Benefits of fungal-to-bacterial quorum quenching as anti-biofouling strategy in membrane bioreactors for wastewater treatment and water reuse.

This study addresses membrane biofouling in membrane bioreactors (MBRs) by exploring fungal-to-bacterial quorum quenching (QQ) strategies. While most research has been focused on bacterial-to-bacterial QQ tactics, this study identified fungal strain Vanrija sp. MS1, which is capable of degrading N-acyl-homoserine lactones (signaling molecules of Gram-negative bacteria). To determine the benefits of fungal over bacterial strains, after immobilization on fluidizing spherical beads in an MBR, MS1 significantly reduced the fouling rate by 1.8-fold compared to control MBR, decreased extracellular polymeric substance levels in the biofilm during MBR operation, and favorably changed microbial community and bacterial network, resulting in biofouling mitigation. It is noteworthy that, unlike Rhodococcus sp. BH4, MS1 enhanced QQ activity when switching from neutral to acidic conditions. These results suggest that MS1 has the potential for the effective treatment of acidic industrial wastewater sources such as semiconductor and secondary battery wastewater using MBRs.

Comparative Analysis of the Gelsemium Alkaloids Metabolism in Human, Pig, Goat, and Rat Liver Microsomes.

AIM: The aim of this study was to investigate the metabolism of Gelsemium elegans in human, pig, goat and rat liver microsomes and to elucidate the metabolic pathways and cleavage patterns of the Gelsemium alkaloids among different species. METHODS: A human, goat, pig and rat liver microsomes were incubated in vitro. After incubating at 37°C for 1 hour and centrifuging, the processed samples were detected by HPLC/Qq-TOFMS was used to detect alcohol extract of Gelsemium elegans and its metabolites. RESULTS: Forty-six natural products were characterized from alcohol extract of Gelsemium elegans and 13 metabolites were identified. These 13 metabolites belong to the gelsemine, koumine, gelsedine, humantenine, yohimbane, and sarpagine classes of alkaloids. The metabolic pathways included oxidation, demethylation and dehydrogenation. After preliminary identification, the metabolites detected in the four species were different. All 13 metabolites were detected in pig and rat microsomes, but no oxidative metabolites of Gelsedine-type alkaloids were detected in goat and human microsomes. CONCLUSION: In this study, Gelsemium elegans metabolic patterns in different species are clarified and the in vitro metabolism of Gelsemium elegans is investigated. It is of great significance for its clinical development and rational application.

Molecular Aspects of the Functioning of Pathogenic Bacteria Biofilm Based on Quorum Sensing (QS) Signal-Response System and Innovative Non-Antibiotic Strategies for Their Elimination.

One of the key mechanisms enabling bacterial cells to create biofilms and regulate crucial life functions in a global and highly synchronized way is a bacterial communication system called quorum sensing (QS). QS is a bacterial cell-to-cell communication process that depends on the bacterial population density and is mediated by small signalling molecules called autoinducers (AIs). In bacteria, QS controls the biofilm formation through the global regulation of gene expression involved in the extracellular polymeric matrix (EPS) synthesis, virulence factor production, stress tolerance and metabolic adaptation. Forming biofilm is one of the crucial mechanisms of bacterial antimicrobial resistance (AMR). A common feature of human pathogens is the ability to form biofilm, which poses a serious medical issue due to their high susceptibility to traditional antibiotics. Because QS is associated with virulence and biofilm formation, there is a belief that inhibition of QS activity called quorum quenching (QQ) may provide alternative therapeutic methods for treating microbial infections. This review summarises recent progress in biofilm research, focusing on the mechanisms by which biofilms, especially those formed by pathogenic bacteria, become resistant to antibiotic treatment. Subsequently, a potential alternative approach to QS inhibition highlighting innovative non-antibiotic strategies to control AMR and biofilm formation of pathogenic bacteria has been discussed.

Optimum quorum quenching bacteria concentration in the better-quality cell entrapping beads to control biofouling in membrane bioreactor.

Quorum quenching (QQ) by cell entrapping beads (CEBs) is known to inhibit biofouling by its biological and physical cleaning effect. Although there are better QQ media reported, due to the ease of fabrication of QQ-CEBs, this study focused on improving the quality of CEBs by comparing two distinct bead-making methods - polyvinyl alcohol-alginate (PVA-alginate) and phase inversion - and on finding the optimum concentration of QQ bacteria in the CEBs. The evaluation of PVA-alginate bead showed better uniformity, and higher mechanical and chemical strength in comparison with the phase inversion bead. Through the operations of two control membrane bioreactors (MBRs) (no bead, vacant bead) and four QQ-MBRs with different Rhodococcus sp. BH4 concentrations (2.5-15 mg cell ml-1) in PVA-alginate CEBs, the maximum QQ effect was observed by 5 mg ml-1 BH4 concentration beads. This implies that an optimum cell concentration of QQ-CEBs is crucial to economically improve MBR performance using QQ.

