Royal Jelly Proteins Research Articles

Biological properties and activities of major royal jelly proteins and their derived peptides

• Royal Jelly (RJ) is a nutritious secretion of honey bees fed to their developing larvae. • As other hive products, such as honey, it is used by humans for decades. • In addition to the nutritional role, Major Royal Jelly Proteins (MRJPs) and their derived peptides present a plethora of valuable health benefits for human. • MRJPs and their derived peptides have anti-inflammatory, immune-modulatory, antimicrobial, anticarcinogenic, hypotensive, hypolipidemic, growth-promoting, wound healing and neuroprotective activities. Royal jelly (RJ) is a complex beehive product that is important for larval development and queen nutrition in the hive and may also have beneficial effects on human health, according to in vitro and in vivo studies. The main proteins in RJ belong to the Major royal jelly proteins family (MRJPs), representing up to 90% of the total proteins. This narrative review aims to compile the results of studies on MRJPs and their derived peptides to understand their biological effects better, their most important activities being antioxidant, antimicrobial, anti-tumor, hypotensive, hypolipidemic, cell growth promoting, wound healing, anti-aging, neuroprotective, anti-inflammatory and immune-modulatory.

Analysis of Major Royal Jelly Proteins Stability and Ligand Binding by Differential Scanning Fluorimetry

Royal jelly (RJ) is produced by the hypopharingeal and mandibular glands of bees. Up to 90% of the RJ proteins are the major royal jelly proteins (MRJPs) that exhibit biological properties. Protein stability is of interest for biotechnology and can be explored by using differential scanning fluorimetry (DSF). We tested the stability by using a fluorescence dye (SYPRO® Orange,1:1000 dilution) in the presence of different ligands for MRJP1 and MRJP2 at 2 µM concentration. The fluorescence intensities (FI) were measured using the Real-Time PCR System and were fitted as a function of temperature according to the Boltzmann equation to determine the melting temperature (Tm) at which the amount of folded and unfolded protein is equal. In general, ligands stabilize the protein when binding to it and increase the Tm. Due to the increased starting fluorescence of the oligomeric MRJP1, the Tm values were only determined for monomeric MRJP1 and MRJP2. ATP, histamine, and thiamine were shown to determine an increase of Tm for the monomeric MRJP1 in 20 mM Tris-HCl, pH 7.4, 150 mM NaCl. For the ligand ATP, the minimum limit of binding starts at 1.5 mM concentration, for the ligand Histamine it starts at 0.6 mM concentration, while for the ligand Thiamine it starts at 3 mM concentration.

Characterization of Various Honey Samples from Different Regions of Morocco Using Physicochemical Parameters, Minerals Content, Antioxidant Properties, and Honey-Specific Protein Pattern

Honey is a bee product relatively expensive; therefore, it has been a target of adulteration by many sweeteners. In this work, we evaluated the good quality, authenticity, and content in bioactive molecules of twenty-two Moroccan honey from different botanical origins and geographical areas. For that, the following analyses were determined: the content in total protein and especially the major royal jelly protein (apalbumin 1), the analysis of total acidity, free acidity, lactonic acidity, pH, ash, Pfund, electrical conductivity, and moisture. In addition, the content of sodium, potassium, calcium, and magnesium, the dosage of polyphenols, flavones, and flavonols, and the antioxidant activities were assessed. All analyzed samples had good antioxidant activities and present a source of antioxidant compounds, the predominant mineral in all honey samples was potassium, and the physicochemical parameters are in line with the standards’ recommended limits. The content of honey samples in total protein and apalbumin 1 ranged between 212 μg/g and 4121.2 μg/g and between 27.4 μg/g and 790.82 μg/g, respectively. Overall, the detection of apalbumin 1 in all honey samples and the results of physicochemical parameters, minerals, bioactive compounds, and antioxidant activities confirm the authenticity and no adulteration of Moroccan honey.

