Manufacturing a Bentley in the traditional way is a time-consuming nod to their brand heritage. Each hand-crafted wheel can take 4.5 hours to finish

Shallaway Youth Choir is bringing a program of music to the Basilica in downtown St. John’s on Remembrance Day. Artistic director Kellie Walsh says the choir will sing about such topics as the Unknown Soldier and schoolchildren who care for Newfoundland graves in London. Where Once They Stood will be performed at 2 p.m. and 7 p.m. on Monday.

Police have reported that a man was fatally electrocuted in Torbay on Saturday evening. Two others are being treated for injuries.

Bombardier control box recovered during second dive to a World War 2 bomber at the bottom of Gander Lake.

Monday marks the first Remembrance Day ceremony at the National War Memorial in St. John's to include the tomb of the Unknown Soldier. We're proud to present again two documentaries that tell a remarkable story.

Large crowds came together at city hall in Corner Brook to lay wreaths and honour veterans on Monday. The ceremony portion was completed with a playing of the Last Post.

The A.C. Hunter Public Library wants people in St. John’s to know that it’s open and accessible, despite how the entrance looks. The main staircase is under repair but there’s plenty of signs directing folks to other entrances. Still, librarians say people are judging this book by its cover and assuming the library is closed.

A Labrador foster parent and head of a group working to better the Labrador foster care system is stressing the importance of keeping Indigenous children with Indigenous parents in their Indigenous communities.

Hundreds congregated in downtown St. John's Monday morning to pay tribute to Newfoundland and Labrador's fallen soldiers, and honoured the dead for the first time at the newly refurbished National War Memorial and tomb of the Unknown Solider. 

Pat White, an entrepreneur who launched EVAS Air and the Gander Flight Training, has died. He was 69 years old.

The 50th Christmas Craft Fair gathered over a hundred vendors and thousands of customers in Torbay over the weekend. One family business was inspired by the early years of the fair and a passion to keep traditional crafts in style.

CBC Newfoundland and Labrador would love to hear your suggestions of folks and organizations who are there when you need them — and all the other times, too.

Nurses' union president Yvette Coffey says more must be done to ensure young nurses stay in the province. Newfoundland and Labrador Health Services says work is underway to match students with job openings.

People using the library in the Arts and Culture Centre in St. John's are being forced to use alternate entrances due to an unsafe staircase in front of the building.

A candidate for the Nova Scotia NDP has apologized and is no longer running for the party in the provincial election after her comments on Israel and the Holocaust drew concern from the Jewish organizations. 

Volunteers will be cutting hair at a salon based inside a Cape Breton food bank. The goal is to provide clients with the same salon experience paying customers would receive.

Independent Elizabeth Smith-McCrossin is back on the campaign trail in Cumberland North, but opponents say the riding would be better represented by an MLA from an official party. 

A Port Hawkesbury woman has long imagined a Remembrance Day with her great-uncle's war medals. It will happen this year for the first time thanks to some sleuthing and the kindness of her great-uncle's foster family.

RCMP are investigating after officers discovered human remains in an abandoned, burned vehicle in Greenhill, N.S.

The only daily TV news package to focus on Nova Scotians and their stories

About a thousand people stood solemnly in the rain at a cenotaph in downtown Halifax on Monday morning to pay tribute to veterans on Remembrance Day.

As Week 3 of the provincial election campaign begins, there are questions about how Nova Scotia's major political parties plan to combat the rising number of opioid overdose deaths. 

The only daily TV news package to focus on Nova Scotians and their stories

The new mayor and council are planning big changes for the Cape Breton Regional Municipality, and one of those is at the top of the staff organizational chart now that chief administrative officer Marie Walsh is set to retire.

The Société de transport de Montréal (STM) has established a non-urgent text line for transit users to signal security issues in the Metro network.

Loved ones, friends, and people from across the city have all come together to support each other at Pastel Rita, the café Jocelyn Després helped so many call home.

The City of Montreal wants to sell more land to community organizations, housing co-ops and other groups to build off-market housing. 

Ten years after their formation, the members of The Brooks discuss how they’ve evolved and how they view their collaboration.

Facing soaring food prices and an escalating housing crisis, Quebec student associations are urgently calling on the provincial government to fund internships required for post-secondary training.

