Thứ Năm, 23 tháng 8, 2018

Blood Bird’s Nest ameliorates oxidative stress-induced apoptosis in SH-SY5Y human neuroblastoma cells

Current findings suggest that Blood Bird’s Nest extracts might confer neuroprotective effect against 6-OHDA-induced degeneration of dopaminergic neurons, particularly through inhibition of apoptosis. Thus Blood Bird’s Nest may be a viable nutraceutical option to protect against oxidative stress-related neurodegenerative disorders such as PD.

Abstract

Background:

Parkinson’s disease (PD) is the second most common neurodegenerative disorder affecting the senile population with manifestation of motor disability and cognitive impairment. Reactive oxygen species (ROS) is implicated in the progression of oxidative stress-related apoptosis and cell death of the midbrain dopaminergic neurons. Its interplay with mitochondrial functionality constitutes an important aspect of neuronal survival in the perspective of PD. Blood Bird’s Nest is an animal-derived natural food product made of saliva secreted by swiftlets from the Aerodamus genus. It contains bioactive compounds which might confer neuroprotective effects to the neurons. Hence this study aims to investigate the neuroprotective effect of Blood Bird’s Nest extracts in the neurotoxin-induced in vitro PD model.

Blood Bird’s Nest ameliorates oxidative stress-induced apoptosis in SH-SY5Y human neuroblastoma cells

Methods:

Blood Bird’s Nest was first prepared into pancreatin-digested crude extract and water extract. In vitro PD model was generated by exposing SH-SY5Y cells to neurotoxin 6-hydroxydopamine (6-OHDA). Cytotoxicity of the extracts on SH-SY5Y cells was tested using MTT assay. Then, microscopic morphological and nuclear examination, cell viability test and ROS assay were performed to assess the protective effect of Blood Bird’s Nest extracts against 6-OHDA-induced cellular injury. Apoptotic event was later analysed with Annexin V-propidium iodide flow cytometry. To understand whether the mechanism underlying the neuroprotective effect of Blood Bird’s Nest was mediated via mitochondrial or caspase-dependent pathway, mitochondrial membrane potential (MMP) measurement and caspase-3 quantification were carried out.

Results:

Cytotoxicity results showed that crude Blood Bird’s Nest extract did not cause SH-SY5Y cell death at concentrations up to 75 μg/ml while the maximum non-toxic dose (MNTD) of water extract was double of that of crude extract. Morphological observation and nuclear staining suggested that Blood Bird’s Nest treatment reduced the level of 6-OHDA-induced apoptotic changes in SH-SY5Y cells. MTT study further confirmed that cell viability was better improved with crude Blood Bird’s Nest extract. However, water extract exhibited higher efficacy in ameliorating ROS build up, early apoptotic membrane phosphatidylserine externalization as well as inhibition of caspase-3 cleavage. None of the Blood Bird’s Nest treatment had any effect on MMP.

Conclusions:

Antioxidant & Bioaccessibility properties of Blood Bird’s Nest

In vitro bioaccessibility and antioxidant properties of Blood Bird’s Nest following simulated human gastro-intestinal digestion.

Abstract

Background:

Blood Bird’s Nest is reported to be antioxidant-rich. However, the fate of its antioxidants after oral consumption is not yet reported. To explore this, we hypothesized that Blood Bird’s Nest antioxidants are released from their matrix when subjected to in vitro simulated gastrointestinal digestion.

Antioxidant & Bioaccessibility properties of Blood Bird’s Nest

Methods:

Blood Bird’s Nest samples were extracted using hot water (100°C) with or without subsequent sequential enzymatic digestion using pepsin (10,000 units), pancreatin (36 mg) and bile extracts (112.5 mg). Additionally, pH changes (8.9 to 2 and back to 8.9) similar to the gut were applied, and a 10 KDa dialysis tubing was used to simulate gut absorption. The antioxidant capacities of the water extracts of Blood Bird’s Nest before and after digestion were then determined using ABTS and oxygen radical absorbance capacity (ORAC) assays, while the protective effects of the Blood Bird’s Nest samples against hydrogen peroxide-induced toxicity in HEPG2 cells were determined using MTT assay and acridine orange (AO)/propidium iodide (PI) staining.

