Inhalation Test Research Articles

Phytochemical analysis and pharmacological activities of essential oils extracted from Zingiber officinale (Roscoe) used in mediterranean diet: in vitro and in silico studies

ABSTRACT For their pungency and tanginess, ginger rhizomes (Zingiber officinale) are often used in cooking. Ginger has long been used in traditional medicine and home remedies to treat pain and inflammation. This research tests Z. officinale (EOZ) essential oils’ chemical composition, insecticidal, and antioxidant properties in vitro and in silico. EOZ yielded 0.69% of root mass with 23 chemicals. The main chemicals in EOZ were α-zingiberene (23.850%), Geranial (14.160%), and (E,E)-α-farnesene (9.980%). EOZ demonstrated noteworthy antioxidant activity in all tests, with an IC50 of 9.53 ± 0.41 mg/mL in the DPPH test and an EC50 of 87.46 ± 3.19 mg/mL in the FRAP system. The results indicate that EOZ has strong efficacy against C. maculatus, even at low concentrations (1.00 μL/100 g). Note that EOZ killed 20 ± 0% and 13.33 ± 4.44% of adult C. maculatus in inhalation and contact tests, respectively. High concentration (20.00 μL/100 g) resulted in 100% adult mortality in inhalation tests and 96.67 ± 4.44% mortality in contact testing. We then used molecular docking to identify EO major component binding modes by targeting insecticidal and antioxidant protein structures 1R20 and 1R4U. This research helps create insecticides and antioxidants from common ginger essential oil.

Particle aerosol generation and potential altering in airflow used for acute/repeated inhalation toxicity testing

Reproducible aerosol generation in combination with stable aerosol properties are essential prerequisites for compliant performance of acute or repeated inhalation toxicity tests of particulate materials according to OECD TG 403, 436, 412, or 413. A frequent problem of powder aerosol generation is the formation of coarse agglomerates with low shear resistance, which are beyond the tolerable size range but not detected by the prescribed aerodynamic measurement techniques by cascade impactor as the measurement conditions cause a disintegration into smaller fragments. But such agglomerates are observed during the transport to the inhalation chambers. These effects particularly apply to high mass concentrations and low-density powders, i.e., pyrogenic oxides. This study describes the transport influence in the airflow on the change of powder aerosols and on their respirability.A simplified short tube set-up was developed for the aerosol transport pre-tests, which allows the determination of the optimal aerosol formation conditions for the inhalation tests. The particles were measured with low shear using laser diffraction measurement or optical particle counters. The calculation of the aerodynamic particle sizes prescribed in the guidelines requires knowledge of the effective particle density of the porous aerosol particles. A practicable method for determining these is presented and described. In the outlook, first low concentration measurements show that clear agglomeration effects can also occur at particle concentrations around 20 mg/m³.

Phytochemical characterization by HS-SPME-GC-MS and exploration of the antifungal, insecticidal and repellent activity of Ptychotis verticillata essential oil

The objective of this investigation was to assess the chemical makeup of essential oil derived from Ptychotis verticillata (PVEO), and to examine its antifungal, insecticidal, and repellent properties. PVEO was extracted through hydrodistillation, and its volatile constituents were analyzed using Headspace Solid-Phase Microextraction Gas Chromatography-Mass Spectrometry. Qualitative and quantitative evaluation of antifungal activity was carried out using the agar diffusion method and the minimum inhibitory concentration (MIC) test against Candida glabrata, Saccharomyces cerevisiae, Aspergillus niger and Penicillium digitatum. We evaluated the repellent potential, as well as the contact and inhalation toxicity of PVEO against Callosobruchus maculatus. The results of the study indicated that the essential oil of P. verticillata was composed mainly of γ-Terpinen (25.86%), β-Cymene (18.70%) O-Cymen-5-ol (16.78) and α-Pinene (12.13%). PVEO showed potent antifungal activity against all strains tested. The results of insecticidal activity of this essential oil were promising in adult C. maculatus. At a dose of 20 ml · dm–3 of air, EO caused maximum mortality with an LC50 value of 5.64 ml · dm–3 for the inhalation test and 3.4 ml · dm–3 for the contact test. In addition, a significant decrease in the number of eggs laid and adult emergence was observed as EO doses increased, reaching a reduction of around 95% at a dose of 20 ml · dm–3 of air. In terms of repellent activity, PVEO also showed encouraging results. It demonstrated an average repellent activity of around 92 ± 10.95%. Furthermore, molecular docking simulations corroborated the in vitro results and demonstrated that specific p-Menthen-3-one compounds formed more robust hydrogen bonding interactions with the target receptors. These experiments underscore PVEO’s effectiveness as a fungicide against the tested fungal strains, demonstrating its role as a bio-insecticide against C. maculatus adults, and its potential as an appealing repellent. This suggests that PVEO could serve as a valuable alternative within integrated pest management strategies.