Protein Analysis of Royal Jelly Bee Apis Mellifera Carpatica as Candidates for Immunotherapy in Reproductive Disorders

Background: The honey bee of the Apis mellifera species is the main type of bee cultivated in almost all countries in the world, including Indonesia. Honey has benefits in various aspects, including food, health, and beauty. Royal jelly is a daily food menu as a food supplement to maintain and increase health and vitality. Purpose: Analyzing the immunogenic and toxinogenic proteins of Royal jelly Apis mellifera carpatica as immunotherapy. Method: Conversion of nucleotides into amino acids. Royal jelly Apis mellifera carpatica, analysis of the three-dimensional structure of Protein of Royal jelly Apis mellifera carpatica, Analysis of Ramachandran Plots of Royal jelly Apis mellifera carpatica, Analysis of epitope and allergen proteins, Analysis of antigens and toxins. Research result: The research results found that six proteins of Royal jelly Apis mellifera carpatica were epitope, antigenic, and non-toxic. Allergens and three Royal jelly proteins, Apis mellifera carpatica, are non-allergenic. In this study, the three-dimensional structure has not been found and opens opportunities for proteomic studies of the Royal jelly Apis mellifera carpatica protein, including protein isolation. Conclusion: The findings of this study can be used as a basis for the use of immunotherapy materials against the protein Royal jelly Apis mellifera carpatica against reproductive disorders.

Identification of insect sources of honey in China based on real-time fluorescent LAMP technology

The main insect sources of Chinese honey are Apis cerana and Apis mellifera. A. cerana honey has a higher nutritional value; therefore, its price is invariably several times higher than that of A. mellifera honey. Driven by economic interests, low-priced A. mellifera honey is often misrepresented as the more valuable A. cerana honey. In this study, Apis Major Royal Jelly Protein 2 (MRJP2) was used as the target gene to design specific primers for two bee species, a rapid real-time fluorescent LAMP method was established, and the specificity and sensitivity of this method were analyzed. Results showed that this method could effectively distinguish between the A. cerana and A. mellifera honey. The detection sensitivity of the A. cerana honey and A. mellifera honey primers were 4 ng/μL and 1 ng/μL, respectively. Eight commercially available honey samples were tested with the method developed in this study, and four adulterated honey samples were identified. The detection method proposed in this study may improve the quality of and safety measures for honey.

Antimicrobial activity of major royal jelly protein 8 and 9 of honeybee (Apis mellifera) venom

• Apis mellifera has major royal jelly protein 8 and 9 (AmMRJP 8 and 9) in its venom. • AmMRJP 8 and 9 inhibit microbial serine proteases. • AmMRJP 8 and 9 exhibit antibacterial and antifungal activities. • AmMRJP 8 and 9 act as antimicrobial agents in honeybee venom. Honeybee venom is a complex mixture of toxic components, including major royal jelly protein (MRJP) 8 and 9. MRJP 8 and MRJP 9 are allergens, and MRJP 8 reduces melittin-induced cell apoptosis. However, their functional roles are poorly understood, and their antimicrobial activities have not been determined. In this study, the antimicrobial role of MRJP 8 and MRJP 9 of honeybee ( Apis mellifera ) venom (AmMRJP 8 and AmMRJP 9) was demonstrated. The presence of AmMRJP 8 and AmMRJP 9 in the secreted venom was observed using antibodies against recombinant AmMRJP 8 and AmMRJP 9 produced in baculovirus-infected insect cells. Recombinant AmMRJP 8 and AmMRJP 9 exhibited an inhibitory activity against microbial serine proteases. Consistent with their inhibitory activity, they induced structural damage by binding to microbial surfaces, resulting in a broad-spectrum antimicrobial activity against bacteria and fungi. They had little effect on hemolysis. Therefore, AmMRJP 8 and AmMRJP 9 could function as antimicrobial agents in honeybee venom.

The effects of sublethal doses of imidacloprid and deltamethrin on honeybee foraging time and the brain transcriptome

AbstractColony collapse disorder (CCD) has occurred in the United States since 2006 and has been reported in many countries with varying levels of severity. Although the cause of CCD is multifactorial, pesticide is a major factor that leads to colony collapse. At sublethal doses, pesticide is known to negatively affect honeybee physiological development and behaviour. Previously, we found the insecticides imidacloprid and deltamethrin significantly reduced honeybee dancing and foraging efficiency. In our experiments, the duration of honeybee imbibing food at the feeder declined and the returning period from the feeder to the hive increased in both insecticide groups compared with the control group. As a follow‐up, we performed a deep RNA‐seq analysis to reveal the gene regulatory mechanisms underlying this altered foraging performance. Genes involved in detoxification were upregulated in both the imidacloprid and the deltamethrin‐treatment groups. Gene members in immune pathways, odorant receptors and major royal jelly protein families were significantly downregulated in the treatment groups compared with the controls. This fluctuating gene expression profile shows that multifaceted aspects of honeybee physiology were affected by the two insecticides, which may lead to inaccurate communication and impaired learning and memory. Our findings reveal candidate molecular mechanisms leading to impaired dance performance in honeybees exposed to insecticides.

