures of expressed breast milk, when available, and swabs from oral, rectal, and inguinal sites were obtained weekly for 12 weeks, or until discharge, transfer, or death. Cultures were analyzed using standard laboratory methods. Clinical information was extracted from medical records. RESULTS: One hundred thirty infants were enrolled from February 2011 to November 2012. Cultures were obtained in 129 patients. The median (interquartile range [IQR]) gestational age was 34.4 weeks (33.1-37.1 weeks). The median (IQR) birth weight was 2157.5 g (1740-3060 g). No infants developed IFIs. Twenty-nine (22%) infants were colonized with yeast. Potential correlates for colonization in univariate analysis included exposure to antenatal steroids, postnatal antibiotics, and receipt of breast milk containing yeast. Potential correlates that remained after multivariable logistic regression included exposure to antenatal steroids and receipt of breast milk containing yeast. In cases in which yeast was recovered from an individual infant and from the breast milk received by that infant, there was only 30% concordance between yeast species. DISCUSSION: Recovery of yeast from breast milk is associated with colonization with yeast in the neonate. Because Candida transmission via breast milk had a 30% concordance, breast milk is only one of several ways colonization occurs. Further study is needed on mechanisms of colonization.

Role of secretin peptide family and their receptors in the hypothalamic control of energy homeostasis.

Secretin family of peptide hormones is a group of structurally related brain-gut peptides that exert their functions via interactions with the class B1 G protein-coupled receptors (GPCRs). Recent researches of these peptides and receptors in metabolism have been an area of intense focus for the development of promising drug targets as therapeutic potentials for metabolic disorders. The fact that agonists of GLP-1, a member in the family, have already started being used as therapeutics clearly indicates the importance and relevance of further research on the clinical applications of these peptides. This review aims to provide an overview of the current understanding regarding the importance of this family of peptides as well as their receptors in metabolism with special focus on their actions in the hypothalamus.

Prognostication of cardiac CT angiography.

The rapidly emerging technique of cardiac computed tomography angiography (CTA) has enabled the anatomical assessment of coronary artery disease. CTA has very good diagnostic accuracy with the ability to detect nonobstructive from obstructive coronary artery disease and provides information on the presence of coronary artery calcification as well as on left ventricular function. Over the last few years, many prognostic studies have reviewed the outcome benefit of different scoring indices in predicting hard cardiac events. The following article will review the most recent literature available on the use of CTA in measuring luminal stenoses, identifying high-risk obstructive CAD, calcium plaque score, and LV function all in different models with their impact on the estimation of clinical risk. More recent data from a large multicenter registry supports the incremental benefit of CAD severity and LVEF as independent predictors of prognosis. Future directions and emerging applications such as the utility of CTA combined with perfusion analysis may lead to a new anatomical-functional diagnostic test that may provide optimal noninvasive assessment of coronary artery anatomy and be superior to invasive coronary angiography.

Molecular cloning and mRNA distribution of pituitary adenylate cyclase-activating polypeptide (PACAP)/PACAP-related peptide in the lungfish.

In this article, we report the isolation of a full-length cDNA clone encoding pituitary adenylate cyclase-activating polypeptide (PACAP)/PACAP-related peptide (PRP) from lungfish Protopterus dolloi. When comparing the deduced amino acid sequences, the lungfish PACAP was found to be highly conserved with other vertebrates; however, the PRP shares only lower levels of sequence identity with known PRP sequences. Consistently in phylogenetic analysis, the lungfish PRP, similar to sturgeon PRP, fails to cluster with other PRPs. In addition to the full-length clone, another cDNA encoding a short precursor that lacks the first 32 amino acids of the PRP was also isolated. Interestingly, similar isoforms were also identified in several nonmammalian vertebrates, and it was suggested that exon skipping of PRP/PACAP transcripts was a mechanism that regulated the expression ratio of PACAP to PRP in nonmammalian vertebrates. By real-time PCR, both long and short PRP/PACAP transcripts were found almost exclusively in the brain, and the short isoform is the more abundant transcript (3.7 times more), indicating that PACAP is the major product produced in lungfish brain. The expression patterns of lungfish and previously studied frog PRP/PACAP suggest that the PRP/PACAP gene in the tetrapod lineage may first express in the central nervous system; in the process of evolution, the functions of these peptides diversified and were later found in other tissues.

Metallo-beta-lactamase-producing Pseudomonas aeruginosa isolated from a large tertiary centre in Kenya.

