Thymoquinone ameliorates Pachycondyla sennaarensis venom-induced acute toxic shock in male rats.

BACKGROUND: For many decades, the sting of Samsun ant (Pachycondyla sennaarensis) has been a serious clinical challenge for the people living in some of the major Middle East and Asian countries. In the present study, the therapeutic potential of Nigella sativa derived plant extract component, thymoquinone (TQ) has been tested against the Samsun ant venom (SAV) at the toxic dose in the rats. METHODS: The adult male rats were divided into four groups (n = 10): control, SAV treated, SAV + TQ treated and TQ alone treated. It was found that the sub-lethal dose of SAV alters not only many of the kidney and liver function markers but also induces oxidative stress in the animals. Moreover, the SAV also disturbs various immunological parameters including expression of PMNs, CD-80, CD-86, interleukins and other cytokines compromising the affected organism towards mild to severe allergic reactions including life-risking anaphylaxis. RESULTS: The plant extract, TQ, effectively restores many of the biochemical and oxidative stress parameters comparable to the normal concomitant with improving the immunological aspects that might attributive in relieving from SAV-induced toxicity and allergic reactions in the affected organism to a greater extent. CONCLUSION: Hence, TQ has an excellent antidote property against SAV-induced toxicities in vivo. Although the study is a vivid indication of the potential therapeutic potential of TQ against the SAV induced in vivo toxicity, yet the actual mechanism of interaction translating the toxicity amelioration warrants further investigations.

In vitro comparison of three common essential oils mosquito repellents as inhibitors of the Ross River virus.

BACKGROUND: The essential oils of Cymbopogon citratus (CC), Pelargonium graveolens (PG) and Vetiveria zizanioides (VZ) are commonly used topically to prevent mosquito bites and thus the risk of infection by their vectored pathogens such as arboviruses. However, since mosquito bites are not fully prevented, the effect of these products on the level of viral infection remains unknown. OBJECTIVES: To evaluate in vitro the essentials oils from Reunion Island against one archetypal arbovirus, the Ross River virus (RRV), and investigate the viral cycle step that was impaired by these oils. METHODS: The essential oils were extracted by hydrodistillation and analyzed by a combination of GC-FID and GC×GC-TOF MS techniques. In vitro studies were performed on HEK293T cells to determine their cytotoxicity, their cytoprotective and virucidal capacities on RRV-T48 strain, and the level of their inhibitory effect on the viral replication and residual infectivity prior, during or following viral adsorption using the reporter virus RRV-renLuc. RESULTS: Each essential oil was characterized by an accurate quantification of their terpenoid content. PG yielded the least-toxic extract (CC50 > 1000 µg.mL-1). For the RRV-T48 strain, the monoterpene-rich CC and PG essential oils reduced the cytopathic effect but did not display virucidal activity. The time-of-addition assay using the gene reporter RRV-renLuc showed that the CC and PG essential oils significantly reduced viral replication and infectivity when applied prior, during and early after viral adsorption. Overall, no significant effect was observed for the low monoterpene-containing VZ essential oil. CONCLUSION: The inhibitory profiles of the three essential oils suggest the high value of the monoterpene-rich essential oils from CC and PG against RRV infection. Combined with their repellent activity, the antiviral activity of the essential oils of CC and PG may provide a new option to control arboviral infection.

Pre-hospital treatment of bee and wasp induced anaphylactic reactions: a retrospective study.

