Proteome and Phosphoproteome of Africanized and European Honeybee Venoms

Proteomics, 2013 Jun 25

Honey bee venom toxins trigger immunological, physiological and neurological responses within victims. The high occurrence of bee attacks involving potentially fatal toxic and allergic reactions in humans and the prospect of developing novel pharmaceuticals make honey bee venom an attractive target for proteomic studies. Using label-free quantification, we compared the proteome and phosphoproteome of the venom of Africanized honeybees with that of two European subspecies, namely A. m. ligustica and A. m. carnica. From the total of 51 proteins, 42 were common to all three subspecies. Remarkably, the toxins melittin and icarapin were phosphorylated. In all venoms, icarapin was phosphorylated at the 205 Ser residue, which is located in close proximity to its known antigenic site. Melittin, the major toxin of honeybee venoms, was phosphorylated in all venoms at the 10 Thr and 18 Ser residues. 18 Ser phosphorylated melittin - the major of its two phosphorylated forms -was less toxic compared to the native peptide.

Eating Drone Larvae Suppresses Blood Glucose, Cholesterol

Apilarnil reduces fear and advances sexual development in male broilers but has no effect on growth

Br Poult Sci, 2013 Jun;54(3):355-361

1. An experiment was conducted to determine the possibility of stimulating sexual development at an early age in male and female broiler chickens by administration of apilarnil, a natural bee product, in the pre-pubertal period.

2. From 28 to 55 d of age, birds were given apilarnil orally. The effects of low (2.5 g/bird) and high (7.5 g/bird) doses of apilarnil on growth performance, testicular weight, secondary sexual characteristics, blood lipids, testosterone and fearful behaviour were evaluated.

3. Apilarnil administration did not cause a positive effect on growth performance of male and female broilers suggesting that apilarnil did not have an anabolic effect.

4. Apilarnil administration suppressed blood glucose and cholesterol.

5. Birds receiving apilarnil remained immobile for a shorter period in a tonic immobility test and showed less home-cage avoidance responses suggesting a lower level of fearfulness.

6. Increases in testicular weight, testosterone concentration and comb growth in males receiving apilarnil implied that it stimulates the sexual maturation at an early age. However, a similar stimulation of secondary sexual characteristics was not observed in females.

Saudis Use Honey Bee Products as Complementary and Alternative Medicine

The use of and out-of-pocket spending on complementary and alternative medicine in Qassim province, Saudi Arabia

Ann Saudi Med, 2013 MAY-JUNE;33(3):282-289.

BACKGROUND AND OBJECTIVES:

The current picture of the Saudis' use of complementary and alternative medicine (CAM) has not yet been developed. The aim of this study was to assess the feasibility of using the international questionnaire to measure use of complementary and alternative medicine (I-CAM-Q) in Saudi Arabia to evaluate the use of and out-of-pocket spending on CAM.

DESIGN AND SETTINGS:

It was a cross-sectional study, conducted in 2011, in primary health care centers in Qassim.

METHODS:

In a multistage sampling technique, 12 primary health care centers were selected randomly in the Al-Qassim province in Saudi Arabia. From each center, 100 attendants were interviewed for a total of 1160 completed questionnaires.

RESULTS:

A total of 74% of subjects had visited CAM providers in 12 months before the survey. This percentage decreased to 47.6% when spiritual healers were excluded. The specific CAM providers who were visited were spiritual healers (26.7%), herbalists (23.2%), providers of honeybee products (14.9%), and hijama (wet cupping; 13%). Chronic illnesses were the main reason for the visits. A total of 50% of subjects were satisfied with their visit. Physicians were the providers of CAM for 11.3% of the participants. More than 75% of the subjects used herbs in the previous 12 months for medical and health reasons, while only 25% used vitamins or minerals. Self-help was used in 26% of the participants. Relaxation (10.3%) was the most common self-CAM practice followed by meditation (6.7%). The subjects spent 350000 (US$) on CAM visits and 300000 (US$) purchasing CAM products.

CONCLUSIONS:

I-CAM-Q can be used in different populations and cultures in the East including Saudi Arabia after customization to overcome its limitations, as the questionnaire was developed in Western societies.

