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Donald Trumps prediction that the country or, at least, the American automobile industry will experience a bloodbath if he is not returned to the presidency was excessive and crude. But over-the-top rhetoric is not new and, for more than 400 licensed medical professionals, its not the most troubling aspect of Trumps public performances.

They see evidence that the former president, at 77, is suffering from probable dementia so they have signed their names, along with hundreds of others, to an online statement asserting that grave concern.

Of course, his detractors claim President Joe Biden, at 81, is the one who shows such signs, and they continue to do so even after Bidens solid delivery of the State of the Union address. The age and fitness of both candidates is a major concern for voters.

But, under the banner of Duty To Warn, organized by Baltimore-based psychologist John Gartner, both clinicians and researchers have joined an effort to defend Biden against dementia claims and warn the nation about what they see as Trumps cognitive decline.

The media never tire of asking, Is [President Joe] Biden too old? Gartner says. Polls showed twice as many people were worried about Bidens cognitive health as Trumps, when Bidens memory lapses are within the normal limits for his age [81]. We all get more forgetful as we age. To say that makes us incompetent or too old is ageism. I would argue that Ive garnered wisdom and judgment from life experience that more than compensates for my memory blips. And I think the same could be said of Biden. Thats why I say Bidens brain is aging, but Trumps brain is dementing.

Mainstream media, says Gartner, push a false equivalency that Biden and Trump are two old men prone to mistakes, presenting equally unappealing choices. In fact, says Gartner, comparing Biden to Trump is comparing apples to rotted oranges. Biden, he notes, had a lifelong struggle with stuttering and was always known for gaffes, while Trumps verbal skills, back in his Manhattan celebrity days, were at a higher level; they are now greatly diminished.

Gartner, a psychotherapist who was an assistant professor for 28 years at the Johns Hopkins University Medical School, has found credible help sounding this alarm. The statement the other professionals signed claims Trump is showing unmistakable signs strongly suggesting dementia, based on his public behavior and informant reports.

The statement cites a decline in Trumps baseline verbal fluency. [Trump] was once highly articulate, with a sophisticated vocabulary, and spoke in polished paragraphs, the experts say. Now, his vocabulary is impoverished, he often has difficulty finishing a thought, sentence or even a word. Typical of dementia patients, he [repeats] and overuses superlatives and filler words.

Duty to Warn has 430,000 followers on X, formerly Twitter. Since last week, a post about Trumps probable dementia has had more than 1.7 million views.

In a long series of testimonials attached to the original post, doctors and other professionals cite Trumps use of incorrect words or jumbled words, suggesting a condition known as paraphasia, a common indicator of dementia. The experts note that Trump frequently makes errors he does not recognize and correct, and mixes up people across generations, mistaking a father for a grandfather.

But Trump has always rambled when he addresses MAGA rallies. So how do Gartner and the dozens of professionals he consulted discern signs of dementia?

When you are literally unable to form words and what youre saying is so incomprehensible that its impossible for someone to understand what you are saying, thats not rambling, says Gartner. Those are serious signs of dementia. If you saw them in a relative, you would run, not walk, to a specialist to get them tested and start thinking about long-term care.

The evidence for dementia in Donald Trump has become overwhelming, writes Dr. Lance Dodes, a psychiatrist and retired professor of the Harvard Medical School. Unlike normal aging, which is characterized by forgetting names or words, Trump repeatedly shows something very different: confusion about reality.

Suzanne Lachman, a New York psychologist, posted this: [Trump will] begin a sentence, and then seemingly forget how the sentence began and invent something in the middle, and then go off on a tangent that results in an incomprehensible word salad. This is behavior we observe frequently in patients who have dementia.

If this guy presented in my office with the symptoms we observe in his public appearances, Id refer him at once to a neurologist for a full workup, wrote John Biggs, a social worker and retired psychotherapist who was based at Sheppard Pratt for 30 years. I wouldnt write him an OK-to-return-to-work note, much less declare him fit to occupy the countrys highest office.

The Alzheimers Society last month published an essay saying diagnosis at a distance is unethical and usually wrong.

Gartner strongly disagrees. Trump and Biden are, next to Taylor Swift, the most exposed people in the country. There are ample ways to observe their behavior and hear their words every day. In real medicine, Gartner argues, thousands of psychiatrists make diagnoses every day in clinical practice based on observation of behavior, history and informant reports. Research shows all of those methods are actually more accurate and reliable than a clinical interview.

The greatest ethical responsibility, he says, is the duty to warn, and at a critical time for the nation.

Originally posted here:
Clinicians defend Biden's fitness, warn of Trump's decline - Baltimore Sun

Fitness influencer Giorgi Tzane Janelidze died on Wednesday after falling into a ravine while filming social media content in Italy.

According to Greek media outlet Ethnos, the accident took place during the 23-year-old's visit to Roghudi Vecchio, Calabria, an ancient mountain village abandoned in the 1970s due to a severe flood. While standing on a balcony without a railing, he slipped and fell into the ravine.

Despite the efforts of firefighters who rushed to the scene, they were unable to save him. Helicopters were eventually called in to retrieve his body due to the depth of the ravine. From there, his body was transported to the town of Saline Joniche, where it was handed over to authorities.

Janelidze's friend, Chris Kogias, who accompanied him on the trip, confirmed news of the death to the outlet.

Tzaneis no longer with us, Kogias, who is also a social media influencer, said. He left us yesterday afternoon during thetripwe made toItaly.Please pray with us for hissoulto rest in peace.

Giorgi Janelidze/Instagram

Janelidze, who was born in Georgia but lived in Greece, has over 100,000 followers on Instagram. In his most recent post, he shared with his followers a video of him holding a bag of his protein powder, which he co-founded with fellowinfluencer Dream Greek.

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The most beautiful person inside and out, the biggest adventure Ive ever had with someone, Janelidzes girlfriend, Elena Margariti, wrote in a since-expired Instagram Story post, per the New York Post. With my man, my whole life.

She continued, Many dreams, a home together, through our difficulties, through our joys, and we made it, us against all odds. So unfairly, so early. I love you so much. We will meet again. My Tzitsi.

See the article here:
Greek Fitness Influencer Falls into Ravine and Dies - PEOPLE

Ryan Andrews is having treatment on a dead leg (Image: PA)

Interim Watford boss Tom Cleverley is hopeful Ryan Andrews and Jamal Lewis will be fit to face Leeds United when the Championship resumes after both players were ruled out of international action.

Right-back Andrews had been named in the England Mens Elite League Squad for away trips to Poland and Czech Republic, but has returned to London Colney for treatment on a dead leg.

The interim Hornets head coach told the club website: Ryan got a dead leg in training and was going to be doubtful for both England games so everyone thought it best for him to return to us for treatment.

Jamal Lewis may return to the Northern Ireland for their game with Scotland on Tuesday (Image: PA)

Lewis is also back with Watford and has been ruled out of Northern Irelands game with Romania tomorrow after being sidelined for last weeks 1-0 victory at Birmingham City with a foot injury, although he may yet return to the national squad for Tuesdays friendly with Scotland.

