Study of antidiabetic and antioxidant activities for 2 species of birch leaves / Монголын эм зүй, эм судлал

Abstract@#The birch leaves were used as a substitute for birch bark, buds and chaga of birch in traditional medicine because the birch leaves are considered to be less toxic. Numerous researches conducted in Russia, Bulgaria, Japan, and China on <i>B.pubescens, B. pendula, B.Rezniczenkoana (Litv) </i> Schischk, <i>B.humilis</i> Schrank, and <i>B.mandshurica</i> Rgl Nakai found that birch barks and leaves contain antioxidants and they have anti-cancer, anti-yeast, antibacterial, anti-inflammatory, liver protective and bile secretion induction properties. The studies conducted on animals with diseases showed that the birch leaves had anti-inflammatory properties on the gastric mucosa during acute stress, as well as anti-biliary and giardiasis. The birch leaf phytopreparations experimentations used on animals showed reduced peripheral tissue insulin resistance and lowered blood sugar. Mongolian traditional medicinal journals noted that the birch barks are used to treat inflammatory acute diseases. Therefore, this study was performed to determine the effects of two species of birch leaves on blood sugar and antioxidant activities in diabetes-induced rats.@*The study materials and methods@#The study was conducted in the Pharmacology Research Laboratory of the Monos Group’s Institute of Pharmacology. 40 WISTAR, non-linear white rats weighing 150-204 g were used in the experiments. Dry extract of birch leaves of the two species (Alloxan monohydrate Tokyo Chemical Industry LTD), IGM-100 3A blood glucose meter (Blood glucose test meter, Infopia LTD, Brussels Belgium) and sugar test (Blood glucose test strip only, province, China) were used for the experiment. Lenzen’s (2008) method was used to induce Alloxan diabetes in the rats and the antioxidant properties were determined by the antioxidant activity kit (Rat Malondialchehyche Elisa KIT, cat. № EKRAT- 0266, Jilin).@*Study Result@#The blood glucose level of the control group with diabetes lowered from 31.5 mmol/l to 17.1 mmol/l in 14 days. As for the <i>B.platyphylla</i> Sukacz group, the blood glucose level reduced to 6.3 mmol/l and the <i>B.hippolytii. </i> Sukacz group’s blood glucose level reduced to 6.9 mmol/l in 14 days.</br> The study results showed that <i>B.hippolytii </i>Sukacz birch leaves and <i>B.platyphilla</i> Sukacz birch leaves’ extracts reduced the maximum level of MDA dilution (4.8 nmol/ml) of B.hippolytii Sukacz and B.platyphilla Sukacz groups by 33.9% and 53.5% respectively. This suggests that the birch leaves had antioxidant effect.@*Conclusion@#<i>B.hippolytii </i>Sukacz birch leaves and <i>B. platyphilla </i> (Sukacz) birch leaves lowered the blood glucose level and had antioxidant properties on diabetes.

Toxicology study of Hippolytii birch (B. Hippolytii. Sukacz) leaves and flat leaved birch (B.Platyphylla. Sukacz) / Монголын эм зүй, эм судлал