Antifungal, Insecticidal, and Repellent Activities of Rosmarinus officinalis Essential Oil and Molecular Docking of Its Constituents against Acetylcholinesterase and β-Tubulin.

The aim of this study was to determine the phytochemical composition and evaluate the antifungal and insecticidal properties of Rosmarinus officinalis essential oil (EO). GC-MS was employed to analyze the phytochemical profile of the EO. The antifungal activity of the EO was assessed by calculating growth inhibition rates for Alternaria alternata, Fusarium oxysporum, and Botrytis cinerea. Repellent capacity and toxicity were evaluated through inhalation and contact tests on Callosobruchus maculatus. Molecular docking techniques were utilized to test the insecticidal and antifungal activities of rosemary EO. The analysis revealed a total of sixteen components in R. officinalis EO, with 1,8-cineole (40.80%) being the major constituent, followed by α-pinene (26.18%) and camphor (19.53%). Antifungal evaluation demonstrated a significant inhibitory impact on the mycelial growth of the tested fungi, with complete inhibition observed against B. cinerea. In terms of insecticidal capacity, the EO induced complete mortality of C. maculatus adults at a concentration of 1 μL/L air, with an inhalation test LC50 value of 0.62 μL/L air. Concentration-dependent reductions were observed in the number of both laid eggs and emerged insects, reaching a 99.36% reduction. The EO also exhibited a moderate effectiveness in repelling insects, with an average repellency rate of 50.83%. In silico analysis identified borneol as the most active molecule against insect acetylcholinesterase (PDB: 6ARY) with a Glide score of -7.254 kcal/mol. α-Caryophyllene showed the highest activity against B. cinerea β-tubulin (PDB: 3N2G) with a Glide score of -7.025 kcal/mol. These findings suggest that the EO derived from Moroccan Rosmarinus officinalis has potential as an effective natural agent against pathogenic fungi and could serve as a sustainable and environmentally friendly alternative as a bioinsecticide.

A new tidal breathing measurement device detects bronchial obstruction during methacholine challenge test

PurposeBronchial hyperresponsiveness (BHR), a hallmark of bronchial asthma, is typically diagnosed through a methacholine inhalation test followed by spirometry, known as the methacholine challenge test (MCT). While spirometry relies on proper patients' cooperation and precise execution of forced breathing maneuvers, we conducted a comparative analysis with the portable nanomaterial-based sensing device, SenseGuard™, to non-intrusively assess tidal breathing parameters. Materials and methodsIn this prospective study, 37 adult participants with suspected asthma underwent sequential spirometry and SenseGuard™ measurements after inhaling increasing methacholine doses. ResultsAmong the 37 participants, 18 were MCT responders, 17 were non-responders and 2 were excluded due to uninterpretable data. The MCT responders exhibited a significant lung function difference when comparing the change from baseline to maximum response. This was evident through a notable decrease in forced expiratory volume in 1 ​s (FEV1) levels in spirometry, as well as in prominent changes in tidal breathing parameters as assessed by SenseGuard™, including the expiratory pause time (Trest) to total breath time (Ttot) ratio, and the expiratory time (Tex) to Ttot ratio. Notably, the ratios Trest/Ttot (∗p ​= ​0.02), Tex/Ttot (∗p ​= ​0.002), and inspiratory time (Tin) to Tex (∗p ​= ​0.04) identified MCT responders distinctly, corresponding to spirometry (∗p ​< ​0.0001). ConclusionsThis study demonstrates that tidal breathing assessment using SenseGuard™ device reliably detects clinically relevant changes of respiratory parameter during the MCT. It effectively distinguishes between responders and non-responders, with strong agreement to conventional spirometry-measured FEV1. This technology holds promise for monitoring clinical respiratory changes in bronchial asthma patients pending further studies.

Refinement of the acute inhalation limit test for inert, nano-sized dusts by an in silico dosimetry-based evaluation: case study for the dissolution of a regulatory dilemma.