Multi-Echo Complex Quantitative Susceptibility Mapping and Quantitative Blood Oxygen Level-Dependent Magnitude (mcQSM + qBOLD or mcQQ) for Oxygen Extraction Fraction (OEF) Mapping.

Oxygen extraction fraction (OEF), the fraction of oxygen that tissue extracts from blood, is an essential biomarker used to directly assess tissue viability and function in neurologic disorders. In ischemic stroke, for example, increased OEF can indicate the presence of penumbra-tissue with low perfusion yet intact cellular integrity-making it a primary therapeutic target. However, practical OEF mapping methods are not currently available in clinical settings, owing to the impractical data acquisitions in positron emission tomography (PET) and the limitations of existing MRI techniques. Recently, a novel MRI-based OEF mapping technique, termed QQ, was proposed. It shows high potential for clinical use by utilizing a routine sequence and removing the need for impractical multiple gas inhalations. However, QQ relies on the assumptions of Gaussian noise in susceptibility and multi-echo gradient echo (mGRE) magnitude signals for OEF estimation. This assumption is unreliable in low signal-to-noise ratio (SNR) regions like disease-related lesions, risking inaccurate OEF estimation and potentially impacting clinical decisions. Addressing this, our study presents a novel multi-echo complex QQ (mcQQ) that models realistic Gaussian noise in mGRE complex signals. We implemented mcQQ using a deep learning framework (mcQQ-NET) and compared it with the existing QQ-NET in simulations, ischemic stroke patients, and healthy subjects, using identical training and testing datasets and schemes. In simulations, mcQQ-NET provided more accurate OEF than QQ-NET. In the subacute stroke patients, mcQQ-NET showed a lower average OEF ratio in lesions relative to unaffected contralateral normal tissue than QQ-NET. In the healthy subjects, mcQQ-NET provided uniform OEF maps, similar to QQ-NET, but without unrealistically high OEF outliers in areas of low SNR, such as SNR ≤ 15 (dB). Therefore, mcQQ-NET improves OEF accuracy by more accurately reflecting realistic Gaussian noise in complex mGRE signals. Its enhanced sensitivity to OEF abnormalities, based on more realistic biophysics modeling, suggests that mcQQ-NET has potential for investigating tissue variability in neurologic disorders.

Oxygen extraction fraction (OEF) values and applications in neurological diseases.

One of the goals of this systematic review is to provide a meta-analysis-derived mean OEF of healthy volunteers. Another aim of this study is to indicate the OEF ranges of various neurological pathologies. Potential clinical applications of OEF metrics are presented. Peer-reviewed studies reporting OEF metrics derived from computed tomography (CT)/positron emission tomography (PET) and/or magnetic resonance imaging (MRI) were considered. Databases utilized included MEDLINE, PubMed, EMBASE, Web of Science, and Google Scholar. The Newcastle-Ottawa scoring system was used for evaluating studies. R Studio was utilized for the meta-analysis calculations when appropriate. The GRADE framework was utilized to assess additional findings. Of 2267 potential studies, 165 met the inclusion criteria. The healthy volunteer meta-analysis included 339 subjects and found a mean OEF value of 38.87 (37.38, 40.36), with a prediction interval of 32.40-45.34. There were no statistical differences in OEF values derived from PET versus MRI. We provided a GRADE A certainty rating for the use of OEF metrics to predict stroke occurrence in patients with symptomatic carotid or cerebral vessel disease. We provided a GRADE B certainty rating for monitoring treatment response in Moyamoya disease. Use of OEF metrics in diagnosing and/or monitoring other conditions had a GRADE C certainty rating or less. OEF might have a role in diagnosing and monitoring patients with symptomatic carotid or cerebral vessel disease and Moyamoya disease. While we found insufficient evidence to support measuring OEF metrics in other patient populations, in many cases, further studies are warranted.

Reducing the virulence of Pseudomonas aeruginosa by using multiple quorum-quenching enzymes.

