Sensitized Guinea Pigs Research Articles

Sinomenine Attenuated Capsaicin-Induced Increase in Cough Sensitivity in Guinea Pigs by Inhibiting SOX5/TRPV1 Axis and Inflammatory Response.

BackgroundChronic cough is a common complaint which affects a large number of patients worldwide. Increased cough sensitivity is a very important cause of chronic persistent cough. However, there are limited clinical diagnosis and treatment for increased cough sensitivity. Transient receptor potential vanilloid-1 (TRPVl) is a member of the transient receptor potential (TRP) family of channels which is very closely associated with respiratory diseases. However, the mechanism through which TRPV1 that influences downstream events is still poorly understood.ResultsCapsaicin induced increase in cough sensitivity by upregulating the protein level of TRPV1, leading to the secretions of Substance P and neurokinin A which stimulated neurogenic inflammation. However, sinomenine, a component of traditional Chinese medicine, significantly attenuated the capsaicin-induced cough by inhibiting the expression of TRPV1 in guinea pigs. In addition, capsaicin increased the expression of SOX5 which mediated the transcriptional upregulation of TRPV1. However, pretreatment with sinomenine reduced the expression of SOX5.ConclusionThese results indicate that capsaicin induced increase in cough sensitivity by activating neurogenic inflammation, while sinomenine attenuated the increase in cough sensitivity by inhibiting the expressions of SOX5 and TRPV1 in guinea pigs. This finding may provide a novel target for the treatment of aggravated cough sensitivity.

Effect of Deer Velvet Antler on Airway Inflammation in Ovalbumin Sensitized Guinea Pigs

Background: Despite better understanding of asthma pathology and adherence to recommended treatments still many patients are experiencing uncontrolled asthma. Deer velvet antler is a traditional animal-based medicine which has shown many pharmacological effects in various diseases including asthma. Objective: To evaluate the effect of deer velvet antler on airway inflammation in ovalbumin sensitized guinea pigs and to compare it with that of dexamethasone. Study Design: Comparative study. Duration: 12 weeks. Methodology: Animals were placed in four groups with six in each. Animals in group I and II were taken as negative and positive control and animals in groups III and IV were treated with deer velvet antler powder and dexamethasone respectively along with induction of allergic airway inflammation by ovalbumin sensitization on day 0 and 14 and challenge on day 25, 26 and 27. On day 28, blood and bronchoalveolar lavage (BAL) fluid samples were taken for total leukocyte count and eosinophil percentage. Results: The study reported that total leukocyte count and eosinophil percentage in blood and bronchoalveolar lavage fluid of deer velvet antler treated guinea pigs was significantly reduced as compared to asthma control, but in comparison with dexamethasone difference was not significant. Conclusion: Deer velvet antler reduces allergic airway inflammation supporting the hypothesis of study.

<p><strong>Interferon gamma and IP 10 mRNA quantitative real time PCR in whole blood culture of guinea pig and cattle to multi-antigen recombinant protein cocktail and PPD of <em>Mycobacterium bovis </em>(3/86Rv)</strong></p>

In vivo tuberculin skin test and in vitro IFNγ assays are the most explored diagnostics for Mycobacterium bovis infection (Bovine Tuberculosis) in animals. However, there are other potential biomarkers like IP10 which may allow diagnosis of tuberculosis. In this study, IFNγ and IP10 mRNA responses in guinea pig and cattle were determined by quantitative RT-PCR in whole blood samples stimulated PPDs and recombinant protein cocktail containing eight purified rCFP2, rESAT6, rCFP10, rMTC28, rMPB53, rMPB63, rMPB64 and rMPB83 proteins. Blood samples were obtained from M. bovis sensitised guinea pigs and tuberculin test positive cattle. Both guinea pigs and cattle blood cultures produced a significant increase in mRNA level of IFNγ and IP10 when stimulated with protein cocktail as compared to that of bovine PPD. Upon different antigenic stimulations, IP10 mRNA responses were followed the similar kinetics to those of IFNγ with high correlation (r = 0.97 for guinea pig and 0.96 for cattle).