Proteome analysis reveals the molecular basis of honeybee brain and midgut response to sulfoxaflor

Sulfoxaflor is a widely used pesticide in agriculture. However, the molecular effects of sublethal sulfoxaflor on honeybees (Apis mellifera L.) remain elusive. Here, the effects of a sublethal dose of sulfoxaflor (0.05 μg/bee) on the brain and midgut proteome response of the honeybee were investigated. Exposure to sublethal sulfoxaflor doses did not cause significant honeybee death, but it induced significant alterations in the brain and midgut proteomes. After sulfoxaflor challenge, 135 and 28 proteins were differentially regulated in the brain and midgut, respectively. The up-regulated proteins were mainly implicated in energy metabolism, neurotransmitter transport and drug metabolism processes, and included in particular enzymes of the citrate cycle and cellular respiration process, such as ATP citrate synthase, malate dehydrogenase, cytochrome b-c1 complex subunits, and NADH dehydrogenase. These findings suggest that honeybees enhance energy metabolism in the midgut and brain to resist sulfoxaflor challenge. Notably, treatment with sulfoxaflor resulted in a 6.8 times increase in expression levels of the major royal jelly protein 1 (MRJP1) in the brain, and knockdown of MRJP1 mRNA expression using RNA interference significantly decreased the survival rate, indicating that MRJP1 may play an important role in sulfoxaflor tolerance. Our data reveals that sulfoxaflor influences multiple processes related to both metabolism and the nervous system, and provides novel insights into the molecular basis of the honeybee brain and midgut response to sublethal dose of sulfoxaflor.

Expression of MRJP3 and HSP70 mRNA Levels in Apis mellifera L. Workers after Dietary Supplementation with Proteins, Prebiotics, and Probiotics.

Simple SummaryThe development of honey bees depends on royal jelly. Some of the main components of this glandular secretion are proteins known as major royal jelly proteins (MRJPs). In addition to these, bees also produce proteins that help balance the body in situations of stress. During periods of food shortage, beekeepers can supplement colony food to maintain the health and productivity of honey bees. Supplementation can be provided with feed that contains, in addition to other components, probiotics and prebiotics, which are important to balance bees’ intestinal microbiota and, consequently, to keep colonies healthy and without food stress. In order to verify if the addition of probiotics and prebiotics in food supplementation is efficient for the health of bees, after supplementation, we analyzed the levels of synthesis of one MRJP and the stress protein HSP70. The results show that, with the addition of probiotics and prebiotics in the supplementation, there was an increase in the synthesis of the analyzed MRJP3 and low synthesis of the stress protein. Therefore, we suggest that these components be added to honey bee supplementation in periods of food shortage, as it will contribute to colony health in apiaries.Royal jelly is an essential substance for the development of bees from larval to adult stages. Studies have identified a group of key proteins in royal jelly, denominated major royal jelly proteins (MRJPs). The group currently consists of nine proteins (MRJP1–MRJP9), with MRJP1 being the most abundant and MRJP3 being used as a microsatellite marker for the selection of queens with a greater production of royal jelly. The diet of bees is mostly composed of proteins, and supplementing this intake to encourage a higher production of their primary product is important for producers. It is estimated that, by adding probiotic and prebiotic organisms to their diets, the benefits to bees will be even greater, both for their immune systems and primary responses to stress. Circumstances that are adverse compared to those of the natural habitat of bees eventually substantially interfere with bee behavior. Stress situations are modulated by proteins termed heat shock proteins (HSPs). Among these, HSP70 has been shown to exhibit abundance changes whenever bees experience unusual situations of stress. Thus, we sought to supplement A. mellifera bee colony diets with proteins and prebiotic and probiotic components, and to evaluate the expression levels of MRJP3 and HSP70 mRNAs using qRT-PCR. The results revealed that differences in the expression of MRJP3 can be attributed to the different types of feed offered. Significant differences were evident when comparing the expression levels of MRJP3 and HSP70, suggesting that protein supplementation with pre/probiotics promotes positive results in royal jelly synthesis carried out by honey bee nurses.

Integrated Approach to Extract and Purify Proteins from Honey by Ionic Liquid-Based Three-Phase Partitioning.