This study was designed to characterize the beta-lactamase content of carbapenem-resistant Pseudomonas aeruginosa isolates recovered during 2006 and 2007 in a large tertiary-care centre in Nairobi, Kenya. Molecular characterization was done using PCR and sequencing, and typing was performed using pulsed-field gel electrophoresis (PFGE). In total, 416 P. aeruginosa isolates were obtained during that period, of which 57 (13.7%) were resistant to carbapenems. All carbapenem-resistant isolates tested positive for metallo-beta-lactamase (MBL) production. All MBL isolates produced VIM-2 with two types of integron structures. PFGE identified three clonally related groups of VIM-2-producing P. aeruginosa, including a pan-resistant clone that was responsible for nosocomial outbreaks during 2006 and 2007 in the intensive-care unit. These findings suggest that continuous molecular surveillance needs to be performed to monitor the spread within the hospital of this pan-resistant strain. This study is the first report of VIM-2-producing P. aeruginosa from the African continent.

Comparison between sevoflurane/remifentanil and propofol/remifentanil anaesthesia in providing conditions for somatosensory evoked potential monitoring during scoliosis corrective surgery.

Somatosensory evoked potential (SSEP) monitoring is an important tool in spinal corrective surgery. Anaesthesia has a significant influence on SSEP monitoring and a technique which has the least and shortest suppressant effect on SSEP while facilitating a fast recovery from anaesthesia is ideal. We compared the effect of sevoflurane/ remifentanil and propofol/remifentanil anaesthesia on SSEPs during scoliosis corrective surgery and assessed patients' clinical recovery profiles. Twenty patients with idiopathic scoliosis receiving surgical correction with intraoperative SSEP monitoring were prospectively randomised to receive sevoflurane/remifentanil anaesthesia or propofol/remifentanil anaesthesia. During surgery, changes in anaesthesia dose and physiological variables were recorded, while SSEP was continuously monitored. A simulated 'wake-up' test was performed postoperatively to assess speed and quality of recovery from anaesthesia. The effects of propofol and sevoflurane resulted in SSEP amplitude variability between 18.0% +/- 3.5% to 28.7% +/- 5.9% and SSEP latency variability within 1.3% +/- 0.4% to 2.6% +/- 1.2%. Patients receiving sevoflurane had faster suppression and faster recovery of SSEP amplitude compared to propofol (P < 0.05), although propofol anaesthesia showed less within-patient variability in Cz amplitude and latency (P < 0.05). On cessation of anaesthesia, time to eye-opening (5.2 vs. 16.5 minutes) and toe movement (5.4 vs. 17.4 minutes) was shorter following sevoflurane (all P < 0.05). These findings indicate that propofol produces a better SSEP signal than sevoflurane. However adjustments in sevoflurane concentration result in faster changes in the SSEP signal than propofol. Assessment of neurological function was facilitated more rapidly after sevoflurane anaesthesia.

Persistent pain in patients following scoliosis surgery.

Chronic or persistent pain is increasingly recognised as a consequence of surgery in a number of different disciplines. The pain often exhibit qualities that differ from the acute post-operative pain and may represent changes in the central nervous system. There is lack of information regarding the incidence of persistent pain in patients following spinal surgery for scoliosis. This study aims to estimate the incidence of persistent pain following spinal surgery for scoliosis in a group of mainly adolescent patients. Questionnaires were distributed to consecutive patients attending the outpatient clinic of a hospital with specialist services in paediatric orthopaedics and spinal surgery. One hundred and five patients out of 122 eligible patients completed the survey. Fifty-two percent had ongoing pain upon hospital discharge either in the primary surgical site and/or in the iliac bone graft site. Approximately 10 and 7% of all patients had back and pelvic pain persisting beyond 12 months, respectively. A small proportion described elements of neuropathic pain. There was a trend suggesting that those who experienced more severe post-operative pain were more likely to develop persistent pain. These data are consistent with those reports that implicate surgery as the trigger for chronic pain.

CCS/CAR/CANM/CNCS/CanSCMR joint position statement on advanced noninvasive cardiac imaging using positron emission tomography, magnetic resonance imaging and multidetector computed tomographic angiography in the diagnosis and evaluation of ischemic heart disease--executive summary.