This case study aims to describe the dilemma faced when exposing rats to very high concentrations of fine, pulverulent materials for acute inhalation studies and to address the regulatory question of whether the effects seen here are relevant to humans and the subject of classification according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS). Many powders match the definition of nanomaterials in the EU; therefore, information on acute inhalation testing of powders up to the GHS cutoff of 5mg/L is required. However, testing rats at such a high aerosol concentration can cause physical obstruction of the airways and even mortality by suffocation. Therefore, to evaluate whether the physical effects on airway obstruction in rats exposed to 5mg/L for 4hours and alternative exposures to 1 and 2mg/L are relevant for humans, an in silico evaluation of aerosol deposition was conducted using the multiple-path particle dosimetry (MPPD) model. For this evaluation, actual exposure conditions for an organic, nano-sized pigment which produced 100% lethality in rats at 5mg/L, but not at 1mg/L, were used to assess the potential for airway obstruction in rats and accordingly in humans. As an indicator of the potential for airway obstruction, the ratio of the diameter of the deposited, aggregated aerosol to airway diameter was calculated for each exposure condition. For rats exposed to 5mg/L for 4h, approximately 75% of tracheobronchial and 22% of pulmonary/alveolar airways were considered vulnerable to significant or complete obstruction (ratios >0.5). In humans, an equivalent exposure resulted in just over 96% of human tracheobronchial airways that received deposited mass to airway diameter ratios between 0.3 and 0.4 (nasal) or 0.4 and 0.5 (oral), with no airways with ratios >0.5. For the pulmonary/alveolar region, ∼88% of the airways following nasal or oral breathing were predicted to have deposited aerosol diameter to airway diameter ratios <0.1, with no airways with ratios >0.5. Thus, the in silico results obtained for rats are in line with the pathological findings of the animal test. The predicted results in humans, however, affirm the hypothesis of a rat-specific high dose effect which does not justify a classification according to GHS.

Development of a non-target strategy for evaluation of potential biological effects of inhalable aerosols generated during purposeful room conditioning using an in vitro inhalation model

Objectives: An integrated in vitro inhalation approach was outlined to estimate potential adverse acute inhalation effects of aerosols from commercial nebulizer applications used for purposeful room conditioning such as disinfection, scenting or others. Aerosol characterization, exposure estimation and evaluation of acute biological effects by in vitro inhalation were included to generate dose-response data, allowing for determination of in vitro lowest observable adverse effect levels (LOAELs). Correlation of these to estimates of human lung deposition was included for quantitative in vitro to in vivo extrapolation approach (QIVIVE) for acute effects during human exposure.Methods: To test the proposed approach, a case study was undertaken using two realistic test materials. An acute in vitro inhalation setup with air-liquid interface A549-cells in an optimized exposure situation (P.R.I.T.® ExpoCube®) was used to expose cells and analysis of relevant biological effects (viability, mitochondrial membrane potential, stress, IL-8 release) was carried out. Results: The observed dose-responsive effects in a sub-toxic dose-range could be attributed to the main component of one test material and its presence in the aerosol phase of the nebulized material. QIVIVE resulted in a factor of at least 256 between the in vitro LOAEL and the estimated acute human lung exposure for this test material.Conclusions: The case-study shows the value of the non-target in vitro inhalation testing approach especially in case of a lack of knowledge on complex product composition. It is expected that approaches like this will be of high value for product safety and environmental health in the future.

Prefatoryin silicostudies andin vitroinsecticidal effect ofNigella sativa(L.) essential oil and its active compound (carvacrol) against theCallosobruchus maculatusadults (Fab), a major pest of chickpea

AbstractTo help discover a reasonable and eco-friendly insecticide, we undertook a study on the insecticidal potential of carvacrol and essential oils extracted by hydrodistillation using a Clevenger apparatus from the plant speciesNigella sativaseeds essential oils ofNigella sativa(EONS) onCallosobruchus maculatusadults. Several tests including contact toxicity, repellent effect, topical contact test, and inhalation effect were conducted. Adults ofC. maculatushave been exposed to the toxic effects of different concentrations of these essential oils as well as with carvacrol. The results obtained showed that both EONS and carvacrol exhibited a moderate repellent effect (class II) onC.maculatusadults. EONS showed the highest toxicity by inhalation test, with an LD50of 13.386 and an LD95of 33.186 μL/cm2, compared to carvacrol (LD50= 21.509 and LD95= 38.877 μL/cm2). The EONS by contact test exhibited more toxic effects, with an LD50of 23.350 µL/100 g and an LD95of 45.315 µL/100 g, compared to carvacrol (LD50= 27.853 µL/100 g and LD95= 45.184 µL/100 g). For the topical contact test results, carvacrol was more toxic, with an LD50of 3.915 and an LD95of 7.696 µL/mL, compared to EONS (LD50= 14.509 and LD95= 25.516 µL/mL). The high toxicity of EONS can be explained by the presence of 25.8% ofo-cymene, 8.53% of cyclofenchene, and 7.71% of beta-pinene, as well as 4.6% of carvacrol, in its chemical composition. Unmitigatedly, these data suggest that the essential oils ofN. sativamay present a raw material for the development of new bio-insecticidal products againstC. maculatus, one of the main pests of stored foodstuffs.