Comparison of Berberine and Dexamethasone on Blood and Bronchial Inflammatory Cells of Ovalbumin Sensitized Guinea Pigs

Objective: To compare the anti-inflammatory effects of berberine and dexamethasone in ovalbumin sensitized guinea pigs. Methods: This experimental controlled study was conducted in April, 2016 at Postgraduate Medical Institute, Lahore. Twenty- four healthy guinea pigs were selected for study. Six of these were assigned randomly in each group; normal control, ovalbumin (OVA) sensitized, berberine treated and dexamethasone treated groups. Airway inflammation was induced on day 0 and 14 by OVA injections via peritoneal route and by inhalation on 25th, 26th and 27th day in each group excluding the normal control. Berberine (1.8 mg/kg) and dexamethasone (20 mg/kg) were introduced via peritoneal route 30 minutes earlier to each trial in berberine treated and dexamethasone treated groups respectively. Results: Total leukocyte count (TLC) in blood sample of berberine treated group (9990±1346 mm3) and dexamethasone treated group (9054±1432 mm3) was significantly low than OVA sensitized group (14261±3151 mm3). TLC of broncho-alveolar lavage (BAL) fluid in berberine treated group (384±26 mm3) and dexamethasone treated group (306±86) mm3 was significantly low than OVA sensitized group (598±110 mm3). Eosinophil percentage in blood of berberine treated (21.50±3.08) and dexamethasone treated (13.33±5.65) groups were significantly low than OVA sensitized group (30.33±6.74), while eosinophil percentage in BAL fluid was 26.00±6.69 and 21.00±7.46 in berberine treated and dexamethasone treated groups respectively with significant difference from OVA sensitized group value of 40.00±7.79. Conclusion: Berberine and dexamethasone both had reduced TLC and eosinophil percentage in both blood and BAL fluid as compared to OVA sensitized group but berberine is less effective than dexamethasone. Keywords: berberine, dexamethasone, asthma, airway inflammation How to cite: Zaidi T.S., Kausar R., Malik M., Sarfraz J., Shafiq A., Chiragh S. Comparison of berberine and dexamethasone on blood and bronchial inflammatory cells of ovalbumin sensitized guinea pigs. Esculapio 202;17(01):34-38

Evaluation of Acute Dermal Irritation, Sensitization and Acute Dermal Toxicity of Leather Cream in Laboratory Animals

Unused leather products attract fungal invasion and deteriorate the product lifespan. The development of antifungal leather cream/polish can solve this array considered that the prepared formulation is skin friendly. Possible adhesion of leather cream to the skin may provide a route for entry of hazardous and harmful ingredients. In this study, leather cream prepared in the laboratory and a marketed product of leather cream were tested for acute dermal irritation in rabbits, skin sensitization in guinea pigs, and acute dermal toxicity in rats. Draize dermal irritation scoring system was used to examine the condition of erythema and edema in rabbits at intervals of 24, 48 and 72 hours. Percentage sensitization rate, sensitization grade, sensitization classifications were accessed in guinea pigs after 24 hours of dermal exposure. Rats were observed daily for 14 days to detect any signs of irritation, change in general behavior, toxicity, morbidity and mortality. Histological examination of rat’s skin was performed at the end of the study. No any kind of dermal responses or any observable signs of systemic toxicity were detected in rabbits treated with either of the leather cream samples. No sensitization, erythema or edema was observed in guinea pigs. In the case of rats, the skin was normal with negative signs of toxicity, erythema or edema. Histological study also failed to show any damage in skin tissue layers of rats applied with test materials. The result provides a base for development of anti-fungal leather cream/polish which can enhance the life of leather and avoid the fungal invasion upon storing for a period of time. © 2020 iGlobal Research and Publishing Foundation. All rights reserved.

