New perspectives on 'Breathomics': metabolomic profiling of non-volatile organic compounds in exhaled breath using DI-FT-ICR-MS.

Breath analysis offers tremendous potential for diagnostic approaches, since it allows for easy and non-invasive sample collection. "Breathomics" as one major research field comprehensively analyses the metabolomic profile of exhaled breath providing insights into various (patho)physiological processes. Recent research, however, primarily focuses on volatile compounds. This is the first study that evaluates the non-volatile organic compounds (nVOCs) in breath following an untargeted metabolomic approach. Herein, we developed an innovative method utilizing a filter-based device for metabolite extraction. Breath samples of 101 healthy volunteers (female n = 50) were analysed using DI-FT-ICR-MS and biostatistically evaluated. The characterisation of the non-volatile core breathome identified more than 1100 metabolites including various amino acids, organic and fatty acids and conjugates thereof, carbohydrates as well as diverse hydrophilic and lipophilic nVOCs. The data shows gender-specific differences in metabolic patterns with 570 significant metabolites. Male and female metabolomic profiles of breath were distinguished by a random forest approach with an out-of-bag error of 0.0099. Additionally, the study examines how oral contraceptives and various lifestyle factors, like alcohol consumption, affect the non-volatile breathome. In conclusion, the successful application of a filter-based device combined with metabolomics-analyses delineate a non-volatile breathprint laying the foundation for discovering clinical biomarkers in exhaled breath.

A loss of function variant in AGPAT3 underlies intellectual disability and retinitis pigmentosa (IDRP) syndrome.

Intellectual disability (ID) and retinal dystrophy (RD) are the frequently found features of multiple syndromes involving additional systemic manifestations. Here, we studied a family with four members presenting severe ID and retinitis pigmentosa (RP). Using genome wide genotyping and exome sequencing, we identified a nonsense variant c.747 C > A (p.Tyr249Ter) in exon 7 of AGPAT3 which co-segregates with the disease phenotype. Western blot analysis of overexpressed WT and mutant AGPAT3 in HEK293T cells showed the absence of AGPAT3, suggesting instability of the truncated protein. Knockdown of Agpat3 in the embryonic mouse brain caused marked deficits in neuronal migration, strongly suggesting that reduced expression of AGPAT3 affects neuronal function. Altogether, our data indicates that AGPAT3 activity is essential for neuronal functioning and loss of its activity probably causes intellectual disability and retinitis pigmentosa (IDRP) syndrome.

Detection of SARS-CoV-2 RNA in exhaled breath and its potential for prevention measures.

Background: To propose infection prevention measures it is essential to understand the dynamics of SARS-CoV-2 shedding, particularly in asymptomatic patients. This report compares the viral load progression in exhaled breath (EB) with the symptom severity. We aim to evaluate the adequacy of symptom assessment regarding the infectivity level of individuals. Methods: We observed infected patients since their first positive test during hospitalization. EB samples were collected on days 1, 3, 5, 7, 10, 12 and 14 of hospitalization using a filter-based device. After extraction, viral loads were quantified with qRT-PCR. The infection trajectory was documented after symptom onset. Case Presentation and Discussion: A 34-year old patient showed mild symptoms, e.g. fever, cough, headache, muscle pain and loss of taste and smell across trajectory of infection (Case 1). The viral loads emitted via exhaling were nearly constant and ranged from 8.6 x 103 and 4.1 x 104 RNA copies per hour. After the infection, the patient developed a pneumonia. The second case of a 65-year old patient depicted an asymptomatic infection trajectory for 14 days after the first diagnosis (Case 2). Nevertheless, the patient exhaled up to 2 x 105 SARS-CoV-2 virus copies hourly, approximately 10 fold higher than measured for Case 1. Conclusion: Symptomatic and asymptomatic COVID-19 patients exhale distinctive amounts of SARS-CoV-2 not necessarily correlating with symptom severity. Particularly, asymptomatic patients might show higher EB viral shedding. Therefore, EB testing should be included in infection prevention measures as it has high potential to reveal the most infectious individuals regardless of their symptoms during infection.

Exome sequencing identifies novel and known mutations in families with intellectual disability.