The purification of value-added compounds by three-phase partitioning (TPP) is a promising alternative to conventional processes since the target compound can be easily recovered from the liquid-liquid interphase. Although this technique has been successfully applied to the recovery of proteins, the minimization of the use of salts and solvents must be pursued to improve the overall process sustainability. Accordingly, we have here investigated the use of biobased glycine-betaine ionic liquids (IL) directly with honey, a carbohydrate-rich matrix, as phase-forming components of TPP systems. These ILTPP systems were applied in the purification of major royal jelly proteins (MRJPs) from honey. The results obtained show that MRJPs mostly precipitate in the ILTPP interphase, with a recovery yield ranging between 82.8% and 97.3%. In particular, MRJP1 can be obtained with a purity level up to 90.1%. Furthermore, these systems allow the simultaneous separation of antioxidants and carbohydrates to different liquid phases. The proposed approach allows the separation of proteins, antioxidants, and carbohydrates from honey in a single step, while using only ILs and a real carbohydrate-rich matrix, thus being sustainable TPP processes.

Major royal jelly proteins alleviate non-alcoholic fatty liver disease in mice model by regulating disordered metabolic pathways.

Nonalcoholic fatty liver disease (NAFLD), the major cause of global chronic hepatic injury, has obtained increasing attention while the current drug treatment still laid safety hazards. Major royal jelly proteins (MRJPs), the water-soluble proteins enriched in royal jelly (RJ), were applied to study its effects on improving NAFLD in the NAFLD mouse model. Herein, we demonstrated that intaking of 250-500 mg/kg/day MRJPs significantly decreased the rate of obesity, dyslipidemia, hepatic steatosis, and insulin resistance. Next, TOF to MRM ("TM") widely targeted metabolomics (untargeted metabolomics + widely targeted metabolomics) was further used to explore the potential mechanism, and we found that 500 mg/kg MRJPs alleviated lipid metabolism, oxidative stress, and inflammation mainly by regulating the metabolisms of alpha-linolenic acid, linoleic acid, arachidonic acid, and biosynthesis of unsaturated fatty acids. Moreover, by detecting multiple oxidative stress factors and inflammatory cytokines, we found that MRJPs indeed exerted antioxidant and anti-inflammatory effects. Together, we demonstrated that MRJPs could mediate the progress of NAFLD through the "multi-component-multi-target-multi-pathway" mechanism, which could be considered as an ideal functional food in alleviating NAFLD. PRACTICAL APPLICATIONS: Royal jelly (RJ) is a bee product with high nutritional value. Major royal jelly proteins (MRJPs) are water-soluble proteins in RJ. Our research showed that MRJPs significantly ameliorated NAFLD induced by a high-fat diet in mice, suggesting that MRJPs could be used as an active ingredient to help improve NAFLD, which was beneficial for the development of related functional foods and the economic value of RJ. Moreover, the metabolic pathways involved in the ameliorative effect of MRJPs were investigated, which provided new ideas for the prevention and treatment of NAFLD.

The C-Terminal Penta-Peptide Repeats of Major Royal Jelly Protein 3 Ameliorate the Progression of Inflammation in Vivo and in Vitro.

Royal jelly (RJ) has been used as a functional foodstuff and in cosmetics for many years. RJ contains various molecules, including major royal jelly proteins (MRJPs), and affords a number of health benefits such as anti-inflammatory activity. As MRJP3 has been reported to possess anti-inflammatory properties by the in vitro analysis, we investigated the anti-inflammatory effects of MRJP3 and its derived peptides both in vitro and in vivo. Expression of both tumor necrosis factor (TNF)-α and interleukin-6 (IL-6) mRNAs in lipopolysaccharides (LPS)-stimulated THP-1 cells was reduced by the addition of MRJP3 or its C-terminal tandem penta-peptide repeats (TPRs) sequence. In the herpes simplex virus type 1 (HSV-1)-induced herpes stromal keratitis (HSK) model mice, the instillation of TPRs reduced the disease scores and the expression levels of TNF-α and IL-6 in HSV-1-infected eyes. In addition, synthetic penta-peptides derived from TPRs reduced the expression of TNF-α and IL-6 both in the THP-1 cell cultures and in the HSK model mice. Our results indicated that MRJP3 TPRs would be useful in controlling inflammation.

