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Study shows SARS-CoV-2 infection, replication and persistence in human brain tissues

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In a recent study published in Nature, researchers investigated the cellular tropism, replication competence, persistence and evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in humans, and associated histopathological changes in infected tissues.

Study: SARS-CoV-2 infection and persistence in the human body and brain at autopsy. Image Credit: Surasak_Photo/Shutterstock

Coronavirus disease 2019 (COVID-19) has reportedly caused multiple organ derangements in the acute period, with a few infected individuals developing persistent symptoms, referred to as PASC (post-acute sequelae of COVID-19). However, the non-respiratory disease burden and the duration of SARS-CoV-2 clearance from non-respiratory tissues such as the brain has not been extensively investigated.

About the study

In the present study, researchers mapped and quantified the distribution, proliferation, and cellular tropism of SARS-CoV-2 in non-respiratory human body tissues such as the brain from the acute COVID-19 period to >7.0 months after the onset of symptoms.

Autopsied brain specimens of 44 unvaccinated and deceased COVID-19 patients were analyzed, and extensive CNS (central nervous system) sampling was performed for 11 individuals between April 26, 2020, and March 2, 2021. Polymerase chain reaction (PCR) analysis was performed to confirm SARS-CoV-2 positivity, and 38 samples were found to be SARS-CoV-2-positive. Three samples (P27, P36, and P37) were seronegative for SARS-CoV-2, and sera were not available for three cases (P3, P4, and P15).

Droplet digital PCR (ddPCR) analysis was performed for SARS-CoV-2 N (nucleocapsid) gene quantification, and ISH (in-situ hybridization) analysis was performed to validate the ddPCR results and to determine SARS-CoV-2 cell tropism. The IF (immunofluorescence) and IHC (immunohistochemistry) analyses were performed further to verify the viral presence within human brain tissues.

Further, subgenomic ribonucleic acid (RNA) was detected using real-time quantitative reverse transcription-PCR (RT– qPCR) analysis, and virus isolation experiments were performed using Vero E6 cells to demonstrate proliferation-capable SARS-CoV-2 among tissues of respiratory and another origin. SARS-CoV-2 S (spike) gene variant diversity and distribution were measured using HT-SGS (high-throughput, single-genome amplification and sequencing) analysis for six individuals.

Among the autopsied specimens, 17, 13, and 14 were categorized as early cases, mid-cases, and late cases based on the day of infection (d) at death within 14 days, between 15 days and 30 days, and beyond 31 days, respectively. Further, image analysis on interventricular septal tissues of 16 individuals was performed to assess the association between SARS-CoV-2 N RNA detected by ddPCR analysis and SARS-CoV-2 S RNA detected by ISH analysis. Furthermore, N protein-targeted ISH assays, IF analysis, and IHC-based analyses were performed to validate SARS-CoV-2 detection and distribution in the CNS.

Results

Among the study individuals, 30% were women, with a median age value of 63 years, and 61% of them suffered from at least three comorbid conditions. The median duration between the onset of symptoms to hospital admission and death was six days and 19 days, respectively, and the median post-mortem duration was 22 hours.

SARS-CoV-2 ribonucleic acid was present at 84 anatomical sites in significantly greater amounts among respiratory tissues than other tissues. SARS-CoV-2 RNA levels among early cases, mid-cases, and late-cases were 2.0 log10 nucleocapsid gene copies for every nanogram RNA, 1.4 log10 nucleocapsid gene copies for every nanogram RNA and 0.7 log10 nucleocapsid gene copies for every nanogram RNA, respectively. SARS-CoV-2 ribonucleic acid was detected in the perimortem sera of 11 and one early cases and mid-cases, respectively.

SARS-CoV-2 ribonucleic acid was persistently present in several tissues of late cases, despite being below detectable levels in sera of any case. SARS-CoV-2 ribonucleic acid was identified within the CNS among 91% (n= 10) cases, including across most brain areas evaluated in five (out of six) late cases. SARS-CoV-2 subgenomic ribonucleic acid was detected across all tissues and in several body fluids, including serum, vitreous humor, and pleural fluid.

