Investigation and management of a large community mumps outbreak among young adults in Toronto, Canada, January 2017-February 2018.

BACKGROUND: In 2017, a mumps outbreak was identified in a cohort of 18-34 year olds in Toronto, Canada. OBJECTIVE: To describe a large community mumps outbreak in an urban centre from January 2017 to February 2018 among young adults. METHODS: A broad range of interventions were implemented in an attempt to reach the target audience; including case and contact management, vaccination clinics at schools and clinicians' offices, school exclusions, bar inspections, traditional communication strategies (including health care provider updates and posters) and newer communication strategies (including three sequential social media campaigns). RESULTS: A total of 143 cases of mumps were identified. Although cases' ages ranged from three to 72 years, most (76%) were 18-34 year olds, many of whom had frequented bars and local food establishments in downtown Toronto. 84% (n=120) of the cases were community-acquired. Only 16% (n=23) of the cases reported exposures in schools and post-secondary school institutions. Of those, 39% (n=56) of cases had an unknown vaccination history; 34% (n=49) were either not vaccinated or partially vaccinated with one dose of measles-mumps-rubella vaccine; and 27% (n=38) had received the recommended two doses of mumps vaccine. Determining vaccination status was a challenge, in part due to the lack of a registry. Vaccination was recommended when subjects were known to have had fewer than two doses of vaccine or had an unknown vaccination status. A social media campaign, emphasizing the risk of social activities if not protected from the mumps, yielded over 500,000 impressions from Facebook and Twitter messages and ads and an impressive engagement rate of between 1% and 10x%. CONCLUSION: This was the largest mumps outbreak in Toronto in over 20 years. Among young adults, ongoing social media and traditional communication campaigns can contribute to the control of community mumps outbreaks. Encouraging vaccine uptake is desirable, but without a vaccine registry it is difficult to assess vaccination coverage among adults. Susceptible cohorts of young adults who were not adequately vaccinated pose a risk for future outbreaks. Given that almost 30% of the mumps cases were fully vaccinated with two doses of mumps-containing vaccine, even two doses may not provide complete protection.

Unlabeled probes for the detection and typing of herpes simplex virus.

BACKGROUND: Unlabeled probe detection with a double-stranded DNA (dsDNA) binding dye is one method to detect and confirm target amplification after PCR. Unlabeled probes and amplicon melting have been used to detect small deletions and single-nucleotide polymorphisms in assays where template is in abundance. Unlabeled probes have not been applied to low-level target detection, however. METHODS: Herpes simplex virus (HSV) was chosen as a model to compare the unlabeled probe method to an in-house reference assay using dual-labeled, minor groove binding probes. A saturating dsDNA dye (LCGreen Plus) was used for real-time PCR. HSV-1, HSV-2, and an internal control were differentiated by PCR amplicon and unlabeled probe melting analysis after PCR. RESULTS: The unlabeled probe technique displayed 98% concordance with the reference assay for the detection of HSV from a variety of archived clinical samples (n = 182). HSV typing using unlabeled probes was 99% concordant (n = 104) to sequenced clinical samples and allowed for the detection of sequence polymorphisms in the amplicon and under the probe. CONCLUSIONS: Unlabeled probes and amplicon melting can be used to detect and genotype as few as 10 copies of target per reaction, restricted only by stochastic limitations. The use of unlabeled probes provides an attractive alternative to conventional fluorescence-labeled, probe-based assays for genotyping and detection of HSV and might be useful for other low-copy targets where typing is informative.

Pre-emptive analgesia and protective premedication. What is the difference?

The management of postoperative pain has been greatly informed by an increasing understanding of the basic science of pain transmission. The idea that analgesia given before the injury would be more effective than the same analgesia given after the injury was named pre-emptive analgesia. The evidence for this phenomenon in postoperative pain management has been very mixed. The methodological problems of such studies, and the difficulties of all the major outcome measures make comparison of the studies available difficult. In the 20 years since the concept was proposed there has been a change in anaesthetic practice that in effect incorporates pre-emptive analgesia with opiates. Evidence for any pre-emptive analgesic with non-steroidal anti-inflammatory drugs is very poor, but the use of local anaesthetic blocks continues to be an area of study. Pre-emptive use of analgesic drugs is not the magic bullet to prevent postoperative pain, but is a strategy of use, among others for managing postoperative pain. Protective analgesia is a strategy that has grown out of the same desire to give drugs before injury to reduce the pain experienced afterwards. In this case the drugs under study have not been primary analgesics, but adjuvant drugs used commonly in the non-acute pain arena. In particular, the drug gabapentin, and to a lesser extent its related drug pregabalin. These drugs have been given by mouth as a pre-medicant, 1 hour before surgery in a variety of operations. A recent meta-analysis of the existing literature shows reduction of postoperative morphine consumption but little reduction in reporting of opiate side effects. Pregabalin, which has a better pharmacokinetic profile, may be a better alternative, and is currently under study. Neither gabapentin nor pregabalin are licensed for use in postoperative pain, and it is unlike that the manufacturers will seek such a licence.