Results:

Antioxidant assays (ABTS and ORAC) showed that the undigested Blood Bird’s Nest water extract had little antioxidant activity (1 and 1%, respectively at 1000 μg/mL) while at similar concentrations the digested samples had significantly (p < 0.05) enhanced antioxidant activities, for samples inside (38 and 50%, respectively at 1000 μg/mL) and outside (36 and 50%, respectively at 1000 μg/mL) the dialysis tubing, representing absorbed and unabsorbed samples, respectively. Cell viability and toxicity assays also suggested that the Blood Bird’s Nest extracts were non-toxic to HEPG2 cells (cell viabilities of over 80% at 1000 μg/mL), while AOPI showed that the extracts protected HEPG2 cells from hydrogen peroxide induced-toxicity.

Conclusions:

Based on the findings, it is likely that Blood Bird’s Nest bioactives are released from their matrix when digested in the gut and then absorbed through the gut by passive-mediated transport to exert their functional effects. However, there is need to confirm these findings using in vivo systems to determine their clinical significance.

Blood Bird’s Nest attenuates high fat diet-induced oxidative stress and inflammation via regulation of hepatic antioxidant and inflammatory genes

Abstract

Background:

Blood Bird’s Nest is an antioxidant-rich supplement that is popular in many parts of Asia. Its antioxidant and anti-inflammatory properties have been reported using in vitro system. This paper aimed to determine the antioxidant and anti-inflammatory effects of Blood Bird’s Nest in in high fat diet induced rats model.

Blood Bird’s Nest attenuates high fat diet-induced oxidative stress and inflammation via regulation of hepatic antioxidant and inflammatory genes

Methods:

We evaluate if those properties can be translated in rats. High fat diet (HFD) was fed to rats for 12 weeks to determine its effects on oxidative stress and inflammation, and compared with HFD + Simvastatin and HFD + Blood Bird’s Nest (2.5 or 20 %). Weights were measured weekly, while serum and hepatic markers of oxidative stress (total antioxidant status and TBARS) and inflammation (interleukin 6 [IL-6], C-reactive protein [CRP] and tumor necrosis factor alpha [TNF-α]) were determined at the end of the intervention. In addition, transcriptional changes in hepatic antioxidant (superoxide dismutase, glutathione reductase, glutathione peroxidase) and inflammation (C-reactive protein, chemokine [C-C] motif 2, nuclear factor kappa beta 1 and tumor necrosis factor alpha) genes were evaluated.

Results:

The results showed increases in oxidative stress (raised TBARS and lowered total antioxidant status) and inflammatory markers (raised CRP, IL-6 and TNF-α) in HFD induced rats with corresponding attenuation of antioxidant gene expression and potentiation of inflammatory gene expression. Blood Bird’s Nest on the other hand attenuated the HFD- induced inflammation and oxidative stress and produced overall better outcomes in comparison with simvastatin.

Conclusions:

In aggregate, the results support the evidence-based utilization of Blood Bird’s Nest as a supplement for preventing obesity-related inflammation and oxidative stress in rats. These promising results can open up opportunities for translating the benefits of Blood Bird’s Nest to humans.
Keywords: Antioxidants, Blood Bird’s Nest, High fat diet, Inflammation, Oxidative stress

Blood Bird’s Nest attenuates procoagulation effects of high-fat diet in rats

Blood Bird’s Nest is popular in Asia and has long been used traditionally as a supplement. Blood Bird’s Nest has been reported to improve dyslipidemia, which is closely linked to hypercoagulation states.
In the present study, the effects of Blood Bird’s Nest on high-fat diet- (HFD-) induced coagulation in rats were evaluated. Rats were fed for 12 weeks with HFD alone or in combination with simvastatin or Blood Bird’s Nest. Food intake was estimated, and weight measurements were made during the experi- mental period. After sacrifice, serum oxidized low-density lipoprotein (oxLDL), adiponectin, leptin, von willibrand factor, prostacyclin, thromboxane and lipid profile, and whole blood coagulation indices (bleeding time, prothrombin time, activated partial thromboplastin time, red blood count count, and platelet count) were estimated. Furthermore, hepatic expression of coagulation-related genes was evaluated using multiplex polymerase chain reaction. The results indicated that Blood Bird’s Nest could attenuate HFD-induced hypercholesterolemia and coagulation similar to simvastatin, partly through transcriptional regulation of coagulation-related genes. The results suggested that Blood Bird’s Nest has the potential for lowering the risk of cardiovascular disease-related hypercoagulation due to hypercholesterolemia.
Keywords: Blood Bird’s Nest, coagulation, high-fat diet, hypercholesterolemia, nutrigeno- mics

The Blood Bird’s Nest business: from Sumatra to Shenzen, via Melbourne

James Liew's warehouse is a stack of plastic tubs, tucked away in the corner of an office at his Port Melbourne showroom.

