sorry for the absence lately, i have been yucky.
Any IBS sufferers out there? yuk! and then add on top of that passing out, dizziness and a whole load of tiredness and thats been me this week.
However, i received an email that made my day/week/month.
Next Bloggers Network awarded me Blogger of the month… How fab! I get this cute badge to put on my blog and they did an interview avec moi and put it on their websites to see.. if you fancy a read about me babbling about blogging its here.
I also had an Email from Modcloth to say Congrats! I've been picked as one of the Style Ambassadors… we've been chatting on facebook ever since! Like minded communities are so much fun!
So as you can see this lot has cheered me up lots and lots this week!
This is what we wore on our girlie shopping day, it was really lovely. She's growing up so fast! And its nice to have girl company.
So folks thats all my news for now, i am off to London this weekend with all the kids (wish me luck on the 4 hour car journey!) to visit the in laws. I'm trying to pack light as packing for 5 is TRICKY!! especially with limited room. I'm gunna try and take as little as possible and remix.. will keep you posted!
Congrats Rachel! Well deserved lovely lady!
Congrats! Congrats! Congrats! That's so awesome!!!
I am so happy for you! Congrats lady! You deserve it.
What a cute outfit! I hope you feel better soon.
Congratulations. I'm not surprised you mom was proud.
Hope you are on the mend soon hun.


text:
When a Camera's Frame Rate is Synced to a Helicopter's Rotor…
By NY - NewsRoom on March 5, 2017
YouTuber Chris Chris captured the above video showing what happens when your camera's frame rate is perfectly synced to the rotation speed of a helicopter's rotor: the blades are frozen at the same angles in each frame, making it look like the helicopter is magically floating around with frozen rotor blades.
40 years ago, Bob Khoury and Warren Steinberg started selling used photo equipment out of a showcase in an Atlanta, Georgia, flea market. Soon they moved to a brick and mortar store which, to incorporate their earlier experience, they called Showcase. The store grew to be the largest in Atlanta and sold photo and video equipment to amateurs and professionals alike and last year they celebrated their 40th anniversary.
Here's an interesting photography first that is set to happen in 2017: scientists are planning to capture the first ever photo of a black hole's event horizon (the boundary of no return that light can't event escape).
Things aren't looking good for the standalone point-and-shoot camera. As smartphone cameras continue to improve, compact camera sales continue to nosedive. A new historical sales chart with 2016 figures shows just how quickly point-and-shoots are dying off.
It's understandable that the great unwashed masses of the larger population might not appreciate contemporary art. But you'd think that photographers, who are creatives in their own right, would appreciate the art and creativity of others in all of its various forms. What I've seen instead is that, when it comes to much contemporary art, most (but not all!) photographers tend to dismiss the work outright.
Lightning is an amazing subject to try and photograph. Dazzling. Unpredictable. Fulfilling. I've been documenting the long arm of Zeus for more than two decades and still love it. First using transparencies and negatives, then digital. There are many ways to be creative when it comes to photographing lightning.
The photographer brand COOPH made this 3-minute video in which photographer Lorenz Holder shares the interesting story behind how he shot a photo that won the prestigious photo contest Red Bull Illume 2016.
I recently rediscovered an old photography technique that allows you to add surreal color to photos that show movement in the frame. The technique seems to be new in the digital domain, but the technique itself has been known since the early era of digital photography.
About a year ago, I was asked if I would like to do a wedding photography at night. The bride Erika had seen a photo of a moon halo I took earlier that year. In that photo, I had two friends that I ran into that night. They were out chasing the northern lights and I asked if I could take a photo of them together with the moon halo.
For landscape photographers, getting your entire scene in focus while keeping things as sharp as possible at the same time can be a challenge.
But if you follow the simple technique laid out by photographer Joshua Cripps in the tutorial above, as he puts it, it becomes "as easy as manually removing a corn syrup-based artificially-flavored confectionary product, from the infantile grasp of a newborn Homo sapien."
Since the introduction of the Fujifilm X-Series line of cameras, reviewers and consumers have struggled to compare them directly to the competition. Fujifilm's is a tightly integrated system, wherein everything is a little bit different.
I initially refused to believe it when this photo came across my feed. My eyes aren't broken! I can see they're strawberries, and they're definitely red. They have to be trolling us with this image, right?
Lomography is selling a new limited edition film called Color Negative F²/400. It's a film with an unusual concept and backstory: it was aged like wine in oak casks for 7 years.
I'm a firm believer that photography is a game of inches. So today I'll share with you what I've learned about mastering autofocus shooting in a variety of difficult situations.
In 2015, photographer David Gaberle walked over 2,200 miles (3,600 kilometers) through some of the world's most metropolitan areas, photographing people in cities such as New York, Tokyo, Hong Kong, Shanghai, Sydney, London and Seoul. He's now turning this project into a book titled Metropolight.
Last year, journalist Esther Honig published a viral series of images showing how photo retouchers in 27 countries around the world "enhanced" a portrait of her according to their cultural preferences. Inspired by that project, the UK medical website Superdrug Online Doctor just published a similar experiment that explores body image.
They say a picture is worth a thousand words. But I wonder, what else do "they" say? In order to find out I've culled together the best quotes on the subject of photography. I hope they inspire you.
Here's an old-ish video that's been making the rounds again lately (viral videos are like viruses — they don't go away very easily). Titled "Camera shutter speed synchronized with helicopter blade frequency," it shows what can happen when your camera is synchronized with the RPM of a helicopter's rotor blades. The resulting footage makes the helicopter look as though it's just floating in the air!
Snapchat is known for popularizing the idea of the self-destructing photo, but did you know that long before Snapchat existed, Polaroid had already offered its own line of self-destructing Polaroid pictures? It was called the Fade to Black line.
I recently wrote an article about 8 Reasons You Should Buy A 50mm f/1.8 Lens and one part was about using it with the "El Bokeh Wall." What's an El Bokeh Wall? It's using some aluminum foil to add beautiful bokeh to the background of your photos. Here's a full tutorial on the technique.