Letter: Topical Honey for Diabetic Foot Ulcers

Dtsch Arztebl Int 2013; 110(21): 373

Local Treatment of Chronic Wounds in Patients With Peripheral Vascular Disease, Chronic Venous Insufficiency and Diabetes by Dr. med. Mike Rüttermann, Dr. med. Andreas Maier-Hasselmann, Brigitte Nink-Grebe, Marion Burckhardt in volume 3/2013

The data regarding different treatments for wounds are still disappointingly scant. No grade B recommendations are available for wound care products, and grade A seems completely out of the question! A grade B recommendation exists only in favor of hydrogel, hyperbaric oxygenation in ulcers that are beyond treatment in diabetic foot syndrome, and integrated care. In view of the many patients and the vast market in wound care products, this is sobering. I was therefore all the more surprised at the recommendation not to use honey for chronic wounds (the context was really generally that of chronic wounds) because honey resulted in more pain in a single study of venous ulcers. Why was a special scenario (venous ulcers) generalized to include all chronic wounds?

Years of experience with manuka honey in patients with diabetic foot syndrome have shown that pain usually does not constitute a problem in these wounds owing to the comorbid polyneuropathy. I would have welcomed a fact-based recommendation, differentiated by evidence level (which exists for a negative recommendation only for venous ulcers). Numerous small, qualitatively moderate to poor studies of honey treatment in diabetic foot syndrome have shown mostly promising results, which are entirely consistent with my own observations. Obviously, high-quality randomized controlled studies of honey treatment in diabetic foot syndrome are absolutely necessary and desirable. But until these have been conducted and reported, the treatment of the diabetic foot syndrome with honey provides an efficient, cost-effective variant with a poor evidence base. This, however, is also the case for all other wound treatments. 

Propolis May Help Treat Hypertension

Propolis Reduces Oxidative Stress in l-NAME-Induced Hypertension Rats

Cell Biochemistry and Function, Early View

The inhibition in the synthesis or bioavailability of nitric oxide (NO) has an important role in progress of hypertension. The blocking of nitric oxide synthase activity may cause vasoconstriction with the formation of reactive oxygen species (ROS). Propolis is a resinous substance collected by honey bees from various plants. Propolis has biological and pharmacological properties. The aim of this study was to examine the effect of propolis on catalase (CAT) activity, malondialdehyde (MDA) and NO levels in the testis tissues of hypertensive rats by Nω-nitro-l-arginine methyl ester (l-NAME).

Rats have received nitric oxide synthase inhibitor (l-NAME, 40 mg kg−1, intraperitoneally) for 15 days to produce hypertension and propolis (200 mg kg−1, by gavage) during the last 5 days. MDA level in l-NAME-treated group significantly increased compared with control group (P < 0.01). MDA level of l-NAME + propolis-treated rats significantly reduced (P < 0.01) compared with l-NAME-treated group. CAT activity and NO level significantly reduced (P < 0.01) in l-NAME group compared with control group. There were no statistically significant increases in the CAT activity and NO level of the l-NAME + propolis group compared with the l-NAME-treated group (P > 0.01).

These results suggest that propolis changes CAT activity, NO and MDA levels in testis of l-NAME-treated animals, and so it may modulate the antioxidant system.

Novel Antimicrobial Peptides Isolated From Wild Bee Venom

Panurgines, Novel Antimicrobial Peptides from the Venom of Communal Bee Panurgus calcaratus

Amino Acids, 2013 Jul;45(1):143-57

Three novel antimicrobial peptides (AMPs), named panurgines (PNGs), were isolated from the venom of the wild bee Panurgus calcaratus. The dodecapeptide of the sequence LNWGAILKHIIK-NH2 (PNG-1) belongs to the category of α-helical amphipathic AMPs. The other two cyclic peptides containing 25 amino acid residues and two intramolecular disulfide bridges of the pattern Cys8-Cys23 and Cys11-Cys19 have almost identical sequence established as LDVKKIICVACKIXPNPACKKICPK-OH (X=K, PNG-K and X=R, PNG-R). All three peptides exhibited antimicrobial activity against Gram-positive bacteria and Gram-negative bacteria, antifungal activity, and low hemolytic activity against human erythrocytes. 

We prepared a series of PNG-1 analogs to study the effects of cationicity, amphipathicity, and hydrophobicity on the biological activity. Several of them exhibited improved antimicrobial potency, particularly those with increased net positive charge. The linear analogs of PNG-K and PNG-R having all Cys residues substituted by α-amino butyric acid were inactive, thus indicating the importance of disulfide bridges for the antimicrobial activity. However, the linear PNG-K with all four cysteine residues unpaired, exhibited antimicrobial activity. PNG-1 and its analogs induced a significant leakage of fluorescent dye entrapped in bacterial membrane-mimicking large unilamellar vesicles as well as in vesicles mimicking eukaryotic cell membrane. On the other hand, PNG-K and PNG-R exhibited dye-leakage activity only from vesicles mimicking bacterial cell membrane.