Cleverley said: Jamal's foot injury is keeping him out of the first of Northern Ireland's two games, but he's optimistic he may be okay for the second game early next week.

From our point of view, we're hopeful they'll both be able to play a part against Leeds, so let's see how they come through next week.

See the article here:
Cleverley 'hopeful' over fitness of injured international pair - Yahoo Eurosport UK

If you'd like to be a Lafayette Police officer, this Saturday you can take the first step.

The Lafayette Police Department is hosting a physical fitness recruitment event on Saturday, March 23 from 8 a.m. until 11 a.m. at the track and field facility at Teurlings High School, 139 Teurlings Drive.

The event will start promptly at 8 a.m. with a physical fitness test, so anyone who wants to participate should be there before 7:45 a.m. to register. Participants should bring water bottles or sports drinks with them.

Participants should wear athletic clothing, including athletic shoes, and be ready to complete push-ups, sit-ups, a 300-meter sprint and an "officer down" drill.

The Lafayette Police Department will have PT instructors on hand and will begin the morning with a warm-up and stretch.

The recruiting staff will also be on hand to answer any questions participants may have regarding employment with the Lafayette Police Department and provide applications. This event is a requirement of the application process.

If you want more information about the application process and the requirements to be a police officer, click here.

More here:
LPD sets physical fitness test for prospective officers - KATC News

One of CrossFits top training camps, PRVN Fitness,announcedtheir roster of 2024 roster athletes in advance of the CrossFit Open 24.3 live announcement at PRVN Fitness HQ in Nashville, TN.

The camp looks to have one of their best lineups ever. Their 15-person roster consists of CrossFit Games veterans, potential 2024 Games rookies, and, of course, six-time champion Tia-Clair Toomey-Orr.

With that being said, lets take a brief look at each of these 15 top PRVN athletes:

[Related: Longtime CrossFit Coach Danny Lesslie Faces $200,000 in Medical Bills for Wifes Cancer Treatment]

The champ is back, and she will lead the charge at PRVN along with her husband and PRVN Fitness co-founder, Shane Orr.

Last year was wild for the Womens Individual division with Toomey-Orr absent. Still, after a tough second-place finish at the 2023 Rogue Invitational just months after giving birth, Toomey-Orr is back to reclaim her spot at the top.

[Related: Interview: CrossFit Athlete Kelly Baker Talks About Raising Awareness for Fertility Issues in Women]

Six-time Games athlete Jay Crouch is coming off his best career finish, breaking into the top 10 at the 2024 CrossFit Games with an eighth-place overall finish.

Crouch made massive improvements throughout the 2023 season, surprising many in the community, and seemingly coming out of nowhere. Crouch will seek a second straight top-10 finish at the Games and show that 2023 wasnt a fluke.

The 2023 Games Rookie of the Year, Olivia Kerstetter, will return as a PRVN athlete and attempt to solidify her position as one of the top in the sport. 2023 was Kerstetters first year in the Individual womens division, and she finished 16th at just 17 years old. What can Kerstetter do now with a year of experience under her belt?

Two years ago, Nick Mathew wasnt even supposed to be at the 2022 CrossFit Games but received an invitation thanks to a backfill that opened. Mathew went on to take two event wins to finish 14th overall.

In 2023, Mathew had a slight setback, finishing 19th, but earned another year of experience that should prove useful in 2024.

After two years in 30th or worse at the Games, fan-favorite Colten Mertens made huge strides in 2023, finishing 18th at the Game and racking up an event win in Ski-Bag along the way.

Mertens is no stranger to hard work and seems to fit in at PRVN. He took the worldwideoverall win in Open 24.1 despite suffering from personal tragedy with the loss of his mother that week.Mertens could potentially have yet another career best in 2024.

In 2023, Sydney kept the Wells name alive at the Games while sister Brooke barely missed out. Sydney had a solid rookie performance, finishing 28th overall with two top-10 event finishes. She will look for more in 2024.

Since Sydney Wells trains at CrossFit East Nashville PRVN alongside Toomey-Orr and the rest of the PRVN crew, Wells has had time to learn from the best. The results should show in 2024.

Another up-and-comer on the PRVN roster is the Brazilian and 2023 Games rookie Kaylan Souza. Souza took over for Guilherme Malheiros and finished 32nd overall at the Games; the highest-placing athlete from Brazil.

At only 23 years old, Souza is still young with plenty of time to improve. Over the next few years, he may be a staple to watch from the Latin America region.

Stanley is the fourth 2023 Games rookie on this list. She finished just one place above her PRVN teammate Sydney Wells in 27th.

Stanley flew under the radar during the 2023 Games because she had no top-10 event finishes but avoided disastrous events. With that middle-of-the-pack consistency, she could continue to rise up the leaderboard each year with slight improvements in each modality.

Four-time CrossFit Games athlete Maddie Sturt competed at the Games for four straight years from 2016-2019, with a best overall finish of 20th in 2018.

Since 2019, however, Sturt has been unable to return, missing out on qualifying for the past three years by just a handful of points each year in heartbreaking fashion. Sturt is consistently one of the top 10 fittest women in the Oceania region, and we should likely expect her back at the Games either this year or the next.

Two-time Games athlete Sydney Michalyshen had a slight setback in 2023, missing Games qualification by 32 points. Still, with Semifinals lineup changes already announced, Michalyshen may take advantage and return to the Games in 2024.

Michalyshen has finished 31st and 25th at the Games and will expect an even higher finish this year. Michalyshen is one of the strongest athletes in the field. If she fixes other areas in her game, shell be dangerous.

In 2023, as an impressive 16-year-old, Lucy McGonigle qualified for the European Semifinal, ultimately finishing in 45th.

Since McGonigle is so young, she will continue developing and improving each year. It would not be surprising to see her competing at the Games before turning 18. McGonigle is a dark horse to watch in 2024.

Luis Oscar Mora was only 40 points away from qualifying for the 2023 Games from the North America West Semifinal. It wouldve been his first ticket there as an individual male.

In 2016, Mora competed in the boys 16-17 age division, finishing fourth overall. Mora is known for great strength and will be a threat in heavy events when he makes it to the Games floor again.

Another athlete who narrowly missed qualifying for the 2023 Games was William Kearney, who failed to qualify by 26 points in the Oceania Semifinal. He finished fourth overall.

Kearneys career so far shows continued improvement in the Open and Quarterfinals, meaning he will likely improve again in 2024. At 24 years old, look out for him to represent Australia at the Games in the next couple of years.

In 2023, Rylee Beebe finished seventh overall at the Games in the Girls 16-17 division and will likely finish higher in 2024.

Beebe is a three-time Games athlete in the teen division. In 2024, as a 17-year-old, Beebe has the opportunity to compete in the Individual division or in the Teen division, depending on her Semifinal performance.

The 2022 Boys 14-15 Games champion and 2023 Boys 16-17 third-place finisher, RJ Mestre, is another of PRVNs elite teen athletes eyeing his Individual division appearance soon.