Characteristics of honey bee physiological proteins extracted from faba bean (Vicia faba L.) honey

Faba bean (Vicia faba L.) is a valuable agricultural crop, widely used in the food industry and environmental protection. In this study, faba bean honey samples were subjected to quantitative and qualitative proteomic analysis. Using a direct mass spectrometry assay, a total of 45 honey bee (Apis mellifera) proteins were identified in faba bean honey, of which 18 were of known honey bee origin, and the others were uncharacterized honey bee proteins. Royal jelly proteins accounted for 39.29% of all known honey bee proteins identified in this study, while major royal jelly protein (MRJP-1) accounted for 47.06% of the royal jelly proteins. Honey bee proteins were separated from acid to basic isoelectric point (pI). Carboxyl ester hydrolase OS=Apis mellifera OX7460 was mostly an acidic protein and separated at pI 4.40. Transferrin OS=Apis mellifera was separated close to the neutral isoelectric point, at pI 6.73. Histone H2B OS=Apis mellifera OX7460 was mostly basic and its pI value was 10.82. The honey bee proteins identified in the faba bean honey confirm its naturalness and valuable enzymatic properties.

Differential Expression of Major Royal Jelly Proteins in the Hypopharyngeal Glands of the Honeybee Apis mellifera upon Bacterial Ingestion.

Simple SummaryTransgenerational immune priming (TGIP) to elicit social immunity in the honeybee Apis mellifera has two axes: the first is the ingested pathogen fragments–vitellogenin (Vg)–queen’s ovary axis for the developing embryo, and the second is the ingested pathogen fragments–Vg–nurse’s hypopharyngeal gland axis for the queen and young larvae through royal jelly. However, the dynamics of the expression of the major royal jelly proteins (MRJPs) in the hypopharyngeal glands of A. mellifera nurse bees after bacterial ingestion must be determined to improve our understanding of the second axis of TGIP. In this study, we investigated the expression patterns of MRJPs 1–7 and defensin-1 in the hypopharyngeal glands and Vg in the fat body of nurse bees fed with live or heat-killed Paenibacillus larvae over 12 h or 24 h by using northern blot analysis. We found that the expression of MRJPs and defensin-1 in the hypopharyngeal glands and Vg in the fat body was significantly induced in nurse bees upon bacterial ingestion, indicating that the differential expression patterns of MRJPs, defensin-1, and Vg were dependent on the bacterial status and timing of bacterial ingestion. We also found that antimicrobial peptide (AMP) genes showed induced expression in young larvae upon bacterial ingestion. In summary, our findings indicate that MRJPs in the hypopharyngeal glands are upregulated along with Vg in the fat body of nurse bees upon bacterial ingestion, providing novel insights into the ingested pathogen fragments–Vg–nurse’s hypopharyngeal gland axis for TGIP.Honeybee vitellogenin (Vg) transports pathogen fragments from the gut to the hypopharyngeal glands and is also used by nurse bees to synthesize royal jelly (RJ), which serves as a vehicle for transferring pathogen fragments to the queen and young larvae. The proteomic profile of RJ from bacterial-challenged and control colonies was compared using mass spectrometry; however, the expression changes of major royal jelly proteins (MRJPs) in hypopharyngeal glands of the honeybee Apis mellifera in response to bacterial ingestion is not well-characterized. In this study, we investigated the expression patterns of Vg in the fat body and MRJPs 1–7 in the hypopharyngeal glands of nurse bees after feeding them live or heat-killed Paenibacillus larvae. The expression levels of MRJPs and defensin-1 in the hypopharyngeal glands were upregulated along with Vg in the fat body of nurse bees fed with live or heat-killed P. larvae over 12 h or 24 h. We observed that the expression patterns of MRJPs and defensin-1 in the hypopharyngeal glands and Vg in the fat body of nurse bees upon bacterial ingestion were differentially expressed depending on the bacterial status and the time since bacterial ingestion. In addition, the AMP genes had increased expression in young larvae fed heat-killed P. larvae. Thus, our findings indicate that bacterial ingestion upregulates the transcriptional expression of MRJPs in the hypopharyngeal glands as well as Vg in the fat body of A. mellifera nurse bees.