redpajama_set_name: RedPajamaCommonCrawl


The general structure of teeth is similar across the vertebrates, although there is considerable variation in their form and position. The teeth of mammals have deep roots, and this pattern is also found in some fish, and in crocodilians.
In most teleost fish, however, the teeth are attached to the outer surface of the bone, while in lizards they are attached to the inner surface of the jaw just by one side. In cartilaginous fish, such as sharks, the teeth are attached by tough ligaments to the hoops of cartilage that form the jaw.
Teeth are among the most distinctive (and long-lasting) features of mammals. Paleontologists use teeth to identify fossil species and determine their relationships. The shape of the animal's teeth are related to its diet. For example, plant matter is very hard to digest, so herbivores have many molars for chewing and grinding. Carnivores, on the other hand, need canines to kill prey and to tear meat.
The term "fish" most precisely describes any non-tetrapod craniate (i.e. an animal with a skull and in most cases a backbone) that has gills throughout life and whose limbs, if any, are in the shape of fins. Unlike groupings such as birds or mammals, fish are not a single clade but a paraphyletic collection of taxa, including hagfishes, lampreys, sharks and rays, ray-finned fish, coelacanths, and lungfish. Indeed, lungfish and coelacanths are closer relatives of tetrapods (such as mammals, birds, amphibians, etc.) than of other fish such as ray-finned fish or sharks, so the last common ancestor of all fish is also an ancestor to tetrapods. As paraphyletic groups are no longer recognised in modern systematic biology, the use of the term "fish" as a biological group must be avoided.
Many types of aquatic animals commonly referred to as "fish" are not fish in the sense given above; examples include shellfish, cuttlefish, starfish, crayfish and jellyfish. In earlier times, even biologists did not make a distinction – sixteenth century natural historians classified also seals, whales, amphibians, crocodiles, even hippopotamuses, as well as a host of aquatic invertebrates, as fish. However, according the definition above, all mammals, including cetaceans like whales and dolphins, are not fish. In some contexts, especially in aquaculture, the true fish are referred to as finfish (or fin fish) to distinguish them from these other animals.


text:
Sharon Nicholson Harrell, DDS
As the founding director of FirstHealth Dental Care Centers, Sharon Nicholson Harrell, DDS, is fulfilling a goal she set for herself in undergraduate school: to care for underserved populations. As an AHEC preceptor, she is encouraging future graduates to do the same.
FirstHealth is a regional dental office with three sites established to provide comprehensive dental care for low-income children up to age 18: Southern Pines in Moore County and Raeford in Hoke County (both Southern Regional AHEC counties) and Troy in Montgomery, a Greensboro AHEC county. Because most local practitioners did not take children or Medicaid patients, the largest unmet need among low-income children in the three counties prior to 1997 was dental care, Harrell explained. "There were many kids with nowhere to go." Local practitioners agreed that FirstHealth should take the lead, and the first site, Southern Pines, opened its doors in October of 1998.
Harrell and her staff precept two to three dental students per year. The students do cleanings, fillings, or restorative care; extractions, patient education, or parent education; and preventive care. "The students get to rotate through several doctors," she said, "giving them a chance to experience different philosophies." Harrell limits the number of students she takes at a time in order to spend quality time with each one. "Several students have come back to the area—some to work with us or to work in private practice and fill in here. It's a win-win for everyone"
Students run the gamut in their range of abilities and comfort level with children, Harrell explained. "Many of those who come to us are not aware of the challenges that lower-income patients face. Two eye-openers for them are patients who don't have phones and who lack transportation and have to come when others can take them." These and other challenges drive home the importance of dental education. Harrell added that other staff members are just as critical in providing students with real-world experiences, from learning about practice scheduling from the receptionists to working chair-side with assistants and hygienists.
"We have three goals for our students. One, to increase competency in pediatric dental procedures. Two, to increase speed; in dental school, students spend three hours with one patient. Here, the students see one patient per hour—that's a three-fold jump. And the third goal is to increase knowledge of public health practice."
A preceptor since 1992, Harrell has been involved with NC AHEC in a number of ways. Her first encounter was as a young provider taking dental education classes at Southern Regional AHEC in 1991. "I had just come from a residency in Maryland and was impressed that I didn't have to to far for quality continuing education," she recalled. She served on the Southern Regional Board of Directors for a three-year term; then, in 2001, she chaired the executive director search committee, which resulted in the selection of Deborah Teasley, PhD, as president and CEO of Southern Regional AHEC.
Harrell's interest in dentistry started in the tenth grade when she began wearing braces on her teeth. She found the instruments on the tray, the rubber bands, and the entire process fascinating. With the unwavering encouragement and support of her parents, both factory laborers from Rockingham, she entered the University of North Carolina at Chapel Hill with a goal of eventually becoming an orthodontist. But in time she realized her true interest lay in public health and in providing a broader range of procedures to underserved populations. She switched to general dentistry while in dental school at UNC. Harrell also received an MPH from UNC and a certificate in advanced general dentistry from the University of Maryland.
The eldest of three sisters, each of whom are professional women, Harrell credits her parents as being her dominant early influence, as well as the driving force behind her twin values of education and hard work. "When I was coming of age in the '70s, my father instilled a can-do attitude so that even without a role model in town, I could achieve my goal of becoming a dentist. He taught us that there had never before been so many opportunities open to young African American females—the sky was the limit." Now a role model herself, Harrell takes that responsibility to heart.
"It's very important to be role models for kids today so that they can see others doing what they aspire to do. I like the opportunity to expose students to real-world dentistry, and I like teaching. The preceptor role allows me to connect with students. It challenges me and keeps me very current. What keeps me motivated is being able to see the progression of students who are more prepared when they enter the real world. We enjoy playing a role in making future graduates feel comfortable in taking care of pediatric patients. We hope their exposure to underserved populations will make them more accepting in their own offices."


text:
Q: What is the Rust equivalent of C++'s shared_from_this? I have an object that I know that is inside an Arc because all the instances are always Arced. I would like to be able to pass a cloned Arc of myself in a function call. The thing I am calling will call me back later on other threads.
In C++, there is a standard mixin called enable_shared_from_this. It enables me to do exactly this
class Bus : public std::enable_shared_from_this<Bus>
{
....
   void SetupDevice(Device device,...)
   {
       device->Attach(shared_from_this());
   }
}

If this object is not under shared_ptr management (the closest C++ has to Arc) then this will fail at run time.
I  cannot find an equivalent.
EDIT:
Here is an example of why its needed. I have a timerqueue library. It allows a client to request an arbitrary closure to be run at some point in the future. The code is run on a dedicated thread. To use it you must pass a closure of the function you want to be executed later.
use std::time::{Duration, Instant};
use timerqueue::*;
use parking_lot::Mutex;
use std::sync::{Arc,Weak};
use std::ops::{DerefMut};

