Assessment of Peripheral Blood Mononuclear Cells Senescence and Endothelial Dysfunction among Adults with High Cardiovascular Risk. / Avaliação da Senescência de Células Sanguíneas Mononucleares Periféricas e na Disfunção Endotelial entre Adultos com Alto Risco Cardiovascular.

BACKGROUND: Cardiovascular diseases (CVD) are one of the leading causes of mortality and morbidity worldwide. Biological aging has been associated with the occurrence of adverse cardiovascular outcomes; however, the underlying mechanism of this process remains unknown. OBJECTIVES: This study sought to evaluate if peripheral blood mononuclear cell (PBMC) senescence and endothelial biomarkers could influence cardiovascular (CV) risk and be suitable markers for the early detection of cardiovascular diseases in adults. METHODS: In this cross-sectional study patients free of CVD were classified as lower (n=32) and higher Interheart Risk (IHR) scores (n=28). PBMC senescence was assessed by estimating the telomerase activity (TA) and detecting the presence of senescent cells and endothelial dysfunction by estimating the concentration of nitrite and nitrate and of total antioxidant capacity (TAC). Statistical analysis was performed with SPSS version 16.0 (SPSS Inc., Chicago, IL). All p-values <0.05 were considered statistically significant. RESULTS: PBMC senescence 0.95 [p-value = 0.0001; 95% CI (0.874-1.026)] was a significant predictor of patients with higher IHR scores with a cut-off value of 21.65 with a sensitivity and specificity of 92% and 88% respectively. PBMC senescence, nitrite and nitrate and TA were found to be independently associated with high IHR scores. CONCLUSION: PBMC senescence, TA and nitrite, and nitrate status are suitable measures to predict high cardiovascular risk in adults with CV risk. Nevertheless, long-term follow-up studies are needed to confirm these findings. (Arq Bras Cardiol. 2021; 116(1):37-47).

FUNDAMENTO: Doenças cardiovasculares (DCV) são uma das principais causas de mortalidade e morbidade em todo o mundo. O envelhecimento biológico tem sido associado à ocorrência de resultados cardiovasculares. Entretanto, o mecanismo subjacente desse processo ainda é desconhecido. OBJETIVOS: Buscamos avaliar se a senescência das células sanguíneas mononucleares periféricas (CSMP) e biomarcadores endoteliais poderiam influenciar o risco cardiovascular (CV) e ser marcadores adequados para a detecção precoce de doenças cardiovasculares em adultos. MÉTODOS: Neste estudo transversal, pacientes livres de DCV foram classificados como baixo (n=32) e alto (n=28) escore de risco intracardaco (IHR) A senescência das CSMP foi avaliada estimando-se a atividade de telomerase (AT) e detectando-se a presença de células senescentes e disfunção endotelial, estimando-se a concentração de nitrito e nitrato e a capacidade antioxidante total (CAT). A análise estatística foi realizada com o software SPSS, versão 16.0 (SPSS Inc., Chicago, IL). Todos os p-valores <0,05 foram considerados estatisticamente significativos. RESULTADOS: A senescência de CSMP de 0,95 [p-valor = 0,0001; 95% IC (0,874-1,026)] foi um indicador significativo de pacientes com escore de IHR mais alto, com um valor de corte de 21,65, com sensibilidade e especificidade de 92% e 88% respectivamente. Identificou-se que a senescência de CSMP, nitrito e nitrato, e AT eram independentemente associadas a um escore de IHR alto. CONCLUSÃO: Os status de nitrito e nitrato e AT, e a senescência de CSMP são medidas adequadas para prever o alto risco cardiovascular em adultos com risco CV. Entretanto devem ser realizados estudos de acompanhamento de longo prazo para confirmar esses achados. (Arq Bras Cardiol. 2021; 116(1):37-47).