Abstract@#Numerous researches conducted in Russia, Bulgaria, Japan, and China on <i>B.pubescens, B. pendula, B.rezniczenkoana (Litv) </i> Schischk, <i>B.humilis</i> Schrank, <i>B.mandshurica</i> Rgl Nakai found that birch barks and leaves contain antioxidants and they have anti-cancer, anti-fungi, antibac- terial and anti-inflammatory properties, protect liver and promote bile secretion. Flat leaved birch (<i>B.platyphylla</i> Sukacz) cortex contains betulin and lupeol of triterpenoids and it’s leaves contain flavonoid and polyphenol compounds. The amounts of compounds found in the cortex are smaller than leaves. Specifically, the amount of flavonoid in leaves is more contained than the that of cortex and leaf buds. In any pharmacology study of new medicines, determination and evaluation of toxicity is the first priority. According to scientific evidences that birch leaves are considered to have less toxins. Not many studies have been conducted on determining toxicity of birch leaves in Mongolia. Therefore, the purpose of this research is to study the species of birches, hippolytii birch (<i>B.hippolytii. </i> Sukacz) and flat leaved birch (<i>B.platyphylla. </i> Sukacz), that were noted to have medical properties in traditional medications and identify their acute toxicity using dry extract and determine mortality dosage (LD₅₀) on animals.@*Research materials and methods@#Evaluation of the acute toxicity of birch leaves was conducted in Pharmacology laboratory of Monos group’s Drug Research Institute between June 19, 2020 and August 10. In this research, 150-204 g of WISTAR breed non-linear 44 white rats were used and 20 g of <i>B.Hippolytii’</i>s dry extract and 20 g of B. <i>Platyphylla</i> ‘s dry extract were injected.</br> The experiments to determine the toxicity of dry extracts of <i>B. hippolytii</i> and <i>B. platyphylla</i> (LD₅₀) were conducted according to Litchfield and Wilcoxon’s method and subcutaneous injects were per formed in the pelvic area of the rats. @*Results of determining acute toxicity level@#The experiments to determine the acute toxicity level of the birch’s dry extracts followed Litchfield and Wilcoxon’s method with 2-stage. LD₅₀ level was determined from the first stage of the research using G.N.Pirshen’s method and the toxicity level was identified using K.K.Sidorov’s toxicity categorization.</br> From the acute toxicity research, no-observed-adverse-effect level (NOAEL), animal daily dosage and human daily dosage (experimental) were determined. LD₅₀ 2950 mg/kg was determined as a result of acute toxicity research of B.hippolytii and B.platyphilla leaves’ dry extract.

Allergenic Pollen Calendar in Korea Based on Probability Distribution Models and Up-to-Date Observations

PURPOSE: The pollen calendar is the simplest forecasting method for pollen concentrations. As pollen concentrations are liable to seasonal variations due to alterations in climate and land-use, it is necessary to update the pollen calendar using recent data. To attenuate the impact of considerable temporal and spatial variability in pollen concentrations on the pollen calendar, it is essential to employ a new methodology for its creation.METHODS: A pollen calendar was produced in Korea using data from recent observations, and a new method for creating the calendar was proposed, considering both risk levels and temporal resolution of pollen concentrations. A probability distribution was used for smoothing concentrations and determining risk levels. Airborne pollen grains were collected between 2007 and 2017 at 8 stations; 13 allergenic pollens, including those of alder, Japanese cedar, birch, hazelnut, oak, elm, pine, ginkgo, chestnut, grasses, ragweed, mugwort and Japanese hop, were identified from the collected grains.RESULTS: The concentrations of each pollen depend on locations and seasons due to large variability in species distribution and their environmental condition. In the descending order of concentration, pine, oak and Japanese hop pollens were found to be the most common in Korea. The pollen concentrations were high in spring and autumn, and those of oak and Japanese hop were probably the most common cause of allergy symptoms in spring and autumn, respectively. High Japanese cedar pollen counts were observed in Jeju, while moderate concentrations were in Jeonju, Gwangju and Busan.CONCLUSIONS: A new methodology for the creation of a pollen calendar was developed to attenuate the impact of large temporal and spatial variability in pollen concentrations. This revised calendar should be available to the public and allergic patients to prevent aggravation of pollen allergy.

Clinical Characteristics of Oral Allergy Syndrome in Children with Atopic Dermatitis and Birch Sensitization: a Single Center Study