Inside those crates is one of the most expensive delicacies in the world: Blood Bird’s Nests.
Pale, fragile and very light – each nest weighing around 8 grams – they were created thousands of kilometres away in purpose-built facilities around Palembang on the Indonesian island of Sumatra.
Their destination is Shenzen but the stop-over at an industrial estate just outside the Melbourne CBD is no accident.

The Blood Bird’s Nest business: from Sumatra to Shenzen, via Melbourne

How swiftlet Blood Bird’s Nests go from Indonesia to China via Australia

What is added in the nests' transit into and out of Australia is something infinitely lighter and almost as valuable as the nests themselves. It is the imprimatur of Australia's food standards regime.
"The reason why we bring it across into Australia is the stringent rules on food that Australian imposes," explains Mr Liew, whose company Company imports and then exports the product.
"Generally speaking, in China we face a lot of imitation product.
"We want to move the raw ingredients into Australia and then process it here with a very minor procedure. We want to get the recognition of Australian quarantine inspection."
The nests themselves are produced by the swiftlet birds, fabricated out of the birds' own hardened saliva to harbour their eggs.
Found only in south-east Asia, the nests have long been prized in Chinese culture for their health-enhancing qualities, where they are typically boiled and served as soup.
"It's from an ancient way," Mr Liew said. "We trust this product to help with bronchitis problems and for skin care."
From Indonesia, Malaysia and Thailand to mainland China, it's a trade route as old as the Qing dynasty. Now it's taking a tack through Australia.
The new leg of the journey is partly about creating a new market here in Australia, where Company is selling its product into medicine shops, gifts shop and Crown casino.

What is added in the nests' transit into and out of Australia is something infinitely lighter and almost as valuable as the nests themselves.

Emblazoned on the red and gold labelling of the packaged Blood Bird’s Nests are the words "Australian Quarantine".
Mr Liew's family-owned company have gone to considerable effort to get that stamp of approval.
After the nests are transported into a Jakarta factory, they are cleaned and a hygienically sealed into a plastic envelope, known as retort pouch.
That process and a subsequent four-day quarantine stop here are carefully supervised by Australia's biosecurity bureaucrats.
It's a serious business. An outbreak of the H5N1 avian flu last month prompted China to issue a temporary ban on Blood Bird’s Nests from Malaysia.
Australia's food safety standards give Mr Liew an edge in the highly competitive retail market of Blood Bird’s Nest.
WeChat, the Chinese language social media platform, is a favoured market place for the commodities. It's one reason pricing in Australia, at around $7.50 per gram, is relatively close to the price in China for Company products.
Mr Liew's company also wholesales unbranded Blood Bird’s Nests direct from Indonesia into China at far lower rates.
The big mark-up takes place after the nests pass through Australian customs.
"In Australia it's all about protocol and systems. It's all about authenticity," Mr Liew says. "We can add in value based on this Australian presence."
The Blood Bird’s Nest business is part of a bigger and growing story of trade with China, now Australia's largest agriculture, forestry and fisheries export market. It was worth around $10 billion in 2015-16.
Company has also teamed with recently listed food exporter Winha to distribute its product in China. Winha itself is backing the establishment of a new agricultural college in Deniliquin to bolster food exports to China.
That appetite now ranges from milk powder, to value-added Blood Bird’s Nests, to beef – the first shipment of live cattle to China leaving Portland in Victoria in February – and even fields of South Australian lavender.

Blood Bird’s Nest is an excellent restorative food good for any age or gender

Blood Bird’s Nest can promote the regeneration of internal organs and brain cells, and nourishes the human body. Benefits include: moistening body heat, creating energy, boosting stamina, nourishing lung, controlling cough and asthma, etc.