The subgenomic ribonucleic acid RT-qPCR analysis and ddPCR analysis findings correlated closely for 1,025 specimens, particularly among 369 respiratory samples, 496 early cases, and 302 specimens showing SARS-CoV-2 positivity by RT-qPCR and ddPCR analyses. SARS-CoV-2 was isolated among Vero E6 cells from 45% (n=25) of specimens from the lymph nodes, heart, adrenal gland, gastrointestinal tissues, and ophthalmic tissues of early cases.

In addition, SARS-CoV-2 was isolated from the P38 thalamus among Vero E6-transmembrane serine protease 2 (TMPRSS2)-T2A-angiotensin-converting enzyme 2 (ACE2) cells. HT-SGS analysis of 46 specimens from six individuals did not show any non-synonymous SARS-CoV-2 genomic diversity in respiratory tissues and other tissues for P18, P19, and P27. Among P27 samples, two SARS-CoV-2 haplotypes, each comprising a synonymous mutation, were detected preferentially among non-respiratory tissues such as the mediastinal lymph nodes and the left and right ventricles.

In P38, residue D80F was detected in all 31 respiratory, but none of the 490 cranial sequences and residue G1219V was limited to the cranial variants. Among intraventricular septum tissues, the mean SARS-CoV-2 N gene copies/nanogram RNA correlated significantly with the median SARS-CoV-2 S RNA-positive cells. In the second ISH assay, SARS-CoV-2 RNA and protein were observed in the cerebellum and hypothalamus of P38, basal ganglia of P40, and cervical spinal cord of P42.

The histopathological analysis findings indicated that 92% (n=35) of cases died due to diffuse alveolar injury or acute pneumonia, and the diffuse alveolar injury cases showed a temporal pattern of progression. Myocardial infiltrates, and paracortical and follicular hyperplasia was observed. However, despite widespread SARS-CoV-2 RNA distribution in the body, negligible evidence of direct SARS-CoV-2 cytopathology or inflammation was observed in non-respiratory tissues.

Overall, the study findings showed that SARS-CoV-2 could infect and replicate in non-respiratory tissues such as the brain early in infection and persist for months (up to 230 days) following symptom onset.

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What’s the greatest holiday gift: lips, hair, skin? Give the gift of great skin this holiday season

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Give the gift of great skin this holiday season

Skinstitut Holiday Gift Kits take the stress out of gifting

Toronto, October 31, 2024 – Beauty gifts are at the top of holiday wish lists this year, and Laser Clinics Canada, a leader in advanced beauty treatments and skincare, is taking the pressure out of seasonal shopping. Today, Laser Clincs Canada announces the arrival of its 2024 Holiday Gift Kits, courtesy of Skinstitut, the exclusive skincare line of Laser Clinics Group.

In time for the busy shopping season, the limited-edition Holiday Gifts Kits are available in Laser Clinics locations in the GTA and Ottawa. Clinics are conveniently located in popular shopping centers, including Hillcrest Mall, Square One, CF Sherway Gardens, Scarborough Town Centre, Rideau Centre, Union Station and CF Markville. These limited-edition Kits are available on a first come, first served basis.

“These kits combine our best-selling products, bundled to address the most relevant skin concerns we’re seeing among our clients,” says Christina Ho, Senior Brand & LAM Manager at Laser Clinics Canada. “With several price points available, the kits offer excellent value and suit a variety of gift-giving needs, from those new to cosmeceuticals to those looking to level up their skincare routine. What’s more, these kits are priced with a savings of up to 33 per cent so gift givers can save during the holiday season.

There are two kits to select from, each designed to address key skin concerns and each with a unique theme — Brightening Basics and Hydration Heroes.

Brightening Basics is a mix of everyday essentials for glowing skin for all skin types. The bundle comes in a sleek pink, reusable case and includes three full-sized products: 200ml gentle cleanser, 50ml Moisture Defence (normal skin) and 30ml1% Hyaluronic Complex Serum. The Brightening Basics kit is available at $129, a saving of 33 per cent.