The union representing dockworkers in Montreal says Canada's labour minister forcing its members back to work at ports in Montreal and Quebec City marks a "dark day for workers' rights."

Quebec provincial police have confirmed that the body found in a Montreal nature park on Oct. 30 was that of kidnapping victim and cryptocurrency influencer Kevin Mirshahi.

Mosquito-borne disease is a major global public health issue. One path toward the development of evidence-based strategies to limit mosquito biting is the study of the mosquito nervous system—in particular, the sensory systems that drive biting behavior. The central nervous system of insects consists of the brain and the ventral nerve cord. Here, we describe a protocol for dissecting, immunofluorescent labeling, and imaging both of these structures in the mosquito. This protocol was optimized for Aedes aegypti and works well on Anopheles gambiae tissue. It has not been tested in other mosquito species, but we anticipate that it would work on a range of mosquitoes, and, if not, our protocol will provide a starting point from which to optimize. Notably, a limited number of antibodies cross-react with Ae. aegypti proteins. This protocol is intended for use with validated antibodies and can also be used to test new antibodies as they are generated. It has been successfully used to visualize protein tags, such as green fluorescent protein, that have been introduced into the mosquito to amplify or detect their presence.

Genetically encoded calcium indicators (GECIs) allow for the noninvasive evaluation of neuronal activity in vivo, and imaging GECIs in Drosophila has become commonplace for understanding neural functions and connectivity in this system. GECIs can also be used as read-outs for studying sleep in this model organism. Here, we describe a methodology for tracking the activity of neurons in the fly brain using a two-photon (2p) microscopy system. This method can be adapted to perform functional studies of neural activity in Drosophila under both spontaneous and evoked conditions, as well as during spontaneous or induced sleep. We first describe a tethering and surgical procedure that allows survival under the microscopy conditions required for long-term recordings. We then outline the steps and reagents required for optogenetic activation of sleep-promoting neurons while simultaneously recording neural activity from the fly brain. We also describe the procedure for recording from two different locations—namely, the top of the head (e.g., to record mushroom body calyx activity) or the back of the head (e.g., to record central complex activity). We also provide different strategies for recording from GECIs confined to the cell body versus the entire neuron. Finally, we describe the steps required for analyzing the multidimensional data that can be acquired. In all, this protocol shows how to perform calcium imaging experiments in tethered flies, with a focus on acquiring spontaneous and induced sleep data.

Sleep studies in Drosophila melanogaster rely mostly on behavioral read-outs to support molecular or circuit-level investigations in this model. Electrophysiology can provide an additional level of understanding in these studies to, for example, investigate changes in brain activity associated with sleep manipulations. In this protocol, we describe a procedure for performing multichannel local field potential (LFP) recordings in the fruit fly, with a flexible system that can be adapted to different experimental paradigms and situations. The approach uses electrodes containing multiple recording sites (16), allowing the acquisition of large amounts of neuronal activity data from a transect through the brain while flies are still able to sleep. The approach starts by tethering the fly, followed by positioning it on an air-supported ball. A multichannel silicon probe is then inserted laterally into the fly brain via one eye, allowing for recording of electrical signals from the retina through to the central brain. These recordings can be acquired under spontaneous conditions or in the presence of visual stimuli, and the minimal surgery promotes long-term recordings (e.g., overnight). Sleep and wake can be tracked using infrared cameras, which allow for the measurement of locomotive activity as well as microbehaviors such as proboscis extensions during sleep. The protocol has been optimized to promote subject survivability, which is an important factor when performing long-term (~16-h) recordings. The approach described here uses specific recording probes, data acquisition devices, and analysis tools. Although it is expected that some of these items might need to be adapted to the equipment available in different laboratories, the overall aim is to provide an overview on how to record electrical activity across the brain of behaving (and sleeping) flies using this kind of approach and technology.

The technique of visualizing axon pathways in the embryonic ventral nerve cord using antibody labeling has been fundamental to our understanding of the genetic and developmental mechanisms underlying nervous system wiring in Drosophila. High-resolution microscopic examination of the ventral nerve cord remains an essential component of many experiments in Drosophila developmental neuroscience. Although it is possible to examine the ventral nerve cord in intact whole-mount embryos, to collect the highest-quality images it is often useful to isolate the nervous system away from the other embryonic tissues through embryo dissection. This protocol describes methods for dissecting ventral nerve cords from Drosophila embryos that have been fixed and stained via immunofluorescence or horseradish peroxidase (HRP) immunohistochemistry. The process of making fine dissection needles for this purpose from electrolytically sharpened tungsten wire is also described. Dissected and mounted ventral nerve cords can be examined and imaged using a variety of microscopy techniques including differential interference contrast (DIC) optics, epifluorescence, or confocal microscopy.