Pharmacological activities of chemically characterized essential oils from Haplophyllum tuberculatum (Forssk.).

The present work aimed at characterizing the phytochemical composition of Haplophyllum tuberculatum essential oil (HTEO), assessing its antifungal activity against various fungal strains, evaluating its insecticidal and repulsive properties against Callosobruchus maculatus, and determine its antioxidant capacity. To this end, Gas chromatography-mass spectrometry analysis detected 34 compounds in HTEO, with β-Caryophyllene being the major constituent (36.94%). HTEO demonstrated predominantly modest antifungal effects, however, it sustains notable activity, particularly against Aspergillus flavus, with an inhibition rate of 76.50% ± 0.60%. Minimum inhibitory concentrations ranged from 20.53 ± 5.08 to 76.26 ± 5.08mg/mL, effectively inhibiting fungal growth. Furthermore, the antifungal, and antioxidant activities of HTEO were evaluated in silico against the proteins Aspergillus flavus FAD glucose dehydrogenase, and beta-1,4-endoglucanase from Aspergillus niger, NAD(P)H Oxidase. Moreover, HTEO displayed strong insecticidal activity against C. maculatus, with contact and inhalation tests yielding LC50 values of 30.66 and 40.28μL/100g, respectively, after 24h of exposure. A dose of 5μL/100g significantly reduced oviposition (48.85%) and inhibited emergence (45.15%) compared to the control group. Additionally, HTEO exhibited a high total antioxidant capacity of 758.34mg AAE/g EO, highlighting its antioxidant potential. Insilico results showed that the antifungal activity of HTEO is mostly attributed to γ-Cadinene and p-Cymen-7-ol, while antioxidant is attributed to α-Terpinyl isobutyrate displayed. Overall, HTEO offers a sustainable and environmentally friendly alternative to synthetic products used to manage diseases.

Control of the Maize Pest Beetle, Anomala denuda (Coleoptera: Scarabaeidae), Using Insecticidal Plants Ricinus communis and Azadirachta indica in a Laboratory Setting

The aim of this study was to identify a natural alternative to synthetic insecticides for controlling the Anomala denuda beetle, a destructive corn pest. The investigation took place at Félix Houphouët-Boigny University's Natural Environments and Biodiversity Conservation Laboratory at a temperature of 17°C. Aqueous extracts of Azadirachta indica leaves and Ricinus communis seed capsules were tested for their effectiveness against the beetle. The reference pesticides, Cypercal 50EC and Viper 42EC, were used as positive control. The tests included three types of toxicity (inhalation, ingestion, and repulsion) with four repetitions each and a four-day exposure period. The ingestion test was the most effective. At 110 g/l, A. indica and R. communis extracts had corrected mortality rates of 87.41 ± 3.21 and 90.28 ± 1.71%, respectively. In the inhalation test, A. indica and R. communis extracts resulted in 80.92 ± 5.88% and 83.14 ± 7.57% mortality rates, respectively, at the same concentration. At the same concentration, R. communis extract repelled 60% of beetles, which was higher than A. indica's 53.3%. The insecticidal effects of the extracts were similar to those of Cypercal in all tests. These results suggest that R. communis and A. indica extracts could be used to control A. denuda pests without relying on synthetic chemical pesticides.

Is pulmonary inflammation a valid predictor of particle induced lung pathology? The case of amorphous and crystalline silicas

Although inflammation is a normal and beneficial response, it is also a key event in the pathology of many chronic diseases, including pulmonary and systemic particle-induced disease. In addition, inflammation is now considered as the key response in standard settings for inhaled particles and a critical endpoint in OECD-based sub-acute/ chronic animal inhalation testing protocols. In this paper, we discuss that whilst the role of inflammation in lung disease is undeniable, it is when inflammation deviates from normal parameters that adversity occurs. We introduce the importance of the time course and in particular, the reversibility of inflammation in the progression towards tissue remodelling and neoplastic changes as commonly seen in rat inhalation studies. For this purpose, we used sub-chronic/ chronic studies studies with synthetic amorphous silicas (SAS) and reactive crystalline silica (RCS) as a source of data to describe the time-course of inflammation towards and beyond adversity. Whilst amorphous silicas induce an acute but reversible inflammatory response, only RCS induces a persistent, progressive response after cessation of exposure, resulting in fibrosis and carcinogenicity in rodents and humans. This suggests that the use of inflammation as a fixed endpoint at the cessation of exposure may not be a reliable predictor of particle-induced lung pathology. We therefore suggest extending the current OECD testing guidelines with a recovery period, that allows inflammation to resolve or progress into altered structure and function, such as fibrosis.