Mestre is one of the favorites to win the Boys 16-17 division in 2024 and then move to the Mens division after. Mestre has good coaches and teachers in Nashville, and hell be sure to make them proud in this upcoming season.

Featured image: @PRVNFitness on Instagram

Continued here:
Get to Know the 2024 PRVN Fitness Athlete Roster - BarBend

Adam Howard, Body Shop Fitness owner, and Bruce Clayton of TriCoach recently presented the Beebe Medical Foundation with a check for $6,000 to support Beebe Healthcares oncology services.

The funds were raised through their Burpees for Beebe challenge, when Howard and Clayton each did 100 burpees a day for 100 days in a row, and their Thanksgiving morning BRUteCamp, a bootcamp-style workout held on Rehoboth Beach. Howard and Clayton collected donations in support of their efforts, raising a total of $6,000 for patients in Sussex County.

We are honored to receive the proceeds from Adam and Bruces efforts again this year! Their annual BRUtecamp has been a tradition for many years, and adding the Burpees for Beebe component really took things up a notch this year. Weve seen this program grow and feel honored to be the annual recipient of the proceeds, said Amy Keller, Beebe Medical Foundation event coordinator. We feel fortunate to have community partners like Adam and Bruce helping us to save and change lives in Sussex County.

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Body Shop Fitness and TriCoach raise $6K for Beebe oncology services - CapeGazette.com

What cant Serena Williams do? Since picking up the tennis racquet at four years old, shes won 23 Grand Slam singles titles and four Olympic gold medals. Not to mention, shes now a mother of two, devoted wife, and entrepreneur with her investment capital firm, Serena Ventures, and Will Perform, a topical pain relief product company, which launched in December 2022.

However, whats currently been at the forefront for Williams is motherhood, as she recently gave birth to her second daughter, Adira River Ohanian, in August 2023. Although shes been happily basking in motherhood with her husband, Alexis Ohanian, Williams did express a desire to get back into the gym to dial up her fitness regimen. Just six months after giving birth to her daughter, we found Williams working on her squats with heavy weights in the gym. In her Instagram Reel filmed in January 2024, she spoke to her fans about how shes getting back into the swing of things and captioned the post, Back into the swing of things #Thisishowyoudoit. She also said, This isnt my usual weight, but this is what Im doing until I get back to where I used to be.

In just one month, she began to see progress as she shared another Reel with her in a sports bra and leggings, letting some of her midriff show, and captioned the video post, feeling confident. Next, we saw her slay the red carpet with a tightly fitted bodycon dress throughout awards season, specifically at the Grammys. For the final reveal, she showed a picture of her in a lavender bikini, holding her newborn, and donning her newly sculpted abs. She captioned the post, Loving yourself is essential. I find that I have to remind myself of that self-love through all different stages in my life. Right now I love that my body is not picture-perfect. I love that I smell like milk that milk sustains @adiraohanian I love getting to know a new version of my body. It is a change, but its a change that has been well worth it. So start this week, knowing that you are loved, and that starts with you. Ok, now Im about to go to the gym .

Aside from consistently staying in the gym and weightlifting, we wanted to share some helpful exercises for getting amazing abs like the star athlete. Check them out below.

Dead squats: If you want to recreate Williamss exact exercise in her Reel, you should master the art of dead squats. This exercise requires the lifter to rest the barbell on the safety pins before executing the lift, ultimately strengthening your core and developing tension (tightening) with your muscles. Although a dead squat isnt a direct ab workout, your abs will be activated and engaged during the movement.

An old-fashioned plank: Although this exercise isnt glamorous or comfortable, routine planks will do the job. To start, lie down and begin in a plank position. Be sure your elbows are aligned underneath your shoulders. Hold for 10 to 20 seconds per set.

Dumbbell side bends: For this exercise, youll need a single medium-weight dumbbell to help apply the necessary tension. This exercise will help shape your abs. To start, stand with your feet apart, keep your back straight, activate your core, and bend to the side as far as possible.

See the article here:
3 Fitness Exercises To Get Sculpted Abs Like Serena Williams - Essence

I recently got my hands on the new Minitailz Dog Health and GPS tracker, announced in January at CES, and promptly affixed it to my dog Beleveder's collar, just in time for an epic afternoon puppy playdate.

Think of it as a smartwatch or fancy fitness tracker for cats and dogs. The Minitailz, from Invoxia, sports an onboard GPS, Wi-fi and LTE-M antenna for real-time location and activity tracking. Priced at $100, the device has a monthly fee of a little over $8 for the cellular connectivity.

Beyond keeping tabs on your fluffy friend's location, the Minitailz provides insights into their exercise habits, as well as common behaviors like barking, drinking, eating, and playing. If worn day and night, it also provides daily wellness reports.

Heart and lung health monitoring are also key features. However, these tools require extended wearing. Over time, the tracker builds a profile of your pet's breathing patterns, resting and maximum heart rates and unique pulse signatures. With this data, it monitors for irregularities and serious health conditions, including atrial fibrillation (AFib).

With only a short amount of time to unbox and set up the Minitailz before Belvedere's ride arrived it was fortuitously delivered the day of his playdate I quickly got it charging and installed the companion Invoxia Petcare app.

Fortunately, the device arrived with a decent amount of battery life, and setup was a breeze. The app asks basic questions about your pet, including age, weight, gender and breed(s).

Belvedere is a mixed-breed dog, and I was able to specify his breakdown of Long-Haired Chihuahua, Miniature Pinscher and Dachshund. With all this info punched in and the device installed on his harness, he was ready to go.

It didn't take long before I received my first notification from the Minitialz: Transportation was detected, and Belvedere was on the move! Using real-time tracking, I was able to check in and see where he and his buddies were headed.

About forty minutes later, I received another update. Transportation had ended, and a pet walk was detected. However, it was the next notification that sent me into a fit of laughter: "Zoomies" had been detected Belvedere had just made a sprint.

A few hours later, I got another update that Belevedere's walk had ended, and he was in the car on his way back home.

Once back safely in my care and completely tuckered out, I popped open the Invoxia app to see what other fun data points and insights might be waiting for me. That's when I discovered that my dog is a big-time trotter.

The Minitailz classifies movement into one of three categories: walking, trotting and running. Belvedere spent only one minute running during his adventure, which was a bit disappointing. But he did manage to get in a solid 34 minutes of trotting and another hour and eleven minutes of walking.

By the way, Invoxia defines trotting as "one of the gaits of dogs. When a dog trots, two of its legs touch the ground simultaneously, followed by the other two, and so on." Here I was thinking that was called prancing.

Somewhere between a sprint and a saunter, the trot is the most sophisticated and graceful of all puppy movements according to me and reserved for only the most regal of Beagles (and mixed-breed pups named Belv).

Of course, activity insights only scratch the surface of what the Minitailz is capable of. And I look forward to testing out the health, behavior and wellness features as soon as I get a proper collar to attach the device to the harness doesn't seem as ideal. Until then, catch Belvedere and me trotting down a sidewalk near you.