BACKGROUND: Intellectual disability (ID) is a phenotypically and genetically heterogeneous disorder. METHODS: In this study, genome wide SNP microarray and whole exome sequencing are used for the variant identification in eight Pakistani families with ID. Beside ID, most of the affected individuals had speech delay, facial dysmorphism and impaired cognitive abilities. Repetitive behavior was observed in MRID143, while seizures were reported in affected individuals belonging to MRID137 and MRID175. RESULTS: In two families (MRID137b and MRID175), we identified variants in the genes CCS and ELFN1, which have not previously been reported to cause ID. In four families, variants were identified in ARX, C5orf42, GNE and METTL4. A copy number variation (CNV) was identified in IL1RAPL1 gene in MRID165. CONCLUSION: These findings expand the existing knowledge of variants and genes implicated in autosomal recessive and X linked ID.

SARS-CoV-2: Viral Loads of Exhaled Breath and Oronasopharyngeal Specimens in Hospitalized Patients with COVID-19.

BACKGROUND: SARS-CoV-2 seems mainly transmissible via respiratory droplets. We compared the time-dependent SARS-CoV-2 viral load in serial pharyngeal swab with exhaled breath (EB) samples of hospitalized COVID-19 patients. METHODS: In this prospective proof of concept study, we examined hospitalized patients who initially tested positive for SARS-CoV-2. Paired oronasopharyngeal swab and EB specimens were taken at different days of hospitalization. EB collection was performed through a simple, noninvasive method using an electret air filter-based device. SARS-CoV-2 RNA detection was determined with real-time quantitative reverse transcription polymerase chain reaction. RESULTS: Of 187 serial samples from 15 hospitalized patients, 87/87 oronasopharyngeal swabs and 70/100 EB specimens tested positive. Comparing the number of SARS-CoV-2 copies, the viral load of the oronasopharyngeal swabs was significantly higher (CI 99%, P<<0,001) than for EB samples. The mean viral load per swab was 7.97 × 106 (1.65 × 102-1.4 × 108), whereas EB samples showed 2.47 × 103 (7.19 × 101-2.94 × 104) copies per 20 times exhaling. Viral loads of paired oronasopharyngeal swab and EB samples showed no correlation. CONCLUSIONS: Assessing the infectiousness of COVID-19 patients merely through pharyngeal swabs might not be accurate. Exhaled breath could represent a more suitable matrix for evaluating infectiousness and might allow screening for superspreader individuals and widespread variants such as Delta.

Needle stick injuries in nurses at a tertiary health care facility.

BACKGROUND: Needle-stick injury (NSI) is a major occupational health and safety issue faced by healthcare professionals globally. This study was aimed to assess the frequency and factors associated with NSIs in nurses of a tertiary health care facility in Lahore, Pakistan. It also focuses on safety measures adopted by these nurses after a needle stick injury. METHODS: This cross-sectional descriptive study was conducted in Ghurki Trust Teaching Hospital, Lahore from October 2009 to January 2010. All nurses have participated in the study with a response rate of 99%. These responses were obtained via a pretested self-administered questionnaire. The data was analysed using SPSS-16. Percentages of the categorical variables were computed and represented in various statistical data presentation forms, for analysis and comparison. Chi-square test was applied as a test of significance with fixing the p-value of 0.05 as significant. RESULTS: Out of 77 nurses who participated in our study, only 33 (42%) nurses were aware of the occupational hazards of their profession when they joined nursing. Needle stick injury was reported by 40 (71.9%) of the nurses in last one year. About 17 (31.5%) were injured at the time of recapping the syringe. The availability of needle cutters in the hospital was reported by 75 (97.4%) nurses while only 46 (60%) of them had undertaken a sharp management training course. Approximately 50 (64.9%) nurses failed to use gloves while administering injections. After getting stuck by a contaminated needle 71 (92%) of the nurses cleaned the wound with a spirit swab, 67 (87%) washed the area with soap and water and 58 (75%) applied a readily available bandage. Only 38 (49%) went on to inform the higher officials about a needle stick injury. Fifty-seven (74%) of the nurses were vaccinated against HBV, and 56 (72.2%) of needle stick injured nurses proceeded for HBV screening, while 53 (68.6%) for HCV and 37(48.5%) for HIV. CONCLUSION: Needle stick injury is the most important occupational health hazard in nurses with alarmingly high rates. Reporting to the concerned authorities, screening of nurses after needle stick injury and promotion of safety measures against it should be greatly encouraged.