The emergence of multidrug-resistant Pseudomonas aeruginosa in healthcare settings poses a tremendous challenge to traditional antibiotic therapy. Pseudomonas aeruginosa utilizes quorum sensing (QS) to coordinate the production of virulence factors and the formation of drug-resistant biofilms. QS is mediated by signal compounds produced by P. aeruginosa as well as signal molecules produced by other non-pseudomonad bacteria. A potential strategy to prevent bacterial pathogenicity is utilizing enzymes to interfere with QS. Here, we used AidC, a quorum-quenching (QQ) enzyme from Chryseobacterium sp. strain StRB126 that can effectively hydrolyze N-(3-oxododecanoyl) homoserine lactone (3OC12-HSL) and N-butanoyl-homoserine lactone (C4-HSL), the major signal molecules synthesized by P. aeruginosa. The exogenous addition of AidC to P. aeruginosa wild-type strain PAO1 cultures significantly reduced the total protease and elastase activities and the production of pyocyanin. In addition, the application of AidC resulted in thin and sparse biofilm formation. Later, we used a metagenomic-derived QQ enzyme, QQ-2, in combination with AidC to attenuate PAO1 virulence when the presence of a non-pseudomonad signal compound, autoinducer-2, aggravated it. These findings suggest that using a combined antimicrobial approach may lead to a more efficacious therapeutic intervention against P. aeruginosa PAO1 infection, as its behavior is modulated in the presence of intraspecies and interspecies signal compounds. ONE-SENTENCE SUMMARY: In this work, the potential of dual enzymes was investigated to interfere with quorum sensing as a novel concept for reducing the virulence of P. aeruginosa, which is influenced by both intra species and interspecies communication.

Rapid Web-Based Recruitment of Patients With Psoriasis: Multinational Cohort Study.

BACKGROUND: Wide-ranging patient recruitment not restricted to the location of the investigator will provide a better representation of the patient population in clinical studies. OBJECTIVE: Our goal was to assess the feasibility of a broad web-based recruitment strategy in an 8-week observational study of 500 study participants with psoriasis and healthy controls from locations remote from the investigator and to assess the cost associated with each participant. METHODS: A decentralized team in Denmark recruited patients with psoriasis and healthy controls using Google and Facebook advertisements and posts to Facebook groups. All individuals were screened via the internet, and patients diagnosed with psoriasis were included. Questionnaires regarding itch and sleep were completed by both groups at inclusion, week 4, and week 8. RESULTS: During a 2-week recruitment period, 12,887 unique advertisement views were registered, and 839 participants were enrolled, of which 507 completed the study (220 with psoriasis and 287 healthy controls) with a retention rate of 60.4%. Participants were recruited from 11 different countries on 4 separate continents, mainly from the United States, Canada, and the United Kingdom. The recruitment rate was 59.9 participants per day, and the conversion rate was 57.2%. Recruitment costs were US $13 per enrolled participant and US $22 per participant completing the study. CONCLUSIONS: It is feasible and rapid to recruit a large number of participants from locations different from the investigator and to retain patients in an observational study with no visits to a clinical site at low costs.

Application of Equal Local Levels to Improve Q-Q Plot Testing Bands with R Package qqconf.

Quantile-Quantile (Q-Q) plots are often difficult to interpret because it is unclear how large the deviation from the theoretical distribution must be to indicate a lack of fit. Most Q-Q plots could benefit from the addition of meaningful global testing bands, but the use of such bands unfortunately remains rare because of the drawbacks of current approaches and packages. These drawbacks include incorrect global Type I error rate, lack of power to detect deviations in the tails of the distribution, relatively slow computation for large data sets, and limited applicability. To solve these problems, we apply the equal local levels global testing method, which we have implemented in the R Package qqconf, a versatile tool to create Q-Q plots and probability-probability (P-P) plots in a wide variety of settings, with simultaneous testing bands rapidly created using recently-developed algorithms. qqconf can easily be used to add global testing bands to Q-Q plots made by other packages. In addition to being quick to compute, these bands have a variety of desirable properties, including accurate global levels, equal sensitivity to deviations in all parts of the null distribution (including the tails), and applicability to a range of null distributions. We illustrate the use of qqconf in several applications: assessing normality of residuals from regression, assessing accuracy of p values, and use of Q-Q plots in genome-wide association studies.

A method establishment and application for biofilm quorum quenching activity assay.