BACKGROUND: Oral allergy syndrome (OAS) is an immunoglobulin E (IgE)-mediated hypersensitivity that occurs frequently in older children with pollen sensitization. This study focused on the clinical characteristics of OAS in children with atopic dermatitis (AD) and birch sensitization. METHOD: s: A total of 186 patients aged 2–18 years with AD and birch sensitization were enrolled in this study between January 2016 and March 2017. Their levels of serum total IgE and birch- and ragweed-specific IgE (sIgE) were measured using ImmunoCAP (Thermo Fisher Scientific, Uppsala, Sweden). Information regarding causative foods and symptoms were obtained via interviews. The patients were divided into 3 groups according to their ages (group 1, 2–6 years; group 2, 7–12 years; and group 3, 13–18 years). RESULTS: Eighty-one of the 186 (43.5%) children with AD who were sensitized to birch pollen were diagnosed as having OAS. The prevalence of OAS in group 1 (the children who had AD and birch sensitization aged 2–6 years) was 36.6%. A greater predominance of men was noted in the non-OAS group (77.1%) compared to the OAS group (60.5%). Apples were the most common causative food in group 2 and 3 while kiwis were the most common cause of OAS in group 1. There was a statistically significant correlation between birch-sIgE levels and the prevalence of OAS (P = 0.000). The cut-off value was 6.77 kUA/L with 55.6% sensitivity and 79.0% specificity (area under the curve 0.653). CONCLUSION: In our study, the prevalence of OAS in children with AD and birch sensitization was 43.5%. Even in the preschool age group, the prevalence of OAS was considerable. Patients with high levels of birch-sIgE were more likely to have OAS. Clinicians should therefore be vigilant about OAS in patients with a high degree of sensitization to birch pollen and even young children if they have birch sensitization.

Oral Allergy Syndrome in Birch Pollen-Sensitized Patients from a Korean University Hospital

BACKGROUND: Oral allergy syndrome (OAS) is a type of allergic reaction that mainly occurs on oral contact with raw fruit, vegetables, or nuts. The most common type of OAS is birch pollen-related food allergy. Although OAS is a common food allergy in adults, only few epidemiologic studies have been reported in Korea. Here we investigate the prevalence and triggers of birch pollen-related food allergy. METHODS: We conducted a retrospective chart review of 1,427 patients who underwent a skin prick test for inhalant allergens at the Asthma and Allergy Clinic in Seoul National University Bundang Hospital from January 2011 to December 2016. RESULTS: Of 1,427 patients, 125 (8.7%) were sensitized to birch pollen. Among them, 20.0% developed OAS, which was the most common food allergy (96.2%). The prevalence of OAS was higher in females, and was 18.2% in birch pollen-sensitized allergic rhinoconjunctivitis patients. Further, 72.0% OAS patients had rhinoconjunctivitis, 20.0% had asthma, and 12.0% had chronic urticaria. Apple (68.0%), peach (56.0%), nuts (36.0%), kiwi (20.0%), persimmon (20.0%), plum (16.0%), and cherry (16.0%) were frequent triggers; however, Chinese yam, kudzu vine, bellflower root, codonopsis, and ginseng were also revealed as triggers. Patients (60.0%) showed OAS with ≥ 3 foods at the same time. Only 3 patients showed mono-sensitivity to birch pollen, while others were multi-sensitized to trees, grasses, weed, or house dust mite allergens. CONCLUSION: OAS was the most common food allergy in birch pollen-sensitized patients. This study revealed the unique triggers of OAS in Korea in addition to well-known triggers.

The Incidence of Delayed-Type Hypersensitivity Reactions to Apples Among Patients Allergic to Birch Pollen

The major apple allergen Mal d 1 cross-reacts with the homologous birch pollen allergen Bet v 1 and causes immunoglobulin E (IgE)-mediated immediate-type allergic reactions. In some patients, delayed-type hypersensitivity to apples may develop within 72 hours without evidence of specific IgE or a positive skin prick test (SPT). The aim of the study was to evaluate the concomitance of delayed-type hypersensitivity reactions and immediate IgE-mediated reactions against high- and low-allergenic apple cultivars in patients with birch pollen allergy. Data were obtained from 45 adults with clinical symptoms of birch pollen allergy. Patients were exposed to apple pulp via atopy patch tests (APTs) and SPTs. Levels of IgE specific to Bet v 1 and Mal d 1 were measured with a radioallergosorbent test. Patients allergic to birch pollen showed the highest rate of positive SPT responses to Golden Delicious apples and the lowest rate to low-allergenic cultivar Grey French Reinette. Among these patients, 9% developed delayed hypersensitivity reactions to either Golden Delicious or Grey French Reinette apples; these reactions manifested clinically as erythema with papules (class ++). Fifty percent of APT-positive patients were concomitantly SPT-negative. Here, we show for the first time the clinical relevance of T cell-driven allergic reactions to apples. APTs may reveal type IV sensitization in patients who are negative for the corresponding type I sensitization tests. Thus, utilization of the APT procedure with fresh apple appears to be a valuable tool for the diagnosis of apple allergy and may improve the accuracy of food allergy diagnoses.

