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Objective: To study associations between fractional exhaled nitric oxide (FeNO) and asthma, airway symptoms, sensitization to common allergens, outdoor pollution and home environment among 380 students in eight junior high schools in two areas in Indonesia., Methods: Data on health and home were collected by a face-to face interview before measuring FeNO and performing skin prick test against common allergens. Exploratory linear mixed and logistic regression models were employed., Results: Geometric mean of FeNO was 17.8 ppb (GSD 2.09) and 139 students (36.6%) had elevated FeNO (>20 ppb). In total, 107 students (28.2%) were sensitized to house dust mite (HDM) (Der p1 or Der f1), 4 (1.1%) to cat and 3 (0.8%) to mold ( Cladosporium or Alternaria ). Moreover, 20 students (5.3%) had diagnosed asthma, 38 (10.0%) had current wheeze, and 107 (28.2%) had current rhinitis. HDM sensitization, diagnosed asthma, current wheeze, and current rhinitis were associated with FeNO. In total, 281 students (73.9%) had mold or dampness, 232 (61.1%) had environmental tobacco smoke (ETS) and 43 (11.3%) had other odor at home. Indoor mold or dampness and other odor at home were associated with FeNO. ETS was negatively associated with FeNO., Conclusion: HDM sensitization and elevated FeNO can be common among children in this part of Indonesia. The high prevalence of elevated FeNO indicate that undiagnosed childhood asthma is common. Dampness, mold and odor at home can be associated with increased FeNO while ETS can be associated with decreased FeNO.

We studied associations between fractional exhaled nitric oxide (FeNO), health and household exposure among school children (N = 348) in Penang, Malaysia. Multiple logistic regression and linear mixed models were applied. Overall, 46.0% had elevated FeNO (>20 ppb) and 10.6% diagnosed asthma. Male gender (p = 0.002), parental asthma or allergy (p = 0.047), cat allergy (p = 0.009) and seafood allergy (p < 0.001), diagnosed asthma (p = 0.001), wheeze (p = 0.001), ocular symptoms (p = 0.001), rhinitis (p = 0.002) and respiratory infections (p = 0.004) were all associated with FeNO. Students exposed to ETS had lower FeNO (p = 0.05). Dampness and mould was associated with wheeze (p = 0.038), especially in wooden homes (interaction p = 0.042) and among students with elevated FeNO (interaction p = 0.024). Cat keeping increased rhinitis (p = 0.041) and respiratory infections (p = 0.008) and modified the dampness associations. In conclusion, FeNO can be associated with ocular and respiratory symptoms. Elevated FeNO, cat keeping and a wooden house can enhance the risk of wheeze when exposed to dampness and mould.

Fungi exposure has been significantly linked to respiratory illness. However, numerous fungi taxa that are potentially allergenic still undocumented and leave a barrier to establishing a clear connection between exposure and health risks. This study aimed to evaluate the association of fungi composition in settled dust with fractional exhaled nitric oxide (FeNO) levels among school children with doctor-diagnosed asthma. A cross-sectional study was undertaken among secondary school students in eight schools in the urban area of Hulu Langat, Selangor, Malaysia. A total of 470 school children (aged 14 years old) were randomly selected and their FeNO levels were measured and allergic skin prick tests were conducted. The settled dust samples were collected and analysed by using metagenomic technique to determine the fungi composition. The general linear regression with complex sampling was employed to determine the interrelationship. In total, 2645 fungal operational taxonomic units (OTUs) were characterised from the sequencing process which belongs to Ascomycota (60.7 %), Basidiomycota (36.4 %), Glomeromycota (2.9 %) and Chytridiomycota (0.04 %). The top five mostly abundance in all dust samples were Aspergillus clavatus (27.2 %), followed by Hyphoderma multicystidium (12.2 %), Verrucoconiothyrium prosopidis (9.4 %), Ganoderma tuberculosum (9.2 %), and Heterochaete