The existence of quorum sensing (QS) and quorum quenching (QQ) plays important roles in biofilm formation. However, direct detection of QS ability is difficult due to the low concentrations of signal molecules inside the biofilm. Therefore, QQ activity is typically used to indicate the attribution of QS/QQ to the biofilm. Nevertheless, current detection methods of QQ activity based on biosensors present undesirable operability and accuracy. In this study, the 96-well plate assay based on a specific biosensor, Agrobacterium tumefaciens A136, and a colorimetric substance, X-gal was established. The reliable fitting results were obtained by standardizing the composition of the A136 X-gal assay solution and optimizing the operating conditions. This method improved the accuracy of QQ activity detection and reduced time and cost consumption. Finally, the 96-well plate assay was successfully applied to detect the QQ activities of biofilm samples and explore possible environmental influencing factors. In general, this study provided a new strategy for understanding the QQ effect in biofilm systems.

The normality assumption on between-study random effects was questionable in a considerable number of Cochrane meta-analyses.

BACKGROUND: Studies included in a meta-analysis are often heterogeneous. The traditional random-effects models assume their true effects to follow a normal distribution, while it is unclear if this critical assumption is practical. Violations of this between-study normality assumption could lead to problematic meta-analytical conclusions. We aimed to empirically examine if this assumption is valid in published meta-analyses. METHODS: In this cross-sectional study, we collected meta-analyses available in the Cochrane Library with at least 10 studies and with between-study variance estimates > 0. For each extracted meta-analysis, we performed the Shapiro-Wilk (SW) test to quantitatively assess the between-study normality assumption. For binary outcomes, we assessed between-study normality for odds ratios (ORs), relative risks (RRs), and risk differences (RDs). Subgroup analyses based on sample sizes and event rates were used to rule out the potential confounders. In addition, we obtained the quantile-quantile (Q-Q) plot of study-specific standardized residuals for visually assessing between-study normality. RESULTS: Based on 4234 eligible meta-analyses with binary outcomes and 3433 with non-binary outcomes, the proportion of meta-analyses that had statistically significant non-normality varied from 15.1 to 26.2%. RDs and non-binary outcomes led to more frequent non-normality issues than ORs and RRs. For binary outcomes, the between-study non-normality was more frequently found in meta-analyses with larger sample sizes and event rates away from 0 and 100%. The agreements of assessing the normality between two independent researchers based on Q-Q plots were fair or moderate. CONCLUSIONS: The between-study normality assumption is commonly violated in Cochrane meta-analyses. This assumption should be routinely assessed when performing a meta-analysis. When it may not hold, alternative meta-analysis methods that do not make this assumption should be considered.

Prolonging the Life Span of Membrane in Submerged MBR by the Application of Different Anti-Biofouling Techniques.

The membrane bioreactor (MBR) is an efficient technology for the treatment of municipal and industrial wastewater for the last two decades. It is a single stage process with smaller footprints and a higher removal efficiency of organic compounds compared with the conventional activated sludge process. However, the major drawback of the MBR is membrane biofouling which decreases the life span of the membrane and automatically increases the operational cost. This review is exploring different anti-biofouling techniques of the state-of-the-art, i.e., quorum quenching (QQ) and model-based approaches. The former is a relatively recent strategy used to mitigate biofouling. It disrupts the cell-to-cell communication of bacteria responsible for biofouling in the sludge. For example, the two strains of bacteria Rhodococcus sp. BH4 and Pseudomonas putida are very effective in the disruption of quorum sensing (QS). Thus, they are recognized as useful QQ bacteria. Furthermore, the model-based anti-fouling strategies are also very promising in preventing biofouling at very early stages of initialization. Nevertheless, biofouling is an extremely complex phenomenon and the influence of various parameters whether physical or biological on its development is not completely understood. Advancing digital technologies, combined with novel Big Data analytics and optimization techniques offer great opportunities for creating intelligent systems that can effectively address the challenges of MBR biofouling.

Patient experience of virtual urogynaecology services during Covid-19 pandemic.