BACKGROUND: Bee and wasp stings are among the most common triggers of anaphylaxis in adults representing around 20% of fatal anaphylaxis from any cause. Data of pre-hospital treatment of bee and wasp induced anaphylactic reactions are sparse. This study aimed to estimate the incidence of bee and wasp induced anaphylactic reactions, the severity of the reactions and to correlate the pre-hospital treatment with the severity of the anaphylactic reaction. METHODS: Retrospective and descriptive study based on data from the Mobile Emergency Care Units (MECUs) in the Region of Southern Denmark (2008 only for Odense and 2009-2014 for the whole region). Discharge summaries with diagnosis related to anaphylaxis according to the International Classification of Diseases 10 (ICD-10) were reviewed to identify bee and wasp induced anaphylactic reactions. The severity of the anaphylactic reaction was assessed according to Sampson's severity score and Mueller's severity score. Treatment was evaluated in relation to administration of adrenaline, glucocorticoids and antihistamine. RESULTS: We identified 273 cases (Odense 2008 n = 14 and Region of Southern Denmark 2009-2014 n = 259) of bee and wasp induced anaphylaxis. The Incidence Rate was estimated to 35.8 cases per 1,000,000 person year (95% CI 25.9-48.2) in the Region of Southern Denmark during 2009-2014. According to Sampson's severity score, 65% (n = 177) of the cases were graded as moderate to severe anaphylaxis (grade 3-5). Almost one third of cases could not be graded according to Mueller's severity score. Adrenaline was administrated in 54% (96/177) of cases with moderate to severe anaphylaxis according to Sampson's severity score, compared to 88% receiving intravenous glucocorticoids (p < 0.001) and 91% receiving intravenous antihistamines (p < 0.001). Even in severe anaphylaxis (grade 5) adrenaline was administered in only 80% of the cases. CONCLUSION: Treatment with adrenaline is not administered in accordance with international guidelines. However, making an assessment of the severity of the anaphylactic reaction is difficult in retrospective studies.

Safety of essential bee venom pharmacopuncture as assessed in a randomized controlled double-blind trial.

ETHNOPHARMACOLOGICAL RELEVANCE: While bee venom (BV) pharmacopuncture use is common in Asia, frequent occurrence of allergic reactions during the treatment process is burdensome for both practitioner and patient. AIM OF THE STUDY: This study compared efficacy and safety in isolated and purified essential BV (eBV) pharmacopuncture filtered for phospholipase A2 (PLA2) and histamine sections, and original BV to the aim of promoting safe BV pharmacopuncture use. MATERIALS AND METHODS: In in vitro, we examined the effect of BV and eBV on nitric oxide (NO) production induced by lipopolysaccharide (LPS) in RAW 264.7 macrophages, and clinically, 20 healthy adults aged 20-40 years were randomly allocated and administered eBV 0.2mL and BV pharmacopuncture 0.2mL on left and right forearm, respectively, and physician, participant, and outcome assessor were blinded to treatment allocation. Local pain, swelling, itching, redness, wheals, and adverse reactions were recorded by timepoint. RESULTS: eBV and BV exhibited similar inhibitory effects on NO production. Also, in comparison between eBV and BV pharmacopuncture administration areas on each forearm, eBV displayed significantly lower local pain at 24h post-administration (P=0.0062), and less swelling at 30min (P=0.0198), 2 (P=0.0028), 24 (P=0.0068), and 48h post-administration (P=0.0253). eBV also showed significantly less itching at 24 (P=0.0119), 48 (P=0.0082), and 96h (P=0.0141), while redness was significantly less at 30min (P=0.0090), 6 (P=0.0005), and 24h (P<0.0001). Time-by-treatment interactions were statistically significant for itching and redness (P<0.001, and P<0.001, respectively), and all original BV pharmacopuncture administered regions showed a tendency toward more severe itching and redness in later measurements. CONCLUSIONS: eBV and BV displayed comparable anti-inflammatory effects, and eBV pharmacopuncture presented less local allergic reactions.

Composición química del aceite esencial de Ocotea cymbarum Kunth (cascarillo y/o sasafrás) de la región Orinoquia / Chemical composition of essential oil of Ocotea cymbarum Kunth (cascarilla and/or sassafras) from the Orinoquía region