Originally posted here:
I put a fitness tracker on my dog here's what happened - Tom's Guide

Blood-feeding induces molecular signatures in I. scapularis hemocytes related to immunity, metabolism, and proliferation

Ticks rely solely on blood as a source of essential metabolites, ingesting ~100 times their body mass per meal19. During feeding, extensive modifications and tissue rearrangements are necessary to accommodate and digest such large volumes of blood19. Considering that hemocytes are the circulating cells in the hemolymph, we hypothesized that these immune cells sense and respond to physiological and microbial exposure during tick infestation on vertebrate hosts. We optimized a protocol to collect hemocytes from I. scapularis nymphs, a clinically relevant stage in the blacklegged tick, and identified three common hemocyte morphotypes reported in the literature (Fig.1a, Supplementary Fig.1)16,17,18. Prohemocytes, considered the stem cell-like hemocyte population, were the smallest cells, with a round or oval shape between 5.7 and 12.5m (average diameter of 8.5m). The cytoplasm was minute (high nuclear/cytoplasmic ratio) and homogeneous, with no apparent protrusions or granules (Supplementary Fig.1a, d). Granulocytes were round or oval shaped cells with diameters ranging from 10.4 to 22.8m (average of 16.7m). The position of the nucleus varied, appearing most often near the periphery of the cell. Their cytoplasm was filled with dark-blue or violet stained granules or vacuoles (Supplementary Fig.1b, d). Plasmatocytes varied in size (ranged from 14.8 to 31.2m, with an average of 21.3m) and shape (oval, ameboid-like, pyriform), and had cytoplasmic protrusions or pseudopodia-like structures. The cytoplasm was clear and had few dark-blue or violet stained granules or vacuoles. The nucleus was located either near the center or the periphery of the cell (Supplementary Fig.1c, d). We then investigated the impact of blood-feeding on hemocytes originating from I. scapularis nymphs. We observed an increased quantity of total hemocytes upon mammalian feeding (Fig.1b). The percentage of plasmatocytes increased in engorged ticks, whereas we noticed a decline in the proportion of prohemocytes and granulocytes in repleted nymphs compared to unfed (Fig.1b). Altogether, we demonstrate the impact of hematophagy on the distribution of tick hemocyte morphotypes.

a Schematic representation of the hemocyte-enriched collection procedure. b Total number of hemocytes (n=40, 25 and 19) and percentages of different morphotypes (prohemocytes, plasmatocytes and granulocytes; n=20, 13 and 17 for all cases) from unfed (ivory), partially fed (light blue) or engorged (dark blue) nymphs. c Functional enrichment analysis of the differentially expressed genes (DEGs) in hemocyte-enriched samples from engorged ticks (blue; Up) compared to unfed ticks (red; Down). Fold enrichment of significant categories (FDR<0.05) is depicted. The number of DEGs per category is shown in parentheses. d The expression of key genes upregulated during feeding in hemocyte-enriched samples from unfed (ivory), partially fed (light blue) and engorged (dark blue) ticks was evaluated by RT-qPCR (n=11, 5 and 7 for all cases; with 4080 pooled ticks per sample). b, d Results are represented as meanSD. At least three independent experiments were performed. Statistical significance was evaluated by Brown-Forsythe ANOVA test, and significant p-values (<0.05) are displayed in the figure. Source data are provided as a Source Data file. 4cl1=4-coumarate-CoA ligase 1; ahcy = adenosylhomocysteinase B; g6pc = glucose-6-phosphatase 2; impdh = inosine-5-monophosphate dehydrogenase 1.

Next, we aimed to examine global transcriptional changes induced by hematophagy through bulk RNA-seq. We collected hemocyte-enriched samples from unfed and engorged nymphs and observed drastic changes in gene expression, with a total of 6134 differentially expressed genes (DEGs) (Fig.1c; Supplementary Data1). Samples from unfed I. scapularis nymphs were enriched for housekeeping genes, including those involved in mRNA transcription (e.g., RNA splicing, mRNA processing, histone modification), protein synthesis (e.g., peptide biosynthetic process, cellular protein modification process, ribosome biogenesis) and membrane receptor signaling pathways (e.g., transmembrane signaling receptor activity, G protein-coupled receptor signaling pathway, protein kinase activity) (Fig.1c; Supplementary Data2). In contrast, hemocyte-enriched samples from engorged ticks exhibited an overrepresentation of gene signatures related to immunity, metabolic pathways, cell proliferation/growth, and arthropod molting/development (Fig.1c; Supplementary Fig.2; Supplementary Data2). Key genes modulated during feeding, including 4-coumarate-CoA ligase 1 (4cl1), adenosylhomocysteinase B (ahcy), glucose-6-phosphatase 2 (g6pc), inosine-5-monophosphate dehydrogenase 1 (impdh), Brahma chromatin remodeling complex subunit osa (osa), runt-related transcription factor 1 (runx) and frizzled-5 (frizzled), were independently validated using RT-qPCR (Fig.1d; Supplementary Fig.3). Notably, the overall expression levels of these genes in hemocyte-enriched samples from partially fed ticks were intermediate compared to those in unfed and engorged ticks, suggesting a transitional phenotype. Collectively, this dataset indicated that I. scapularis hemocytes exhibit a dynamic genetic program during hematophagy.

To uncover whether the transcriptional changes observed through bulk RNA-seq are accompanied with heterogeneity among hemocytes, we performed scRNA-seq. We collected hemocyte-enriched samples from (1) unfed nymphs, (2) engorged nymphs fed on uninfected mice, and engorged nymphs fed on mice infected with either (3) the rickettsial pathogen A. phagocytophilum or (4) the Lyme disease spirochete B. burgdorferi. After stringent quality controls, we profiled a total of 20,432 cells (unfed = 4630; engorged uninfected = 6000; engorged A. phagocytophilum-infected = 6287; engorged B. burgdorferi-infected = 3515), with a median of 744 unique molecular identifiers (UMIs), 261 genes and 6.2% of mitochondrial transcripts per cell across conditions (Supplementary Fig.4). Consistent with the bulk RNA-seq results, the principal component analysis (PCA) identified distinct distributions between unfed and fed conditions, reinforcing the notion that significant cellular and/or transcriptional changes occur following a blood meal (Supplementary Fig.5).

Following unsupervised clustering, we identified seven clusters in unfed ticks and thirten clusters in engorged nymphs (Supplementary Fig.6). Based on similarities in marker gene profiles, two clusters in the unfed and two clusters in the engorged datasets were merged (Supplementary Data3, 4). Thus, six and twelve clusters remained, respectively, with each cluster expressing a unique set of cell type-defining genes (Fig.2ac, Supplementary Data57). One cluster in each dataset showed high expression of gut-associated genes cathepsinB, cathepsinD and boophilinH2 (Fig.2ac, Supplementary Data57) and two additional clusters in the engorged dataset had gene expression profiles indicative of cuticle and salivary glands tissues (Fig.2ac, Supplementary Data5 and 7). Therefore, these clusters were excluded from subsequent analysis. We assigned putative functions to the top 50 DEGs per cluster using information for tick genes in VectorBase, the sequence homology to D. melanogaster in FlyBase and the presence of functionally annotated protein domains in InterPro (Fig.2ce, Supplementary Data6, 7). We also performed functional enrichment analysis based on gene ontology (GO) using the entire list of DEGs for each cluster (Supplementary Data8, 9). Altogether, we defined five hemocyte clusters from unfed and nine clusters in engorged I. scapularis nymphs.