Chemical Characterization, Antioxidant, Insecticidal and Anti-Cholinesterase Activity of Essential Oils Extracted from Cinnamomum verum L.

This study is aimed at evaluating the potential of the essential oil of Cinnamomum verum (EOCV) as an antioxidant, as an insecticide against Callosobruchus maculatus and for its anti-acetylcholinesterase activity. To this end, EOCV was extracted via hydrodistillation from this plant, and the identification of the phytochemicals was performed using gas chromatography–mass spectrometry (GC–MS). The antioxidant power was determined via in vitro tests, the insecticidal ability was tested via exposing C. maculatus to EOCV, and molecular docking was used to evaluate the anti-cholinesterase ability. The results of these GC–MS analyses show that the main composition of EOCV comprises Cinnamaldehyde dimethyl acetal (64.50%), cinnamicaldehyde (35.04%) and α-Copaene (0.11%). The insecticidal potential of the studied OEs, determined by using the inhalation test, and expressed as the concentration of EOs required for the death of 50% of the insects (LC50) and that required the death of 95% of adults (LC95) after 96 h of exposure, was 3.99 ± 0.40 and 14.91 ± 0.10 μL/L of air, respectively. In the contact test, 96 h of exposure gave an LC50 and LC95 of 3.17 ± 0.28 and 8.09 ± 0.05 μL/L of air, respectively. A comparison of the antioxidant activity of EOCV to that of ascorbic acid via DPPH free radical scavenging ability and Ferric Reducing Antioxidant Power (FRAP) revealed the IC50 and EC50 values of EOCV to be much higher than that obtained for ascorbic acid, and the molecular docking simulation revealed Coumarin, Piperonal, Cinnamaldehyde dimethyl and alpha-Copaene as possessing potential inhibitory activities against human acetylcholinesterase. However, further experimental validation is needed to enhance the prospects of this study.

Predicting the tolerability of inhalation therapy as a stage of a personalized approach in the treatment of children with cystic fibrosis

Background. Considering the features of basic therapy for chronic inflammatory bronchopulmonary process in cystic fibrosis (CF), which includes almost daily inhalation of pharmacological drugs (mucolytics, hypertonic saline (HS), antibiotics), evaluation of their tolerability is relevant in practice. The purpose of the study is to create a prognostic scale convenient for use in practical medicine, which would predict development of bronchial hyperreactivity syndrome in children with CF, based on clinical and anamnestic data, the results of molecular genetic research and inhalation tests with increasing concentrations of HS to prescribe personalized treatment. Materials and methods. The results of clinical and anamnestic, microbiological, biochemical, molecular and genetic studies and inhalation tests with increasing concentrations of HS were analyzed in 40 children with CF. The creation of the predictive algorithm was based on the application of correlation analysis, receiver operating characteristic analysis, binary logistic regression, Wald and Kullback method. Results. Two prognostic models have been created that can be used at different stages of providing medical care. Model 1 was built for the ambulatory stage of providing medical care to patients with CF, considering clinical and anamnestic data, molecular genetic predictors, as well as clinical symptoms during inhalation tests with increasing concentrations of HS. This model also can be used in young children who cannot perform spirometry to assess their external breathing function. For the highly specialized medical care, model 2 was created, which considers indicators of spirometric data of inhalation tests with increasing concentrations of HS and the level of exhaled nitric oxide. Conclusions. The developed models make it possible to calculate with high probability the risk of developing bronchial hyperreactivity to perform a personalized selection of HS and choose preventive brocholytic therapy as needed.