shearii (7.2 %). The regression results indicated that A. clavatus, Brycekendrickomyces acaciae, Candida parapsilosis, Hazslinszkyomyces aloes, H. multicystidium, H. shearii, Starmerella meliponinorum, V. prosopidis were associated in increased of FeNO levels among the asthmatic group at 0.992 ppb (95 % CI = 0.34-1.68), 2.887 ppb (95 % CI = 2.09-3.76), 0.809 ppb (95 % CI = 0.14-1.49), 0.647 ppb (95 % CI = 0.36-0.94), 1.442 ppb (95 % CI = 0.29-2.61), 1.757 ppb (95 % CI = 0.59-2.87), 1.092 ppb (95 % CI = 0.43-1.75) and 1.088 ppb (95 % CI = 0.51-1.62), respectively. To our knowledge, this is a new finding. The findings pointed out that metagenomics profiling of fungi could enhance our understanding of a complex interrelation between rare and unculturable fungi with airway inflammation., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Indoor microbial exposure is associated with asthma, but the health effects of indoor metabolites and chemicals have not been comprehensively assessed., Methods: We collected classroom dust from 24 junior high schools in three geographically distanced areas in Malaysia (Johor Bahru, Terengganu and Penang), and conducted culture-independent high-throughput microbiome and untargeted metabolomics/chemical profiling., Results: 1290 students were surveyed for asthma symptoms (wheeze). In each centre, we found significant variation in the prevalence of wheeze among schools, which could be explained by personal characteristics and air pollutants. Large-scale microbial variations were observed between the three centres; the potential protective bacteria were mainly from phyla Actinobacteria in Johor Bahru, Cyanobacteria in Terengganu and Proteobacteria in Penang. In total, 2633 metabolites and chemicals were characterised. Many metabolites were enriched in low-wheeze schools, including plant secondary metabolites flavonoids/isoflavonoids (isoliquiritigenin, formononetin, astragalin), indole and derivatives (indole, serotonin, 1 H -indole-3-carboxaldehyde), and others (biotin, chavicol). A neural network analysis showed that the indole derivatives were co-occurring with the potential protective microbial taxa, including Actinomycetospora , Fischerella and Truepera , suggesting these microorganisms may pose health effects by releasing indole metabolites. A few synthetic chemicals were enriched in high-wheeze schools, including pesticides (2(3 H )-benzothiazolethione), fragrances (2-aminobenzoic acid, isovaleric acid), detergents and plastics (phthalic acid), and industrial materials (4,4-sulfonyldiphenol)., Conclusions: This is the first association study between high-throughput indoor chemical profiling and asthma symptoms. The consistent results from the three centres indicate that indoor metabolites/chemicals could be a better indicator than the indoor microbiome for environmental and health assessments, providing new insights for asthma prediction, prevention and control., Competing Interests: Conflicts of interest: The authors disclose no conflicts of interest., (Copyright ©The authors 2022.)

Little is known on respiratory effects of indoor chemicals in the tropics. We investigated associations between asthma and respiratory infections in Malaysian students and chemical exposure at home and at school. Moreover, we investigated differences in home environment between the three main ethnic groups in Malaysia (Malay, Chinese, Indian). Totally, 462 students from 8 junior high schools in Johor Bahru participated (96% participation rate). The students answered a questionnaire on health and home environment. Climate, carbon dioxide (CO 2 ), volatile organic compounds (VOC), formaldehyde and nitrogen dioxide (NO 2 ) were measured inside and outside the schools. Multilevel logistic regression was applied to study associations between exposure and health. Totally 4.8% were smokers, 10.3% had wheeze, 9.3% current asthma, and had 18.8% any respiratory infection in the past 3 months. Malay students had more dampness or mould (p < 0.001), more environmental tobacco smoke (ETS) (p < 0.001) and more cats (p < 0.001) at