Amplicon DNA melting analysis for mutation scanning and genotyping: cross-platform comparison of instruments and dyes.

BACKGROUND: DNA melting analysis for genotyping and mutation scanning of PCR products by use of high-resolution instruments with special "saturation" dyes has recently been reported. The comparative performance of other instruments and dyes has not been evaluated. METHODS: A 110-bp fragment of the beta-globin gene including the sickle cell anemia locus (A17T) was amplified by PCR in the presence of either the saturating DNA dye, LCGreen Plus, or SYBR Green I. Amplicons of 3 different genotypes (wild-type, heterozygous, and homozygous mutants) were melted on 9 different instruments (ABI 7000 and 7900HT, Bio-Rad iCycler, Cepheid SmartCycler, Corbett Rotor-Gene 3000, Idaho Technology HR-1 and LightScanner, and the Roche LightCycler 1.2 and LightCycler 2.0) at a rate of 0.1 degrees C/s or as recommended by the manufacturer. The ability of each instrument/dye combination to genotype by melting temperature (Tm) and to scan for heterozygotes by curve shape was evaluated. RESULTS: Resolution varied greatly among instruments with a 15-fold difference in Tm SD (0.018 to 0.274 degrees C) and a 19-fold (LCGreen Plus) or 33-fold (SYBR Green I) difference in the signal-to-noise ratio. These factors limit the ability of most instruments to accurately genotype single-nucleotide polymorphisms by amplicon melting. Plate instruments (96-well) showed the greatest variance with spatial differences across the plates. Either SYBR Green I or LCGreen Plus could be used for genotyping by T(m), but only LCGreen Plus was useful for heterozygote scanning. However, LCGreen Plus could not be used on instruments with an argon laser because of spectral mismatch. All instruments compatible with LCGreen Plus were able to detect heterozygotes by altered melting curve shape. However, instruments specifically designed for high-resolution melting displayed the least variation, suggesting better scanning sensitivity and specificity. CONCLUSION: Different instruments and dyes vary widely in their ability to genotype homozygous variants and scan for heterozygotes by whole-amplicon melting analysis.

A single-tube nucleic acid extraction, amplification, and detection method using aluminum oxide.

A disposable 0.2-ml polymerase chain reaction (PCR) tube modified with an aluminum oxide membrane (AOM) has been developed for the extraction, amplification, and detection of nucleic acids. To assess the dynamic range of AOM tubes for real-time PCR, quantified herpes simplex virus (HSV) DNA was used to compare AOM tubes to standard PCR tubes. AOM PCR tubes used for amplification and detection of quantified HSV-1 displayed a crossing threshold (C(T)) shift 0.1 cycles greater than PCR tube controls. Experiments with HSV-1-positive cerebrospinal fluid (CSF) examined the extraction, amplification, and detection properties of the AOM tubes compared to the Qiagen DNA blood mini kit. The AOM extraction, amplification, and detection of HSV-1 in CSF displayed differences of less than one C(T) when compared to Qiagen-extracted samples. Experiments testing the AOM method using clinical CSF samples displayed 100% concordance with reported results. AOM tubes have no adverse effects on amplification or fluorescence acquisition by real-time PCR and can be effectively used for the extraction, amplification, and detection of HSV from CSF. The AOM single tube method is a fast, reliable, and reproducible technique for the extraction, amplification, and detection of HSV in CSF.

Increased sensitivity of bacterial detection in cerebrospinal fluid by fluorescent staining on low-fluorescence membrane filters.

A membrane-filter-based, fluorescent Gram stain method for bacterial detection in cerebrospinal fluid samples was developed and evaluated as a rapid, sensitive alternative to standard Gram stain protocols. A recently developed, modified version of the aluminium oxide membrane Anopore with low-fluorescence optical properties showed superior performance in this application. Other aspects of the fluorescent Gram stain system that were evaluated include membrane filter selection, strategies to reduce fluorescence fading and the effect of patient blood cells on bacterial detection in the fluorescently stained cerebrospinal fluid samples. The combination of the membrane filter's bacteria-concentrating ability and absolute retention along with high-contrast, fluorescent Gram discriminating dyes enabled rapid bacterial detection and Gram discrimination, with a 1-1.5 order of magnitude increase in the bacterial concentration limit of detection.

A retrospective observational study of pre-operative sickle cell screening.