Promising Insecticidal Properties of Essential Oils from Artemisia aragonensis Lam. and Artemisia negrei L. (Asteraceae) by Targeting Gamma-Aminobutyric Acid and Ryanodine Receptor Proteins: In Vitro and In Silico Approaches

Artemisia negrei (A. negrei) and Artemisia aragonensis (A. aragonensis) are in the family Asteraceae, which has been used in traditional medicine. The use of plant-derived insecticides has become a promising strategy to reduce the harmful effects of synthetic insecticides and overcome the bio-resistance of pest insects to insecticides. In this regard, the purpose of the current study was to determine the chemical composition and evaluate insecticidal effects of essential oils (EOs) extracted from A. negrei (EON) and A. aragonensis (EOA). Notably, all chemical constituents present in the EOs were identified through GC-MS analysis, whilst the insecticidal properties against Callosobruchus maculatus Fab. (C. maculatus) were investigated by use of in vitro an in silico approaches. The obtained results showed that both tested EOs present a significant insecticidal effect against C. maculatus, which increased significantly upon the dose used in both contact and inhalation tests. The lethal concentrations (LC50) for the inhalation test were found to be 2.1 and 2.97 μL/L, while in the contact test they were 2.08 and 2.74 μL/L of air for EON and EOA, respectively. At 5 μL/L of air, the spawn reduction rate was 88.53 % and 77.41%, while the emergence reduction rate was 94.86% and 81.22% by EON and EOA, respectively. With increasing doses of up to 20 μL/L of air, the reduction in individual emergence reached 100% by the two oils tested after 36 h of treatment. In addition, Molecular docking (MD) simulations supported the in vitro findings and indicated that certain identified components in EOA and EON exhibited stronger hydrogen bonding interactions with the target receptors. Interestingly, the prediction of ADMET properties indicates that the molecules investigated have great pharmacokinetic profiles with no side effects. Taken together, our findings suggest that EOA and EON may exert both potential contact and inhalation insecticidal actions and could be used as an alternative tool for the control of this major insect pest of stored products.

Bioequivalence study of ipratropium bromide inhalation aerosol using PBPK modelling.

Systemic pharmacokinetic (PK) studies can reflect the overall exposure of orally inhaled drug Products (OIDPs) in the blood after inhalation into the lung and can be used to evaluate the bioequivalence of test and reference products. The aim of this article is: (1) to study the PK characteristics and bioequivalence of ipratropium bromide (IB) inhalation aerosol, reference and test products in healthy Chinese subjects; (2) to establish a physiologically based pharmacokinetic (PBPK) model and verify the accuracy of the model in predicting bioequivalence; (3) attempt to use the model to predict the regional distribution of particles in the lung after inhalation, and discuss the effect of gastrointestinal drug absorption of IB on systemic exposure. The study involved two clinical studies. Clinical study-1 (registration number: CTR20201284) was used with non-clinical data to construct and validate a PBPK model in the B2O simulator, a web-based virtual drug development platform. This model assessed different test and reference products' bioequivalence. Results were compared to a second clinical study (Clinical study-2: registration number CTR20202291). The particles' regional distribution in the lung and the gastrointestinal absorption effect on systemic exposure were discussed based on the simulation results. The established PBPK model successfully simulated the in vivo PK characteristics of IB inhalation aerosol, with r 2 close to 1. Gastrointestinal absorption had a negligible effect on systemic exposure. Particles accumulated in the alveolar area were cleared within an hour, followed by particles in the bronchioles and bronchi. This model provided a reliable method for exploring the correlation between in vitro and in vivo PK studies of IB inhalation aerosols. According to the simulation results, the test and reference products were bioequivalent.

Allergy and immunotoxicology in preventive and clinical medicine from theory to practice: Occupational allergy and isocyanate-induced asthma

Genetic and environmental factors and their interactions cause diseases and deteriorate health (Genetic and Environmental Interaction). Exposure to environmental factors plays a major role in the deterioration of health in the workplace.Occupational asthma (OA) is a common disorder in the workplace. Approaches to OA are well described and discussed in "Japanese Guideline for Diagnosis and Management of Occupational Allergic Diseases" by the Japanese Society of Occupational and Environmental Allergy. According to the guideline, OA and work-aggravated asthma comprise work-related asthma, and OA can be further divided into two disease entities: sensitizer-induced OA and irritant-induced OA. The guidelines also describe diagnostic and therapeutic strategies for OA. Since a definitive diagnosis of OA requires a comprehensive decision based on a detailed interview on clinical symptoms related to employment status and clinical tests, including inhalation tests of suspected substances as needed, the possibility of OA should be considered as the first step toward diagnosis of the patient. Otherwise, OA may not be diagnosed. Therapeutic strategies include exposure avoidance, environmental arrangements in the workplace, utilization of social resources for workers, and conventional pharmacotherapy for asthma.Artificially synthesized small compounds are used in various industries and can cause allergies. For example, isocyanates are small compounds in the -NCO group, which have been toxicologically studied. It was later shown that isocyanate could cause various nontoxic adverse health effects, including allergic reactions. Since small agents with low molecular weights bind to proteins, detecting their specific immunoglobulin E (IgE) antibodies targeting small compounds is generally difficult. In contrast, isocyanate-specific IgE antibodies are detectable in individuals with isocyanate allergies.Suspecting OA is essential in cases exposed to newly synthesized compounds, or to those that are already known but applied to new uses, which can be better understood and predicted by studying the health effects of isocyanates.Academic interest in various issues related to allergies, immunology, and toxicology in the workplace includes clinical medicine, epidemiology, and epigenetics related to environmental exposure. Further advanced research in these areas is necessary and promising.