Avaliação da Senescência de Células Sanguíneas Mononucleares Periféricas e na Disfunção Endotelial entre Adultos com Alto Risco Cardiovascular / Assessment of Peripheral Blood Mononuclear Cells Senescence and Endothelial Dysfunction among Adults with High Cardiovascular Risk

Resumo Fundamento Doenças cardiovasculares (DCV) são uma das principais causas de mortalidade e morbidade em todo o mundo. O envelhecimento biológico tem sido associado à ocorrência de resultados cardiovasculares. Entretanto, o mecanismo subjacente desse processo ainda é desconhecido. Objetivos Buscamos avaliar se a senescência das células sanguíneas mononucleares periféricas (CSMP) e biomarcadores endoteliais poderiam influenciar o risco cardiovascular (CV) e ser marcadores adequados para a detecção precoce de doenças cardiovasculares em adultos. Métodos Neste estudo transversal, pacientes livres de DCV foram classificados como baixo (n=32) e alto (n=28) escore de risco intracardaco (IHR) A senescência das CSMP foi avaliada estimando-se a atividade de telomerase (AT) e detectando-se a presença de células senescentes e disfunção endotelial, estimando-se a concentração de nitrito e nitrato e a capacidade antioxidante total (CAT). A análise estatística foi realizada com o software SPSS, versão 16.0 (SPSS Inc., Chicago, IL). Todos os p-valores <0,05 foram considerados estatisticamente significativos. Resultados A senescência de CSMP de 0,95 [p-valor = 0,0001; 95% IC (0,874-1,026)] foi um indicador significativo de pacientes com escore de IHR mais alto, com um valor de corte de 21,65, com sensibilidade e especificidade de 92% e 88% respectivamente. Identificou-se que a senescência de CSMP, nitrito e nitrato, e AT eram independentemente associadas a um escore de IHR alto. Conclusão Os status de nitrito e nitrato e AT, e a senescência de CSMP são medidas adequadas para prever o alto risco cardiovascular em adultos com risco CV. Entretanto devem ser realizados estudos de acompanhamento de longo prazo para confirmar esses achados. (Arq Bras Cardiol. 2021; 116(1):37-47)

Abstract Background Cardiovascular diseases (CVD) are one of the leading causes of mortality and morbidity worldwide. Biological aging has been associated with the occurrence of adverse cardiovascular outcomes; however, the underlying mechanism of this process remains unknown. Objectives This study sought to evaluate if peripheral blood mononuclear cell (PBMC) senescence and endothelial biomarkers could influence cardiovascular (CV) risk and be suitable markers for the early detection of cardiovascular diseases in adults. Methods In this cross-sectional study patients free of CVD were classified as lower (n=32) and higher Interheart Risk (IHR) scores (n=28). PBMC senescence was assessed by estimating the telomerase activity (TA) and detecting the presence of senescent cells and endothelial dysfunction by estimating the concentration of nitrite and nitrate and of total antioxidant capacity (TAC). Statistical analysis was performed with SPSS version 16.0 (SPSS Inc., Chicago, IL). All p-values <0.05 were considered statistically significant. Results PBMC senescence 0.95 [p-value = 0.0001; 95% CI (0.874-1.026)] was a significant predictor of patients with higher IHR scores with a cut-off value of 21.65 with a sensitivity and specificity of 92% and 88% respectively. PBMC senescence, nitrite and nitrate and TA were found to be independently associated with high IHR scores. Conclusion PBMC senescence, TA and nitrite, and nitrate status are suitable measures to predict high cardiovascular risk in adults with CV risk. Nevertheless, long-term follow-up studies are needed to confirm these findings. (Arq Bras Cardiol. 2021; 116(1):37-47)

Pharmacologic downregulation of protein arginine methyltransferase1 expression by adenosine dialdehyde increases cell senescence in breast cancer.

We investigated the role of protein arginine methylation (PAM) in estrogen receptor (ER)-positive breast cancer cells through pharmacological intervention. Tamoxifen (TAM) or adenosine dialdehyde (ADOX), independently, triggered cell cycle arrest and down-regulated PAM, as reduced protein arginine methyltransferase1 (PRMT1) mRNA and asymmetric dimethylarginine (ADMA) levels. Synergistic effect of these compounds elicited potent anti-cancer effect. However, reduction in ADMA was not proportionate with the compound-induced down-regulation of PRMT1 mRNA. We hypothesized that the disproportionate effect is due to the influence of the compounds on other methyltransferases, which catalyze the arginine dimethylation reaction and the diversity in the degree of drug-protein interaction among these methyltransferases. In silico analyses revealed that independently, ADOX or TAM, binds with phosphatidylethanolamine-methyltransferase (PEMT) or betaine homocysteine-methyl transferase (BHMT); and that the binding affinity of ADOX with PEMT or BHMT is prominent than TAM. These observations suggest that in breast cancer, synergistic effect of ADOX + TAM elicits impressive protective function by regulating PAM; and plausibly, restoration of normal enzyme activities of methyltransferases catalyzing arginine dimethylation could have clinical benefits.