Effect of MeJA combined with high temperature stress in treatment for accumulation of triterpenoids in birch suspension cells / 中草药

Objective: To investigate the effect of MeJA combined with high temperature stress in the treatment for the accumulation of triterpenoids in the birch (Betula platyphylla) suspension cells. Methods: After MeJA (25, 50, 100, and 150 μmol/L)and high temperature (50℃ for 2 h) treatment, the cell growth, viability, content of MDA, the activity of defense enzyme, total triterpenoids content, and the gene expression levels of triterpenoids synthesis were measured. Results: The combination of high temperature stress and MeJA treatment had a more powerful positive effect on the synthesis of triterpenoids than single MeJA or high temperature treatment in birch cells. Moreover, the concentration of total triterpenoids had the highest level when adding 150 μmol/L MeJA after the high temperature processing, was up to 76.6 mg/g, which was 81.3%, 159.9% and 13.1% higher than those in the blank control, individual MeJA treatment or the heat treatment alone respectively. Meanwhile, the gene expression levels of SS, SE, BPW, and BPY, related to the triterpenoids synthesis, had an increase about 297.1%, 83.7%, 1 032.6%, and 282.4% compare to the control. The MeJA after high temperature treatment enhanced the activity of SOD and PAL compared with the control, inhibited the cell growth and viability. Conclusion: The treatment of MeJA after high temperature affects the cell growth, viability, and activity of defense enzyme, regulates the genes expression level of triterpenoids synthesis, and eventually could make cells to produce the triterpenoids substance effectively.

Mimotopes for Api g 5, a Relevant Cross-reactive Allergen, in the Celery-Mugwort-Birch-Spice Syndrome

PURPOSE: In the celery-mugwort-birch-spice syndrome, a significant proportion of IgE is directed against high molecular weight (HMW) glycoproteins, including the celery allergen Api g 5. BIP3, a monoclonal antibody originally raised against birch pollen, recognizes HMW allergens in birch and mugwort pollens, celery, and Apiaceae spices. Our aim was to generate mimotopes using BIP3 for immunization against the HMW allergens relevant in the celery-mugwort-birch-spice cross reactivity syndrome. METHODS: Mimotopes were selected from a random-peptide display library by BIP3 and applied in IgE inhibition assays. The 3 phage clones with the highest inhibitory capacity were chosen for immunization of BALB/c mice. Mouse immune sera were tested for IgG binding to blotted birch pollen extract and used for inhibiting patients' IgE binding. Furthermore, sera were tested for binding to Api g 5, to horseradish peroxidase (HRP) as a second glycoprotein, or to non-glycosylated control allergen Phl p 5 in ELISA, and the specific Api g 5-specific IgG titers were determined. RESULTS: Three rounds of biopanning resulted in phage clones exhibiting 7 different sequences including 1 dominant, 1-6-cyclo-CHKLRCDKAIA. Three phage clones had the capacity to inhibit human IgE binding and induced IgG to the HMW antigen when used for immunizing BALB/c mice. The induced BIP3-mimotope IgG reached titers of 1:500 specifically to Api g 5, but hardly reacted to glycoprotein HRP, revealing a minor role of carbohydrates in their epitope. CONCLUSIONS: The mimotopes characterized in this study mimic the epitope of BIP3 relevant for Api g 5, one of the cross-reactive HMW allergens relevant in the celery-mugwort-birch-spice syndrome. BIP3 mimotopes may be used in the future for hyposensitization in this clinical syndrome by virtue of good and specific immunogenicity.