Interpretation of murine local lymph node assay (LLNA) data for skin sensitization: Overload effects, danger signals and chemistry-based read-across

There is a large body of information on testing of chemicals for skin sensitization in the murine local lymph node assay (LLNA), in which potency is quantified by the EC3 value, derived from dose-response data. This information finds use in risk assessment and regulatory classification, and also in assessing the performance of non-animal methods. However, some LLNA results are not straightforward to interpret, and in some cases published EC3 values are questionable. These cases usually arise where the dose–response does not show a monotonic increasing pattern but is bell-shaped, or shows a decrease in response with increasing dose over the whole dose range tested. By analogy with a long-recognised phenomenon in guinea pig sensitization, this is referred to as the overload effect. Here a mechanistic rationale is presented to explain the overload effect, and at the same time to explain the production of danger signals even when the sensitizer is non-irritant. Some illustrative examples are presented where the overload effect can lead to misinterpretation of LLNA results, and chemistry-based read-across is applied to reinterpret the data.

Prospects for the use of a drug based on a complex of fatty acid esters in models of laboratory animals in vivo

In this work we carried out studies of the first obtained micellar composition of fatty acid esters with allantoin on a water basis to determine its local irritant effect and therapeutic efficacy in the treatment of burns. It was found that there was a sequential change in the phases of the burn wound process in all groups. During the observation period, wound suppuration was not observed; there were 4 cases of a complicated course of the wound process and a fatal outcome in the control group. When conducting studies of local irritant effects, the assessment was carried out in points, in the experimental group it was on average 2 points as for the animals of the control group. The conducted studies allow us to conclude that the drug, when applied to the skin according to the interstate standard GOST ISO 10993-10—2011, has a weak degree of response to irritation in guinea pigs. The application of a permissive dose of the drug to sensitized guinea pigs does not cause a response from the skin, which indicates the absence of allergenic properties in this composition.

Purification and biological analysis of specific antigens (ESAT6/CFP10) from Mycobacterium tuberculosis

The pathogenesis of Mycobacterium tuberculosis (Mtb) is related to its low molecular weight proteins mainly ESAT6 and CFP10 that are highly specific and potentially useful for the diagnosis of tuberculosis. This research focused on isolation, purification, and characterization of low molecular weight proteins from Mtb. Cultures of Mtb were inactivated by heating at 68 °C for 90 min and 100 °C for 3 hrs, respectively. Inactivated cultures were filtered and the proteins in the supernatant fluid precipitated with two rounds of ammonium sulfate, at 4 °C. The collected precipitates were dialyzed and subjected to gel chromatography (G-50) and the obtained fractions were analyzed for protein concentrations and molecular weight. ESAT6 and CFP10 protein complex in the purified fraction was confirmed by Western blotting. Guinea pig sensitization assay was used for estimating the potency of the purified fraction compared to the standard PPD. The maximum amount of low molecular weight proteins were precipitated by 20% ammonium sulfate. SDS-PAGE analysis revealed protein bands of approximately 10-15 kDa. The purity of the proteins was ≥95%, as confirmed by SDS-PAGE. The presence of the ESAT-6/CFP10 complex was confirmed by Western blot analysis. The purified fractions showed no cross-reaction with BCG or M. avium strain. ESAT-6/CFP-10 purified by the ammonium sulfate method appeared to be suitable for the development of a diagnostic kit for the detection of Mtb.

Total dose defines the incidence of percutaneous IgE/IgG1 mediated immediate-type hypersensitivity caused by papain.

Assessment of human health risk requires an understanding of antigen dose metrics associated with toxicity. Whereas assessment of the human health risk for delayed-type hypersensitivity is understood, the metrics remain unclear for percutaneous immediate-type hypersensitivity (ITH) mediated by IgE/IgG1. In this work, we aimed to investigate the dose metric for percutaneous ITH mediated by IgE/IgG1 responses. Papain, which causes ITH via percutaneous sensitization in humans, was used to sensitize guinea pigs and mice. The total dose per animal or dose per unit area was adjusted to understand the drivers of sensitization. Passive cutaneous anaphylaxis (PCA) and enzyme-linked immunosorbent assay (ELISA) for papain-specific IgG1 enabled quantification of the response in guinea pigs. In mice, the number of antigen-bearing B cells in the draining lymph nodes (DLN) was calculated using flow cytometry papain-specific IgG1 and IgE levels were quantified by ELISA. PCA positive test rates and the amounts of antigen-specific antibody corresponded with total dose per animal, not dose per unit area. Furthermore, the number of B cells taking up antigen within DLN also correlated with total dose. These findings indicate that the total antigen dose is the important metric for percutaneous IgE/IgG1-mediated ITH.