Hemocyte-enriched samples pooled from individual unfed (n=90) or engorged uninfected, A. phagocytophilum- or B. burgdorferi-infected I. scapularis ticks (n=50 for each) were collected immediately post-detachment from the host. t-Distributed Stochastic Neighbor Embedding (t-SNE) plot clustering of samples collected from (a) unfed (4,630 cells) and (b) engorged (15,802 cells) nymphs. The engorged t-SNE includes cells from uninfected (6000 cells), A. phagocytophilum-infected (6287 cells) and B. burgdorferi-infected (3515 cells) ticks. c Dot plot of the top 5 marker genes present in clusters from engorged ticks based on average gene expression. Color intensity demarks gene expression level, while the size of the dot indicates the percentage of cells within individual clusters expressing the corresponding gene. d Heatmap depicting the expression of marker genes for hemocyte subtypes from engorged ticks. The top 20 marker genes per cluster based on gene expression are included, with representative genes highlighted. e The top 50 marker genes from each hemocyte cluster were manually annotated using the VectorBase, FlyBase, and UniProt publicly available databases. The percentage of predicted functional categories, such as ncRNA/pseudogenes (yellow), protein synthesis (black), secreted/extracellular matrix (blue), unknown (orange), actin/cell rearrangement (brown), detoxification (white), cell proliferation/differentiation (grey), metabolism (green), hormone-related (purple), and immunity (red) are shown. f Pseudotime analysis using slingshot defined six hemocyte lineages (indicated by arrows) in engorged ticks. Tick images in (a, b) were created with BioRender.com.

The molecular features and differentiation process of I. scapularis hemocytes is currently unknown. Thus, based on GO enrichment and marker gene profiles, we were able to characterize clusters of hemocytes shared by both unfed and engorged ticks (Supplementary Figs.7, 8, Supplementary Data57). The Immune 1 cluster showed high expression of genes related to phagocytosis or cytoskeleton organization; coagulation and agglutination functions, such as lectins (hemocytin, techylectin-5A), chitin-binding and clotting proteins; and secreted proteins related to immunity, such as astakine, microplusin, mucins and cystatin domain peptides (Fig.2ce, Supplementary Data57). The Immune 2 cluster displayed genes encoding secreted proteins involved in immunity, such as antimicrobial peptides (AMPs) and clotting related peptides (Fig.2ce, Supplementary Data57). The Proliferative 1 cluster was enriched with mitochondrial genes, characteristic of stem cells in ancient arthropods, such as crayfish20. This cell cluster also had high expression of genes related to actin polymerization, cell proliferation and differentiation (Fig.2ce, Supplementary Data57). The Proliferative 2 cluster displayed a high percentage of transcription factors, RNA binding proteins and genes related to actin dynamics. Marker genes for this cluster included several genes involved in hormone-related responses, suggesting they may be responsive to ecdysteroids synthesized after a blood meal (Fig.2ce, Supplementary Data57). Lastly, both datasets had a Transitional cluster indicative of intermediate subtypes (Fig.2ce, Supplementary Data57).

Four hemocyte clusters were only observed in engorged I. scapularis ticks. The Immune 3 cluster displayed an enrichment in secreted proteins and genes related to immune functions. This cluster was enriched for chitinases, matrix and zinc metalloproteinases, peptidases, and actin binding proteins, which have roles related to wound healing or tissue rearrangement. Several glycine-rich proteins (GRPs), commonly associated with antimicrobial properties or structural proteins, were also present (Fig.2ce, Supplementary Data5, 7). The Immune 4 cluster showed an overrepresentation of genes related to protein degradation, immune function, and cell proliferation (Fig.2ce, Supplementary Data5, 7). Thus, we posit that the Immune 4 cluster represents an intermediate state between the Immune 2 and the Proliferative 2 clusters. Two clusters displayed an enrichment for genes related to metabolic functions. The Metabolism 1 cluster represented genes involved in sulfonation of proteins, lipids and glycosaminoglycans, transmembrane solute transporter, nucleotide, and protein metabolism (Fig.2ce, Supplementary Data5, 7). The Metabolism 2 cluster displayed genes related to detoxification, histamine binding, lipid metabolism, methionine and juvenile hormone metabolism (Fig.2ce, Supplementary Data5, 7).

Based on these findings, we predicted hemocyte ontogeny using pseudotime analysis, which orders hemocyte clusters based on their gene expression profiles, enabling us to infer developmental lineages (Fig.2f)21. We found six trajectories considering the cluster Proliferative 1 as a stem cell-like subpopulation. Lineage 1 and 2 ended with the Immune 2 and Proliferative 2 clusters, respectively, with the Immune 4 cluster serving as an intermediate state. Lineages 3 and 4 gave rise to the Immune 3 and Immune 1 clusters, respectively. Finally, two lineage trajectories ended with the metabolic clusters, Metabolism 1 and Metabolism 2. Overall, our findings suggest the presence of an oligopotent subpopulation that differentiates into more specialized subtypes involved in immune and metabolic functions, a process evoked by hematophagy.

The impact of bacterial infection on subtypes of tick hemocytes remains elusive. Thus, we collected hemocytes from I. scapularis nymphs fed on uninfected, A. phagocytophilum- or B. burgdorferi-infected mice and determined morphotype percentages. During infection with the rickettsial agent A. phagocytophilum, a relative decrease in prohemocytes and increase in plasmatocytes was noted (Fig.3a). Conversely, only a slight decrease in the proportion of prohemocytes was observed during infection with the Lyme disease spirochete B. burgdorferi (Fig.3a). No difference in total hemocyte numbers was observed across infection conditions (Supplementary Fig.9). However, partitioning the engorged scRNA-seq datasets by treatments revealed a reduction in the Transitional cluster with an expansion in the Metabolism 1 cluster during B. burgdorferi infection (Fig.3b, c). We next analyzed transcriptional changes at the cellular level in engorged uninfected nymphs compared to engorged infected ticks. Hemocyte clusters were grouped according to three molecular programs: Immune (Immune 1-4), Proliferative (Proliferative 1-2 and Transitional) and Metabolism (Metabolism 1-2). During A. phagocytophilum infection, we identified 177 DEGs within the Proliferative clusters, 53 DEGs in the Metabolism clusters, and 5 DEGs among the Immune clusters (Fig.3d, Supplementary Data10). Conversely, during B. burgdorferi infection, we detected 244 DEGs within the Proliferative clusters, 81 DEGs among the Metabolism clusters, and 12 DEGs associated with the Immune clusters (Fig.3d, Supplementary Data10). In total, we identified 188 and 257 unique DEGs across all hemocyte clusters during A. phagocytophilum and B. burgdorferi infection, respectively. Notably, 11 genes were differentially expressed during both bacterial infections and shared amongst all cluster groupings, in which 36.4% (4 out of 11) were marker genes of the Immune 1 cluster (Fig.3e). Collectively, our results defined specific hemocyte subpopulations and genes that are differentially expressed in I. scapularis in response to A. phagocytophilum or B. burgdorferi infection.