Hydration Heroes is a mix of hydration essentials and active heroes that cater to a wide variety of clients. A perfect stocking stuffer, this bundle includes four deluxe products: Moisture 15 15 ml Defence for normal skin, 10 ml 1% Hyaluronic Complex Serum, 10 ml Retinol Serum and 50 ml Expert Squalane Cleansing Oil. The kit retails at $59.

In addition to the 2024 Holiday Gifts Kits, gift givers can easily add a Laser Clinic Canada gift card to the mix. Offering flexibility, recipients can choose from a wide range of treatments offered by Laser Clinics Canada, or they can expand their collection of exclusive Skinstitut products.

 

Brightening Basics 2024 Holiday Gift Kit by Skinstitut, available exclusively at Laser Clincs Canada clinics and online at skinstitut.ca.

Hydration Heroes 2024 Holiday Gift Kit by Skinstitut – available exclusively at Laser Clincs Canada clinics and online at skinstitut.ca.

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Here is how to prepare your online accounts for when you die

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LONDON (AP) — Most people have accumulated a pile of data — selfies, emails, videos and more — on their social media and digital accounts over their lifetimes. What happens to it when we die?

It’s wise to draft a will spelling out who inherits your physical assets after you’re gone, but don’t forget to take care of your digital estate too. Friends and family might treasure files and posts you’ve left behind, but they could get lost in digital purgatory after you pass away unless you take some simple steps.

Here’s how you can prepare your digital life for your survivors:

Apple

The iPhone maker lets you nominate a “ legacy contact ” who can access your Apple account’s data after you die. The company says it’s a secure way to give trusted people access to photos, files and messages. To set it up you’ll need an Apple device with a fairly recent operating system — iPhones and iPads need iOS or iPadOS 15.2 and MacBooks needs macOS Monterey 12.1.

For iPhones, go to settings, tap Sign-in & Security and then Legacy Contact. You can name one or more people, and they don’t need an Apple ID or device.

You’ll have to share an access key with your contact. It can be a digital version sent electronically, or you can print a copy or save it as a screenshot or PDF.

Take note that there are some types of files you won’t be able to pass on — including digital rights-protected music, movies and passwords stored in Apple’s password manager. Legacy contacts can only access a deceased user’s account for three years before Apple deletes the account.

Google

Google takes a different approach with its Inactive Account Manager, which allows you to share your data with someone if it notices that you’ve stopped using your account.

When setting it up, you need to decide how long Google should wait — from three to 18 months — before considering your account inactive. Once that time is up, Google can notify up to 10 people.

You can write a message informing them you’ve stopped using the account, and, optionally, include a link to download your data. You can choose what types of data they can access — including emails, photos, calendar entries and YouTube videos.

There’s also an option to automatically delete your account after three months of inactivity, so your contacts will have to download any data before that deadline.

Facebook and Instagram

Some social media platforms can preserve accounts for people who have died so that friends and family can honor their memories.

When users of Facebook or Instagram die, parent company Meta says it can memorialize the account if it gets a “valid request” from a friend or family member. Requests can be submitted through an online form.

The social media company strongly recommends Facebook users add a legacy contact to look after their memorial accounts. Legacy contacts can do things like respond to new friend requests and update pinned posts, but they can’t read private messages or remove or alter previous posts. You can only choose one person, who also has to have a Facebook account.

You can also ask Facebook or Instagram to delete a deceased user’s account if you’re a close family member or an executor. You’ll need to send in documents like a death certificate.

TikTok

The video-sharing platform says that if a user has died, people can submit a request to memorialize the account through the settings menu. Go to the Report a Problem section, then Account and profile, then Manage account, where you can report a deceased user.

Once an account has been memorialized, it will be labeled “Remembering.” No one will be able to log into the account, which prevents anyone from editing the profile or using the account to post new content or send messages.

X

It’s not possible to nominate a legacy contact on Elon Musk’s social media site. But family members or an authorized person can submit a request to deactivate a deceased user’s account.

Passwords

Besides the major online services, you’ll probably have dozens if not hundreds of other digital accounts that your survivors might need to access. You could just write all your login credentials down in a notebook and put it somewhere safe. But making a physical copy presents its own vulnerabilities. What if you lose track of it? What if someone finds it?