// inline me keeper cos not on github
pub struct MeKeeper<T> {
    them: Mutex<Weak<T>>,
}

impl<T> MeKeeper<T> {
    pub fn new() -> Self {
        Self {
            them: Mutex::new(Weak::new()),
        }
    }
    pub fn save(&self, arc: &Arc<T>) {
        *self.them.lock().deref_mut() = Arc::downgrade(arc);
    }
    pub fn get(&self) -> Arc<T> {
        match self.them.lock().upgrade() {
            Some(arc) => return arc,
            None => unreachable!(),
        }
    }
}
// -----------------------------------

struct Test {
    data:String,
    me: MeKeeper<Self>,
}

impl Test {

    pub fn new() -> Arc<Test>{
        let arc = Arc::new(Self {
            me: MeKeeper::new(),
            data: "Yo".to_string()
        });
        arc.me.save(&arc);
        arc
    }

    fn task(&self) {
        println!("{}", self.data);
    }

    // in real use case the TQ and a ton of other status data is passed in the new call for Test
    // to keep things simple here the 'container' passes tq as an arg

    pub fn do_stuff(&self, tq: &TimerQueue) {
        // stuff includes a async task that must be done in 1 second

        //.....
        let me = self.me.get().clone();
        tq.queue(
            Box::new(move || me.task()),
            "x".to_string(),
            Instant::now() + Duration::from_millis(1000),
        );
    }

}

fn main() {
    // in real case (PDP11 emulator) there is a Bus class owning tons of objects thats
    // alive for the whole duration
    let tq = Arc::new(TimerQueue::new());
    let test = Test::new();
    test.do_stuff(&*tq);
    // just to keep everything alive while we wait
    let mut input = String::new();
    std::io::stdin().read_line(&mut input).unwrap();
}

cargo toml
[package]
name = "tqclient"
version = "0.1.0"
edition = "2018"

# See more keys and their definitions at https://doc.rust-lang.org/cargo/reference/manifest.html

[dependencies]
timerqueue = { git = "https://github.com/pm100/timerqueue.git" }
parking_lot = "0.11"

A: There is no way to go from a &self to the Arc that self is stored in. This is because:

*

*Rust references have additional assumptions compared to C++ references that would make such a conversion undefined behavior.

*Rust's implementation of Arc does not even expose the information necessary to determine whether self is stored in an Arc or not.

Luckily, there is an alternative approach. Instead of creating a &self to the value inside the Arc, and passing that to the method, pass the Arc directly to the method that needs to access it. You can do that like this:
use std::sync::Arc;

struct Shared {
    field: String,
}

impl Shared {
    fn print_field(self: Arc<Self>) {
        let clone: Arc<Shared> = self.clone();

        println!("{}", clone.field);
    }
}

Then the print_field function can only be called on an Shared encapsulated in an Arc.

A: having found that I needed this three times in recent days I decided to stop trying to come up with other designs. Maybe poor data design as far as rust is concerned but I needed it.
Works by changing the new function of the types using it to return an Arc rather than a raw self. All my objects are arced anyway, before they were arced by the caller, now its forced.
mini util library called mekeeper
use parking_lot::Mutex;
use std::sync::{Arc,Weak};
use std::ops::{DerefMut};

pub struct MeKeeper<T> {
    them: Mutex<Weak<T>>,
}

impl<T> MeKeeper<T> {
    pub fn new() -> Self {
        Self {
            them: Mutex::new(Weak::new()),
        }
    }
    pub fn save(&self, arc: &Arc<T>) {
        *self.them.lock().deref_mut() = Arc::downgrade(arc);
    }
    pub fn get(&self) -> Arc<T> {
        match self.them.lock().upgrade() {
            Some(arc) => return arc,
            None => unreachable!(),
        }
    }
}

to use it
pub struct Test {
    me: MeKeeper<Self>,
    foo:i8,
}

impl Test {
    pub fn new() -> Arc<Self> {
        let arc = Arc::new(Test {
            me: MeKeeper::new(),
            foo:42
        });
        arc.me.save(&arc);
        arc
    }
}

now when an instance of Test wants to call a function that requires it to pass in an Arc it does:
 fn nargle(){
     let me = me.get();
     Ooddle::fertang(me,42);// fertang needs an Arc<T>
 }

the weak use is what the shared_from_this does so as to prevent refcount deadlocks, I stole that idea.
The unreachable path is safe because the only place that can call MeKeeper::get is the instance of T (Test here) that owns it and that call can only happen if the T instance is alive. Hence no none return from weak::upgrade

redpajama_set_name: RedPajamaStackExchange


— Do not use them!
" I've been a customer since 2012 and have both the electricity and gas plans. I've retained all mailed correspondence and recently discovered (January 2019) that I had no updated plan agreement or renewal contract information since 2016. In January I received notice that Stream had not charged me the gas supply fee for the 11/15/18 - 12/17/18 period and subsequently, I would be charged for that past period and current period on my next invoice. The double charge "seemed reasonable" based on similar them usage for both periods. While I had no problem paying what I should have been previously charged, it would have been appreciated to have the option for payment options or payment breakouts, considering it was Stream's error. THEN... I received my bill for the 1/17/19 - 2/18/19 period and was SHOCKED at the gas supply charge was no longer fixed, but was now variable, month-to-month at 125% higher rather than my last agreed upon contract. Having had not received ANY documentation to advise of this disparity, I spoke with one representative for 15 minutes, and then a supervisor for 10 minutes with no resolution for any type of remediation.
1. I asked the supervisor to email me all of the correspondence notices that were supposedly sent to me previously, but he advised that they could only be mailed. In this technological age, this is unacceptable.
2. The first representative advised me that I could've renewed my plans via phone or online after my plan expired in 2017. And had I received notification of such options, I most certainly would have done so to maintain the fixed rates that I had had since 2012. (Most consumers opt for stable billing of fixed rates rather than month-to-month variable rates that increased by 125%). There was NO letter or call to confirm what was clearly a drastic change of plan history. Repeating myself twice to the supervisor about NOT receiving any correspondence, and the billing error made by Stream, the call concluded with nothing but an offer to renew for only ONE plan option of 12 months.