The aim of this study was to determine the ethnic mix of those patients being pre-operatively screened for sickle cell disease in a London teaching hospital and to determine the rate of carriage of sickle haemoglobin amongst those tested. We retrospectively studied 1879 patients undergoing surgery over a 2-month period. Two hundred and thirteen (11%) were screened for sickle cell disease and of these, 12 (5%) tested positive for sickle cell trait (HbAS). There were no patients homozygous for sickle cell disease (HbSS) or with haemoglobin SC disease (HbSC). Screening rates varied widely in different ethnic groups from 0% of the Chinese population to 85.2% of the Afro-Caribbean population. We conclude that at present there is no coherent pre-operative screening policy for sickle cell disease in our institution. Sickle cell disease poses unique anaesthetic risks and with a rapidly expanding 'mixed race' population high-risk patients are difficult to identify phenotypically. We propose a universal screening policy be implemented in high-risk areas.

Evaluation of a needle-free injection system for local anaesthesia prior to venous cannulation.

We evaluated a single-use, disposable, carbon-dioxide-powered, needleless injector (J-Tip, National Medical Products Inc., CA, USA), which is claimed to deliver a virtually painless, subcutaneous injection. Seventy-two patients undergoing various types of surgery had a large-bore intravenous cannula inserted prior to induction of general anaesthesia. Three minutes beforehand, a subcutaneous injection of 0.3 ml of 1% plain lidocaine was administered. Subjects were randomly allocated to receive the lidocaine either by the needleless injector or from a conventional syringe and a 25 G needle. Pain scores were recorded on injection of the lidocaine and on insertion of the cannula. There was significantly less pain on injection with the needleless injector than with the 25 G needle (p < 0.001) but, surprisingly, there was more pain on cannulation (p < 0. 001). We conclude that the device certainly delivers a less painful subcutaneous injection than a 25 G needle, but perhaps provides less effective skin anaesthesia for venous cannulation at sites where the subcutaneous space is small; its use might be better suited to areas where the subcutaneous space is deeper.

The Objective Structured Clinical Exam - practical aspects.

Assessment of clinical skills in addition to factual knowledge has become part of the examination system in both undergraduate and postgraduate examinations. There is an increasing interest in the concept of competence and how it can be measured, particularly for the process of revalidation of practitioners. The Objective Structured Clinical Examination (OSCE) has been introduced for the assessment of clinical skills into postgraduate anaesthetic examinations in the United Kingdom. As with any examination, the issues regarding setting a standard are under continual review. The application of this exam format to the measurement of competency as well as skills has been the subject of scrutiny during the period reviewed. The literature suggests that competence is a much more complex construct than skill and that OSCE exams will be a contributory part but more work is required to establish assessment methods which will produce a global rating of competence. The role of the OSCE format in education as well as assessment has been examined and it is suggested that formative assessment can be made, and peer review learning brought about using an OSCE performed in teams.

The role of CCR7 in TH1 and TH2 cell localization and delivery of B cell help in vivo.

Subsets of murine CD4+ T cells localize to different areas of the spleen after adoptive transfer. Naïve and T helper 1 (TH1) cells, which express the chemokine receptor CCR7, are home to the periarteriolar lymphoid sheath, whereas activated TH2 cells, which lack CCR7, form rings at the periphery of the T cell zones near B cell follicles. Retroviral transduction of TH2 cells with CCR7 forces them to localize in a TH1-like pattern and inhibits their participation in B cell help in vivo but not in vitro. Thus, differential expression of chemokine receptors results in unique cellular migration patterns that are important for effective immune responses.

Placental transfer of fentanyl in early human pregnancy and its detection in fetal brain.

We have investigated the transfer of fentanyl across the early human placenta in 38 women (8-14 weeks' gestation) undergoing termination of pregnancy. After administration of a bolus dose of fentanyl 2 micrograms kg-1 at induction of anaesthesia, maternal blood n = 38), placenta (n = 38), amniotic fluid (n = 38) and fetal brain (n = 7) samples were collected and assayed for fentanyl by radioimmunoassay. Fentanyl was detected in all placental and fetal brain samples but not in amniotic fluid. There was a rapid decrease in fentanyl concentrations in maternal serum after the bolus but placental concentrations had not started to decline 30 min later. There was no difference in placental drug concentrations at different gestational ages. These data suggest that there is rapid transfer of fentanyl to the fetus in early pregnancy and that the drug remains in fetal tissue for some time after the initial dose is given to the mother.

I.v. clonidine: does it work as a hypotensive agent with inhalation anaesthesia?