Determination of total protein, trans- 10-Hydroxy-2-Decenoic Acid (10-HDA) and major royal jelly proteins in royal jelly produced at different harvest times in queenless and queenright colonies

Two of the most important ingredients to add functional properties to royal jelly are 10-HDA and major royal jelly proteins (MRJPs). The effect of royal jelly (RJ) production and the effect of royal jelly harvest time (24, 48 and 72 hours) on 10-HDA, the total protein content of RJ, and molecular weights of major proteins in RJ were investigated in queenright and queenless colonies. RJ production colonies were divided into 2 groups as queenless and queenright where the queen was confined with frames. Subgroups were formed within each group (queenless and queenright) according to harvest time (24, 48 and 72 hours). 80 larvae were transferred to each colony. In this study, as the harvest time increased, total protein (TP) ratio decreased (p<0.05); TP ratio in RJs harvested at 24, 48 and 72 hours was determined as 18.4±1.24%, 15.2±0.80%, 10.6±0.27%, respectively. As the harvest time increased, 10-HDA decreased. It was determined 10-HDA rates in RJ harvested at 24, 48 and 72 hours respectively; 2.52±0.109%, 2.20±0.110%, 2.00±0.112%. MRJP1, MRJP2, MRJP3 and MRJP5 were found from the MRJP family, and their molecular weights were determined as 53 kDa, 46.5 kDa, 66.8 kDa, 80.9 kDa, respectively. As a result, the early harvested (24 and 48 hours) RJ had higher values in terms of TP and trans 10-HDA than the royal jelly harvested at 72 hours.

Changes in chemical composition and antioxidant activity of royal jelly produced at different floral periods during migratory beekeeping

Over 90% of global royal jelly (RJ), a functional food with various health benefits, is produced in China mainly by migratory beekeeping of a high RJ-producing honeybee (RJB) strain. To explore quality changes of RJ produced by migratory RJBs at different floral periods, we performed metabolomics and proteomics analysis and assessed RJ antioxidant activity. Overall, the RJ metabolic and proteomic profiles were observed to vary with floral periods. Minor sugars (raffinose, erlose, and sucrose) and major RJ protein 5 (MRJP5) were identified among the discriminating components mainly contributing to the altered profiles. Water, crude protein, and the trans-10-hydroxy-2-decenoic acid (10-HDA) content fulfill the requirements of the International Organization for Standardization regardless of floral periods. Notably, the 10-HDA content increased 11.05%–19.65% during tea blooming. Moreover, changes in antioxidants resulted in significant difference in RJ antioxidant activity. The integrated omics data provide a detailed view of chemical composition for RJ quality evaluation.

Suppressing crucial oncogenes of leukemia initiator cells by major royal jelly protein 2 for mediating apoptosis in myeloid and lymphoid leukemia cells.

Relapse of leukemia and drug resistance are still the major obstacles to therapy due to leukemia-initiating stem/progenitor cells (LICs); thus, targeting them using safe compounds is crucial. Here, we evaluated the anti-leukemic effect of royal jelly (RJ) components, which had a higher safe concentration (EC100 values) than the chemotherapeutic drug doxorubicin (DOX). The RJ-protein fraction 50 (PF50, precipitated at 40-50% ammonium sulfate saturation) and its constituents, major RJ protein (MRJP) 2 and its isoform X1, exhibited the highest growth inhibitory effect against myeloid NFS-60 and lymphoid Jurkat cell lines. MRJP2 has a nanosize, which may be the reason for its higher anti-leukemic activity than its isoform. These RJ proteins, particularly MRJP2, suppressed LIC-associated oncogenes (GATA2 and Evi-1) and eliminated CD34+ LICs, in contrast to the low anti-LIC efficacy of DOX. MRJP2 demonstrated higher apoptotic activity than its isoform by upregulating p53 and p21-mediated cell cycle arrest. This study also reported the potent inhibitory effect of RJ-proteins on matrix metallopeptidase 10 (metastatic marker) and histone deacetylase 8 (mediates LIC survival) activities. Thus, MRJP2 can be considered a promising novel therapeutic agent for both myeloid and lymphoid leukemia.

Dermatan sulfate and chondroitin sulfate from Lophius litulon alleviate the allergy sensitized by major royal jelly protein 1.