home as compared to Chinese or Indian students. Wheeze was associated with ethnicity (p = 0.02; lower in Indian), atopy (p = 0.002), current smoking (p = 0.02) and recent indoor painting at home (p = 0.03). Current asthma was associated with ethnicity (p = 0.001; lower in Chinese) and para-dichlorobenzene in classroom air (p = 0.008). Respiratory infections were related to atopy (p = 0.002), ethylbenzene (p = 0.02) and para-dichlorobenzene (p = 0.01) in classroom air. Para-dichlorobenzene is used in Asia against insects. In conclusion, chemical emissions from recent indoor painting at home can increase the risk of wheeze. In schools, para-dichlorobenzene can increase the risk of current asthma and respiratory infections while ethylbenzene can increase the risk of respiratory infections., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Introduction: Resistance to corticosteroid is an essential mechanism in uncontrolled asthma as the corticosteroid is the mainstay of therapy. There are recent reports that epigenetic factors play a crucial role in the regulation of steroid action. Overexpression of P glycoprotein (P-gp) and reduced expression of Histone Deacetylase 2 (HDAC2) have been linked to regulating the steroid action in other diseases like Nephrotic Syndrome (NS). However, their role in uncontrolled asthma is still not clear and warrants further investigation. We evaluated the expression and activity of P-gp and HDAC2 in patients with Controlled Asthma (CA) and Uncontrolled Asthma (UA)., Methods: A total of 60 CA (mean age 51.72±17.02 years, male=38), and 38 of UA (mean age=53.55±11.90 years, male=17) were recruited. The level of control was defined according to (Global Initiative for Asthma) GINA 2016 criteria. The mRNA expression of HDAC2 and P-gp was studied by quantitative real-time Polymerase Chain Reaction (PCR), the functional activity of P-gp was evaluated by a commercially available kit via flow cytometry, and HDAC2 enzymatic activity was measured by commercially available kit by Enzyme-Linked Immunosorbent Assay (ELISA)., Results: P-gp expression and the functionality were significantly higher in the UA group of patients as compared to the CA group of patients (p<0.005), moreover HDAC2 expression was significantly reduced in UA patients as compared to CA patients, (p<0.005). The enzymatic activity of HDAC2 was also significantly reduced in UA patients as compared to CA patients (p<0.005)., Conclusion: P-gp overexpression and HDAC2 under expression play an essential role in uncontrolled asthma by impairing the response to corticosteroid., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background : Almost one third of the world population suffers from allergic conditions. Respiratory symptoms are common in Malaysian children but there are few studies on fractional exhaled nitric oxide (FeNO), inclusive of field clinical test for asthma among children in Malaysia. The aim was to provide insight on factors related to level of FeNO among students in Terengganu, Malaysia. Methods : In total, 487 randomly selected students from eight secondary schools participated (13-14 years old). A Standardized questionnaire was used to obtained information on doctors' diagnosed asthma, current asthma and respiratory symptoms. FeNO measurement and skin prick test (SPT to common allergen) were conducted. Results : The geometric mean FeNO was 16.7 ppb. Totally, 38.4% of students had elevated FeNO level (>20 ppb) and 40.3% had had positive SPT to house dust mites allergens (HDM), Dermatophagoides pteronyssinus (Der p 1), Dermatophagoides farinae (Der f 1) or Felis domisticus (cat). Male gender, height, parental history of allergy, self-reported allergy, and atopy were associated with FeNO. In particular, a combination of sensitization to HDM or cat and elevated FeNO were associated with doctor-diagnosed asthma and self-reported allergy to food, pollen and cat. Conclusion : Asthma, respiratory symptoms and sensitization to HDM and cat are common among students and presence of elevated FeNO levels indicate ongoing airway inflammation.