Introducción: en algunas poblaciones de la Orinoquia colombiana y/o venezolana venden, en plazas de mercado, los aceites de cascarillo y sasafrás; estos aceites son empleados para aliviar dolencias como artritis reumática, afecciones respiratorias y tratar picaduras de insectos y animales ponsoñosos. Los productos se obtienen de la especie Ocotea cymbarum Kunth, la cual tiene una abundante sinonimia botánica como Alseodaphne cymbarum,Licaria cymbarum, Misanteca cymbarum, Nectandra barcellensis, Nectandra cymbarum, Nectandra elaiophora, Nectandra oleífera y Ocotea barcellensis. Objetivo: realizar un análisis comparativo de las características fisicoquímicas del aceite esencial de cascarillo y/o sasafrás (posible Ocotea cymbarum Kunth). Métodos: la composición química relativa de los aceites obtenidos de madera y corteza fue determinada mediante cromatografía de gases acoplada a espectrometría de masas (CG-EM), el cálculo y comparación de índices de retención en columnas de polaridad ortogonal y la comparación de los espectros de masas por impacto electrónico con los de las bases de datos NIST08.L, Wiley9.L. Resultados: fueron determinados 45 compuestos en las muestras analizadas y más del 90 por ciento de la composición relativa establecida para cada aceite corresponde a compuestos monoterpénicos principalmente alcanfor y alfa-terpineol. Al realizar un análisis de las rutas biosintéticas que conducen a la generación de los principales compuestos detectados en los aceites analizados, se evidenció que las especies de donde se obtuvieron estos aceites, presentan diferentes grados de evolución micromolecular. Conclusión: los aceites de cascarillo y sasafrás provienen de especies vegetales con diferentes grados de evolución química. Este resultado refuerza la evidencia de la alta diversidad biológica de la familia Lauraceae. El trabajo fue realizado con el apoyo financiero del Laboratorio de Productos Naturales Vegetales del departamento de Química de la Universidad Nacional de Colombia sede Bogotá(AU)

Introduction: cascarilla and sassafras oils are sold in market places of some settlements in the Colombian and/or Venezuelan Orinoquía. These oils are used to alleviate conditions such as rheumatoid arthritis and respiratory disorders, and to treat the bites of insects and poisonous animals. The oils are obtained from the species Ocotea cymbarum Kunth, which displays abundant botanical synonymy as Alseodaphne cymbarum, Licaria cymbarum,Misanteca cymbarum, Nectandra barcellensis, Nectandra cymbarum, Nectandra elaiophora, Nectandra oleífera and Ocotea barcellensis. Objective: carry out a comparative analysis of the physicochemical characteristics of essential oil of cascarilla and/or sassafras (Ocotea cymbarum Kunth).Methods: the relative chemical composition of oils obtained from wood and bark was determined by gas chromatography coupled with mass spectrometry (GC-MS). Estimation and comparison of retention indices was conducted with orthogonal polarity columns. Comparison of electron impact mass spectra was performed with the databases NIST08.L and Wiley9.L. Results: forty-five compounds were determined from the samples analyzed. More than 90 percent of the relative composition established for each oil corresponds to monoterpenic compounds, mainly camphor and alpha-terpineol. Analysis of the biosynthetic pathways leading to the generation of the main compounds detected in the oils studied, showed that the species from which the oils were obtained exhibit varying degrees of micromolecular evolution. Conclusion: cascarilla and sassafras oils are obtained from plant species at varying stages of chemical evolution. This result constitutes additional evidence of the great biological diversity of the Lauraceae family. The study was conducted with financial support from the Natural Plant Products Laboratory of the Chemistry Department at the National University of Colombia at Bogotá(AU)

Human scFv antibodies (Afribumabs) against Africanized bee venom: Advances in melittin recognition.

Africanized Apis mellifera bees, also known as killer bees, have an exceptional defensive instinct, characterized by mass attacks that may cause envenomation or death. From the years 2000-2013, 77,066 bee accidents occurred in Brazil. Bee venom comprises several substances, including melittin and phospholipase A2 (PLA2). Due to the lack of antivenom for bee envenomation, this study aimed to produce human monoclonal antibody fragments (single chain fragment variable; scFv), by using phage display technology. These fragments targeted melittin and PLA2, the two major components of bee venom, to minimize their toxic effects in cases of mass envenomation. Two phage antibody selections were performed using purified melittin. As the commercial melittin is contaminated with PLA2, phages specific to PLA2 were also obtained during one of the selections. Specific clones for melittin and PLA2 were selected for the production of soluble scFvs, named here Afribumabs: prefix: afrib- (from Africanized bee); stem/suffix: -umab (fully human antibody). Afribumabs 1 and 2 were tested in in vitro and in vivo assays to assess their ability to inhibit the toxic actions of purified melittin, PLA2, and crude bee venom. Afribumabs reduced hemolysis caused by purified melittin and PLA2 and by crude venom in vitro and reduced edema formation in the paws of mice and prolonged the survival of venom-injected animals in vivo. These results demonstrate that Afribumabs may contribute to the production of the first non-heterologous antivenom treatment against bee envenomation. Such a treatment may overcome some of the difficulties associated with conventional immunotherapy techniques.