The objective of the present study was to explore the desensitization effect of dermatan sulfate (DS) and chondroitin sulfate (CS) from Lophius litulon (Ll) on mice sensitized by major royal jelly protein 1 (MRJP1). First, the affinity between six glycosaminoglycans and the MRJP1 polyclonal antibody was measured by the ELISA method. Lophius litulon dermatan sulfate (Ll DS) and Lophius litulon chondroitin sulfate (Ll CS) were selected due to their highest binding affinity. Second, the molecular docking method was used to explore the interaction between Ll DS and MRJP1 and Ll CS and MRJP1. The results showed that Ll DS and Ll CS combined with MRJP1 successfully, which meant a potential function of relieving the MRJP1-caused allergy. Finally, the MRJP1-sensitized mice model was established and confirmed that Ll DS and Ll CS had the desensitization ability to relieve MRJP1-induced allergic symptoms. To validate the conclusion, the relief of allergic symptoms in mice was observed. The production of total IgE, MRJP1-specific IgE and histamine was measured. The desensitization mechanism was further studied by measuring cytokines (IL-4 and IFN-γ) from splenocytes stimulated with MRJP1 in vitro. Based on in vivo and in vitro experiments, it was confirmed that Ll DS and Ll CS have the ability to alleviate MRJP1-induced allergic symptoms, which proposes a potential candidate material against IgE-mediated food allergy.

Major Royal Jelly Proteins

Günümüzde fonksiyonel gıdalara olan ilgi giderek artmaktadır. Arı ürünlerinden biri olan arı sütü de fonksiyonel gıdalar arasında yer alır. Arı sütü genç yaştaki (5-15 gün) işçi arılar tarafından salgılanır ve kolonide işçi arı larvasının ana arı larvasına dönüşmesini sağlar, kısaca kast belirleyici gıda olarak da bilinir. Arı sütünün içeriğinde temel olarak, su (%60 -70), protein (%9-18), lipit (%3-8), karbonhidrat (%7-18), kül (%0.8-3),10-hidroksi-2-dekenoik asit (10-HDA) (>%1.4), az miktarda vitamin, tuz ve serbest amino asit bulunur. Şu anki bilgilere göre, arı sütüne fonksiyonel özellik veren en önemli iki içerik, 10-HDA ve major arı sütü proteinleridir. Arı sütündeki toplam proteinin %82-90’ını major arı sütü proteinleri (MASP) oluşturur. MASP ailesi 10 üyeden oluşur. MASP ailesinde ilk keşfedilen ve en bol olan MASP1 üzerine son yıllarda çok sayıda araştırmalar yapılmıştır. Yapılan araştırmalar, MASP ailesinin genel olarak ömür uzunluğu, anti-tümör ve antioksidan etkisi, bağışıklık düzenleyici etkilerinin olduğunu ortaya koymuştur. Bu derlemede, arı sütünde bağışıklık proteinleri olarak da adlandırılan MASP ailesi ayrıntılı bir biçimde ele alınacaktır

Proteomic profiling of royal jelly produced by Apis mellifera L. exposed to food containing herbicide-based glyphosate

Royal jelly (RJ) is rich in protective elements associated with collective immune defenses in the hive of Apis mellifera. Exposure of bees to glyphosate-based herbicides causes ultrastructural changes in the hypopharyngeal glands and a reduction in the production of RJ. However, the effects of glyphosate-based herbicides on the protein composition of RJ and consequences for the hive are unknown. Thus, we performed proteomic profiling of royal jelly produced in hives of A. mellifera exposed to food containing 1,5 μL of Roundup® (2.16 mg. g−1 of glyphosate). The production of RJ was carried out in six hives, following the method of artificial production of queens. The combs containing 80 grafting cells were introduced into the hives, and the collection of royal jelly was performed after 72 h. Two treatments were determined based on hive feeding and the hive as the experimental unit: Control and “Roundup®”. Royal jelly from the Roundup® treatment hives was compared to the Control hives. Proteins with differences in expression were identified by mass spectrometry. Only the proteins present in all three biological replicates were considered in the differential abundance analysis, using Student's t-test (p-value < 0.05, two-tailed). Hives that received food containing Roundup®, analysis showed alterations in protein profile in the RJ produced therein. In total, 24 proteins were identified, and the accumulation of Major royal jelly protein 3 (MRJP3) was downregulated, showing a significant reduction in hives exposed to food containing Roundup® in relation to control hives (t = 0.0017). MRJP3 acts analogously to polyclonal antigen-antibody reactions, performing functions related to immunity in bees of different ages and castes. To the best of our knowledge, this is the first study to demonstrate changes in the proteomic profile of RJ caused by glyphosate-based herbicides, indicating its negative effects on the nutrition and social immunity of bees.