Indoor microbial diversity and composition are suggested to affect the prevalence and severity of asthma by previous home microbiome studies, but no microbiome-health association study has been conducted in a school environment, especially in tropical countries. In this study, we collected floor dust and environmental characteristics from 21 classrooms, and health data related to asthma symptoms from 309 students, in junior high schools in Johor Bahru, Malaysia. The bacterial and fungal composition was characterized by sequencing 16s rRNA gene and internal transcribed spacer (ITS) region, and the absolute microbial concentration was quantified by qPCR. In total, 326 bacterial and 255 fungal genera were characterized. Five bacterial (Sphingobium, Rhodomicrobium, Shimwellia, Solirubrobacter, Pleurocapsa) and two fungal (Torulaspora and Leptosphaeriaceae) taxa were protective for asthma severity. Two bacterial taxa, Izhakiella and Robinsoniella, were positively associated with asthma severity. Several protective bacterial taxa including Rhodomicrobium, Shimwellia and Sphingobium have been reported as protective microbes in previous studies, whereas other taxa were first time reported. Environmental characteristics, such as age of building, size of textile curtain per room volume, occurrence of cockroaches, concentration of house dust mite allergens transferred from homes by the occupants, were involved in shaping the overall microbial community but not asthma-associated taxa; whereas visible dampness and mold, which did not change the overall microbial community for floor dust, was negatively associated with the concentration of protective bacteria Rhodomicrobium (β = -2.86, p = 0.021) of asthma. The result indicates complex interactions between microbes, environmental characteristics and asthma symptoms. Overall, this is the first indoor microbiome study to characterize the asthma-associated microbes and their environmental determinant in the tropical area, promoting the understanding of microbial exposure and respiratory health in this region., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

OBJECTIVE: To examine the associations between endotoxin and (1,3)-β-glucan concentrations in office dust and respiratory symptoms and airway inflammation among 695 office workers in Malaysia. METHODS: Health data were collected using a questionnaire, sensitisation testing and measurement of fractional exhaled nitric oxide (FeNO). Indoor temperature, relative air humidity (RH) and carbon dioxide (CO₂) were measured in the offices and settled dust was vacuumed and analysed for endotoxin and (1,3)-β-glucan concentrations. Associations were analysed by two level multiple logistic regression. RESULTS: Overall, 9.6% of the workers had doctor-diagnosed asthma, 15.5% had wheeze, 18.4% had daytime attacks of breathlessness and 25.8% had elevated FeNO (≥25 ppb). The median levels in office dust were 11.3 EU/mg endotoxin and 62.9 ng/g (1,3)-β-glucan. After adjusting for personal and home environment factors, endotoxin concentration in dust was associated with wheeze ( P = 0.02) and rhinoconjunctivitis ( P = 0.007). The amount of surface dust ( P = 0.04) and (1,3)-β-glucan concentration dust ( P = 0.03) were associated with elevated FeNO. CONCLUSION: Endotoxin in office dust could be a risk factor for wheeze and rhinoconjunctivitis among office workers in mechanically ventilated offices in a tropical country. The amount of dust and (1,3)-β-glucan (a marker of indoor mould exposure) were associated with Th2 driven airway inflammation.

Few health studies exist on dampness and mould in schools in the tropics. We studied associations between fraction of exhaled nitric oxide (FeNO), respiratory symptoms and airway infections among students and dampness and fungal DNA in schools in Malaysia. A total of 368 randomly selected students from 32 classrooms in 8 secondary schools in Penang, Malaysia, participated (58% participation rate). Information on current respiratory symptoms and the home environment was collected by a standardised questionnaire. FeNO was measured by NIOX MINO (50ml/min). The classrooms were inspected and dust was collected by vacuuming on special filters and was analysed for five fungal DNA sequences by quantitative PCR. Linear mixed models and 3-level multiple logistic regression (school, classroom, student) were applied adjusting for demographic data and the home environment. Totally 10.3% reported doctor's diagnosed asthma, 15.1% current wheeze, 12.4% current asthma, 37.3% daytime breathlessness, 10.2% nocturnal breathlessness, 38.9% airway infections and 15.5% had pollen or furry pet