a Hemocyte morphotypes (prohemocytes, plasmatocytes and granulocytes) in I. scapularis nymphs fed on A. phagocytophilum- (Ap, pink) or B. burgdorferi- (Bb, green) infected mice compared to uninfected [(-), dark blue] (n=16 and 12; n=14 and 14, respectively). Results are presented as meanSD. A minimum of two independent experiments were conducted. Statistical significance was determined using an unpaired two-tailed t test with Welchs correction, and significant p-values (<0.05) are displayed in the figure. b t-Distributed Stochastic Neighbor Embedding (t-SNE) plot clustering of cells collected from the hemolymph of uninfected (6000 cells), A. phagocytophilum- (6287 cells) or B. burgdorferi-infected (3515 cells) I. scapularis nymphs. c Percentage of hemocyte clusters in uninfected, A. phagocytophilum- or B. burgdorferi-infected ticks. d Venn diagram illustrating the number of differentially expressed genes (DEGs) between groups of hemocyte clusters during A. phagocytophilum (top) or B. burgdorferi (bottom) infection compared to uninfected ticks. Hemocyte clusters were categorized into three molecular programs: Immune (Immune 1-4), Proliferative (Proliferative 1-2 and Transitional) and Metabolism (Metabolism 1-2). DEGs were determined using pairwise comparisons against uninfected. e Heatmap representing the average expression patterns of DEGs altered during infection and shared between all 3 cluster groups. For each DEG, the mean average across all experimental conditions was centered at zero for each hemocyte group. The # symbol indicates Immune 1 marker genes. Source data are provided as a Source Data file. Tick images in (b) were created with BioRender.com. (-)=Uninfected. Anaplasma=A. phagocytophilum. Borrelia=B. burgdorferi.

The preceding findings suggested that the Immune 1 hemocyte cluster represented a subpopulation of cells that responded to bacterial infections in ticks (Fig.3d). Therefore, our focus shifted to two marker genes from the Immune 1 hemocyte cluster: hemocytin and astakine (Fig.4a, Supplementary Data6, 7). Hemocytin is homologous to hemolectin in D. melanogaster and von Willebrand factors of mammals22,23. Hemocytin encodes for a large multidomain adhesive protein involved in clotting, microbial agglutination and hemocyte aggregation22,23,24,25. Conversely, astakine is a cytokine-like molecule present in chelicerates and crustaceans and is homologous to vertebrate prokineticins26,27,28,29. Astakine also induces hemocyte proliferation and differentiation of immune cells26,27,28,29. Hemocytin and astakine were broadly expressed in the Immune 1 cluster of I. scapularis hemocytes (Fig.4a). Validating our scRNA-seq results, we confirmed the expression of hemocytin and astakine in hemocyte-enriched samples obtained from unfed ticks using RNA-FISH, illustrating that these genes serve as markers for the Immune 1 cluster (Fig.4b, Supplementary Fig.10). Furthermore, we noted an upregulation of hemocytin and astakine in hemocyte-enriched samples collected from engorged ticks, a pattern also observed in other markers of the Immune 1 cluster (Fig.4c, Supplementary Data11). These findings suggested that blood-feeding expanded this hemocyte subtype and/or upregulated the expression of its marker genes in I. scapularis ticks.

a Expression patterns of hemocytin (left) and astakine (right) on t-Distributed Stochastic Neighbor Embedding (t-SNE) plots of hemocyte-enriched samples from engorged nymphs. Their highest expression is denoted in the Immune 1 cluster (outlined). b RNA FISH image of I. scapularis hemocytes labeled for hemocytin (hmc, green), astakine (astk, red), and nuclei (DAPI). White scale bars indicate a length of 50m. Refer to Supplementary Fig.10 for control images. c RT-qPCR evaluation of hemocytin (hmc; n=9 and 6) and astakine (astk; n=9 and 6) expression in hemocyte-enriched samples collected from unfed (ivory) or engorged (dark blue) ticks (with 4080 pooled ticks per sample). di Ticks were subjected to microinjection with clodronate (CLD) or empty liposomes (Control) and subsequently fed on either (d-f, hi) uninfected or (g) A. phagocytophilum-infected mice. d Total hemocyte counts (n=9 and 9) and (e) morphotype percentages (prohemocytes, plasmatocytes and granulocytes; n=8 and 9 in all cases) were assessed in the hemolymph of individual ticks. f RT-qPCR analysis of hemocytin (hmc; n=18 and 20) and astakine (astk; n=18 and 20) expression, and (g) A. phagocytophilum load in individual ticks (n=28 and 20). Bacterial quantification was based on the expression of A. phagocytophilum 16s rRNA (Ap16S) gene. h Weight measurements of engorged nymphs (n=34 and 25). i Percentage of nymphs that molted to adults. Results are represented as (ch) meanSD or as (i) a percentage from the total. A minimum of two independent experiments were conducted. Statistical significance was evaluated by (cf) an unpaired two-tailed t test with Welchs correction, (gh) two-tailed MannWhitney U test or (i) by a Fisher exact test, and significant p values (<0.05) are displayed in the figure. Source data are provided as a Source Data file.

Hemolectin is a known marker of phagocytic plasmatocytes in Drosophila30,31,32, and phagocytic hemocytes expressing hemocytin are present in crustaceans20,33,34. Therefore, we explored the phagocytic potential of the Immune 1 cluster by employing clodronate liposomes (CLD), which have been used to deplete phagocytic hemocytes in flies, mosquitoes, and, more recently, ticks35,36,37,38. We found that CLD treatment led to a 36% reduction in the total number of hemocytes in I. scapularis nymphs (Fig.4d). We observed an increase in the proportion of granulocytes along with a decrease in the percentages of prohemocytes and plasmatocytes compared to ticks treated with empty liposomes (Fig.4e). Interestingly, the expression of both Immune 1 marker genes, hemocytin and astakine, also decreased after CLD treatment, implying the Immune 1 cluster is phagocytic (Fig.4f). Taken together, CLD treatment effectively reduced the number of phagocytic hemocytes in I. scapularis, particularly impacting prohemocytes, plasmatocytes and cells associated with the Immune 1 cluster.

Phagocytosis serves as a pivotal immune mechanism against invading microbes. Previous studies have demonstrated that the depletion of phagocytic immune cells can influence the survival of arthropods following infection with either Gram-positive or Gram-negative bacteria35,36,37,38. However, the impact of depleting phagocytic hemocytes on the acquisition of tick-borne bacteria remains unclear. Thus, we further investigated the effects of CLD treatment during A. phagocytophilum infection, as this intracellular microbe is known to interact with I. scapularis hemocytes shortly after uptake39. Our findings revealed a reduction in A. phagocytophilum load in engorged ticks after injection with CLD compared to controls (Fig.4g), with no differences in tick attachment (Supplementary Fig.11), suggesting that phagocytic hemocytes promote either the acquisition or proliferation of A. phagocytophilum during blood-feeding.