Age-Related Features of Mast Cell Reactivity in Allergies

Introduction: During last few years, a novel scientific sphere has developed and is successfully growing,associated with the study of age-related changes in immune and allergic reactions at the cellular level.Although there is a fact that mast cells are the subject of many researches, many aspects of their physiologyand pathology remain unsolved. Moreover, clarification in the age-related features of mast cell reactivitymakes a great contribution to the expansion of our view of point on the characteristics of the clinic, etiologyand pathogenesis of allergies in ontogenesis.Materials and Method: We used 443 guinea pigs of both sexes of different ages, previously experienced 10-14 day-quarantine control under conditions of vivariums. Animals were constantly monitored, body weightwas periodically determined, and rectal temperature was measured. Sensitization and anaphylactic reactionswere reproduced by conventional method. Normal horse serum was used as an allergen. The resolving doseof a specific allergen was administered once intravenously or intraperitoneally 14-16 days after the startof the first sensitizing dose of the allergen. Collected digital data processed on electronic microcalculatorRokwell international Model 44 (USA).Results: With age, the rate of filling mast cells with granules increases. In guinea pigs of an older age, ascompared to newborns, the rates of filling mast cells with granules are 1.3-2.2 times greater (p <0.05), whichis consistent with the literature. Spontaneous mast cell degranulation was observed in intact animals. Theirfrequency ranged from 5.4 to 8.6% and, in principle, did not depend on age. A comparative analysis of thefrequency of degranulation, depending on age, showed that the frequency of degranulation in sensitizednewborns is slightly lower than in older animals. However, with age in sensitized guinea pigs, the frequencyof degranulated mast cell forms increased and the mast cell reactivity in one month aged pigs becameapproximately the same as in mature animals.Conclusion: In general, the functional activity of mast cells increases with age, and by the age of one monthreaches the level of functional activity. In the development of specific therapy, a decrease in the titer ofhomocytotropic antibodies and the percentage of mast cell degranulation is vital.

Thermal homeostasis impacts the sensitivity of guinea pigs to QT-prolonging drugs

Thermal homeostasis impacts the sensitivity of guinea pigs to QT-prolonging drugs

АЛЛЕРГИЗИРУЮЩИЕ И РАЗДРАЖАЮЩИЕ СВОЙСТВА НОВЫХ СЕЛЕКТИВНЫХ ПРЕПАРАТОВ НА ОСНОВЕ ЭКЗОСОМАЛЬНЫХ И ХИТОЗАНОВЫХ ЧАСТИЦ

Each drug, before being introduced to the pharmaceutical market, undergoes a mandatory set of preclinical and clinical studies, including those for allergenicity. As a result of the work carried out from 2015 to 2020 at the Department of Therapy and Pharmacology, selective preparations based on exosomal (Exazit) and chitosan (Chitasit) nanoparticles have been developed, and their physicochemical and pharmacological and therapeutic properties have been analyzed. This article presents the results of studying the allergenic and irritating properties of these drugs. To study the allergic effect after oral administration of drugs, 6 experimental and 6 control groups of guinea pigs weighing 250–300 g were formed. Testing of the sensitizing properties of drugs was carried out on days 10, 15 and 30 from the start of administration. The results of the conducted studies have established that the conjunctival test of sensitized guinea pigs, throughout the experiment, did not differ from the reaction in control animals. During a visual assessment of the state of the conjunctiva, cornea and eyelids of the eyes of experimental animals, it was found that the studied drugs did not cause irritation of the conjunctiva both immediately after instillation and throughout the entire experiment. The setting of the provocative nasal test did not cause hyperemia of the mucous membrane, sneezing and increased mucus secretion in animals. The percentage of degranulated mast cells in the experimental groups is less than 10%. Thus, as a result of the study of the allergenic properties of Exazit and Chitasit in guinea pigs, it was found that there is no allergenic effect that irritates the conjunctiva of the eyes and mucous membranes, the drugs do not cause degranulation of mast cells in rats beyond the permissible limits.