text:
Maoberry (Antidesma bunius) ameliorates oxidative stress and inflammation in cardiac tissues of rats fed a high-fat diet
Arunwan Udomkasemsab1,
Chattraya Ngamlerst2,
Poom Adisakwattana3,
Amornrat Aroonnual1,
Rungsunn Tungtrongchitr1 and
Pattaneeya Prangthip1Email authorView ORCID ID profile
BMC Complementary and Alternative Medicine201818:344
Received: 9 August 2018
Backgound
Chronic fat-rich diets consumption is increased risk associated with cardiovascular diseases (CVD). Prevention or reduction the progression of cardiac tissue deterioration could benefit in CVD. This study aimed to examine the effects of maoberry (Antidesma bunius), a antioxidant-rich tropical fruit, supplementation on oxidative stress and inflammation in cardiac tissues of rats fed a high-fat diet (HFD).
The male rats orally received HFD with maoberry extract doses of 0.38, 0.76 or 1.52 g/kg or simvastatin (10 mg/kg) for 12 weeks. At the end of the experimental period, the rats were fasted, euthanized and harvested for the hearts.
Significantly reduced oxidative stress (malondialdehyde levels) and enhanced antioxidant capacity (ferric-reducing activities) in cardiac tissues of the rats were found. Maoberry extract remarkably ameliorated the expressions of genes involved with pro-inflammatory such as the tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), vascular cell adhesion molecule-1 (VCAM-1), monocyte chemoattractant protein-1 (MCP-1) and endothelial nitric oxide synthase (eNOS).
Our findings suggest that maoberry extract has remarkable effects on preventing progression of cardiac tissue deterioration at least through lowering oxidative stress and inflammation.
Cardiac tissue
Maoberry
Cardiovascular diseases (CVD) leading to heart failure are the main cause of mortality worldwide. Recent studies have indicated that a systemic inflammatory process can leads to the malfunctioning of the cardiac endothelium. Various mechanisms such as oxidative stress and inflammation are involved in cardiac pathogenesis. Therefore, the prevention or reduction of cardiac tissue deterioration to prevent CVD progression is considerable interested [1, 2].
Hypercholesterolemia is considered to be the hallmark of early CVD and is usually observed before vascular lesions appear. Previous studies have reported increased rates of reactive oxygen species (ROS) production in patients with hyperlipidaemia [3]. This may be attributed to fat-rich diets that increase the expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase genes [4], which is the key enzyme responsible for ROS production [5], and that contributes to increased ROS formation in cells [6]. Overproduced ROS can react with nitric oxide (NO) to form peroxynitrite (ONOO−), a reactive short-lived peroxide, resulting in the inactivation of endothelial NO synthase (eNOS) [7], which lowers NO production and impairs vasodilatation, both of which can cause endothelial dysfunction [8–10]. Furthermore, oxidative stress contributes to CVD pathogenesis through inflammatory reactions. The lipid oxidation process originates from uncontrolled ROS overproduction, causing necrotic cell death, a major driver of inflammation [11]. In response to inflammation, monocytes and macrophages secrete pro-inflammatory cytokines such as the tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6), stimulate the vasculatures to produce inflammatory mediators such as vascular cell adhesion molecule-1 (VCAM-1) and release chemokines such as monocyte chemoattractant protein-1 (MCP-1). All of these regulate the migration and infiltration of monocytes/macrophages into vascular inflammatory sites [2, 12]. The abundance of ROS causes low density lipoprotein (LDL) oxidation activating inflammatory cytokines, mediators and chemokines from infiltrating and resident macrophages. Both ROS and inflammation play a critical role in CVD occurrence and development [13].
Previous studies examined the beneficial effects of polyphenols, presenting in natural extracts of fruits, vegetables, soy, cocoa, tea and wine, and reported that these compounds can have biological effects on hearts [10, 14, 15]. The consumption of polyphenols as supplements appears to have various health benefits. Polyphenols present in red grape juice and red wine can markedly reduce NADPH oxidase activity and gene expression in human peripheral blood neutrophils and endothelial cells [15]. Patients with diabetes who daily consumed pomegranate as polyphenol supplements for 4 weeks showed a reduction in free radical-induced lipid peroxidation, notably through radical scavenger receptor activities [14]. Moreover, improvements in the regulation of pro-inflammatory molecules such as TNF-α, IL-6, VCAM-1 and MCP-1 [16–18].
Maoberry (Antidesma bunius) is a wild plant naturally growing throughout north-eastern Thailand. Maoberry fruits are very popular and are used in commercial products with healthy nourishment in Thailand. The fruits are considered to possess the ability to cure several ailments such as parched tongue, lack of appetite, indigestion, high blood pressure and diabetes [19, 20], possibly because of high polyphenol levels and antioxidant activity. Antidesma spp. contains several compounds such as phenolics, flavonoids, ascorbic acid and total proanthocyanidin [20–22]. Antidesma spp. also possesses strong antioxidant activity against oxidative damage with a variety of assays [21, 22].
Previously, we proposed maoberry extract benefit on atherogenic risk factors including lipid profiles, inflammation and oxidative stress in bloods [23]. To further understand the beneficial effects of maoberry extract on cardiovascular disease, we examined the effects of the maoberry extract on cardiac tissues in a hypercholesterol animal model. Oxidative stress and the specific molecules involved in endothelial damage were investigated. The results may provide additional data and encourage the application of natural extracts as an alternative for preventing damage in cardiac tissue.
Maoberry preparation
Dark-purple maoberry cultivars (Antidesmabunius spp.) were purchased from local orchard area of Khok Si Suphan District with Geocode of 4715, Sakon Nakhon province, North-eastern Thailand, during July and August in year 2016. The identification of maoberry cultivars were inspected confirmed by Assistant Dr. Prof. Pornprapha Chunthanom, Faculty of Natural Resources, Rajamangala University of Technology Isan, Sakon Nakhon campus, Sakon Nakhon, Thailand. After identifying, the fruits were washed, homogenized and concentrated via rotary evaporation method at 45 °C to a 40% concentration (v/v). The extracted juice was then aliquoted and preserved in opaque tubes at − 20 °C until used. A voucher specimen of maoberry has been deposited in Faculty of Natural Resources, Rajamangala University of Technology Isan, Sakon Nakhon campus.
Animals and experimental settings