allergy. The geometric mean of FeNO was 19.9ppb and 45% had elevated FeNO (>20ppb). Boys had higher levels of FeNO. Chinese had less daytime breathlessness than Malay (OR=0.30: p<0.001). Indoor carbon dioxide levels were low (380-720ppm). Dampness was observed in 18% of the classrooms and was associated with respiratory infections (OR=3.70; 95% CI 1.14-12.1) and FeNO (p=0.04). Aspergillus versicolor DNA was detected in 67% of the classrooms. Higher numbers of Aspergillus versicolor DNA in classroom dust were associated with wheeze (p=0.006), current asthma (p=0.002), respiratory infections (p=0.005) and elevated FeNO levels (p=0.02). In conclusion, respiratory symptoms were common among the students and the high FeNO levels indicate ongoing airway inflammation. Building dampness and the mould Aspergillus versicolor in schools in Malaysia can be risk factors for impaired respiratory health among the students., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objective: There are few studies on fractional exhaled nitric oxide (FeNO) and respiratory symptoms among adults in tropical areas. The aim was to study associations between FeNO and selected personal factors, respiratory symptoms, allergies, office characteristics and indoor office exposures among office workers (n = 460) from a university in Malaysia., Methods: Information on health was collected by a questionnaire, skin prick test and FeNO measurement. Temperature, relative air humidity, carbon monoxide and carbon dioxide were measured in the offices. Settled dust was vacuumed in the offices and analyzed for endotoxin, (1,3)-β-glucan and house dust mites allergens, namely Dermatophagoides pteronyssinus (Der p 1) and Dermatophagoides farinae (Der f 1). Two-level linear mixed models and multiple logistic regression were used to analyze the associations., Results: One-fourth (25.9%) of the office workers had elevated FeNO level (≥ 25 ppb) and 61.5% had HDM, cat, seafood or pollen allergy. Male gender (p < 0.001), current smoking (p = 0.037), height (p < 0.001) and atopy (p < 0.001) were associated with FeNO. The amount of vacuumed dust was associated with FeNO among atopic subjects (p = 0.009). Asthma and rhinitis symptoms were associated with FeNO (p < 0.05), especially among atopic subjects. In particular, a combination of atopy and elevated FeNO were associated with doctor-diagnosed asthma (p < 0.001), rhinitis (p < 0.001) and airway symptoms last 12 months (p < 0.001)., Conclusion: Gender, smoking, height and atopy are important risk factors for elevated FeNO levels. A combination of allergy testing and FeNO measurement could be useful in respiratory illness epidemiology studies and patient investigations in tropical areas.

A prevalence study was conducted among office workers in Malaysia (N= 695). The aim of this study was to examine associations between asthma, airway symptoms, rhinitis and house dust mites (HDM) and cat allergy and HDM levels in office dust. Medical data was collected by a questionnaire. Skin prick tests were performed for HDM allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae) and cat allergen Felis domesticus. Indoor temperature and relative air humidity (RH) were measured in the offices and vacuumed dust samples were analyzed for HDM allergens. The prevalence of D. pteronyssinus, D. farinae and cat allergy were 50.3%, 49.0% and 25.5% respectively. Totally 9.6% had doctor-diagnosed asthma, 15.5% had current wheeze and 53.0% had current rhinitis. The Der p 1 (from D. pteronyssinus) and Der f 1 (from D. farinae) allergens levels in dust were 556 ng/g and 658 ng/g respectively. Statistical analysis was conducted by multilevel logistic regression, adjusting for age, gender, current smoking, HDM or cat allergy, home dampness and recent indoor painting at home. Office workers with HDM allergy had more wheeze (p= 0.035), any airway symptoms (p= 0.032), doctor-diagnosed asthma (p= 0.005), current asthma (p= 0.007), current rhinitis (p= 0.021) and rhinoconjuctivitis (p< 0.001). Cat allergy was associated with wheeze (p= 0.021), wheeze when not having a cold (p= 0.033), any airway symptoms (p= 0.034), doctor-diagnosed asthma (p= 0.010), current asthma (p= 0.020) and nasal allergy medication (p= 0.042). Der f 1 level in dust was associated with daytime breathlessness (p= 0.033) especially among those with HDM allergy. Der f 1 levels were correlated with indoor temperature (p< 0.001) and inversely correlated with RH (p< 0.001). In conclusion, HDM and cat allergies were common and independently associated with asthma, airway symptoms and rhinitis. Der f 1 allergen can be a risk factor for daytime breathlessness.