Efficacy and safety of topical Trikatu preparation in, relieving mosquito bite reactions: a randomized controlled trial.

INTRODUCTION: Trikatu is composed of dried fruits of Piper nigrum L and Piper retrofractum Vahl, and dried rhizomes of Zingiber officinale R. Although this preparation has been used to relieve pruritis, pain, and inflammation for a long time, there is no clinical evidence to confirm its efficacy and safety. Therefore, we performed a double-blind, within person-randomized controlled study of 30 healthy volunteers to determine efficacy and safety of topical Trikatu on mosquito bite reactions. METHODS: All subjects were bitten by Aedes aegypti laboratory mosquitoes on their forearms and they were randomly assigned arms to apply either Trikatu or reference product on the mosquito bite papule. The main outcome was the difference of papule size reduction at 30 min, measured by a caliper, between the Trikatu and reference arms. Pruritis, redness, pain, and patient satisfaction were assessed at 15, 30, 60, 180, and 360 min as secondary outcomes. RESULTS: There were no significant differences between treatment and reference arms on any outcome at any time of measurement. CONCLUSION: Trikatu did not show additional effects for relieving mosquito bite reaction as compared with the reference product containing camphor, menthol, and eucalyptus. For further study, it is very important to consider a proper selection of subjects, comparator product, and concentration of extract when Trikatu preparation is investigated.

The healing effects of a topical phytogenic ointment on insect bite hypersensitivity lesions in horses.

Insect bite hypersensitivity (IBH) is the most common cause of pruritus in horses and is a serious welfare issue for affected animals. In this study, the effect of a topical phytogenic ointment on the healing of cutaneous lesions was investigated in a double-blind trial involving 26 horses with I B H. The number of lesions and their total surface area were recorded on days 0, 7, and 21 in horses treated for 3 weeks with either verum or placebo ointment. After unblinding of treatment assignment, the horses that had been treated with the placebo ointment received the verum preparation for an additional 3 weeks and the number of lesions and their total surface area were again recorded. This part of the study was not blinded. The number of lesions and the total surface area decreased in both treatment groups (no significant difference). Owners also scored the degree of discomfort suffered by their horses as a result of IBH lesions, and at the end of the 3-week period this score was significantly lower in the verum than in the placebo group (P = 0.04). When placebo-treated horses subsequently received the verum ointment, their wound severity score also decreased significantly (P < 0.01). Daily application of an ointment (verum or placebo) does not cure IBH, but use of the phytogenic ointment led to a decrease in the owner-assessed discomfort suffered by horses.

Bioactivity-guided fractionation and GC/MS fingerprinting of Angelica sinensis and Angelica archangelica root components for antifungal and mosquito deterrent activity.

Bioassay-guided fractionation of the chloroform extract from the roots of Angelica sinensis led to isolation and characterization of (Z)-ligustilide using direct-bioautography with Colletotrichum species. The structure of (Z)-ligustilide was confirmed by (1)H and (13)C NMR spectroscopy and GC/MS. (Z)-Ligustilide deterred the biting of two mosquito species more effectively than DEET. Three different A. sinensis accessions and one Angelica archangelica root oil were evauated by GC and GC/MS, and the dominant component in A. sinensis was 61-69% (Z)-ligustilide. Two other prominent compounds in A. sinensis oils were 5.7-9.8% (E)-3-butylidene phthalide and 1.5-2.3% (Z)-3-butylidene phthalide. The main constituents that comprised A. archangelica oil were monoterpene hydrocarbons such as 24.5% alpha-pinene, 13.8% delta-3-carene, 10.1% beta-phellandrene, 8.8% p-cymene, 8.4% limonene, and 6.3% sabinene. Phthalides and monoterpene hydrocarbons were determined to be good systematic markers or chemical fingerprints for A. sinensis and A. archangelica root oils. Chemical fingerprinting by GC/MS of A. sinensis also confirmed the misidentification of one A. archangelica sample sold in the Chinese market.

Randomized controlled trial of topical aspirin in the treatment of bee and wasp stings.