Five-week-old male Sprague Dawley rats, weighing 160–180 g, were obtained from the National Laboratory Animal Center at the Salaya Campus, Mahidol University. Seventy-eight rats were housed according to the rules and regulations of the Animal Care Ethical Committee of Laboratory Animal Science Center, Faculty of Tropical Medicine, Mahidol University (Approval no. FTM-ACUC 011/2018). All procedures were conducted according to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health. Two rats were housed per plastic cage in a controlled room (temperature, 25 °C ± 2 °C; relative humidity, 55% ± 10%; a 12-h light–dark cycle). After acclimatization for 1 week with free access to standard diet and drinking water, the rats were randomly separated into two groups with the same average weight. One group (n = 12; ND group) was fed a standard diet (3.90 kcal/g) containing 20.3% protein, 5% fat and 66% carbohydrate for 16 weeks, whereas the other group (n = 60; HFD group) were fed a high-fat diet (HFD; 5.40 kcal/g) containing 20.2% protein, 58.3% fat and 21.5% carbohydrate. Lard was the main fat component in HFD. After 4 weeks of feeding, the HFD rats were randomly divided into the following five subgroups (12 rats each) on the basis of the types of treatments they received (no differences in mean weight): HFD with distilled water (1.52 ml/kg, HF subgroup); HFD with maoberry extract (0.38 g/kg, ML subgroup, 0.76 g/kg, MM subgroup or 1.52 g/kg, MH subgroup) and 10 mg/kg simvastatin (STAT) by oral gavage every other day for 12 weeks.
At the end of the experimental period, the rats were fasted for 12 h, following which they were euthanized by CO2 inhalation. The hearts were harvested and weighed. Cross-sections obtained from the middle of the heart were fixed on filter paper using 10% formalin buffer for at least 2 days. The remaining parts were snap frozen in liquid nitrogen and stored at − 80 °C. Each frozen heart part was exsanguinated by perfusion with cold normal saline (0.9%) and was placed in 100 mg/ml phosphate-buffered saline containing heparin. After cardiac tissues were completely lysed, they were homogenized on ice using a sonicator (Sonic Inc., Stratford, CT, USA) and were centrifuged at 10,000×g for 5 min at 4 °C. The supernatants were collected and stored at − 80 °C for further analysis.
Determination of protein in cardiac tissues
The Bradford method (BioRad, Hercules, CA, USA) is rapid and accurate for estimating protein concentration. Following the manufacturer's protocol with slight modifications, the dye reagent was prepared by diluting one part of dye reagent concentrate with four parts of distilled, de-ionized water. Bovine serum albumin (BSA) was used as the standard. The linear range of the assay for BSA is 0–2 mg/ml. Next, 160 μl of each standard and sample was pipetted into 96-well plate, and 40 μl of the diluted dye reagent was added to each well. The sample and reagent were thoroughly mixed using a microplate mixer. The final solution was incubated at room temperature for at least 5 min. Absorbance was measured at 450 and 595 nm. The eq. Y = 3.663X + 0.6762 (r2 = 0.9919) was used, and the results were recorded in g/L.
Determination of total phenolic contents (TPC)
Total phenolic contents were determined using the Folin–Ciocalteu colorimetric method according to Baba and Malik (2015) [24] with slight modifications. In brief, 10 μl of cardiac tissue samples was blended with 150 μl of distilled water, mixed with 25 μl of Folin–Ciocalteu reagent and incubated for 3 min. Subsequently, 100 μl of 20% (w/v) sodium carbonate was added to each sample. The absorbance was measured at 650 nm after 1-h incubation in the dark at room temperature. A calibration curve was generated using gallic acid (10–100 μg/ml) solutions and the eq. Y = 0.0035X + 0.046 (r2 = 0.9999). The results were recorded as milligram gallic acid equivalences (mg GAE)/g protein. The solutions were assayed in duplicate.
Determination of total flavonoid contents (TFC)
Total flavonoid contents were determined using the aluminium chloride colorimetric method [24] with some modifications. In brief, 1.5 μl of cardiac tissue sample was combined with 30 μl of methanol, thoroughly mixed with 120 μl of distilled water and treated with nine μl of 5% NaNO2 solution. After 5-min incubation, nine μl of 10% AlCl3 solution was added, and the mixtures were allowed to stand for 6 min. Next, 60 μl of 1 mol/L NaOH solution was added, and the final mixture was incubated for 15 min. Absorbance was measured at 410 nm. The calibration curve was generated using solutions of quercetin (100–1000 μg/ml) and the eq. Y = 8E–0.5X − 0.0025 (r2 = 0.9961). The results were presented in milligram quercetin equivalence (mg QE)/g protein. The solutions were assayed in duplicate.
Oxidative stress in the heart
Thiobarbituric acid reactive substance (TBARS) assay was used for determining the level of malondialdehyde (MDA) by referring to lipid peroxidation from ROS. In brief, 100 μl of cardiac supernatant was thoroughly mixed with 350 μl of stock solution that contained 100 μl of 10% sodium dodecyl sulphate lysis solution and 150 μl of Thiobarbituric acid (TBA) reagent (25 ml of 10% acetic acid for 130 mg TBA) and was heated for 60 min at 95 °C. The absorbance of the supernatant was measured at 532 nm. MDA (#10009202, Cayman Chemical Company, Ann Arbor, MI, USA) was used as the standard. The standard curve (0–0.06 μmol/L of MDA) was derived using the following equation: Y = 0.0162X + 0.0097 (r2 = 0.9973). The results were presented as nmol/g protein.
Ferric reducing antioxidant power (FRAP) assay
Ferric-reducing activities in the heart were determined using a modified FRAP assay [25]. In brief, the FRAP reagent was prepared in the dark using 300 mM sodium acetate buffer (pH 3.6), 10 mM 2,4,6-tri (2-pyridyl)-s-triazine solution in 40 mMHCl and 20 mM FeCl3 solution in a ratio of 10:1:1. The fresh working solution was warmed at 37 °C before use. Ten microliters of the cardiac tissue supernatant were allowed to react with 300 μl of the FRAP reagent. After incubation at 37 °C for 4 min, the absorbance of the reaction mixture was measured at 593 nm. A calibration curve was generated using the standard solutions of trolox (0–1000 μmol/L) and the eq. Y = 0.0011X − 0.0057 (r2 = 0.9917). The results were presented as μmol trolox equivalence (TE)/g protein.
Histopathological analysis of cardiac tissues
Formalin-fixed cross-sections that were obtained from the middle of the heart were dehydrated and embedded in a paraffin–polyisobutylene mixture (Leica Biosystems, Harbourfront Centre, Singapore); 4-μm-thick sections were cut and prepared for haematoxylin and eosin (H&E) staining. The cardiac morphology was assessed using the Olympus BX-53 light microscope with a camera attachment (Tokyo, Japan) and the CellSens computer-based image analysis software (Olympus, Bangkok, Thailand).
Real-time polymerase chain reaction