INTRODUCTION AND HYPOTHESIS: Due to increasing burden on outpatient services, there is a drive from NHS policy makers to utilise virtual clinics to help curb unsustainable demand. During the COVID-19 pandemic, urogynaecology clinics were converted to telephone consultation (TC). We used this opportunity to evaluate patient perspective and identify which patients may be best suited to TC. METHODS: Postal questionnaires were sent to patients following urogynaecology TCs in May to June 2020. Clinical outcome data were obtained from electronic records. The survey combined three validated tools: QQ-10, Patient Enablement Index (PEI) and NHS Friends and Family Test (NHS-FFT). Qualitative and quantitative data were analysed. RESULTS: Of the 308 patients contacted, 165 responded (54%). Eighty-six percent of patients described their experience of TC as "very good" or "good" (NHS-FFT). Positive themes included convenience, thoroughness and feeling at ease in terms of communicating intimate symptoms. QQ-10 results demonstrated a mean value score of 77 and a mean burden score of 17 (range 0-100); 72% of patients "strongly" or "mostly" agreed to repeat TC. Following TC, 22% of patients were discharged, 72% required follow-up and 37% needed face-to-face (F2F) consultation. Post-operative patients and those with lower urinary tract symptoms benefited most, whereas many prolapse patients required F2F consultation. CONCLUSIONS: We report the largest qualitative and quantitative study of patient experience of TC in urogynaecology. TC is a convenient, acceptable and effective medium for conducting patient care. TC can support patients in communicating intimate symptoms with health professionals.

Biofilm formation in xenobiotic-degrading microorganisms.

The increased presence of xenobiotics affects living organisms and the environment at large on a global scale. Microbial degradation is effective for the removal of xenobiotics from the ecosystem. In natural habitats, biofilms are formed by single or multiple populations attached to biotic/abiotic surfaces and interfaces. The attachment of microbial cells to these surfaces is possible via the matrix of extracellular polymeric substances (EPSs). However, the molecular machinery underlying the development of biofilms differs depending on the microbial species. Biofilms act as biocatalysts and degrade xenobiotic compounds, thereby removing them from the environment. Quorum sensing (QS) helps with biofilm formation and is linked to the development of biofilms in natural contaminated sites. To date, scant information is available about the biofilm-mediated degradation of toxic chemicals from the environment. Therefore, we review novel insights into the impact of microbial biofilms in xenobiotic contamination remediation, the regulation of biofilms in contaminated sites, and the implications for large-scale xenobiotic compound treatment.

Quorum sensing unveils the sludge floccule-assisted stabilization of aerobic granules in granule-dominated sequencing batch reactors.

Floccules are another major form of microbial aggregates in aerobic granular sludge systems. Previous studies mainly attributed the persistence of floccules to their relatively faster nutrient uptake and higher growth rate over aerobic granules; however, they failed to unravel the underlying mechanism of the long-term coexistence of these two aggregates. In this work, the existence and function of the floccules in an aerobic granule-dominated sequencing batch reactor were investigated from the view of quorum sensing (QS) and quorum quenching (QQ). The results showed that though the floccules were closely associated with the granules in terms of similar community structures (including the QS- and QQ-related ones), they exhibited a relatively higher QQ-related activity but a lower QS-related activity. A compatible proportion of floccules might be helpful to maintain the QS-related activity and keep the granules stable. In addition, the structure difference was demonstrated to diversify the QS- and QQ-related activities of the floccules and the aerobic granules. These findings could broaden our understanding of the interactions between the coexistent floccules and granules in aerobic granule-dominated systems and would be instructive for the development of the aerobic granular sludge process.

Research Progress and Hopeful Strategies of Application of Quorum Sensing in Food, Agriculture and Nanomedicine.

Quorum sensing (QS) regulates the expression of several genes including motility, biofilm development, virulence expression, population density detection and plasmid conjugation. It is based on "autoinducers", small molecules that microorganisms produce and release in the extracellular milieu. The biochemistry of quorum sensing is widely discussed and numerous papers are available to scientists. The main purpose of this research is to understand how knowledge about this mechanism can be exploited for the benefit of humans and the environment. Here, we report the most promising studies on QS and their resulting applications in different fields of global interest: food, agriculture and nanomedicine.

The protective effects of NE 52-QQ57 against interleukin-33-induced inflammatory response in activated synovial mast cells.

Cytokines-mediated immunity is essential for the pathological development of rheumatoid arthritis (RA). Inhibition of signaling has suggested a potential remedial approach to RA. G protein-coupled receptor 4 (GPR4) has been proven to possess a broad range of physiological functions, but its function in synovial mast cells and RA is less reported. In this study, the protective effects of NE 52-QQ57, a GPR4 antagonist, against interleukin (IL)-33-challenged inflammatory response in activated synovial mast cells were investigated. We report that IL-33 amplified GPR4 expression in HMC-1 mast cells. The GPR4 antagonist NE 52-QQ57 alleviated IL-33-caused secretions of IL-17, interferon-γ, and tumor necrosis factor-α in HMC-1 mast cells. Furthermore, we note that NE 52-QQ57 reduced IL-33-induced expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9. Also, NE 52-QQ57 inhibited cyclooxygenase 2 and prostaglandin E2 expression in IL-33-challenged cells. Also, NE 52-QQ57 ameliorated IL-33-induced oxidative stress by reducing mitochondrial reactive oxygen species and 4-hydroxynonenal. Mechanistically, NE 52-QQ57 mitigated IL-33-induced activation of the p38/nuclear factor-κB signaling pathway. We conclude that targeting GPR4 might be a promising strategy for RA treatment.