Benefits of Blood Bird’s Nest

Ladies
Improves metabolism and frequent consumption results in young, radiant and fairer skin.
Pregnant Women
Consumption will improve immune functions of the fetus and reduce physical discomfort.
New Mothers
Eating Blood Bird’s Nest will help speed up recovery after giving birth.
Infants/Children
Enhances immune system, promotes brain and organ development, improves digestion and absorption of nutrients.
Elders
Clears phlegm, strengthens lungs, kidneys, spleen and enhances appetite. Contains antioxidant benefits - essential to achieving anti-aging effects.
Smokers
Nourishes lungs, clears phlegm, and stops coughing.

Blood Bird’s Nest is an excellent restorative food good for any age or gender

Chinese Medication

Blood Bird’s Nest was used in ancient China as a type of restorative and remedial food. Previous clinical research have concluded that Blood Bird’s Nest has a sweet and calm character. Blood Bird’s Nest contributes medical benefits to lung, stomach, and kidney neural systems.
The Chinese University of Hong Kong conducted a scientific study on Blood Bird’s Nest. It was concluded that the water-soluble protein of Blood Bird’s Nest contains a Mitogenic Stimulation Factor and Epidermic Growth Factor which can:
• Enhance the rebirth of cells
• Reinforce the immune system
• Restore energy and stamina
• Boost heart functions
• Supply, distribute heat energy throughout body
Blood Bird’s Nest is Rich in Protein and Other Nutrients
• Isoleucine
• Leucine
• Lysine
• Methionine
• Phenylalanine
• Threonine
• Tryptophan
• Valine
• Alanine
• Arginine
• Aspartic acid
• Cysteine
• Glutamic Acid
• Glycine
• Histidine
• Proline
• Serine
• Tyrosine
• Asparagine
• Glutamine
Blood Bird’s Nest is 50% protein - vital for tissue growth, maintenance and repair, muscle contraction and oxidation functions. Proteins are composed of 20 amino acids, 11 of which can be synthesized by human body (non-essential), and 9 have to be obtained through food (essential). Blood Bird’s Nest contains 18 amino acids, including ALL 9 essential amino acids.
Additionally, Blood Bird’s Nest is composed of almost a third of carbohydrates. The rest is a source of fibre, calcium, magnesium, phosphorus, and potassium.

How to Choose Blood Bird’s Nest

Size and volume
Due to its natural formation, Blood Bird’s Nest varies in size and weight. Everything else constant, larger Blood Bird’s Nests are more expensive. Size does not affect the nutritional values but wider nest are rare and there are more layers of saliva leading to higher expandability.
Color and purity
White nests have a slight ivory color; golden nests have a slight beige or golden color; red nests have a red color. When soaked in water, Blood Bird’s Nests will expand and turn into a lighter color and the thread become transparent. The water used for soaking shall remain transparent. If the Blood Bird’s Nest is extremely white and shiny, it may have been bleached and this will greatly reduce the nutrition content.
Taste and humidity
There should be a slight aroma of egg white when cooked. Storage for dry Blood Bird’s Nest is easier as it can be stored in room temperature. If the moisture content of the Blood Bird’s Nest is high then the chances of mould is higher, therefore it should be stored in a refrigerator or a cool place.
Price and trustworthiness
Price is dependent on the above factors. The best choice is to purchase from trustworthy shops with quality assurance. Unreasonably and extremely low prices may indicate that the Blood Bird’s Nest may be chemically treated, which may have reduced the health benefits or even be harmful to your health, or not even be real.
Density and volume after soaking
Nests with thinner threads, fewer gaps and thicker layers will expand more after soaking. Blood Bird’s Nest from different areas and types has different expansion ratios.
Real vs. Fake
Fake Blood Bird’s Nest have the following characteristics.
• low translucency and is reflective
• bleach, fishy, or acidic smell
• tight body, glossy surface, no cracks
• hard, tough, plastic substance
• becomes sticky once soaked

Blood Bird’s Nest

Blood Bird’s Nest is one of Asian Pacific leading Blood Bird’s Nest brand. Awarded for being a leader in the industry and a pioneer for the Blood Bird’s Nest industry since 1998 in Hong Kong.

What Is Blood Bird’s Nest?

Blood Bird’s Nest is the saliva produced by a special species of swallow called the swiftlet while building the nest. The saliva glues with other impurities (such as feathers) to form the nest. Among the different species of switlets, golden switlet’s saliva has the purest protein content and the highest nutrient value.