Peroxiredoxin of Arthrospira platensis derived short molecule YT12 influences antioxidant and anticancer activity.

This study demonstrates both the antioxidant and anticancer potential of the novel short molecule YT12 derived from peroxiredoxin (Prx) of spirulina, Arthrospira platensis (Ap). ApPrx showed significant reduction in reactive oxygen species (ROS) against hydrogen peroxide (H2 O2 ) stress. The complementary DNA sequence of ApPrx contained 706 nucleotides and its coding region possessed 546 nucleotides between position 115 and 660. Real-time quantitative reverse transcription polymerase chain reaction analysis confirmed the messenger RNA expression of ApPrx due to H2 O2 exposure in spirulina cells at regular intervals, in which the highest expression was noticed on Day 20. Cytotoxicity assay was performed using human peripheral blood mononuclear cells, and revealed that at 10 µM, the YT12 did not exhibit any notable toxicity. Furthermore, ROS scavenging activity of YT12 was performed using DCF-DA assay, in which YT12 scavenged a significant amount of ROS at 25 µM in H2 O2 -treated blood leukocytes. The intracellular ROS in human colon adenocarcinoma cells (HT-29) was regulated by oxidative stress, where the YT12 scavenges ROS in HT-29 cells at 12.5 µM. Findings show that YT12 peptide has anticancer activity, when treated against HT-29 cells. Through the MTT assay, YT12 showed vital cytotoxicity against HT-29 cells. These finding suggested that YT12 is a potent antioxidant molecule which defends ROS against oxidative stress and plays a role in redox balance.

Arthrospira platensis transglutaminase derived antioxidant peptide-packed electrospun chitosan/ poly (vinyl alcohol) nanofibrous mat accelerates wound healing, in vitro, via inducing mouse embryonic fibroblast proliferation.

In this present study, we have carried out the antioxidant function of transglutaminase (TG) identified from Arthrospira platensis (Ap) transcriptome. The antioxidant peptide ML11 (MLRSIGIPARL) has been predicted from the transglutaminase core domain and the peptide's free radical scavenging potential was evaluated and it shows that it functions on a dose dependent manner. The ML11 peptide cell toxicity was analysed in the human blood leucocytes which resulted no cytotoxic activity in any of the cell population. Moreover, the nanofibre mat encapsulated with antioxidant peptide ML11 was prepared by electrospinning technique. The antioxidant peptide ML11 encapsulated mat showed increase in fibre diameter compared to the chitosan polyvinyl alcohol blended mat. The change in the crystalline behaviour of both chitosan and polyvinyl alcohol polymer to the amorphous nature was determined by X-ray diffraction at the broad band between 20 and 30° (2θ°). FTIR revealed the functional groups which present in the polymer as well as the interaction between their components of chitosan (CS) and polyvinyl alcohol (PVA). The fibre retains the antioxidant activity due to the peptide encapsulated by scavenging the intracellular ROS that was confirmed by flowcytometry and fluorescence microscopy. The ML11 peptide encapsulated mat showed no cytotoxicity in the NIH-3T3 mouse embryonic fibroblast cells. Also, ML11 peptide encapsulated fibre showed potential wound healing activity in NIH-3T3 cells. Taken altogether, the study indicates that the wound healing potential of the ML11 peptide encapsulated nano fibre mat may be used as biopharmaceutical drug.

Design and characterization of a novel Arthrospira platensis glutathione oxido-reductase-derived antioxidant peptide GM15 and its potent anti-cancer activity via caspase-9 mediated apoptosis in oral cancer cells.