Effect of Aeration Intensity on Performance of Lab-Scale Quorum-Quenching Membrane Bioreactor.

Biofouling is one of the main drawbacks of membrane bioreactors (MBRs). Among the different methods, the quorum-quenching (QQ) technique is a novel method as it delays biofilm formation on the membrane surface through disruption of bacterial cell-to-cell communication and thus effectively mitigates membrane biofouling. QQ bacteria require a certain concentration of dissolved oxygen to show their best activities. Despite the importance of the amount of aeration, there have not been enough studies on aeration condition utilizing the separate determination of pure QQ effect and physical cleaning effect. This research aimed to find the optimum aeration intensity by separation of the two effects from QQ and physical cleaning. Three bead type conditions (no bead, vacant bead, and QQ beads) at three aeration intensities (1.5, 2.5, and 3.5 L/min representing low, medium, and high aeration intensity) were applied. From the results, no QQ effect and small QQ effect were observed at low and high aeration, while the greatest QQ effect (48.2% of 737 h improvement) was observed at medium aeration. The best performance was observed at high aeration with QQ beads having a 1536 h operational duration (303% improvement compared to the no bead condition); however, this excellent performance was attributed more to the physical cleaning effect than to the QQ effect.

Layered Antibiofouling Composite Membrane for Quenching Bacterial Signaling.

Bacterial quorum quenching (QQ) media with various structures (e.g., bead, cylinder, hollow cylinder, and sheet), which impart biofouling mitigation in membrane bioreactors (MBRs), have been reported. However, there has been a continuous demand for membranes with QQ capability. Thus, herein, we report a novel double-layered membrane comprising an outer layer containing a QQ bacterium (BH4 strain) on the polysulfone hollow fiber membrane. The double-layered composite membrane significantly inhibits biofilm formation (i.e., the biofilm density decreases by ~58%), biopolymer accumulation (e.g., polysaccharide), and signal molecule concentration (which decreases by ~38%) on the membrane surface. The transmembrane pressure buildup to 50 kPa of the BH4-embedded membrane (17.8 h ± 1.1) is delayed by more than thrice (p < 0.05) of the control with no BH4 in the membrane's outer layer (5.5 h ± 0.8). This finding provides new insight into fabricating antibiofouling membranes with a self-regulating property against biofilm growth.

Estimating changes in air pollutant levels due to COVID-19 lockdown measures based on a business-as-usual prediction scenario using data mining models: A case-study for urban traffic sites in Spain.

In response to the COVID-19 pandemic, governments declared severe restrictions throughout 2020, presenting an unprecedented scenario of reduced anthropogenic emissions of air pollutants derived mainly from traffic sources. To analyze the effect of these restrictions derived from COVID-19 pandemic on air quality levels, relative changes in NO, NO2, O3, PM10 and PM2.5 concentrations were calculated at urban traffic sites in the most populated Spanish cities over different periods with distinct restrictions in 2020. In addition to the changes calculated with respect to the observed air pollutant levels of previous years (2013-2019), relative changes were also calculated using predicted pollutant levels for the different periods over 2020 on a business-as-usual scenario using Multiple Linear Regression (MLR) models with meteorological and seasonal predictors. MLR models were selected among different data mining techniques (MLR, Random Forest (RF), K-Nearest Neighbors (KNN)), based on their higher performance and accuracy obtained from a leave-one-year-out cross-validation scheme using 2013-2019 data. A q-q mapping post-correction was also applied in all cases in order to improve the reliability of the predictions to reproduce the observed distributions and extreme events. This approach allows us to estimate the relative changes in the studied air pollutants only due to COVID-19 restrictions. The results obtained from this approach show a decreasing pattern for NOx, with the largest reduction in the lockdown period above -50%, whereas the increase observed for O3 contrasts with the NOx patterns with a maximum increase of 23.9%. The slight reduction in PM10 (-4.1%) and PM2.5 levels (-2.3%) during lockdown indicates a lower relationship with traffic sources. The developed methodology represents a simple but robust framework for exploratory analysis and intervention detection in air quality studies.