Analysis of Immunogenicity of Acanthopanax senticosus Injection

The cause of the immunogenicity of Acanthopanax senticosus injection has been studied. The method used for the preparation of the injection includes boiling the plant material in water, precipitation with ethanol, and filtration to produce the extract. In the current study, this extract was treated with saturated ammonium sulfate to collect the protein from the extract. Indicators of systemic allergy, including total immunoglobulin (IgE), interleukin -4 (IL-4), and interferon-gamma (IFN-γ) levels in the antiserum of Guinea pigs sensitized with this protein extract were used to determine the immunogenicity of the protein extract. Testing was performed using Western blotting to identify IgE in antiserum binding in Guinea pigs sensitized by injection. The target protein band was further identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The protein extract from A. senticosus caused positive reactions and significantly increased the levels of total IgE, IL-4, and IFN-γ in the serum of sensitized Guinea pigs. Western blotting showed that a group of proteins with a molecular weight of 60 000 in the A. senticosus protein extract acted as the major immunogenic protein by binding to serum IgE of Guinea pigs sensitized with A. senticosus injection. The protein was a pyruvate kinase homologous protein with high sequence similarity to the pyruvate kinase protein of Vitis vinifera. It was concluded that the allergic reaction caused by the injection of A. senticosus may be related to the interaction of these macromolecular proteins within the body.

Safety and immunogenicity of a novel quadrivalent subunit influenza vaccine in animal models

ABSTRACT Background: Compared with trivalent influenza vaccines, quadrivalent influenza vaccines are expected to provide wider protection against influenza B virus infections. We developed a novel quadrivalent subunit influenza vaccine which was distinct from the influenza vaccines available on the market in production process. In this research, we evaluated the safety and immunogenicity of the quadrivalent subunit influenza vaccine in animal models. Methods: In toxicity assessment, 40 SD rats were randomly assigned to be intramuscularly injected with 1.0 ml of the tested vaccine (33 μg/ml) or 0.9% sodium chloride solution. In irritation assessment, eight rabbits were randomly assigned to receive 0.5 ml of tested vaccine or phosphate buffer solution intramuscularly. Thirty-two guinea pigs were randomly assigned to be intramuscularly injected with high-dose tested vaccine (0.5 ml), low-dose tested vaccine (0.05 ml), ovalbumin, or 0.9% sodium chloride solution, respectively, for sensitization assessment. In immunogenicity assessment, 50 BALB/c mice were equally randomized to receive one dose of tested vaccine, two doses of tested vaccine with an interval of 14 days, 0.5 ml of trivalent subunit influenza vaccine, 0.5 ml of monovalent subunit influenza vaccine, or 0.5 ml of phosphate buffer solution. Orbital blood was collected before and 28 and 42 days after administration of the injections for detecting influenza antibody titers. Results: No abnormal toxicity and irritation in rats and rabbits showed in the gross autopsy and histopathological examinations. The results of sensitization in guinea pigs indicated that no obvious allergic symptoms observed in the high-dose and low-dose vaccine groups within 30 min after twice provocations, and the result of sensitization evaluation was negative. Vaccine induced significant immune responses in mice with 100% seroconversion rates at 28 and 42 days after the first dose. The geometric mean titers (GMTs) of hemagglutination inhibition (HI) antibodies at day 28 in one-dose quadri–vaccine and two-dose quadri–vaccine groups were comparable to those in the tri–vaccine or mono-vaccine groups for shared influenza strains. However, the GMTs of HI antibodies against H1N1 (P = 0.025) and BV (P = 0.049) at day 42 in one-dose quadri–vaccine group were significantly lower than those in the tri–vaccine or mono-vaccine groups. The GMTs of HI antibodies against H1N1, H3N1, BY, and BV at day 28 and day 42 were comparable between one-dose quadri–vaccine and two-dose quadri–vaccine groups. Conclusions: The quadrivalent subunit influenza vaccine was safe and immunogenic in animal models. One dose of the vaccine could elicit a satisfactory antibody response in mice.