Glutathione oxido-reductase (GR) is a primary antioxidant enzyme of most living forms which protects the cells from oxidative metabolism by reducing glutathione (GSH) from its oxidized form (GSSG). Although the antioxidant role of the enzyme is well characterized, the specific role of conserved N' peptide sequence in antioxidant mechanism remains unclear. In this study, we have identified an RNA sequence encoding GR enzyme from spirulina, Arthrospira platensis (Ap) and the changes in its gene expression profile was analysed during H2O2 stress. Results showed that H2O2 (10 mM) stimulated the expression of ApGR throughout the timeline of study (0, 5, 10, 15 and 20 days) with highest expression at 5th day post-exposure which confirmed the antioxidant role of ApGR in spirulina during H2O2 induced oxidative stress. A dithiol containing short antioxidant peptide, 39GGTCVIRGCVPKKLM53 (GM15) from ApGR was predicted and its radicals (superoxide and hydroxyl radical) scavenging potential was confirmed by in vitro cell-free assays. GM15 (12.5 µM) reduced the intracellular generalized oxidative stress level, as measured using DCFDA assay in H2O2 exposed leucocytes without affecting any of the cellular population. Further, the biomedical application of the radical scavenging property of GM15 was validated in oral carcinoma (KB) cells where GM15 exhibited significant cytotoxicity. Also, GM15 exhibited heterogenous effects on intracellular oxidative stress level in KB cells: at lower concentration (6.25 µM), the peptide reduced oxidative stress whereas, at higher concentration (25 µM) it increased the intensity of oxidative stress. GM15 (25 µM) induced caspase-9 mediated apoptosis in KB cells along with membrane disruption and DNA degradation which are confirmed by propidium iodide (PI) internalization and comet assays, respectively. Overall, the study shows that GM15 peptide i) scavenges superoxide, hydroxyl radicals, and influences intracellular oxidative stress, and ii) has anti-cancer effect in oral cancer cells.

Pharmacological inhibition of guanosine triphosphate cyclohydrolase1 elevates tyrosine phosphorylation of caveolin1 and cellular senescence.

The role of 2,4-diamino-6-hydroxypyrimidine (DAHP), on cellular-senescence remains unclear as differential effects of DAHP have been reported in cardiovascular and cerebrovascular systems. We investigated the effect of pharmacologically-induced guanosine-triphosphate-cyclohydrolase1 (GTPCH1)-inhibition, through DAHP, on cellular-senescence in experimentally-induced diabetic and non-diabetic Wistar rats. Cellular-senescence was evaluated through senescence-associated events, namely, cell-cycle-arrest of peripheral blood mononuclear cells (PBMNCs); myocardial DNA fragmentation, total antioxidant capacity (TAC), telomerase-activity, nicotinamide adenine dinucleotide (NAD+)-content and tyrosine14-phosphorylation of caveolin1 (pY14) in similarly-aged, pubertal Wistar rats with streptozotocin (STZ) and/or DAHP. Oxidative stress (OS) indices such as myocardial biopterin concentrations (tetrahydrobiopterin-BH4 and dihydrobiopterin-BH2) and plasma total nitrite and nitrate (NOx) were determined. DAHP, per se, exhibited distinct senescence; in addition, in STZ+DAHP (the cardiomyopathy model), there was a marked accumulation of cells in G0G1 phase, as evidenced through flow-cytometry analysis, as-well-as fragmented DNA, than the respective controls suggesting the DAHP-mediated onset of senescence in circulating cells and the myocardium, with or without STZ. Concentrations of BH4 and BH2, and NOx were impaired in STZ and/or DAHP, indicating elevated OS in the treatment groups. In the independent treatment groups or the combination treatment, typical senescence indicators including myocardial telomerase-activity, NAD+-content and TAC were significantly reduced, while there was a marked elevation in the concentrations of pY14 as compared to the respective controls, reinforcing the occurrence of senescence in PBMNCs and the myocardium. We postulate that DAHP promotes early onset of cellular-senescence, potentially through OS-mediated cellular events in diabetic or non-diabetic models.

Forskolin attenuates doxorubicin-induced accumulation of asymmetric dimethylarginine and s-adenosylhomocysteine via methyltransferase activity in leukemic monocytes.

Doxorubicin (DOX) is an antitumor drug, associated with cardiomyopathy. Strategies to address DOX-cardiomyopathy are scarce. Here, we identify the effect of forskolin (FSK) on DOX-induced-asymmetric-dimethylarginine (ADMA) accumulation in monocytoid cells. DOX-challenge led to i) augmented cytotoxicity, reactive-oxygen-species (ROS) production and methyltransferase-enzyme-activity identified as ADMA and s-adenosylhomocysteine (SAH) accumulation (SAH-A). However, except cytotoxicity, other DOX effects were decreased by metformin and FSK. FSK, did not alter the DOX-induced cytotoxic effect, but, decreased SAH-A by >50% and a combination of three drugs restored physiological methyltransferase-enzyme-activity. Together, protective effect of FSK against DOX-induced SAH-A is associated with mitigated methyltransferase-activity, a one-of-a-kind report.