There are few studies on associations between respiratory health and allergens, fungal and bacterial compounds in schools in tropical countries. The aim was to study associations between respiratory symptoms in pupils and ethnicity, chemical microbial markers, allergens and fungal DNA in settled dust in schools in Malaysia. Totally 462 pupils (96%) from 8 randomly selected secondary schools in Johor Bahru, Malaysia, participated. Dust was vacuumed from 32 classrooms and analysed for levels of different types of endotoxin as 3-hydroxy fatty acids (3-OH), muramic acid, ergosterol, allergens and five fungal DNA sequences. Multiple logistic regression was applied. Totally 13.1% pupils reported doctor's diagnosed asthma, 10.3% wheeze and 21.1% pollen or pet allergy. Indian and Chinese children had less atopy and asthma than Malay. Carbon dioxide levels were low (380-690 ppm). No cat (Fel d1), dog (Can f 1) or horse allergens (Ecu cx) were detected. The levels of Bloomia tropicalis (Blo t), house dust mite allergens (Der p 1, Der f 1, Der m 1) and cockroach allergens (Per a 1 and Bla g 1) were low. There were positive associations between levels of Aspergillus versicolor DNA and daytime breathlessness, between C14 3-OH and respiratory infections and between ergosterol and doctors diagnosed asthma. There were negative (protective) associations between levels of C10 3-OH and wheeze, between C16 3-OH and day time and night time breathlessness, between cockroach allergens and doctors diagnosed asthma. Moreover there were negative associations between amount of fine dust, total endotoxin (LPS) and respiratory infections. In conclusion, endotoxin at school seems to be mainly protective for respiratory illness but different types of endotoxin could have different effects. Fungal contamination measured as ergosterol and Aspergillus versicolor DNA can be risk factors for respiratory illness. The ethnical differences for atopy and asthma deserve further attention.

While there is a large variation of prevalence of asthma symptoms worldwide, what we do know is that it is on the rise in developing countries. However, there are few studies on allergens, moulds and mycotoxin exposure in schools in tropical countries. The aims were to measure selected fungal DNA, furry pet allergens and mycotoxins in dust samples from schools in Malaysia and to study associations with pupils' respiratory health effects. Eight secondary schools and 32 classrooms in Johor Bahru, Malaysia were randomly selected. A questionnaire with standardized questions was used for health assessment in 15 randomly selected pupils from each class. The school buildings were inspected and both indoor and outdoor climate were measured. Dust samples were collected by cotton swabs and Petri dishes for fungal DNA, mycotoxins and allergens analysis. The participation rate was 96% (462/480 invited pupils), with a mean age of 14 yr (range 14-16). The pupils mostly reported daytime breathlessness (41%), parental asthma or allergy (22%), pollen or pet allergy (21%) and doctor-diagnosed asthma (13%) but rarely reported night-time breathlessness (7%), asthma in the last 12 months (3%), medication for asthma (4%) or smoking (5%). The inspection showed that no school had any mechanical ventilation system, but all classrooms had openable windows that were kept open during lectures. The mean building age was 16 yr (range 3-40) and the mean indoor and outdoor CO(2) levels were 492 ppm and 408 ppm, respectively. The mean values of indoor and outdoor temperature and relative humidity were the same, 29°C and 70% respectively. In cotton swab dust samples, the Geometric Mean (GM) value for total fungal DNA and Aspergillus/Penicillium (Asp/Pen) DNA in swab samples (Cell Equivalents (CE)/m(2)) was 5.7*10(8) and 0.5*10(8), respectively. The arithmetic mean (CE/m(2)) for Aspergillus versicolor DNA was 8780, Stachybotrys chartarum DNA was 26 and Streptomyces DNA was 893. The arithmetic means (pg/m(2)) for the mycotoxins sterigmatocystin and verrucarol were 2547 and 17, respectively. In Petri dish dust samples, the GM value for total fungal DNA and Asp/Pen DNA (CE/m(2) per day) was 9.2*10(6) and 1.6*10(6), respectively. The arithmetic mean (CE/m(2) per day) for A. versicolor DNA was 1478, S. chartarum DNA was 105 and Streptomyces DNA was 1271, respectively. The GM value for cat (Fel d1) allergen was 5.9 ng/m(2) per day. There were positive associations between A. versicolor DNA, wheeze and daytime breathlessness and between Streptomyces DNA and doctor-diagnosed asthma. However, the associations were inverse between S. chartarum DNA and daytime breathlessness and between verrucarol and daytime breathlessness. In conclusion, fungal DNA and cat allergen contamination were common in schools from Malaysia and there was a high prevalence of respiratory symptoms among pupils. Moreover, there were associations between levels of some fungal DNA and reported respiratory health in the pupils., (© 2010 John Wiley & Sons A/S.)