Antimicrobial Properties of Honey

Am J Ther, 2013 Jun 18

Honey has been widely accepted as food and medicine by all generations, traditions, and civilizations, both ancient and modern. For at least 2700 years, honey has been used by humans to treat a variety of ailments through topical application, but only recently have the antiseptic and antimicrobial properties of honey been discovered. Honey has been reported to be effective in a number of human pathologies. Clinical studies have demonstrated that application of honey to severely infected cutaneous wounds rapidly clears infection from the wound and improves tissue healing. 

A large number of in vitro and limited clinical studies have confirmed the broad-spectrum antimicrobial (antibacterial, antifungal, antiviral, and antimycobacterial) properties of honey, which may be attributed to the acidity (low pH), osmotic effect, high sugar concentration, presence of bacteriostatic and bactericidal factors (hydrogen peroxide, antioxidants, lysozyme, polyphenols, phenolic acids, flavonoids, methylglyoxal, and bee peptides), and increase in cytokine release, and to immune modulating and anti-inflammatory properties of honey; the antimicrobial action involves several mechanisms. Despite a large amount of data confirming the antimicrobial activity of honey, there are no studies that support the systemic use of honey as an antibacterial agent.

Demand for Manuka Honey Exports to Indonesia

Radio New Zealand, 6/21/2013

A Maori business consultancy firm says there's huge potential for Maori business in Indonesia and it may be able to break through by exporting manuka honey.

Members of the Poutama Trust recently attended a trade mission to Jakarta.

Chief executive Richard Jones says manuka honey has taken off in China, but so far not in Indonesia, despite high demand.

He says a large Indonesian pharmaceutical company, Kalbe Farma, is interested in importing the product and samples of manuka honey, energy bars, gels and lozenges were sent over this week…

Review: Me, the Bees and Cancer

Review by Wyndham Wise, Associate Editor, Canadian MovieDatabase, 6/20/2013

(June 20, 2013 - Toronto, Ontario) John Board is veteran Canadian 1st AD/associate producer with an impressive resume that dates back to 1970 and Paul Almond’s Act of the Heart. He has worked with David Cronenberg on seven of his films (The Brood, Videodrome, The Dead Zone, The Fly, Dead Ringers, Naked Lunch and M. Butterfly), and his other credits include the Oscar-nominated Atlantic City and the Genie-winning The Grey Fox.

Board is also a passionate advocate of homeopathic medicine, and in 2002 he compiled something call he calls his ‘Hollywood Survival Kit,’ a handy briefcase full of remedies, herbs and ointments, which he claims, enthusiastically and repeatedly, has something to cure anything. In fact, Board is so convinced the first scene in his documentary Me, the Bees and Cancer (which opens tomorrow, June 21 at the Royal in Toronto) is devoted to a rap song that he wrote and performs in front of his house – sort of an advertisement in rap extoling the virtues of his ‘Survival Kit.’

Two-years ago Board was diagnosed with Hodgkin’s lymphoma, a fairly common form of cancer resulting in the growth of a tumor in his right armpit. With a grand of seed money from the $1,000 Feature Film Challenge, he sought to document his own cancer alternative – bee sting or venom therapy. The result is an intensely personal and at times hard-to-watch documentary…

Video: Report on Medicinal Benefits of Bee Products – Honey, Propolis, Venom, Royal Jelly

Euronews, 6/18/2013

Honey has been used as a medicine since time immemorial; as a remedy for digestive problems, bad circulation, breathing difficulties and – of course – as the ultimate elixir for a sore throat.

But honey is also an effective antiseptic and antibiotic, with the power to ward off wound infections, reduce inflammation and promote healing.

The medicinal molten gold includes germ-killing inhibins, which slow down the production of bacteria, and the protein defensin-1, which helps stimulates the immune system.

Besides honey, bees also produce other useful substances, such as propolis. This is a yellow, waxy resin bees use to seal the cells in their hive. They collect it from the sap of trees, and when these trees have strong antibiotic properties – as the poplar, willow and birch do – the propolis transfers these antibiotic benefits to the hive, and thus to the honey itself.

As Professor Henri Joyeux explains, the uses of propolis are endless: “We realized that propolis could have had antiseptic effects, as an antibiotic or anti-inflammatory. And when the bees leave the hive, we will are able to extract it, and purify it. This purification is such that one can produce a small alcoholic extract, which can help, because it is quite thick, fat, and contains considerable vitamins, minerals, and has a very interesting effect on bronchoalveolar respiration. For example, if you have a child suffering from a little asthma, you put 5 drops of propolis in his breakfast, you mix it, and it’ll solve the problem immediately.”