The Drosophila embryonic central nervous system has been used for decades as a model for understanding the genetic regulation of axon guidance and other aspects of neural development. Foundational studies using antibody staining to examine the embryonic ventral nerve cord in wild-type and mutant animals led to the discovery of evolutionarily conserved genes that regulate fundamental aspects of axon guidance, including midline crossing of axons. The development of the regular, segmentally repeating structure of axon pathways in the ventral nerve cord can illustrate basic principles of axon guidance to beginning students and can also be used by expert researchers to characterize new mutants, detect genetic interactions between known genes, and precisely quantify variations in gene function in engineered mutant lines. Here, we describe a protocol for collecting and fixing Drosophila embryos and visualizing axon pathways in the embryonic ventral nerve cord using immunofluorescence or immunohistochemical staining methods. As embryogenesis in Drosophila takes ~24 h to complete, a 1-d collection yields embryos representing all stages of development from newly fertilized through ready-to-hatch larvae, allowing investigation of multiple developmental events within a single batch of collected embryos. The methods described in this protocol should be accessible to introductory laboratory courses as well as seasoned investigators in established research laboratories.

Mosquitoes take blood meals from a diverse range of host animals and their host associations vary by species. Characterizing these associations is an important element of the transmission dynamics of mosquito-vectored pathogens. To characterize mosquito host associations, various molecular techniques have been developed, which are collectively referred to as blood meal analysis. DNA barcoding has diverse biological applications and is well-suited to mosquito blood meal analysis. The standard DNA barcoding marker for animals is a 5' fragment of the cytochrome c oxidase I (COI) gene. A major advantage of this marker is its taxonomic coverage in DNA sequence reference databases, making it feasible to identify a wider range of mosquito host species than with any other gene. However, the COI gene contains high sequence variation at potential priming sites between vertebrate orders. Coupled with the need for primer sequences to be mismatched with mosquito priming sites so that annealing to mosquito DNA is inhibited, it can be difficult to design primers suitable for blood meal analysis applications. Several primers are available that perform well in mosquito blood meal analysis, annealing to priming sites for most vertebrate host taxa, but not to those of mosquitoes. Because priming site sequence variation among vertebrate taxa can cause amplification to fail, a hierarchical approach to DNA barcoding-based blood meal analysis can be applied. In such an approach, no single primer set is expected to be effective for 100% of potential host species. If amplification fails in the initial reaction, a subsequent reaction is attempted with primers that anneal to different priming sites, and so on, until amplification is successful.

Mosquito species vary in their host associations. Although some species are relative generalists, most specialize, to varying extents, on particular types of host animals. Mosquito host associations are among the most important factors that influence the transmission dynamics of mosquito-vectored pathogens, and understanding these associations can provide insight on how such pathogens move within ecosystems. Characterization of the host associations of mosquito species requires applying blood meal analysis to the largest possible sample size of mosquito blood meals. Processing large samples of mosquito blood meals can be time-consuming, especially when chain-termination sequencing is used, necessitating individual processing of each specimen. Various methods and commercially available kits and products are available for extracting DNA from mosquito blood meals. The hot sodium hydroxide and Tris (HotSHOT) method is a rapid and inexpensive method of DNA extraction that is compatible with the recovery of DNA from mosquito blood meals preserved on QIAcard Flinders Technology Associates (FTA) Classic Cards (FTA cards). FTA cards allow nucleic acids found in blood meals to be preserved easily, even in field conditions. DNA prepared using this method is suitable for polymerase chain reaction (PCR)-based blood meal analysis.