A mini review on immune role of chemokines and its receptors in snakehead murrel Channa striatus.

Chemokines are ubiquitous cytokine molecules involved in migration of cells during inflammation and normal physiological processes. Though the study on chemokines in mammalian species like humans have been extensively studied, characterization of chemokines in teleost fishes is still in the early stage. The present review provides an overview of chemokines and its receptors in a teleost fish, Channa striatus. C. striatus is an air breathing freshwater carnivore, which has enormous economic importance. This species is affected by an oomycete fungus, Aphanomyces invadans and a Gram negative bacteria Aeromonas hydrophila is known to cause secondary infection. These pathogens impose immune changes in the host organism, which in turn mounts several immune responses. Of these, the role of cytokines in the immune response is immense, due to their involvement in several activities of inflammation such as cell trafficking to the site of inflammation and antigen presentation. Given that importance, chemokines in fishes do have significant role in the immunological and other physiological functions of the organism, hence there is a need to understand the characteristics, activities and performace of these small molecules in details.

Polycystic ovarian syndrome-associated cardiovascular complications: An overview of the association between the biochemical markers and potential strategies for their prevention and elimination.

Polycystic ovarian syndrome (PCOS) is associated with multiple cardiovascular risk factors (CVRF) including endothelial dysfunction (ED) and presence of metabolic syndrome (MS). The probable reason suggested for elevated CVRF in PCOS is oxidative stress (OS), which is an integral factor in cardiometabolic complications (CMC) seen in PCOS women. The interrelated mechanisms by which CVRF instigate clinical manifestation plays a crucial role in identification of a strategy to treat different comorbidities in PCOS. The existing treatment for PCOS mostly focuses on management of individual disorders, however, therapeutic strategies or novel targets to address cardiovascular complications in PCOS deserve extensive analysis.

Caveolin1/protein arginine methyltransferase1/sirtuin1 axis as a potential target against endothelial dysfunction.

Endothelial dysfunction (ED), an established response to cardiovascular risk factors, is characterized by increased levels of soluble molecules secreted by endothelial cells (EC). Evidence suggest that ED is an independent predictor of cardiac events and that it is associated with a deficiency in production or bioavailability of nitric oxide (NO) and/or an imbalance in the relative contribution of endothelium-derived relaxing and contracting factors. ED can be reversed by treating cardiovascular risk factors, hence, beyond ambiguity, ED contributes to initiation and progression of atherosclerotic disease. Majority of cardiovascular risk factors act by a common pathway, oxidative stress (OS), characterized by an imbalance in bioavailability of NO and reactive oxygen species (ROS). Enhanced ROS, through several mechanisms, alters competence of EC that leads to ED, reducing its potential to maintain homeostasis and resulting in development of cardiovascular disease (CVD). Influential mechanisms that have been implicated in the development of ED include (i) presence of elevated levels of NOS inhibitor, asymmetric dimethylarginine (ADMA) due to augmented enzyme activity of protein arginine methyl transferase-1 (PRMT1); (ii) decrease in NO generation by endothelial nitric oxide synthase (eNOS) uncoupling, or by reaction of NO with free radicals and (iii) impaired post translational modification of protein (PTM) such as eNOS, caveolin-1 (cav1) and sirtuin-1 (SIRT1). However, the inter-related mechanisms that concur to developing ED is yet to be understood. The events that possibly overlay include OS-induced sequestration of SIRT1 to caveolae facilitating cav1-SIRT1 association; potential increase in lysine acetylation of enzymes such as eNOS and PRMT1 leading to enhanced ADMA formation; imbalance in acetylation-methylation ratio (AMR); diminished NO generation and ED. Here we review current literature from research showing interdependent association between cav1-PRMT1-SIRT1 to the outcomes of experimental and clinical research aiming to preserve endothelial function with gene- or pharmaco-therapy.

Effect of protein arginine methyltransferase-1 inhibition on hypoxia-induced vasoconstriction.