Reports on Dipteran model organisms have demonstrated that hemocytes play roles beyond immunity, including tissue communication, clearing apoptotic cells during molting and development, and serving as vehicles for molecules40,41,42,43,44. However, whether hemocytes have non-immune roles in ticks remains unknown. Our sequencing results revealed an enrichment in functions related to metabolism, cell proliferation, and development after a blood meal, suggesting that hemocytes may participate in the feeding or molting processes in ticks (Figs.1, 2). To investigate this further, we recorded these physiological processes after injecting ticks with CLD. Although no differences in attachment were observed (Supplementary Fig.11), engorged ticks treated with CLD weighed significantly less compared to the control treatment, indicating that phagocytic hemocytes are required for proper hematophagy (Fig.4h). Furthermore, the number of nymphs that successfully molted to adults was significantly lower in CLD-treated ticks (Fig.4i). Collectively, these results indicated that phagocytic hemocytes pleiotropically impacted tick immunity, feeding, and molting in I. scapularis. Importantly, the observed correlation between changes in physiological parameters and the reduced expression of the Immune 1 markers, hemocytin and astakine, during CLD treatment suggested a potential regulatory role for these genes in various aspects of tick physiology, prompting us to explore this hypothesis further.

We found that hematophagy induces hemocyte proliferation and differentiation (Fig.1b) while upregulating astakine expression (Fig.4c). Thus, we investigated whether astakine was directly implicated in the proliferation or differentiation of I. scapularis hemocytes. We microinjected increasing amounts of recombinant astakine (rAstk) in unfed nymphs and observed a dose-dependent increase in the total number of hemocytes (Fig.5a). Specifically, we measured a decrease in the percentage of prohemocytes and an increase in plasmatocytes (Fig.5b). These findings matched alterations observed during normal blood-feeding (Fig.1b). Interestingly, we also detected an increase in tick IDE12 cell numbers in vitro following treatment with rAstk, supporting a role of astakine in inducing cell proliferation in hemocyte-like cells (Supplementary Fig.12). To corroborate these results, we then silenced astakine by microinjecting unfed nymphs with small-interfering RNA (siRNA) before feeding on uninfected mice. We utilized siRNA to knockdown gene expression, as genome editing through CRISPR has only recently been introduced for adult germline manipulation and possesses low efficiency in ticks45. We recovered 41% less hemocytes and observed an increase in the percentage of prohemocytes with a decrease in plasmatocytes from engorged ticks when astakine was silenced (Fig.5ce). Therefore, we determined that astakine acts on hematopoietic processes in the ectoparasite I. scapularis.

a Total hemocyte counts in the hemolymph of unfed I. scapularis nymphs subjected to microinjection with increasing amounts of rAstk (orange) or BSA (grey) as a control (n=21, 14, 14 and 24). b Percentage of hemocyte morphotypes (prohemocytes, plasmatocytes and granulocytes; n=10 and 10 in all cases) in the hemolymph of unfed nymphs following microinjection with 5ng rAstk (orange) compared to BSA controls (grey). ch Ticks were subjected to microinjection with astk siRNA (si-astk; blue) or scrambled RNA (sc-astk; grey) and subsequently fed on either (cg) uninfected or (h) A. phagocytophilum-infected mice. c Efficacy of astk silencing (n=20 and 16), (d) total number of hemocytes (n=9 and 8) and (e) percentage of hemocyte morphotypes in individual ticks (n=12 and 10 in all cases). f Weight measurements of engorged nymphs (n=29 and 19). g Percentage of nymphs that molted to adults. h RT-qPCR assessment of astk silencing efficiency (n=16 and 15) and A. phagocytophilum load (n=18 and 14) in individual infected ticks. Bacterial quantification was based on the expression of A. phagocytophilum 16s rRNA (Ap16S) gene. Results are represented as (af, h) meanSD or as (g) a percentage from the total. A minimum of two independent experiments were conducted. Statistical significance was evaluated by (a) one-way ANOVA with Dunnetts multiple comparisons test; (bf, h) an unpaired two-tailed t test with Welchs correction or (g) by a Fisher exact test, and significant p-values (<0.05) are displayed in the figure. Source data are provided as a Source Data file. rAstk = recombinant astakine; BSA = bovine serum albumin.

Our previous results showed alterations in the percentage of hemocytes by the CLD treatment, which influenced bacterial infection and tick physiology. Given that astakine also alters hemocyte composition in ticks, we then investigated whether decreasing the expression of astakine affects A. phagocytophilum infection, hematophagy or ecdysis in I. scapularis. Accordingly, we observed a significant reduction in weight, molting success, and A. phagocytophilum burden in ticks silenced for astakine compared to the control treatment, without differences in tick attachment (Fig.5fh; Supplementary Fig.13). Corroborating these findings, A. phagocytophilum load was also lower in astakine-silenced tick cells in vitro (Supplementary Fig.14). Overall, we demonstrated that astakine regulates hemocyte composition in I. scapularis, which affects not only bacterial infection but also feeding and ecdysis, supporting a pleiotropic role of hemocytes in ticks.

The broad expression of hemocytin detected in I. scapularis hemocytes (Fig.4a), and its homology to a Drosophila gene used as a marker of phagocytic immune cells, prompted us to explore whether hemocytin could be involved in hemocyte proliferation or differentiation, phagocytosis or immune signaling in ticks. Surprisingly, no differences were observed in hemocyte numbers or subtype proportions between ticks injected with siRNA targeting hemocytin, nor in the phagocytic capacity of hemocytin-silenced tick cells (Supplementary Figs.15, 16). We then asked whether hemocytin could act as an immune pathway regulator in I. scapularis. We focused on the IMD and the c-Jun N-terminal kinase (JNK) pathways, as previous reports indicated the importance of these molecular networks during bacterial infection of ticks5,6.