Serum Immunoglobulin G Testing against Pigeon Egg in Stable Fibrotic Hypersensitivity Pneumonitis

Introduction: The accuracy of serum immunoglobulin (Ig) G testing for diagnosis of stable bird-related fibrotic hypersensitivity pneumonitis (HP) is controversial. Furthermore, avian serum, extracts, or feathers were employed as antigens in bird-related HP; however, the usage of egg whites has not been reported. We investigated the utility of IgG testing against pigeon egg whites in patients with stable bird-related fibrotic HP. Methods: Patients having a positive inhalation test for pigeon antigen and a histological investigation with diagnostic confidence of fibrotic HP greater than moderate confidence were included. The control group consisted of patients with interstitial lung diseases (ILDs) other than HP. To select patients in the stable phase, patients with fibrotic HP were excluded if they were clinically considered to be in the acute exacerbation or acute phase. The IgG testing against pigeon egg whites by enzyme-linked immunosorbent assay and the commercialized anti-pigeon IgG testing by fluorescence enzyme immunoassay were investigated. Results: In this study, 37 patients with stable bird-related fibrotic HP and 32 patients with ILDs other than HP participated. Serum IgG testing for pigeon egg whites revealed that the control group’s optical density was 0.147 and the group with bird-related fibrotic HP had a mean value of 0.207 (p = 0.011). IgG testing in bronchial alveolar lavage fluid was not significantly higher in the bird-related fibrotic HP group than in controls (p = 0.42). No significant difference in area under the curve between an IgG testing against pigeon egg whites and a commercialized anti-pigeon IgG testing was observed (p = 0.24). Test accuracy for stable bird-related fibrotic HP ranged from 62% to 76% sensitivity and 59–66% specificity. Conclusion: IgG testing to identify the inciting antigen in patients with stable bird-related fibrotic HP had relatively low accuracy.

THE ROLE OF HEREDITARY PREDISPOSITION (POLYMORPHIC MARKERS OF GLUTATHIONE-S-TRANSFERASE, CATALASE, ENDOTHELIAL NITROGEN OXIDE SYNTHASE GENES) AND SOME ADVERSE ENVIRONMENTAL FACTORS IN THE DEVELOPMENT OF BRONCHO-OBSTRUCTIVE PATHOLOGY IN CHILDREN LIVING IN RADIOACTIVELY CONTAMINATED AREAS.

summarizing the results of many years of research by the authors on the influence of gene polymorphisms encoding xenobiotic biotransformation enzymes (GSTТ1, GSTM1, GSTР1), antioxidant protection (С^262Т of the catalase gene), endothelial nitric oxide synthase (4a/4b VNTR polymorphism of the eNOS gene), and some environmental factors on the occurrence of broncho-obstructive disorders and the development of bronchial asthma in children, residents of radioactively contaminated areas. The examined school-aged children were residents of radioactively RCA who had no clinical signs of respiratory pathology. Deletion polymorphism of catalase gene (CAT C^262T), polymorphism of glutathione-S-transferase gene (GSTТ1, GSTM1, GSTР1) and the polymorphism in the 4th intron (4a/4b) of the eNOS gene were studied in the molecular genetics laboratory of the State Institution «Reference Center for Molecular Diagnostics of Public Health Ministry of Ukraine». Molecular genetic studies were performed by polymerase chain reaction. The study of the ventilation lung capacity was carried out by the method of computer spirometry based on the data of the «flow-volume» loop analysis. A pharmacological inhalation test with a bronchodilator drug which affects the β2-adrenergic receptors of the lungs was used to detect early changes in the ventilatory lung capacity - bronchial hyperreactivity. One of the leading mechanisms, due to which the implementation of hereditary predisposition to bronchial asthma in children living in radioactively contaminated areas is the polymorphism of certain genes of glutathione-S-transferase, catalase, endothelial nitric oxide synthase. With such polymorphic variants of the GST genes, isoforms of enzymes with reduced activity are produced, which limits their ability to effectively neutralize free radicals, which are formed in excess when free radical oxidation processes are activated due to the constant intake of radionuclides with a long half-life into the body of children. Unfavorable factors that increase the risk of developing broncho-obstructive disorders and the likelihood of their implementation in the form of bronchial asthma in children, residents of radioactively contaminated areas, have been identified. It has been established that among them the leading role is played by hereditary predisposition to this disease. On the part of the child, such negative factors were unfavorable conditions of intrauterine development, the presence of signs of exudative-catarrhal diathesis, manifestations of allergies and frequent respiratory diseases from the first months of life.