BACKGROUND: Over the past few decades, advanced imaging modalities with excellent diagnostic capabilities have emerged. The aim of the present position statement was to systematically review existing literature to define Canadian recommendations for their clinical use. METHODS: A systematic literature review to 2005 was conducted for positron emission tomography (PET), multidetector computed tomographic angiography and magnetic resonance imaging (MRI) in ischemic heart disease. Papers that met the criteria were reviewed for accuracy, prognosis data and study quality. Recommendations were presented to primary and secondary panels of experts, and consensus was achieved. RESULTS: Indications for PET include detection of coronary artery disease (CAD) with perfusion imaging, and defining viability using fluorodeoxyglucose to determine left ventricular function recovery and/or prognosis after revascularization (class I). Detection of CAD in patients, vessel segments and grafts using computed tomographic angiography was considered class IIa at the time of the literature review. Dobutamine MRI is class I for CAD detection and, along with late gadolinium enhancement MRI, class I for viability detection to predict left ventricular function recovery. Imaging must be performed at institutions and interpreted by physicians with adequate experience and training. CONCLUSIONS: Cardiac imaging using advanced modalities (PET, multidetector computed tomographic angiography and MRI) is useful for CAD detection, viability definition and, in some cases, prognosis. These modalities complement the more widespread single photon emission computed tomography and echocardiography. Given the rapid evolution of technology, initial guidelines for clinical use will require regular updates. Evaluation of their integration in clinical practice should be ongoing; optimal use will require proper training. A joint effort among specialties is recommended to achieve these goals.

Latent membrane protein 1 suppresses RASSF1A expression, disrupts microtubule structures and induces chromosomal aberrations in human epithelial cells.

Epstein-Barr virus (EBV) infection is closely associated with nasopharyngeal carcinoma (NPC) and can be detected in early premalignant lesions of nasopharyngeal epithelium. The latent membrane protein 1 (LMP1) is an oncoprotein encoded by the EBV and is believed to play a role in transforming premalignant nasopharyngeal epithelial cells into cancer cells. RASSF1A is a tumor-suppressor gene commonly inactivated in many types of human cancer including NPC. In this study, we report a novel function of LMP1, in down-regulating RASSF1A expression in human epithelial cells. Downregulation of RASSF1A expression by LMP1 is dependent on the activation of intracellular signaling of NF-kappaB involving the C-terminal activating regions (CTARs) of LMP1. LMP1 expression also suppresses the transcriptional activity of the RASSF1A core promoter. RASSF1A stabilizes microtubules and regulates mitotic events. Aberrant mitotic spindles and chromosome aberrations are reported phenotypes in RASSF1A inactivated cells. In this study, we observed that LMP1 expression in human epithelial cells could induce aberrant mitotic spindles, disorganized interphase microtubules and aneuploidy. LMP1 expression could also suppress microtubule dynamics as exemplified by tracking movements of the growing tips of microtubules in live cells by transfecting EGFP-tagged EB1 into cells. The aberrant mitotic spindles and interphase microtubule organization induced by LMP1 could be rescued by transfecting RASSF1A expression plasmid into cells. Downregulation of RASSF1A expression by LMP1 may facilitate its role in transformation of premalignant nasopharyngeal epithelial cells into cancer cells.

Secretin: a pleiotrophic hormone.

Secretin holds a unique place in the history of endocrinology and gastrointestinal physiology, as it is the first peptide designated as a hormone. During the last century since its first discovery, the hormonal effects of secretin in the gastrointestinal tract were extensively studied, and its principal role in the periphery was found to stimulate exocrine secretion from the pancreas. Recently, a functional role of secretin in the brain has also been substantiated, with evidence suggesting a possible role of secretin in embryonic brain development. Given that secretin and its receptors are widely expressed in multiple tissues, this peptide should therefore exhibit pleiotrophic functions throughout the body. The present article reviews the current knowledge on the central and peripheral effects of secretin as well as its therapeutic uses.

Endogenous release of secretin from the hypothalamus.

Previous studies demonstrated that secretin could be released from the cerebellum, where it exerts a facilitatory action on the GABAergic inputs into the Purkinje neurons. In the present article, we provide evidence of the endogenous release of secretin in the hypothalamus and the mechanisms underlying this release. Incubation of the hypothalamic explants with KCl induces the release of secretin to 4.35 +/- 0.45-fold of the basal level. This K+-induced release was tetrodotoxin and cadmium sensitive, suggesting the involvement of voltage-gated sodium and calcium channels. The use of specific blockers further revealed the involvement of L-, N-, and P-type high voltage-activated (HVA) calcium channels. Results present in the current article provide further and more solid evidence of the role of secretin as a neuropeptide in the mammalian central nervous system.