All PCR- and DNA-based blood meal analyses require host DNA from a mosquito blood meal to be effectively preserved between the time when the specimen is collected and the extraction of DNA. As soon as a mosquito ingests blood from a host animal, digestion of host cells and cellular components within the blood meal by enzymes in the mosquito midgut begins to degrade the host DNA templates that are the targets of polymerase chain reaction (PCR) amplification. Without effective preservation, host DNA is typically undetectable by PCR 48 h after feeding, because of digestion. Preservation methods for mosquito blood meals vary in their efficacy, and the logistics of fieldwork can limit the options for preservation of blood meals and maintenance of the integrity of host DNA. This protocol describes a method of blood meal preservation that is effective, convenient, and amenable to fieldwork in remote locations where cryopreservation at –20°C or –80°C may not be feasible. It uses a Flinders Technology Associates (FTA) card, which is a chemically treated card that lyses cells and allows nucleic acids to be preserved. This method is also expected to preserve the DNA or RNA of pathogens present within the engorged mosquito abdomen, including RNA viruses.

The insect eggshell is a multifunctional structure with several important roles, including generating an entry point for sperm via the micropyle before oviposition, serving as an oviposition substrate attachment surface, and functioning as a protective layer during embryo development. Eggshell proteins play major roles in eggshell tanning and hardening following oviposition and provide molecular cues that define dorsal–ventral axis formation. Precise eggshell formation during ovarian follicle maturation is critical for normal embryo development and the synthesis of a defective eggshell often gives rise to inviable embryos. Therefore, simple and accurate methods for identifying eggshell proteins will facilitate our understanding of the molecular pathways regulating eggshell formation and the mechanisms underlying normal embryo development. This protocol describes how to isolate and enrich eggshells from mature oocytes of Aedes aegypti mosquitoes and how to extract their eggshell proteins for liquid chromatography with tandem mass spectrometry (LC–MS/MS) proteomic analysis. Although this methodology was developed for studying mosquito eggshells, it may be applicable to eggs from a variety of insects. Mosquitoes are ideal model organisms for this study as their ovarian follicle development and eggshell formation are meticulously regulated by blood feeding and their follicles develop synchronously throughout oogenesis in a time-dependent manner.

In insects, oocyte resorption (oosorption) or follicular atresia is one of the key physiological processes and evolutionary strategies used to optimize reproductive fitness. Mosquitoes are ideal model organisms for studying egg maturation in arthropods, as their follicle development is initiated only following the ingestion of a blood meal, followed by a carefully orchestrated series of hormonally regulated events leading to egg maturation. A cohort of approximately 100 follicles per mosquito ovary begin developing synchronously. However, a significant fraction of follicles ultimately undergo apoptosis and oosorption, especially when available resources from the blood meal are limited. Therefore, simple, rapid, and reliable techniques to accurately evaluate follicular atresia are necessary to understand mechanisms underlying follicle development in insects. This protocol describes how to detect apoptotic follicle cells within the Aedes aegypti mosquito ovaries using a commercially available fluorescent-labeled inhibitor of caspases (FLICA). Caspases are key players in animal apoptosis. In this assay, the FLICA reagent enters the intracellular compartment of follicles in dissected mosquito ovaries and covalently binds to active caspases. The bound reagent remains within the cell and its fluorescent signal can be observed by confocal microscopy. Although this method was specifically developed for visualizing apoptotic ovarian follicles during Ae. aegypti mosquito egg development, it should be applicable to other mosquito tissues that undergo caspase-mediated program cell death in a time-dependent manner.

A Two-Day Festival in the historic Arts & Industries Building brings community, artists and scholars together for a “Culture Lab”

An unlikely friendship with Guy Burgess, the infamous British double-agent, brought unexpected joy to Stanley Weiss

As The Meg hits theaters, dive into what we really know about this chompy predator

The ancient burial structures, strikingly similar all across Europe and the mediterranean, have puzzled scientists and historians for centuries

Tammy Roberts, Home Base's executive director, said before these services launched, there was "no place for youth to go" other than the emergency room or to RCMP cells. 

With president-elect Donald Trump promising to drill in Alaska's Arctic National Wildlife Refuge, the Gwich'in are preparing to fight for the Porcupine caribou herd.

Wanita Johnson, who celebrates her birthday this weekend in Whitehorse, still has a sharp memory for how the Second World War helped shape her life.

Yukon appeal court Justice Karen Horsman granted a temporary stay of the requirement on Friday, the latest development in an ongoing legal battle over the property between the territorial government, cabin-owner Len Tarka and tenant Eric DeLong.