Chemical Composition and Evaluation of Antifungal and Insecticidal Activities of Essential Oils Extracted from Jambosa caryophyllus (Thunb.) Nied: Clove Buds.

Jambosa caryophyllus has been used in traditional phytotherapy as a treatment against infections. In the present work, essential oils extracted from clove buds (Jambosa caryophyllus ) (EO-JC) were investigated for their composition, antifungal, and insecticidal properties. Extraction of EO-JC was performed by use of hydrodistillation using a Clevenger-type apparatus, and the EOs were analyzed by gas chromatography coupled with mass spectrometry (GC-MS). Antifungal activity of EO-JC was evaluated by the use of solid-state diffusion (disc method) and microdilution to determine the minimum inhibitory concentration (MIC), against three strains of fungus, Aspergillus niger, Aspergillus flavus, and Fusarium oxysporum. Insecticidal activity of EO-JC against the cowpea weevil, Callosobruchus maculatus, was determined to assess utility of EO-JC to control this pest. Several exposures including inhalation and contact were used to determine lethality, as well as the repulsion test was conducted at concentrations of 4, 8, 16, and 32 μL EO-JC. Characterization of EO-JC by GC/MS revealed 34 compounds accounting for 99.98% of the mass of the extract. The predominant compound was eugenol (26.80%) followed by β-caryophyllene (16.03%) and eugenyl acetate (5.83%). The antifungal activity of EO-JC on solid media exhibited inhibitions in the range of 49% to 87%, and MIC was between 3.125 and 7.80 μg EO-JC/mL. Insecticidal activity, as determined by the use of the inhalation test, and expressed as the LD50 and LD95 after 96 hours of exposure was 2.32 and 21.92 μL/L air, respectively. In the contact test, a 96-hour exposure resulted in LD50 and LD95 of 5.51 and 11.05 μL/L of air, respectively. EO-JC exhibited insecticidal activity against fungi and pest chickpea weevil.

Respiratory and heart rate dynamics during peripheral chemoreceptor deactivation compared to targeted sympathetic and sympathetic/parasympathetic (co-)activation.

The importance of peripheral chemoreceptors for cardiorespiratory neural control is known for decades. Pure oxygen inhalation deactivates chemoreceptors and increases parasympathetic outflow. However, the relationship between autonomic nervous system (ANS) activation and resulting respiratory as well as heart rate (HR) dynamics is still not fully understood. In young adults the impact of (1) 100% pure oxygen inhalation (hyperoxic cardiac chemoreflex sensitivity (CHRS) testing), (2) the cold face test (CFT) and (3) the cold pressor test (CPT) on heart rate variability (HRV), hemodynamics and respiratory rate was investigated in randomized order. Baseline ANS outflow was determined assessing respiratory sinus arrhythmia via deep breathing, baroreflex sensitivity and HRV. Baseline ANS outflow was normal in all participants (23±1years, 7 females, 3 males). Hyperoxic CHRS testing decreased HR (after 60±3 vs before 63±3min-1, p=0.004), while increasing total peripheral resistance (1053±87 vs 988±76dyne*s+m2/cm5, p=0.02) and mean arterial blood pressure (93±4 vs 91±4mmHg, p=0.02). HRV indicated increased parasympathetic outflow after hyperoxic CHRS testing accompanied by a decrease in respiratory rate (15±1vs 19±1min-1, p=0.001). In contrast, neither CFT nor CPT altered the respiratory rate (18±1 vs 18±2min-1, p=0.38 and 18±1 vs 18±1min-1, p=0.84, respectively). Changes in HR characteristics during deactivation of peripheral chemoreceptors but not during the CFT and CPT are related with a decrease in respiratory rate. This highlights the need of respiratory rate assessment when evaluating adaptations of cardiorespiratory chemoreceptor control.