The molecular mechanism involved in the atherogenic protection of maoberry extract was determined by analyzing the mRNA levels of endothelial nitric oxide synthase (eNOS), tumor necrosis factor- alpha (TNF-α), interleukin-6 (IL-6), scavenger receptor CD36, vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1). Total RNA was isolated from frozen cardiac specimens using Trizol reagent (Invitrogen, Carlsbad, CA, USA, Cat. No. 15596–026) following the manufacturer's instructions. The NanoDrop spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, USA) was used to determine the concentration and purity of the extracted RNA. Total RNA (2 μg) was reverse transcribed into cDNA using random oligo (dT)18 primers (Thermo Fisher Scientific Inc., MA, USA) and reverse transcriptase enzyme (Thermo Fisher Scientific Inc., MA, USA). Polymerase chain reaction (PCR) using cDNA templates were performed in 10-μl reaction mixtures that contained 0.3 μl of each specific primer (10 μM), five μl of LightCycler® 480 SYBR Green I Master (Cat. No.04707516001) and 3.4 μl of RNase-free water. The reactions were run using the LightCycler® 480 Real-Time PCR detection system (Roche, Indianapolis, IN USA) with the following conditions: 95 °C for 5 min in the pre-incubation phase, 95 °C for 10 s, 58 °C for 10 s and 72 °C for 10 s and 45 cycles at the amplification phase. All primers (Table 1) were synthesized by Pacific Science Co., Ltd. (Bangkok, Thailand). The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the internal control to normalize for differences in quantity and quality between the RNA samples. The fold differences in the expression of different mRNAs were calculated and compared with those in that of the ND group using the 2−ΔΔCt calculation as follows [26]:
$$ \Delta \Delta \mathrm{Ct}=\Delta \mathrm{Ct}\left(\mathrm{treatment}\ \mathrm{group}\right)-\Delta \mathrm{Ct}\left(\mathrm{normal}\ \mathrm{group}\right) $$
$$ \Delta \mathrm{Ct}=\mathrm{Ct}\left(\mathrm{target}\ \mathrm{gene}\right)-\mathrm{Ct}\left(\mathrm{reference}\ \mathrm{gene}\right) $$
The primer details used for real-time PCR analysis
Forward sequence (5′ to 3′)
Reverse sequence (5′ to 3′)
Amplicon length (bp)
GenBank accession number
GGA TTC TGG CAA GAC CGA TTA C
GGT GAG GAC TTG TCC AAA CAC T
NM_021838.2
Vcam-1
GGA GCC TGT CAG TTT TGA GAA TG
TTG GGG AAA GAG TAG ATG TCC AC
TNF-α
ACT GAA CTT CGG GGT GAT TG
GCT TGG TGG TTT GCT ACG AC
ATATGTTCTCAGGGAGATCTTGGAA
GTGCATCATCGCTGTTCATACA
AGGAAGTGGCAAAGAATAGCAG
ACAGACAGTGAAGGCTCAAAGA
Mcp-1
GGC CTG TTG TTC ACA GTT GCT
TCT CAC TTG GTT CTG GTC CAG T
GCA AGT TCA ACG GCA CAG
GCC AGT AGA CTC CAC GAC AT
The statistical analysis software SPSS (version 18.0; SPSS Inc., IBM, Chicago, IL, USA) was used to perform one-way analysis of variance and post-hoc tests for multiple comparisons. A p value of < 0.05 was considered to be significant. All values were expressed as mean ± standard deviation of two determinations. The GraphPad Prism software version 5.0 (La Jolla, CA, USA) was used for curve fitting.
Total phenolic and flavonoid contents in cardiac tissues
No mortality, illness and alterations in appearance or behavior were observed in all rats, both during and after 12 weeks of maoberry extract administration. A significant increase in both total phenols (p < 0.01 in the ML and MM subgroups and p < 0.001 in the MH subgroup) and total flavonoids (p < 0.01 in the MM subgroup and p < 0.001 in the ML and MH subgroups) were observed in cardiac tissues of rats that were fed HFD and different concentrations of maoberry extract compared with those of rats that were fed HFD with distilled water (Fig. 1a and b).
Total phenolic content (a), total flavonoid content (b), malondialdehyde level (c) and Ferric reducing antioxidant power (FRAP) level (d) in the cardiac tissue. Data are expressed as mean ± SD, n = 12. ND: standard diet, HF: high fat diet, ML, MM, MH: high fat diet with Mao Luangcrude extract 0.38 or 0.76 or 1.52 g/kg, respectively, STAT: high fat diet with simvastatin 10 mg/kg.*p < 0.05, **p < 0.01, ***p < 0.001 represent significant differences when compared with the ND group. #p < 0.05, ## p < 0.01, ### p < 0.001 represent significant differences when compared with the HF group
Oxidative stress in cardiac tissues
Oxidative stress was evaluated on the basis of the production of MDA (a biomarker of lipid peroxidation) using the TBARS assay. After 12 weeks of treatment, MDA levels were significantly higher in the HF group than in the ND group (p < 0.001), confirming the establishment of oxidative stress in myocytes associated with hypercholesterolemia. Furthermore, MDA levels were significant lower in all rats that were fed HFD with maoberry extract (p < 0.05 in the ML and MH subgroups and p < 0.01 in the MM subgroup) or simvastatin (p < 0.01) than in rats that were fed HFD with distilled water (Fig. 1c).
Antioxidant capacity in cardiac tissue
The FRAP level markedly decreased in the HF group compared with that in ND group (p < 0.01). In contrast, the FRAP levels significantly increased (p < 0.01 in the MM subgroup and p < 0.001 in the ML and MH subgroups) in rats that were fed HFD with maoberry extract or simvastatin (p < 0.05) compared with rats that were fed HFD with distilled water (Fig. 1d).
Histopathological changes in cardiac tissue
Microscopic images of H&E-stained cardiac tissues of rats in the ND group (Figs. 2a and 3a) revealed benign, blunt-looking, arranged cardiac muscles with no abnormal pathological findings. Alternatively, histological alterations were observed in cardiac tissues of rats in the HF group. Pathological alterations such as myocyte hypertrophy, enhancement of fat droplets and accumulation of mononuclear cells associated with inflammation were evident in the HF group compared with the ND group (Figs. 2b and 3b).
Light microscopic images of H&E-stained transverse-sections of cardiac tissues of rats after 12 weeks of Maoberry. For comparison, images showed different fat droplets in the different treatment groups (magnification × 100). a ND: rats fed a standard diet, b HF: rats fed a high-fat diet with distilled water, c ML, d MM and e MH: rats fed a high-fat diet with Maoberry extract at 0.38, 0.76 and 1.52 g/kg, respectively, f STAT: rats fed a high-fat diet with 10 mg/kg simvastatin. Arrows represent fat droplets. Panels enlarge the area of pathological alterations (magnifiacation × 400)
Pathological changes in transverse-sections of cardiac tissues of rats after 12 weeks of treatment showing amounts of mononuclear cell infiltration in different treatments. a ND: a standard diet, b HF: a high-fat diet with distilled water, c ML, d MM and e MH: a high-fat diet with Maoberry extract at 0.38, 0.76 and 1.52 g/kg, respectively, f STAT: a high-fat diet with 10 mg/kg simvastatin. H&E staining (magnification× 100). Arrows represent mononuclear cell infiltration. Panels enlarge the area of pathological alterations (magnifiacation × 400)
After 12 weeks of treatment, mild myocytic hypertrophy and a decrease in the number of fat droplets (Fig. 2c-e) and mononuclear cells (Fig. 3c-e) were evidently observed in Maoberry-treated rats, particularly those of the MH group, compared with those in HF group.
Real-time PCR analysis of cardiac tissues
To understand the effects of maoberry extract on HFD-induced inflammation and vascular dysfunction, mRNA levels were examined by real-time PCR analysis. The mRNA levels of inflammatory cytokines such as TNF-α and IL-6 were significantly elevated in the HF group (p < 0.01 and p < 0.001 respectively), whereas the levels were markedly decreased in rats that were fed HFD with different concentrations of maoberry extract or simvastatin (Fig. 4a and b).