BACKGROUND: The New South Wales Poisons Information Centre (NSW PIC) has been recommending the use of topical aspirin paste for bee and wasp stings since the early 1980s. Anecdotal evidence from calls suggested it was effective in reducing the swelling and duration of pain, but a literature search found no evidence to support this. OBJECTIVE: The objective of this study was to assess the effectiveness of advice given by a PIC to apply topical aspirin for the treatment of bee and wasp stings. METHODS: Patients were recruited from callers to the NSW PIC who reported a bee or wasp sting. They were randomly assigned, using a 2:1 ratio, to two different treatment advices: to apply an ice pack (control group), or to apply an ice pack and topical aspirin paste (treatment group). Initial follow-up was within 24-48 hours. Primary outcome was the presence of swelling at 12 hr. Secondary outcomes included the presence of pain at 12 hr, the presence of itchiness, and duration of redness. RESULTS: There were 37 patients who received treatment advice and 19 in the control group. Of the 37 patients advised to apply aspirin, 21 (57%) had no swelling at 12 hr compared with 14 of the 19 (74%) patients with ice alone (difference -17%; 95% CI: -47-12%; p = 0.26). Eighty-one percent (30/37) of patients advised to apply aspirin had no pain at 12 hr compared with (18/19) 95% of the others (-14%; 95% CI: -39-14%; p = 0.34). The median duration of redness was 6 hr [interquartile range (IQR): 2-48 hr] in those advised to apply aspirin paste compared with 2 hr (IQR: 0-10 hr) in those that only applied ice (p = 0.04). CONCLUSIONS: Topical aspirin paste was not effective in reducing the duration of swelling or pain in bee and wasp stings, and significantly increased the duration of redness. Symptoms rapidly subsided with ice alone as treatment.

Review of sangre de drago (Croton lechleri)--a South American tree sap in the treatment of diarrhea, inflammation, insect bites, viral infections, and wounds: traditional uses to clinical research.

OBJECTIVE: The objective of this review is to provide an overview of the pharmacologic evidence that may or may not support clinical and ethnomedical uses of the sap of sangre de drago (dragon's blood; Croton lechleri Müll. Arg.). Data sources used were BIOSIS, EMBASE, PubMed, TOXLIT, International Pharmaceutical Abstracts, manual searches, papers on file from peer-reviewed journals, textbooks available at Armana Research, Inc., and researchers in the field of South American botanical medicine. CONCLUSIONS: The results of in vitro and in vivo studies largely support the majority of ethnomedical uses of sangre de drago including the treatment of diarrhea, wounds, tumors, stomach ulcers, herpes infection, the itching, pain and swelling of insect bites, and other conditions. Clinical studies of sangre de drago products have reported positive results in the treatment of traveler's and watery diarrhea and the symptoms of insect bites. Because the sap has shown low toxicity and preparations used in clinical studies were well tolerated, further clinical and pharmacologic studies are anticipated. Acknowledgment of the diversity in the chemical makeup of the sap from one geographic area to another and the recent characterization of alkaloid chemotypes of sangre de drago will require that materials developed for clinical use are standardized.

The efficacy of Prrrikweg gel in the treatment of insect bites: a double-blind, placebo-controlled clinical trial.

OBJECTIVE: This study was conducted to examine the efficacy of Prrikweg gel, a homeopathic after-bite gel, in relieving the effects of mosquito bites, in particular itching and erythema. DESIGN: A double-blind, randomized, placebo-controlled clinical trial. SETTING: London School of Hygiene and Tropical Medicine. SUBJECTS: 100 healthy volunteers. METHODS: All subjects were bitten under laboratory conditions by Aedes aegypti mosquitoes at one spot on the ventral aspect of the left forearm and another on a corresponding position on the right forearm. One spot was treated with the homeopathic after-bite gel and the other with a placebo gel. MAIN OUTCOME MEASURES: Itching was assessed on a 5-point discrete rating scale at 0, 0.5, 1, 26.5, and 48 h post-bite to compare the itch-relieving efficacy of the two treatments. Erythema development was assessed by photographing the bite sites, measuring length and width of the erythema with a calliper, and comparing the ratio of the erythema surface at baseline T(0) to the mean erythema surface at 0.5, 1, 26.5, and 48 h post-bite (T mean) for the two treatments. RESULTS: Testing erythema development by comparing the ratio T(0)/T (mean, after-bite gel) and the ratio T(0)/T (mean, placebo gel) gave a two-tailed p = 0.098(95% Cl, -0.031-0.361) in favour of the after-bite gel. There was not a statistically significant difference between the itch relief provided by the two treatments (two-tailed p = 0.424; 95 percent Cl, -0.541-0.191). The correlation between itching and erythema was significant (r = 0.46; p < 0.001). CONCLUSIONS. There are strong indications that the homeopathic after-bite gel reduces erythema development following mosquito bites. The homeopathic mother tinctures of Echinacea angustifolia DC., Ledum palustre L., Urtica urens L. as well as the Hamamelis extract in this gel, whether alone or in combination, are the biologically active ingredients. The homeopathic after-bite gel was not demonstrated to relieve itching; however, based on the correlation between erythema and itching, an effect on itching is not inconceivable.