In a double-blind, randomized, placebo-controlled study, 41 patients received clonidine 3 micrograms kg-1 or placebo at induction of isoflurane and nitrous oxide in oxygen anaesthesia. Metoprolol was also given to achieve a systolic arterial pressure of 80 mm Hg. Requirements for metoprolol were significantly less in the clonidine group (P < 0.00035), with no significant difference in mean arterial pressures over time. It would appear that clonidine is an i.v. hypotensive agent worthy of consideration, but data during the recovery period are required to comment further on the safety of this technique.

A comparison of the vasodilatation produced by two topical anaesthetics.

The vasoactive properties of EMLA (eutectic mixture of local anaesthetic) and amethocaine were compared using Doppler colour ultrasound to measure vein diameter in 20 male volunteers. EMLA or amethocaine cream were applied over veins on the dorsum of the hand and vein diameter was measured prior to application of the preparations and at 1, 1.5 and 2 h after removal. Visual analogue scores for skin colour and vasodilatation were performed at 1 h. The results showed no significant difference in vein diameter between the EMLA and amethocaine groups. However, the amethocaine-treated hands were significantly more erythematous at 1 h (p < 0.00002).

Placental transfer of fentanyl in early human pregnancy.

To investigate the transfer of fentanyl across the early human placenta, we have collected samples of maternal blood and fetal fluids and/or blood, simultaneously, between 5 and 22 min following an intravenous bolus of fentanyl (1.5 microg/kg) to the mother. The pregnancies were between 6 and 16 weeks of gestation and scheduled for elective termination of pregnancy under general anaesthesia. Total fentanyl concentration was determined by radioimmunoassay in 11 pairs of first trimester maternal serum and fetal coelomic fluid samples, 14 pairs of maternal serum and amniotic fluid samples, seven series of first trimester maternal serum and coelomic and amniotic fluid samples, and 10 series of early second trimester maternal and fetal sera and amniotic fluid samples. Fentanyl was not detected in coelomic fluid samples at any gestational age and in amniotic fluid samples collected after 12 weeks of gestation. Measurable concentrations of fentanyl were found in maternal serum collected within 15 min after the initial bolus and in fetal serum collected between 10 and 12 min later. These findings indicate that fentanyl is transferred across the early placenta into the amniotic cavity and fetal blood circulation but not into the exocoelomic cavity. The distribution of this molecule inside the early gestational sac is probably influenced by the increased binding by maternal and fetal sera, its short half-life of distribution and the specific biology of the fetal fluid formation and composition.

Maternal endocrine activation during pregnancy alters neurobehavioral state in primate infants.

OBJECTIVES: The purpose of this study was to investigate whether maternal endocrine activation during pregnancy would affect the neurobehavioral state of primate offspring in a manner similar to that observed in human infants from pregnancies involving maternal substance abuse or maternal stress. METHOD: Twenty-two rhesus monkey (Macaca mulatta) infants were derived from females administered either adrenocorticotrophic hormone (ACTH), which increased the mother's endocrine activity, or saline solutions for 14 consecutive days during mid-pregnancy. On days 15 and 30 postpartum, infants underwent brief separations from their mothers and were videotaped for later evaluation of neurobehavioral state. RESULTS: Infants from mothers administered ACTH spent significantly more time in a drowsy state than did controls (p < .04), and the increased drowsiness tended to be most pronounced during the postseparation period, when acute stress was highest. In contrast, controls remained in a more active alert state (p < .03), presumably searching for their mother, a species-typical adaptive response to maternal separation. Female infants spent more time in distressed state than did males on day 15, and the proportion of time in distressed state decreased in all infants after administration of .2 ml of 24% sucrose solution. CONCLUSION: The results demonstrate that neurobehavioral state alterations are found in infants from mothers with increased endocrine activity during pregnancy. Neurobehavioral state disorganization can have an adverse impact on the human infant's concurrent and subsequent occupational performance. These findings establish the usefulness of the nonhuman primate model for advancing knowledge on early contributions to the development of human infant occupational behavior.

Dreams, images and emotions associated with propofol anaesthesia.

One hundred and twelve patients scheduled for day case varicose vein surgery were randomly allocated to one of three groups: total intravenous anaesthesia with propofol, propofol induction followed by inhalational anaesthesia with nitrous oxide and isoflurane or thiopentone induction followed by inhalational anaesthesia with nitrous oxide and isoflurane. Assessments were made in the recovery room of the incidence of dreaming, the content of the dreams and the emotional status of the patients. The groups differed significantly in reporting that they had been dreaming: patients who underwent total intravenous anaesthesia reported the most dreaming and patients who received thiopentone the least. However, despite the large number of case reports of sexual imagery following propofol anaesthesia and despite the two groups who had received propofol experiencing significantly greater happiness upon recovery than the thiopentone group, there were no appreciable differences in the sexual content of the dreams. Each group had only a small number of dreams even remotely related to sex.