TNF-α, IL-6, VCAM-1, MCP-1, CD36 and eNOS mRNA levels from cardiac tissues of each groupby Real-time PCR detection. The expressions of TNF-α, IL-6, VCAM-1, MCP-1, CD36 and eNOS mRNA in HF group were significantly higher than those in the ND group. The expressions of IL-6, VCAM-1, MCP-1, and eNOS mRNA in Maoberrytreated groups were significantly lower than those in the HF group. Each bar represents the relative mean ± SD, n = 12 each. *p < 0.05, **p < 0.01 and ***p < 0.01 represent significant differences when compared with the ND group. #p < 0.05, ##p < 0.01 and ###p < 0.001 represent significant differences when compared with the HF group. ND: a standard diet, HF: a high fat diet, ML, MM, MH: a high fat diet with Maoberry extract 0.38 or 0.76 or 1.52 g/kg, respectively, STAT: a high fat diet with simvastatin 10 mg/kg
Moreover, mRNA levels of VCAM-1, MCP-1, CD36 and eNOS were significantly higher in cardiac tissues of the HF group (p < 0.001, except eNOS, which was p < 0.01) than those of the ND group (Fig. 4c-f). Supplementing with different concentrations of maoberry extract or simvastatin significantly reduced the upregulation of VCAM-1, MCP-1 and eNOS mRNA levels compared with supplementing HFD with distilled water. However, significant downregulation in CD36 gene expressions was observed only in the STAT group.
According to our previous studies [23], maoberry extract doses of 0.38, 0.76 or 1.52 g/kg for 12 weeks may have tremendous beneficial effects on risk factors of cardiovascular disease such as atherogenic indices, inflammation and oxidative stress in peripheral blood and spleen histopathology.
The principle of maoberry dose use calculated according to American Heart Association recommendation to consume fruits 4 servings per day for long-term benefits to health and heart [27]. Four portions of fresh maoberries equal to 0.38 g per 1 kg body weight. Two and four times of maoberry extracts are 0.76 and 1.52 g/kg body weight, respectively. Previously, we examined nutritive values of maoberry extract and its antioxidant activities based on oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays [23]. Gallic acid, epicatechin, catechin, and cyanidin-3-O-glucoside were analysed by HPLC and reported as the major polyphenolic components in fourteen maoberry cultivars from northeast Thailand [28]. Catechin, procyanidin B1, and procyanidin B2 analysed by HPLC are the major flavonoid compounds in fifteen cultivars from Northeast Thailand as well [29]. These polyphenols and flavonoids are often known to be antioxidants and may be involved in and contribute to the antioxidant activity of maoberry extract.
We previously reported that maoberry extract could reduce the number of immune cells associated with inflammation and the platelet population connected to the vascular blockage [23]. There are many finding on major active compounds from maoberry (Antidesma bunius) on oxidative stress and inflammation. Cyanidin-3-glucoside showed to inhibit free radicals and inhibit the expression of important cytokines in inflammation including nuclear factor-kappa B (NF-kB) in human vascular endothelial cells with dose-dependent manner [30]. Catechin showed to significantly reduce lipid peroxidation (MDA) in drug inducing cardiotoxicity in rats. Significant decrease in NF-kB, tumor necrosis factor alpha and inducible nitric oxide synthase were also present in rats treated with catechin [31]. Gallic acid available in maoberry was reported to be one of the most active dietary antioxidants in humans [32]. One of observation from Medical University of Vienna, Austria evidenced that administration of gallic acid in the amount of daily consumption could reduce oxidative DNA damage, oxidized-LDL and C-reactive protein in plasma of patients with type 2 diabetes mellitus [33].
The current study further demonstrates that maoberry extract contributes to prevent deterioration in cardiac tissue by improving the oxidative stress status and down regulating the expression of inflammatory cytokines and chemokines.
Oxidative stress, which occurs because of an imbalance between excess free radical formation and low antioxidant defense, has unspecific damage to the structure and functions of cells [34]. In the present study, a high fat diet (HFD) consumption appeared to increase lipid peroxidation in cardiac tissues, demonstrated by increased MDA levels in the tissues. MDA is a by-product of lipid peroxidation and is accepted as a vital marker of oxidative stress [35]. In previous studies in which animals were fed HFD, supplementation with lard induced oxidative stress, which originates from the upregulated expression of NADPH oxidase (up to threefold) [4, 8, 34]. After maoberry extract administration, numerous polyphenols in the extract pass through the stomach, are hydrolyzed and absorbed in the small intestine. Some polyphenols enter into the blood circulation and reach the organs [36]. In this study, although polyphenol levels in tissues depend on the amount of uptake and secretion by specific tissues, significantly higher levels of total flavonoids and total phenols were observed in the cardiac tissue of rats consuming maoberry extract. This meant that the cardiac tissue can uptake polyphenols from maoberry extract.
Furthermore, the results of the FRAP assay demonstrated the role of maoberry extract that reduces ferric iron (Fe3+) to ferrous iron (Fe2+) in the cardiac tissue. Altogether, these findings confirm that cardiac tissues can absorb polyphenolic substances, which are responsible for radical scavenging and reducing activities [37]. Similarly, polyphenols from other sources such as pomegranate [14], tea, cacao and red grape juice [15] are potential antioxidants that exert their effects by radical scavenging, which inhibits lipid peroxidation, Fe3+ reduction and downregulating NADPH oxidase activity [12, 15, 38].
Moreover, oxidative stress-induced LDL oxidation increases the expression levels of scavenger receptors and pro-inflammatory cytokines, mainly secreted by macrophages, and upregulates the expression of inflammatory adhesion molecules on endothelial cells [12, 34, 39]. In the current study, along with histopathological alterations, significantly elevated mRNA levels of TNF-α, IL-6, VCAM-1, CD36 and MCP-1 were observed in cardiac tissues of the HF group. Thus, it appears that the progression of endothelial dysfunction is associated with inflammation in the endothelium [40]. Inflammation reduced after 12 weeks of treatment with maoberry extract. A significant decrease in mRNA expression levels of pro-inflammatory cytokines and chemokines, such as TNF-α, IL-6 and MCP-1 and adhesion molecules, such as VCAM-1 was also observed. These results are buttressed by the apparent reduction in the number of lymphocytic cells in histological cardiac tissues of rats fed HFD with maoberry extract. In agreement with previous studies, the protective effects of polyphenols in maoberry extract may prevent endothelial dysfunction and inflammation via the downregulation of mRNA