After transfection with siRNA targeting hemocytin, we found a decrease in JNK phosphorylation in hemocytin-silenced tick cells, without alteration in Relish cleavage (Fig.6a, b). To complement our findings, we overexpressed hemocytin in tick cells through CRISPR activation (CRISPRa). CRISPRa has been widely used to enhance the expression of an endogenous locus, employing a catalytically inactive Cas9 (dCas9) fused with transcription activators and single guide RNAs (sgRNAs) that direct the modified enzyme to the promoter region of a gene of interest46. However, so far none of the CRISPRa effectors have been tested in tick cell lines and no endogenous promoter has been identified for genetic expression of sgRNAs. We optimized reagents and developed a protocol to up-regulate hemocytin in ISE6 cells using two rounds of nucleofection with different expression plasmids. The first plasmid expressed dCas9-VPR paired with neomycin resistance under the control of the CMV promoter. The second plasmid expressed either a sgRNA specific to the promoter region of hemocytin (hmc-sgRNA) or a control sgRNA (ctrl-sgRNA) driven by an endogenous RNA polymerase III promoter (Fig.6c). Tick ISE6 cells were used as a platform for CRISPRa given the lower expression of hemocytin compared to IDE12 cells (Supplementary Fig.17). Strikingly, we detected a 277% increase in the expression of hemocytin in dCas9+ cells transfected with the hmc-sgRNA compared to dCas9+ cells transfected with the ctrl-sgRNA (Fig.6d). Importantly, we also noticed elevated levels of both jun (the transcription factor for the JNK pathway) and jnk expression (Fig.6d).

a, b Tick cells were transfected with either hmc siRNA (si-hmc) or scrambled RNA (sc-hmc). a Efficiency of hmc silencing in IDE12 cells (n=12 and 11). b Representative western blot (left) of N-Rel and p-JNK during treatment with sc-hmc (lane 1) or si-hmc (lane 2). N-Rel and p-JNK protein expression was quantified (right) in si-hmc (blue) or sc-hmc (grey) IDE12 cells (n=4). Data were normalized to the scrambled control, with N-Rel values relative to Actin and p-JNK values to JNK. A representative blot from four experiments is shown. Uncropped blots containing the molecular weight markers are supplied in the Source data. c Overview of CRISPRa-mediated hmc overexpression in ISE6 cells. d RT-qPCR analysis of hmc (left; n=9 and 10), jnk (middle; n=9 and 9) and jun (right; n=10 and 10) expression in dCas9+ ISE6 cells transfected with either a single guide RNA (sgRNA) specific to the promoter region of hemocytin (hmc-sgRNA, blue) or a random sgRNA (ctrl-sgRNA, grey). eh Ticks were subjected to microinjection with hmc siRNA (si-hmc; blue) or scrambled siRNA (sc-hmc; grey) and subsequently fed on either (eg) uninfected or (h) A. phagocytophilum-infected mice. e RT-qPCR analysis of hmc (left; n=19 and 18), relish (middle; n=17 and 18) and jun (right; n=17 and 18) expression in engorged ticks. f Weight measurements of engorged nymphs (n=20 and 23). g Percentage of nymphs that molted to adults. h RT-qPCR assessment of hmc silencing efficiency (n=11 and 10) and A. phagocytophilum load (n=11 and 10) in individual infected ticks. Bacterial quantification was based on the expression of A. phagocytophilum 16s rRNA (Ap16S) gene. Results are represented as meanSD. A minimum of two independent experiments were performed. Statistical significance was evaluated by (a, b, d, e) an unpaired two-tailed t-test with Welchs correction; (f, h) a two-tailed MannWhitney U test or (g) by a Fisher exact test, and significant p-values (<0.05) are displayed in the figure. Source data are provided as a Source Data file. N-Rel = cleaved Relish; p-JNK = phosphorylated JNK; JNK = c-Jun N-terminal kinase.

To corroborate our results in vivo, we microinjected unfed ticks with a scrambled control or siRNA targeting hemocytin and allowed them to feed on nave mice. Upon repletion, we measured the expression of jun and relish. Consistently, we found that reduction in hemocytin expression led to a decrease in jun levels in ticks, without affecting relish expression (Fig.6e), uncovering a role for hemocytin in the activation of the JNK pathway in I. scapularis.

Finally, building on our previous findings demonstrating that blood-feeding and infection alter the expression of hemocytin in ticks (Fig.4c, Supplementary Data9), coupled with the documented roles of hemocytin as an agglutinating factor and the JNK pathway in enhancing organismal growth and metabolism in insects47, we delved deeper into the relationship between hemocytin expression and physiological parameters in I. scapularis. We detected a decrease in weight and molting success to adulthood in hemocytin-silenced ticks that fed on uninfected mice compared to scrambled controls, with no differences in tick attachment (Fig.6f, g; Supplementary Fig.18). Additionally, we noted that silencing hemocytin increased A. phagocytophilum load both in vivo and in vitro (Fig.6h; Supplementary Fig.19), supporting an antimicrobial role for hemocytin. Collectively, our findings indicated that tick hemocyte subtypes and their associated marker genes play a critical role in I. scapularis immunophysiology.

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Tick hemocytes have a pleiotropic role in microbial infection and arthropod fitness - Nature.com

A competition described as a "global fitness race" is building up a mass following among UK gym-goers.

Hyrox, which combines running with a full-body workout, is the latest fitness craze among those who want to "go one better" than a Parkrun but have had enough of "hardcore obstacle courses" like Tough Mudder, said health writer Peta Bee in The Times. And the hype about the exercise challenge is "huge", in part because the target market isn't triathlon or marathon types, but rather "regular gym-goers seeking an outlet for a previously untapped competitive streak".

Launched in 2017 by German former athlete Christin Ttzke, Hyrox has gained a global following, said the BBC, with a "1,000% increase in participation in the past five years".

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The premise of the race is simple. Participants run 1km and then complete a functional fitness exercise. "Rinse and repeat eight times, and you have yourself a Hyrox race," said GQ.

The exercises comprise a 1km sprint on a SkiErg, a full-body cardio machine; a 50m weighted sled push; a 50m sledge pull; 80m of burpee broad jumps; 1,000m of rowing; a 200m farmer's carry (using a kettlebell); 100m of sandbag lunges; and 100 wall ball throws.

There are no qualification entry requirements or finishing time restrictions. And each Hyrox race consists of waves of starters every 10 minutes throughout the day, so "there's no fear of finishing in 'last' place", said GQ. It is an "own-pace, own-race event" where amateurs and pros take part side by side.

More than 175,000 people are due to take part in 65 Hyrox races this year, some of which are in major arenas, such as Birmingham's NEC and London ExCel.Entries to a race in May at London Olympia "sold out months ago", said Bee in The Times, with some 12,500 people expected to take part.

Hyrox founder Ttzke has even bigger ambitions for the competition, telling The Times that he wanted to see it "become the gym-goer's equivalent of the London Marathon for runners".

The hashtag '#hyroxlondon' has garnered "millions of views on TikTok and tens of thousands of Instagram posts", said the BBC.Along with social media, another key reason why Hyrox is popular is that people don't have to compete alone; many are in pairs or teams of four.

It is also within the realms of possibility for most regular gym-goers. The functional stations "aren't highly technical and don't require brute strength but are pitched so that even people with little circuit training experience can have a stab at them", said Bee.

The race has become particularly popular with 35- to 39-year-olds, with the average participant at the latest London Hyrox race aged 37. But there is no upper age limit to participate Hyrox said its oldest finisher was 74 "and still going strong".

Hyrox classes are also increasing in popularity as more gym-goers become interested in participating in the races, as well as the opportunity the classes offer to train in groups while using a range of equipment.

The Gym Group first offered Hyrox classes in March last year at one of its venues and has since expanded into 14 gyms across London. One of the chain's trainers, Jenni Tardiff, told the BBC that Hyrox classes "really quickly became the most popular class in the gym".

"It started off with members who maybe knew about Hyrox and then it just exploded into everybody," she said.

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Hyrox: the new fitness trend taking over gyms - The Week

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