Bee therapy’s most prized product, however, is still royal jelly; a concentrated honey made exclusively for queen bees...

Major Royal Jelly Protein Exists Mainly as Apisin in Royal Jelly

Quantification of Major Royal Jelly Protein 1 in Fresh Royal Jelly by Indirect Enzyme-Linked Immunosorbent Assay

Biosci Biotechnol Biochem, 2013 Jun 7

Rabbit polyclonal antibody produced by a major royal jelly protein 1 (MRJP1) specific peptide reacted only with a MRJP1. Indirect ELISA with the antibody revealed a MRJP1 level of 4.12-4.67 g/100 g in different company's royal jelly, which almost agreed with that of a hexametric form of MRJP1 (apisin) measured by high performance liquid chromatography. These results suggest that MRJP1 exists mainly as apisin in royal jelly.

First Case of Folliculitis Associated with Granuloma as a Complication of Bee Sting Acupuncture

Granulomatous Inflammation with Chronic Folliculitis as a Complication of Bee Sting Acupuncture

Indian J Dermatol Venereol Leprol, 2013 Jul-Aug;79(4):554

Sir,

Bee sting therapy is one of the traditional herbal medical procedures that has been widely used for the treatment of chronic recalcitrant neuralgia and arthralgia in Asian and Middle Eastern countries, and is also called apitherapy. There are two methods of this therapy, one is stinging directly to the treatment site [Figure 1] and the other is injecting the artificially extracted venom from a bee. The substrates that compose the venom of a bee have anti-inflammatory and analgesic effect. The bee leaves poison sac and neural plexus when stinging, and the retained sting materials at the treatment site may induce inflammation and granulomatous reaction.

A 50-year-old man visited our clinic complaining of erythematous papules and nodules with tingling sensation on his back [Figure 2] which showed wax and wane for the past 3 months, despite he had been treated with intralesional injection of steroid in a private clinic. He had no specific history but received bee sting acupuncture for several years in an oriental medical clinic due to his back pain. The skin lesions occurred after 2-3 years from the apitherapy procedures, directly at the sites that received the acupuncture. Histopathologic findings of biopsy specimen taken from the back showed perifollicular dense inflammatory infiltration composed of lymphocytes, plasma cells, and histiocytes [Figure 3]a and b. Around the perifollicular infiltration, ruptured granulomatous reaction composed of epitheloid cells and lymphocytes was shown [Figure 3]c. There was no giant cell or foreign body in the whole specimen. Special stains, Gomori methenamine silver (GMS) and diastase-periodic acid-Schiff (d-PAS) revealed negative findings. The patient was diagnosed as chronic folliculitis and granuloma caused by bee sting acupuncture treatment. For anti-inflammatory effect, he was treated with oral and topical antibiotics for a short period. The lesions showed slight improvement but the patient was not able to be followed-up…

Folliculitis is an inflammation in the perifollicular area that presents as erythematous papules and pustules on the trunk, neck and groin area. The causes of folliculitis are mostly infection, bacterial organisms, fungi such as Pityrosporum, viruses, and demodex. Histopathologic findings of chronic folliculitis differs from acute folliculitis, that it shows perifollicular infiltration composed of lymphoid cells, histiocytes and plasma cells rather than neutrophils.

Granulomatous inflammation is a distinctive inflammatory pattern characterized by granulomas, containing epitheloid histiocytes. Of the five histological types of granuloma, foreign body granulomatous reaction is divided into two subtypes, allergic and non-allergic. Foreign body type giant cells and foreign body surrounded by numerous histiocytes and lymphocytes are characteristically visualized in the non-allergic type. Besides, granuloma composed of histiocytes is seen in the allergic type, and phagocytosis of foreign body is not seen. [2] The granuloma of this patient is suggested to be an allergic type foreign body granulomatous reaction due to remained poison sac or neural plexus in the skin, not due to the stinger which should have been discovered in the serial section of the specimen.

Although several reports had been published in Korean literatures, there are only five reports of complications of bee sting acupuncture in English literatures, since apitherapy is usually performed in Asian and Middle Eastern countries. Only one case of the five English literatures is a dermatologic complication. Therefore, we report the first case of chronic folliculitis associated with granuloma as a complication of bee sting acupuncture therapy, and no reports of chronic folliculitis combined with granulomatous reaction as in our case have been reported.