Allergic sensitization increases contractile responses to 5-HT in guinea pig aorta.

Epidemiological and clinical studies suggest that asthma is associated with adverse cardiovascular outcomes, but its mechanism is uncertain. 5-Hydroxytryptamine (5-HT) is a mediator involved in asthma and in cardiovascular functioning. Thus, in the present study, we explored whether allergic sensitization in guinea pigs modifies 5-HT-induced contractile responses and 5-HT2A receptor expression in thoracic aorta rings. We found that sensitization produced a significant increase of 100 microM 5-HT-induced contractions of aorta rings (~27 % greater contraction than in non-sensitized animals, p<0.05). Preincubation with 10 nM ketanserin (a 5-HT2A receptor antagonist) reduced by ~30 % (p=0.003) and ~36 % (p=0.005) the area under the curve of 5-HT-induced contractions in aortas from non-sensitized and sensitized animals, respectively. There were no differences between sensitized and non-sensitized animals with respect to mRNA (qPCR) and protein (Western blot) expression of 5-HT2A receptor in thoracic aortas. We concluded that in this guinea pig model of asthma, allergic sensitization is not confined to airways, but also affects arterial contractile responses to 5-HT; changes in the expression of the 5-HT2A receptor appear not to be involved in this phenomenon.

RESEARCH OF ALLERGIZING PROPERTIES OF DERMATOLOGICAL FILM WITH ALOE EXTRACT LIQUID AND ACTOVEGIN

A study of allergenic properties of the developed dermatological film with liquid aloe extract and actovegin was conducted. The experiments were carried out by the method of cutaneous samples and by the method of epicutaneous sensitization in guinea pigs. The results were evaluated using the scale of S.V. Suvorov. As a result of the work done, allergenic properties have not been identified.

Potential effect of one-alpha-hydroxy vitamin D3 (alfacalcidol) on experimentally induced allergic asthma

Background: Alfacalcidol is a vitamin D analog mainly used in the management of osteoporosis. Apart from the well-known role of vitamin D in bone and calcium metabolism, an immune-modulator role of vitamin D in allergic diseases was suggested. Aim: To evaluate the potential effect of alfacalcidol either alone or in combination with salbutamol or/and prednisolone and its role in the protection and management of experimentally induced allergic asthma. Materials and methods: Male guinea pigs were divided into two groups: group A constituted normal guinea pigs (received saline) and group B constituted ovalbumin-sensitized guinea pigs, which were divided into eight subgroups: group B1 (control): groups from B2 to B4 was treated with salbutamol, prednisolone, alfacalcidol, respectively, for 1 week, while group B5 was treated with salbutamol+prednisolone; group B6 was treated with salbutamol+alfacalcidol; group B7 was treated with prednisolone+alfacalcidol; and group B8 was treated with salbutamol, prednisolone, and alfacalcidol for 1 week. Twenty-four hours after the last dose, the animals were subjected to both: (a)pharmacological studies, (b) histopathological studies (group A, group B1, B3, and B4). Results: (a) Pharmacological studies: sensitization of guinea pigs caused a significant increase in amplitude of histamine-induced contractions of isolated tracheal smooth muscles in comparison to that of the normal one. Treatment of sensitized guinea pigs with salbutamol, prednisolone, or alfacalcidol produced a significant reduction in the amplitude of histamine-induced contractions in comparison to sensitized nontreated guinea pigs. Addition of alfacalcidol to salbutamol or prednisolone in the treatment of sensitized guinea pigs caused more decrease in the amplitude of histamine-induced contractions. However, the highest reduction in the amplitude of contractions of isolated tracheal strips was produced by the treatment of guinea pigs with a combination of salbutamol, prednisolone, and alfacalcidol. (b) Histopathological examinations: treatment of sensitized guinea pigs with alfacalcidol resulted in an anti-remodeling effect, while prednisolone showed better anti-inflammatory effect, when it was compared with that of the control group. Conclusion: Alfacalcidol was shown to possess anti-inflammatory and anti-remodeling effects in allergic asthma. It was superior to prednisolone regarding the anti-remodeling effect. So, alfacalcidol could be beneficial in the management of allergic asthma.