Hypoxia is defined as a decrease in the oxygen supply to a level below physiological levels which is insufficient to maintain cellular function, in the presence of unrestricted coronary inflow. It is one of the leading causes of global mortality and morbidity, due to its association with the pathology of cancer, cardiovascular disease and stroke. The common feature in these pathologies is the limitation of oxygen availability that participates in the development of these conditions. The pulmonary response to hypoxia, when hypoxia is localized, is hypoxic pulmonary vasoconstriction (HPV). HPV is a physiological and self-regulatory mechanism by which pulmonary capillary blood flow is automatically adjusted to alveolar ventilation for maintaining the optimal balance of ventilation and perfusion. In pathological conditions, HPV occurs as an acute episode during progressive critical illness or as a sustained response with vascular remodeling and pulmonary hypertension. Inspite of the hypoxia-induced shift in the redox status to a more oxidized state, the endothelium-dependent mediators of HPV that cause vasoconstrictor response to hypoxia include nitric oxide (NO), endothelin-1 and angiotensin-II. Indeed, in chronic hypoxia, due to the enhanced reactive oxygen species (ROS) generation, inhibition of endothelial nitric oxide synthase (eNOS) activity and reduced nitric oxide (NO) production there is an imbalance in the vasoconstriction-vasodilation status toward constriction. It is our hypothesis that, in hypoxic stress, a key player in initiating this imbalance is the enzyme, protein arginine methyltransferase-1 (PRMT1) which indirectly affects eNOS activity by increased production of asymmetric dimethylarginine (ADMA), a NOS-inhibitor. Thus, pharmacological inhibition of PRMT1 should restore the cellular and vascular homeostasis in hypoxic conditions.

Oxidative stress and cardiovascular complications in polycystic ovarian syndrome.

Polycystic ovarian syndrome (PCOS) is a complex endocrine condition which is associated with metabolic and cardiovascular complications. It is elevated to a metabolic disorder with significant long term health ramification due to the high prevalence of insulin resistance (IR), impaired glucose tolerance, type 2 diabetes (T2D), dyslipidemia and numerous cardiovascular risk factors in PCOS women. This article concentrates on the recent developments in the regulation of oxidative stress (OS) in PCOS and on the association between PCOS and CVD outcomes. The prognostic events that define the severity of PCOS and involvement of cardiovascular risk in PCOS include endothelial dysfunction (ED) and impaired cardiac structure. Fact is that, in PCOS women, the circulating biomarkers of OS are in abnormal levels that are independent of overweight, which depicts the participation of OS in the pathophysiology of this common derangement. In addition, hyperglycemia (HG) per se, promotes reactive oxygen species (ROS) generation in PCOS. When the destructive ROS outbalances the concentration of physiological antioxidants, OS occurs. The resultant OS, directly stimulates hyperandrogenism and causes extensive cellular injury, DNA damage and/or cell apoptosis. To further the burden, the total serum antioxidant level in PCOS women is compromised, which diminishes the body's defense against an oxidative milieu. Thus, it is evident that OS regulates several cellular mechanisms in PCOS. Improving our understanding about the regulation of OS, critical role of ROS and protein biomarkers in PCOS should lead to novel therapeutic strategies in addressing PCOS-induced CVD. Besides, it is possible that the beneficial effects of dietary or therapeutic antioxidants have significant clinical relevance in PCOS.

Bioactive proteins from Solanaceae as quorum sensing inhibitors against virulence in Pseudomonas aeruginosa.

Cell-to-cell communication or quorum sensing (QS) is a generic event in bacteria that is used to coordinate gene expression among local populations. The phenomenon of QS depends on the fact that presence of sufficient bacteria ascertains a threshold level of autoinducer concentration that allows bacteria to sense a critical cell mass and to activate or repress target genes. Thus, QS has been an attractive target for the development of anti-infective strategies that are not based on the use of antibiotics. Several anti-QS approaches have been demonstrated including natural products from plant-based secondary metabolites. However, the role of plant bioactive proteins as an anti-QS peptide is yet to be deciphered. Against a backdrop of ever-increasing antibiotic resistant pathogens, there is a strong need for development of alternative therapeutic strategies. Thus, our hypothesis is that bioactive proteins from the plant family Solanaceae are quorum quenching molecules that can be exploited to develop a therapeutic strategy against virulence. We presume that bioactive proteins will inactivate or inhibit or degrade QS signals from bacteria to prevent cell-to-cell communication and thus inhibit development of virulence in Pseudomonas aeruginosa. Further, the use of proteins as quorum quenchers will delay the bacteria to develop resistance against these quenching molecules.