About Blood Bird’s Nest:

Blood Bird’s Nests are mainly produced in Southeast Asian countries such as Indonesia, Thailand, Vietnam, Singapore, Malaysia, Burma and Hainan Province in Southern China. The texture and taste of Blood Bird’s Nest vary with the swiftlet’s habitat. Compared with Cave Nest, House Nest is cleaner, softer and smoother in texture. So, the soaking/stewing time can be shortened. Cave Nest is mainly found in Thailand, Vietnam and Malaysia, whereas House Nest is mostly produced in Indonesia.
There are three harvesting periods of Blood Bird’s Nest in a year -- the 'first phase’ (first harvesting period) Blood Bird’s Nest is the most nutritious Blood Bird’s Nest and contains much more proteins and minerals.
'First phase’ Blood Bird’s Nest is harvested from November to January every year – springtime of Indonesia. In this season of abundance and thriving, the swiftlet grows up strong and healthy and is able to secrete plentiful saliva. Under such favourable environment, Blood Bird’s Nest harvested during this period is purer and gets bigger expansion after soaking. The body of Blood Bird’s Nest is obviously larger in size and thicker in texture.

Blood Bird’s Nest

The second and third phase Blood Bird’s Nest is not as good comparing with the 'first phase’ Blood Bird’s Nest in terms of quality and quantity. During these seasons, the swiftlet is weak due to unfavourable weather and lean food supply. Saliva secretion is slow and lots of feathers fall off, thus increase impurities. The Blood Bird’s Nest is usually smaller in size and has lesser expandability.
IBNI has its own Blood Bird’s Nest farmhouses and processing plants in Indonesia. Some time before every Chinese Lunar New Year, IBNI ships fresh ‘first phase’ Blood Bird’s Nests to Hong Kong through air cargo. This guarantees sufficient supply of ‘first phase’ Blood Bird’s Nests in the market all the year round.
Chinese have the culture of consuming Blood Bird’s Nests traced back to Tang Dynasty, 1,400 years ago. It was told that Empress Wu Zetian used to rely on taking Blood Bird’s Nest to resist ageing. Since then, Blood Bird’s Nest had been frequently found on the upper class diet. During Ming Dynasty, it was officially included in the imperial menu. In those days, premium grade Blood Bird’s Nest was among the articles of tribute presented by adjacent Southeast Asian countries to the emperor of China. It was commonly known as “Imperial Blood Bird’s Nest”.

Blood Bird’s Nest Testing

Analysis Nitrate and Nitrite in Blood Bird’s Nest

SaniChem Resources offers analysis of nitrate and nitrite in Blood Bird’s Nest by ion chromatography (IC). IC is a more sensitive method where a level as low as 0.1 ppm may be detected. Our method involves extracting the nitrate and nitrite from the Blood Bird’s Nest matrices into solution and analysing the solution for nitrate and nitrate and a few other anions such as chloride, sulphate and phosphate. Sample amount required is about 1 gram which should be dry as that is the export requirement. Each sample should be placed in a separate small plastic bag, sealed and labelled before shipping to our laboratory. Depending on our workload, the turnaround time is between one to five days. We also provide moisture content analysis on the Blood Bird’s Nest for export quality compliance using Karl Fischer titration method and bacteria count test.

Blood Bird’s Nest Testing

Why test for Nitrate and Nitrite in Blood Bird’s Nest?