In Vitro Antibacterial Effect and Sensitization Evaluation of Tea-Tree Oil Preparation

In order to promote the development and utilization of tea tree oil in china, The tea tree oil which was extracted from terpinen-4-ol typed Melaleuca alternifolia introduced and selected by Guangxi Forestry Science Institute, was tested in antibacterial activity, local irritation and sensitization research. The results demonstrated that the tea tree oil preparation to the selected staphylococcus aureus, epidermis staphylococcus aureus, escherich-ia coli, pseudomonas aeruginosa and candida albicans has good antibacterial activity. There were no irritating reations observed in the dermal irritation test in rabbits and 0.5% tea tree oil has no sensitization in guinea pigs. This study confirms that tea tree oil has significant antibacterial activity, low -irritating and hypo-allergenic properties, promising a broad development value.

Plasma based targeted metabolomic analysis reveals alterations of phosphatidylcholines and oxidative stress markers in guinea pig model of allergic asthma

Bronchial asthma is one of the most common, chronic respiratory diseases, characterized by reversible airway obstruction, eosinophil and Th2 infiltration, airway hyperresponsiveness and airway remodelling; with many cells and mediators involved. Metabolomics is a relatively new field in “omics” sciences enabling the identification of metabolome for better diagnostics and studying of diseases phenotype.The aim of this study was to investigate the role of targeted metabolomics study for better understanding of the bronchial asthma pathophysiology and finding potential biomarkers in experimental models of eosinophilic inflammation.Plasma level of 185 metabolites was measured with the AbsoluteIDQ™ p180 kit in guinea pigs with experimentally-induced allergic inflammation (n = 15) compared to naïve non-sensitised and non-challenged controls (n = 18).Of the 185 metabolites identified in plasma, 22 were significantly different and changed in ovalbumin sensitised animals. Plasma level of 13 phosphatidylcholines with saturated and unsaturated long-chain fatty acids, total phosphatidylcholines count, carnitine, symmetric dimethylarginine and its ratio to total unmodified arginine, and kynurenine to tryptophan ratio were found to be decreased, while phospholipase A2 activity indicator, tryptophan, taurine and ratio of methionine sulfoxide to unmodified methionine were found to be increased in sensitised guinea pigs compared to naïve controls.Targeted metabolomic analysis revealed significant differences in plasma metabolome of sensitised guinea pigs. Our observations point to the activation of inflammatory and immune pathways, as well as the involvement of oxidative stress.

Gamma-poly glutamate/gelatin composite hydrogels crosslinked by proanthocyanidins for wound healing

A novel multifunctional poly(γ-glutamic acid) (γ-PGA)/gelatin hydrogel has been developed and used as a wound dressing. An ideal wound dressing should effectively provide a moist environment, absorb wound exudates and protect the wound from foreign microbes. Water soluble γ-PGA salts of sodium and calcium forms were chosen for their good biocompatibility, biodegradability and water absorption capacity. Oligomeric proanthocyanidins (OPCs), naturally occurring plant metabolites and potent antioxidants, were investigated as a non-toxic crosslinking agent in this study. The effects of hydrogels on the degree of crosslinking, swelling, in vitro degradation, mechanical properties and radical scavenging activity were systemically evaluated. A cell viability assay demonstrated that these OPCs crosslinked γ-PGA/gelatin (PGO) hydrogels were not cytotoxic to L929 fibroblasts. Dermal irritation and skin sensitization tests were examined using a guinea pig model; the hydrogels were considered to be neither allergic nor a dermal sensitizer in guinea pigs. Lastly, an in vivo wound healing model in rats was used to study the effects of the hydrogels on wound healing for 21 days. PGO hydrogels formed by both Na and Ca salts could accelerate wound contraction and re-epithelialization, in which Na-PGO hydrogel was significantly better than the untreated control group. The findings suggest that PGO hydrogels are promising wound dressing materials for the treatment for wound healing.