A placebo controlled clinical trial investigating the efficacy of a homeopathic after-bite gel in reducing mosquito bite induced erythema.

A randomised, placebo controlled clinical trial was conducted to examine the efficacy of a homeopathic after-bite gel in the symptomatic relief of mosquito bites. Sixty eight healthy volunteers were bitten under laboratory conditions by Aedes aegypti mosquitoes at three spots, on the ventral aspect of the forearm. One bite was treated with the homeopathic after-bite gel, another bite with a placebo gel which was identical in appearance and smell to the homeopathic after-bite gel, and the third bite remained untreated. Immediately after the bites and 1, 3, 6, 26 and 31 hours post-bite, the length and width of the erythema were measured with a calliper, and photographs were taken of the bite sites from which the size of the erythema was subsequently determined. This was followed by assessment of the extent of itching with a verbal analogue scale, and finally treatment took place. For each spot the total erythema was calculated as the area under the plotted curve of the erythema at different time points (mm2*h) and the total sum of the itch scores was determined. For the bites treated with the homeopathic after-bite gel the median total erythema was 10.500 mm2*h. For the spots treated with the placebo gel and the untreated spots the median total erythema was 12.900 mm2*h and 13.300 mm2*h, respectively. The difference between the spots treated with the homeopathic after-bite gel and the untreated spots came close to significance (two-tailed P = 0.06), which was not the case for the difference between the spots treated with the homeopathic after-bite gel and the spots treated with placebo gel (P = 0.13). After pooling the data of a very similar previous pilot study and the present study (ntotal = 83), the homeopathic after-bite gel was significantly superior to no treatment (two-tailed P = 0.003) as well as to placebo gel (two-tailed P = 0.03). Comparing itching after the three treatments, no significant differences could be demonstrated. The extent of itching was positively correlated with the area of the erythema (r = 0.63). Treatment of mosquito bites with the homeopathic after-bite gel will reduce the erythema compared to no treatment. Comparison with the placebo gel suggests it is the plant extracts which are the active components of this gel.

[The rebirth of the H1-antagonists]. / Renaissance des anti-H1.

Anti H1s of the third generation, of which the first was Terfenadine, possess not only blocking effects on H1 receptors to histamine, but also have a "cromone-like" action on the membranes of cells that have been, or not, activated by specific stimuli (antigens). They act on the early phase of immediate hypersensitivity. These anti H1s of the new generation also have wider indications: asthma, perennial or pollen rhinitis, dermatoses such as urticaria and atopic dermatitis. Their tolerance has been remarkably improved with regard to the central effect (very reduced), such as absence of an atropine effect.

Reduction of side effects from rush-immunotherapy with honey bee venom by pretreatment with terfenadine.

In a double-blind placebo-controlled trial on 52 patients with bee venom allergy we studied the effect of a pretreatment with terfenadine 120 mg twice daily on the occurrence of local and systemic allergic side effects from rush-immunotherapy. Large local reactions were significantly reduced by terfenadine pretreatment (P less than 0.01), while systemic side effects were observed with similar frequencies in both groups. Analysis of individual systemic allergic manifestations showed that cutaneous symptoms like itching (P less than 0.025) and urticaria/angioedema (P less than 0.05) were significantly reduced, while respiratory or cardiovascular symptoms were not influenced. A lower consumption of additional anti-allergic medication was found during terfenadine pretreatment (P less than 0.05). Pretreatment with antihistamines during immunotherapy may thus be helpful in the management of patients with cutaneous side effects.