Instead, consider a password manager that has an emergency access feature. Password managers are digital vaults that you can use to store all your credentials. Some, like Keeper,Bitwarden and NordPass, allow users to nominate one or more trusted contacts who can access their keys in case of an emergency such as a death.

But there are a few catches: Those contacts also need to use the same password manager and you might have to pay for the service.

___

Is there a tech challenge you need help figuring out? Write to us at onetechtip@ap.org with your questions.

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Pediatric group says doctors should regularly screen kids for reading difficulties

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The Canadian Paediatric Society says doctors should regularly screen children for reading difficulties and dyslexia, calling low literacy a “serious public health concern” that can increase the risk of other problems including anxiety, low self-esteem and behavioural issues, with lifelong consequences.

New guidance issued Wednesday says family doctors, nurses, pediatricians and other medical professionals who care for school-aged kids are in a unique position to help struggling readers access educational and specialty supports, noting that identifying problems early couldhelp kids sooner — when it’s more effective — as well as reveal other possible learning or developmental issues.

The 10 recommendations include regular screening for kids aged four to seven, especially if they belong to groups at higher risk of low literacy, including newcomers to Canada, racialized Canadians and Indigenous Peoples. The society says this can be done in a two-to-three-minute office-based assessment.

Other tips encourage doctors to look for conditions often seen among poor readers such as attention-deficit hyperactivity disorder; to advocate for early literacy training for pediatric and family medicine residents; to liaise with schools on behalf of families seeking help; and to push provincial and territorial education ministries to integrate evidence-based phonics instruction into curriculums, starting in kindergarten.

Dr. Scott McLeod, one of the authors and chair of the society’s mental health and developmental disabilities committee, said a key goal is to catch kids who may be falling through the cracks and to better connect families to resources, including quicker targeted help from schools.

“Collaboration in this area is so key because we need to move away from the silos of: everything educational must exist within the educational portfolio,” McLeod said in an interview from Calgary, where he is a developmental pediatrician at Alberta Children’s Hospital.

“Reading, yes, it’s education, but it’s also health because we know that literacy impacts health. So I think that a statement like this opens the window to say: Yes, parents can come to their health-care provider to get advice, get recommendations, hopefully start a collaboration with school teachers.”

McLeod noted that pediatricians already look for signs of low literacy in young children by way of a commonly used tool known as the Rourke Baby Record, which offers a checklist of key topics, such as nutrition and developmental benchmarks, to cover in a well-child appointment.

But he said questions about reading could be “a standing item” in checkups and he hoped the society’s statement to medical professionals who care for children “enhances their confidence in being a strong advocate for the child” while spurring partnerships with others involved in a child’s life such as teachers and psychologists.

The guidance said pediatricians also play a key role in detecting and monitoring conditions that often coexist with difficulty reading such as attention-deficit hyperactivity disorder, but McLeod noted that getting such specific diagnoses typically involves a referral to a specialist, during which time a child continues to struggle.

He also acknowledged that some schools can be slow to act without a specific diagnosis from a specialist, and even then a child may end up on a wait list for school interventions.

“Evidence-based reading instruction shouldn’t have to wait for some of that access to specialized assessments to occur,” he said.

“My hope is that (by) having an existing statement or document written by the Canadian Paediatric Society … we’re able to skip a few steps or have some of the early interventions present,” he said.

McLeod added that obtaining specific assessments from medical specialists is “definitely beneficial and advantageous” to know where a child is at, “but having that sort of clear, thorough assessment shouldn’t be a barrier to intervention starting.”

McLeod said the society was partly spurred to act by 2022’s “Right to Read Inquiry Report” from the Ontario Human Rights Commission, which made 157 recommendations to address inequities related to reading instruction in that province.

He called the new guidelines “a big reminder” to pediatric providers, family doctors, school teachers and psychologists of the importance of literacy.

“Early identification of reading difficulty can truly change the trajectory of a child’s life.”

This report by The Canadian Press was first published Oct. 23, 2024.

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