Nitrate (NO3-) and nitrite (NO2-) are naturally occurring ions that are ubiquitous in the environment. Both are products of the oxidation of nitrogen by micro-organisms in plants, soil or water and, to a lesser extent, by electrical discharges such as lightning. Nitrate is the more stable form of oxidized nitrogen but can be reduced by microbial action to nitrite, which is moderately reactive chemically. Studies have shown that nitrate are found in many types of food especially meat products and vegetables. In fact it was shown that up to 90% of human dietary intake of nitrate come from vegetables which may contain up to two grams of nitrate per kilogram of vegetables. Nitrite, on the other hand are generally found in baked food and certain types of vegetables at much lower level at 2-4 mg per kg food. However, about 20% of nitrate may be converted to nitrite in the mouth by action of saliva and bacteria and more will be converted in the stomach. The toxicity of nitrate is generally due to its reduction to nitrite. Nitrite may react with some amine compounds to form N-nitroso compounds which were found to cause cancer in animal study and thus are suspected carcinogens. Based on many studies, health regulating bodies such as World Health Organization (WHO) has issue the allowable daily intake (ADI) of nitrate and nitrite to a maximum of 3.7 mg and 0.07 mg per Kg body weight respectively. Thus the allowable daily intake of a 60 Kg human is 222 mg for nitrate and 4.2 mg for nitrate per day. The WHO allowable limit for nitrite in food is 34 ppm and under the Malaysian Standard for Blood Bird’s Nest, the allowable limit is set at 30 ppm for nitrite.
China has recently adopted an allowable limit of 30 ppm (mg/Kg) of nitrite in imported Blood Bird’s Nest. This limit had been agreed with the Malaysian Government recently. Recent study by the Department of Veterinary Services, Johor(a state in Malaysia) reported that most Blood Bird’s Nest produced in Johor contain levels of nitrites in excess of the allowable limit. Their finding is summarised in the table below.
It is interesting to note that the percentage of uncleaned Blood Bird’s Nest containing 0-20 ppm nitrite was 57% while 32% contained 0-160 ppm nitrate. Upon cleaning, the percentage of 0-20 ppm nitrite increased to 62% and 0-160 ppm nitrate level was 81%. Nitrate and Nitrite are naturally occurring ions that can be found abundantly in the environment, including food. Studies have shown that Nitrate and nitrite are found in many types of food such as vegetables, fruit and meat product.

Nitrate and Nitrite

Nitrite is often used as a food additive for preservation and coloring purposes and the prevention of bacterial growth. Although it is widely used to cure meat, nitrite, in the presence of secondary amines in food product or in digestive system, will be converted into Nitroso Compounds and can cause health problems, including cancer, if ingested in large amounts. However, Nitrite will not pose serious threats to human health if it is used in accordance with the standards.
Nitrate which always present as sodium nitrate is the more stable form but tend to be oxidized to Nitrite in the existence of bacterial activity and oxygen. Thus, both nitrate and nitrite must be monitored to ensure the quality and safety of meat products.
Nitrite in red Blood Bird’s Nest may be formed naturally. Though, there are patches of circumstances found that a few sellers attempted to turn white Blood Bird’s Nest into red Blood Bird’s Nest to persuade customers who believe the blood-red nests are more nutritious thus increase the higher prices and demand from consumer for the red Blood Bird’s Nest. Unfortunately, adulterated Blood Bird’s Nests often feature excessive nitrite levels.

Heavy Metal

Heavy metal comes from an external sources. Swiflet birds travel around to acquire foods and drinks. During the process, there are a lot of possibility for the birds to bring along contaminant to their nests. Other than bacteria, heavy metal is one of a main concern among Blood Bird’s Nest consumer because most of heavy metal is very well known to cause cancer.

Bacteria

Since swiflet bird made the nest using its saliva, there are very high probability for bacterial growth especially the nest will be going to be a place where the birds live which we can expect fecal from the birds to present. Because of that, hygienic factor of the Blood Bird’s Nest is the main priority. Cleaning process must be done very thoroughly to make sure the Blood Bird’s Nests are clean and edible without affecting health of consumer.

Protein analysis

Protein analysis seems to be unnecessary because protein is not the main nutrient aimed by the consumer here. Raw Blood Bird’s Nest contain a quite high amount of protein. A high quality Blood Bird’s Nest should contain a high amount of protein too.
Though, there is a research found that after processing, the protein level decrease so much due to the cleaning process by using hydrogen peroxide, H2O2 as bleaching agent. It does not only eliminate all the bacteria and heavy metals, it also removes the essential sialic acid, minerals, epidermal growth factor and antioxidants. Through protein analysis, we can confirm no bleaching agent was used during the processing and the nutrient level is preserved to the optimum.

Moisture

Some Blood Bird’s Nest processor spray water to their processed Blood Bird’s Nest for easier packaging. While the Blood Bird’s Nest already clean, the high water content in the Blood Bird’s Nest provide a good environment for yeast and molds growth. This will force them to use preservative to prevent the growth thus increasing the level of nitrite, which when consumed in high amount will cause cancer.