Texas Man Uses Bee Venom Therapy to Treat MS

Man Hopes to Sting Away Effects of MS

By Kimberly Sutton, The Courier, 6/16/2013

THE WOODLANDS — The fatigue from Alan Swor’s multiple sclerosis was so bad that he had to stop and rest while brushing his teeth or while shaving.

However, the 48-year-old resident of The Woodlands is feeling much better and the fatigue is almost gone since starting a treatment regimen that involves stinging himself with bees. It’s called Bee Venom Therapy.

“It hurts,” Swor said about the stings.

Swor turned to Bee Venom Therapy because his doctor’s prescriptions were not effective.

“Nothing that the doctor told me was helping. I was using all kinds of drugs to treat MS symptoms,” he said.

His sister, Brenda Vozzo, is his health care provider and assists him by picking up bees from the Montgomery County Beekeeper Association three times per week and stinging him with about 30 bees during each treatment. Beekeepers are donating the bees to Swor at no cost, he said.

A friend told Swor about Bee Venom Therapy and he began to research it. He found Alan Lorenzo’s website, www.beewelltherapy.com. Lorenzo lives in Stanford, Conn., and flew to Houston to train Swor and his sister.

“You need to know what to do and where to sting yourself,” Lorenzo said. “You just can’t sting yourself anywhere and expect it to work,”

Lorenzo is certified in Apitherapy, using bees and products of the hive to treat health conditions. He is a member of the American Apitherapy Society.

“It’s been called America’s best-kept medicine secret,” Lorenzo said.

Bee Venom Therapy is the therapeutic application of honeybee venom through live bee stings to bring relief and healing for various spinal, neural, joint or musculo-skeletal ailments, according to Lorenzo’s website.

Lorenzo said bee venom is antibacterial, antiviral and kills cancer cells.

“Only during the first stage, if you sting a cancer tumor with a bee sting, it disappears,” he said.

Conditions that respond to the treatment are Parkinson’s disease, Alzheimer’s, depression, fibromyalgia, bursitis, tendinitis, lupus and many more, Lorenzo said…

Toronto Documentary About Bee Sting Therapy Cost Less Than $1,000 to Make

Longtime Toronto filmmaker John Board, 78, endures more than 1,200 stings in all in his documentary

The Star, 6/15/2013

Would you be brave enough to clasp a writhing, angry honeybee in tweezers and let it sting you? How about 30 stings at a time — several times a week over the course of an entire summer — in and around your armpit?

Longtime Toronto assistant director , 78, endures more than 1,200 stings in all in his documentary Me, the Bees and Cancer, a film he made in 2012 for less than $1,000 as one of five features funded through the 1K Wave Challenge, launched by director Ingrid Veninger and The Royal cinema’s Stacey Donen.

“If I’m on a TV movie or series, I would probably spend $1,000 on a lunch,” said Board to help put his meager budget in context. “A thousand dollars a day is barely taking care of your food on a smallish production.”

Diagnosed with cancer after he discovered a lump under his arm, Board chose to treat his illness with homeopathy, healthy diet and regular doses of bee venom — an ancient and medically controversial practice involving bee hive products called apitherapy — on and around his tumor…

Me the Bees and Cancer premieres at the Royal Cinema, 608 College St. W. June 21.

Indian Stingless Bee Propolis Demonstrates Anticancer Activity

Anticancer Activity of Indian Stingless Bee Propolis: An in vitro Study

Evid Based Complement Alternat Med, 2013;2013:928280

Indian stingless bee propolis has a complex chemical nature and is reported to possess various medicinal properties. In the present study, anticancer activity of the ethanolic extract of propolis (EEP) was explored by testing the cytotoxic and apoptotic effect in four different cancer cell lines, namely, MCF-7 (human breast cancer), HT-29 (human colon adenocarcinoma), Caco-2 (human epithelial colorectal adenocarcinoma), and B16F1 (murine melanoma), at different concentrations.

Cytotoxicity was evaluated by MTT assay and Trypan blue dye exclusion assay. EEP at a concentration of 250 μg/mL exhibited ≥50% mortality in all cell lines tested (i.e., IC50 value). EEP revealed a concentration and time dependent cytotoxic effect. Apoptosis was estimated by differential staining (ethidium bromide/acridine orange) and TUNEL (deoxynucleotidyl transferase-dUTP nick end labeling) assay. Light microscopy and atomic force microscopy demonstrated morphological features of apoptosis in all the cell lines after treatment with 250 μg/mL EEP for 24 h.

Thus, early onset of apoptosis is the reason for anticancer activity of Indian stingless bee propolis. Further, the antioxidant potential of Indian stingless bee propolis was demonstrated to substantiate its anticancer activity.