Component-resolved diagnosis using microarray for diagnosing hypersensitivity to raw fruits in birch pollen sensitized children

PURPOSE: Recently, component-resolved diagnosis (CRD) using microarray technology has been introduced to the field of clinical allergy. This study was aimed to investigate the clinical usefulness of microarray-based IgE detection for diagnosing clinical raw fruit allergy in birch pollen-sensitized children. METHODS: Thirty-one children with allergic disease who had been sensitized to pollen were studied. A pollen-sensitized patient was defined as having an allergen-specific history with concomitant positive skin-prick tests (SPTs) to natural allergen extracts or positive allergen-specific IgE. All subjects underwent SPTs for pollen and fruit. In all subjects, specific IgE to pollen and fruit were measured by ImmunoCAP. Specific IgE antibodies to allergen components were determined by a customized allergen microarray (ISAC). RESULTS: Thirteen of the 31 patients (41.9%) had a history of fruit hypersensitivity with positive SPTs. Measuring IgE to allergen components by ISAC, all the 13 patients with fruit hypersensitivity were positive to at least one of Mal d 1, Pru p 1, Pru p 3, Act d 8, and Act d 2 compared to 12 of the 13 patients (92.3%) who had at least 1 positive IgE to fruits (apple, peach, and kiwi) using ImmunoCAP. The sensitivity of ISAC microarray was 100.0% for the diagnosis of fruit hypersensitivity, but its specificity was 27.7% (5/18). The sensitivity of ImmunoCAP was 92.3%, and its specificity was 83.3%. CONCLUSION: The sensitivity of allergen components tested using microarray for the diagnosis of clinical fruit hypersensitivity in children with pollen allergy was high; however, its specificity was low.

Identification of immunoglobulin E binding components of two major tree pollens, birch and alder

PURPOSE: Pollinosis is one of the major allergic diseases caused by airborne pollens. Alder and birch pollens are the major sensitizing tree pollens in this country. The immunoglobulin E (IgE) reactivity to each pollen allergen is known to be variable according to the region. We determined the major IgE binding components of these tree pollens in sera of adult patients with allergic rhinitis. METHODS: Allergic rhinitis patients, of whom specific IgE level to birch and/or alder pollens (>10 kU/L by ImmunoCAP) were included. The protein bands of two pollen extracts were determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and their IgE-binding components were identified by IgE immunoblot analysis. The binding specificity and cross-reactivity between two pollens were evaluated by IgE enzyme linked immunosorbent assay (ELISA) inhibition test. RESULTS: Six IgE binding components were found in birch pollens in which two (14 kDa and 17 kDa) were major components. Two IgE binding components were found in alder pollens in which the 17 kDa was a major component. The IgE binding component to the major allergen component of 17 kDa was observed in 90.3% of the study subjects sensitive to alder pollens and 72.7% of them sensitive to birch pollens. The ELISA inhibition tests showed significant inhibitions with additions of birch/alder pollen extracts. CONCLUSION: We identified two major IgE binding components (17 kDa and 14 kDa) from birch pollens and one component (17 kDa) from alder pollens. Significant cross reactivity was noted between these two pollens.

Introduction to Distribution and Ecology of Sterile Conks of Inonotus obliquus

Inonotus obliquus is a fungus that causes white heart rot on several broad-leaved species. This fungus forms typical charcoal-black, sterile conks (chaga) or cinder conks on infected stems of the birche (Betula spp). The dark brown pulp of the sterile conk is formed by a pure mycelial mass of fungus. Chaga are a folk remedy in Russia, reflecting the circumboreal distribution of I. obliquus in boreal forest ecosystems on Betula spp. and in meridional mountain forests on beech (Fagus spp.) in Russia, Scandinavia, Central Europe, and Eastern Europe. Distribution at lower latitudes in Western and Southern Europe, Northern America, Asia, Japan, and Korea is rare. Infected trees grow for many years without several symptoms of decline. The infection can penetrate through stem injuries with exterior sterile conks developing later. In the Czech Republic, cinder conk is found on birches inhabiting peat bogs and in mountain areas with a colder and more humid climate, although it is widespread in other broad leaved species over the Czech Republic. The most common hosts are B. pendula, B. pubescens, B. carpatica, and F. sylvatica. Less frequent hosts include Acer campestre, Acer pseudoplatanus, Alnus glutinosa, Alnus incana, Fraxinus excelsior, Quercus cerris, Q. petraea, Q. robur, Q. delachampii, and Ulmus sp.