expression levels of TNF-α, IL-6, VCAM-1 and MCP-1 [18, 34, 39] by suppressing the activation of the transcription factor NF-κB in endothelial cells [41, 42]. In contrast, LDL oxidation enhances the expression of scavenger receptors such as CD36 in macrophages, which uptakes the excess modified lipid molecules, thus leading to the formation of foam cells, a key player in early inflammation-induced atherogenesis [43]. In the present study, we also found apparently high mRNA levels of CD36 in the HF group compared with those in the ND group, probably owing to the upregulation of structurally defined oxidized molecules, which serve as high affinity ligands for CD36 [43]. Although no significant difference was noted between the Mao Luang-treated groups and the HF group, mRNA expression levels of CD36 demonstrated a tendency to decline. Therefore, the beneficial effects of maoberry extract may be related to reduced CD36 expression and functions and suppressed oxLDL modification [39, 43]. Moreover, polyphenols in maoberry extract may suppress or downregulate the expression of other scavenger receptors that recognize oxidation-specific oxLDL, including SRA-1, SRA-2, MARCO, SR-B1, LOX-1 and PSOX [12].
Endothelial dysfunction is characterized by the impairment of endothelium-dependent relaxation owing to decreased vascular NO bioavailability caused by oxidative stress. eNOS is the major enzyme that is responsible for NO production. Lower eNOS levels or the lack of a substrate or cofactor such as L-arginine and tetrahydrobiopterin (BH4), which is affected by ROS, can lead to uncoupled eNOS formation, resulting in decreased NO levels and increased ONOO− production. The overproduction of ONOO− can enhance BH4 oxidation, leading to BH4 deficiency, thus generating a vicious circle. Furthermore, eNOS exposure to ONOO− leads to the uncoupling of eNOS and is a crucial mechanism that contributes to vascular dysfunction [40, 44]. In the current study, we observed higher mRNA levels of eNOS in the HF group than in the ND group. After 12 weeks of treatment with maoberry extract, a reduction in upregulated mRNA levels of eNOS was observed, particularly in the ML (p < 0.05) and MM (p < 0.01) subgroups compared with HF group. This is in agreement with the results of a study on human umbilical vein endothelial cells, where in Genistein [45], a soy polyphenol, improved vascular reactivity by increasing eNOS expression. Genistein acutely stimulates eNOS synthesis in vascular endothelial cells. In contrast, Lund et al. [46] revealed that soy isoflavone did not affect eNOS expression in hyperlipidemic rabbits. These findings suggest that eNOS can be regulated by genomic and nongenomic factors [45]. Generally, regulatory systems allow living cells to change biochemical processes or gene expression programs automatically in response to alterations in the intracellular and/or extracellular environment. Therefore, in this study maoberry extract may affect on the modulation of eNOS expression into the normal range. Furthermore, during inflammation, TNF families alone could reduce eNOS mRNA half-life from 48 h to 3 h. [47]. Consequently, decreased mRNA levels of eNOS may indicate positive autoregulation related to inflammation, resulting in increased mRNA transcription and translation of eNOS [48], which were found in HF group. In contrast, maoberry extract may function as a free radical scavenger because of which eNOS and other substrates or cofactors can take effective action in NO production; negative feedback may control eNOS transcription and translation caused the decrease in eNOS mRNA expression [48]. Although the molecular mechanism remains unclear and complicated, the present study may be the first one to indicate that maoberry extract may prevent cardiac tissue deterioration.
The current study reported for the first time that oral administration of maoberry extract had significant beneficial effects in cardiac tissues by reducing oxidative stress and enhancing antioxidant activity, thereby preventing progression of cardiac tissue deterioration involved in inflammation. In accordance with the outcomes of this study, maoberry extract ameliorates oxidative stress and inflammation in cardiac tissues of rats fed with a high-fat diet by at least through: (1) acting as a radical scavenger and Fe3+ reduction (2) reducing MDA levels that are associated with lipid peroxidation inside tissues and enhancing cardiac antioxidative capacity, (3) counteracting against the upregulation of inflammatory cytokines (TNF-α, IL-6, VCAM-1 and MCP-1) to reduce inflammation and (4) downregulating eNOS mRNA expression associated with positive and negative autoregulation.
Although the precise molecular mechanisms by which maoberry extract protects cardiac tissue deterioration remains unclear, this study may profoundly demonstrate that maoberry extract treatment may be a congenial alternative application against in damage progression of cardiac tissue.
Bovine serum albumin
eNOS:
Endothelial NO synthase
FRAP:
Ferric reducing antioxidant power
Glyceraldehyde-3-phosphate dehydrogenase
H&E:
Haematoxylin and eosin
HFD:
A high-fat diet
High-fat diet
LDL:
Monocyte chemoattractant protein-1
MDA:
Malondialdehyde
MH:
High-fat diet with maoberry extract 1.52 g/kg
ML:
NADPH:
Nicotinamide adenine dinucleotide phosphate
ND:
Normal diet
ONOO− :
Peroxynitrite
High-fat diet with simvastatin
TBA:
Thiobarbituric acid
Trolox equivalence
TFC:
Total flavonoid contents;TBARS, Thiobarbituric acid reactive substance
VCAM-1:
Vascular cell adhesion molecule-1
This research acknowledges to the Faculty of Tropical Medicine for supporting the equipment and facilities.
This research was supported by the Faculty of Tropical Medicine in the study design, collection, analysis and in writing the manuscript. The research was also supported by and Thailand Research Fund (TRF) under Grand No.MRG6180101 in the study design, analysis and interpretation of data.
The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.
AU made a contribution in the experimentation, analyzing and drafting of the manuscript. CN made a contribution in the experimentation. PA, AA, RT contribution in designing the experimentation, PP takes the entire responsibility of the manuscript. All authors read and approved the final manuscript.
PP did her PhD (Nutrition) in The Institute of Nutrition, Mahidol University. She is working as Assistant Professor at the Department of Tropical Nutrition and Food Science, Mahidol University, Bangkok, Thailand.
This study makes use of rats under rules and regulations of the Animal Care Ethical Committee of Laboratory Animal Science Center, Faculty of Tropical Medicine, Mahidol University (Approval no. FTM-ACUC 011/2018).
Department of Tropical Nutrition and Food Science, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok, Thailand
Institute of Nutrition, Mahidol University, Nakhon Pathom, Thailand
Department of Helminthology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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redpajama_set_name: RedPajamaGithub


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