PA Strain Research Articles

Self-healing, piezoresistive and temperature responsive behaviour of chitosan/polyacrylic acid dynamic hydrogels

Flexible electronics have introduced new challenges for efficient human–machine interactions. Hydrogels have emerged as prominent materials for electronic wearable applications due to their exceptional mechanical deformability and lightweight characteristics combined in some cases with conductive properties, and softness. Additionally, bio-interphases require multisensory response to stress, strain, temperature, and self-healing capacity. To mimic these properties, this work developed interpenetrated hydrogel networks composed of chitosan (CHI) and polyacrylic acid (PAA), combined with Fe (III) ions and varying amounts of NMBA (0–0.25 %), to achieve tailored conductivity (0.8–2.5 mS/cm), self-healing, self-standing and mechanical properties (E = 11.7–110 Pa and fracture strain = 64.9–1923 %) suitable for strain sensor applications. The results revealed a significant influence of the restrictive effect on the mobility of uncrosslinked chain segments, caused by Fe ions and NMBA, on the piezoresistance (GF 2.1–1.3) and self-healing capability of the gels.Interestingly, a transparent/turbid transition, driven by microphase separation that is characteristic of systems with high dynamic interactions, was encountered for the first time in these hydrogels. This transition was analyzed in relation to external temperature, water content, pH, and the influence of Fe ions and NMBA. The simultaneous sensitivity of these materials to temperature and pH, along with their piezoresistive and self-healing behaviour, can be highly valuable for multifunctional sensors in a wide range of applications.

Highly diverse dynamics of Pseudomonas aeruginosa colonization from initial detection in cystic fibrosis patients: A 7-year longitudinal genetic diversity study

In cystic fibrosis (CF), Pseudomonas aeruginosa (Pa) is a major pathogen that can persistently colonize patients. Genetic studies showed a high diversity of Pa, the success of widespread or ‘international’ clones and described epidemic clones in CF and Epidemic High-Risk (ERH) clones. Here, we characterized Pa genetic diversity over time after first colonization in CF patients, with the aim of accurately describing the dynamics of colonization in a context of scarce longitudinal studies including the first isolated Pa strain. Results represent the first genotyping data based on multilocus sequence typing (MLST) available for CF Pa in France. Forty-four CF patients with a first Pa colonization were included; 265 strains collected over 7 years in these patients were genotyped by multiplex rep-PCR, MLST, pulsed-field gel electrophoresis and/or whole genome sequencing. Forty-one sequence types were identified: 4 were unknown, 22 never previously reported for CF patients, and 6 corresponded to widespread clones colonizing 16 patients (36%). Unrelated strains were identified in 41 patients (93%). Twenty-six patients (59%) presented a recurrence during the study period. No specific clones were associated with transient, recurrent or persistent colonization. Our longitudinal study revealed that 9 of the 26 patients with recurrence (35%) harbored strains of different genotypes. Great genetic diversity was observed among initial Pa isolates excluding any cross-transmission. Persistent colonization may appear more complex than expected, imitating persistence, with successive colonization events by unrelated Pa.

617. Pharmacodynamics of Ceftolozane-Tazobactam (C/T) as Monotherapy and in Combination with Tobramycin or Fosfomycin Against P.aeruginosa with C/T MICs at or Above 4mg/L

Abstract Background Ceftolozane (C/T) is widely used in clinical practice to treat difficult and/or multidrug resistant Pseudomonas aeruginosa (Pa). However, clinical data is absent on the use of C/T therapy to treat Pa strains with C/T MICs ≥ 4mg/L, that is outside the wild type population. To address this, we simulated 7-day courses of C/T against 8 strains of Pa C/T MICs 4-64mg/L as monotherapy and in combination with tobramycin (T) or fosfomycin (F) using an in vitro pharmacokinetic model. Methods A single compartment dilutional in vitro model was used, volume central chamber 360mL, inoculum 106 CFU/mL. Differing drugs t½ were simulated using add back. Pa strain C/T MICs were 4-64mg/L, T 0.25-8mg/L and F 4-1024mg/L. C/T doses of 2-1g, 8h (Cmax 112-32mg/L, t½ 2.5-1h), T doses 7mg/Kg 24h (Cmax 22mg/L, t½ 2.5h) and F doses 4g 8h (Cmax 250mg/L, t½ 2.5h) were simulated for 7 days. Endpoints were changes in bacterial load (log CFU/ml) and population profiles (growth on C/T containing media at MICx4 and MICx8 concentrations). Results C/T alone resulted in rapid clearance of all strains after 4-12h exposure, with the antibacterial effect being poorest with strains with the highest C/T MICs. Regrowth occurred with 7/8 strains after 24-48h, counts reaching 6-8 logs by 7 days. Combination of C/T plus T increased the rate of initial clearance and suppressed regrowth with all strains. Combination of C/T plus F did not increase the rate of initial clearance except in the strains with the highest C/T MICs but resulted in suppression of regrowth with final bacterial counts of 3-4 logs at day 7. Changes in C/T population profiles occurred after 7 days exposure with growth on C/T MICx8 containing media in 6/8 strains exposed to C/T alone, but 0/8 strains exposed to C/T plus T and 1/8 strains exposed to C/T plus F. Conclusion Combinations of C/T plus T or F resulted in long term suppression of Pa bacterial load and minimised emergence of resistance in Pa strains with C/T MICs on or just above the epidemiological cut off value (MIC 4mg/L-64mg/L). Disclosures Alasdair P. MacGowan, MD, GSK: Grant/Research Support|InfectoPharm: Grant/Research Support|Merck: Advisor/Consultant|Shionogi: Advisor/Consultant|Venatorx: Advisor/Consultant.

The Effectiveness of Ultraviolet-C (UV-C) Irradiation on the Viability of Airborne Pseudomonas aeruginosa.

Pseudomonas aeruginosa (Pa) is the predominant bacterial pathogen in people with cystic fibrosis (CF) and can be transmitted by airborne droplet nuclei. Little is known about the ability of ultraviolet band C (UV-C) irradiation to inactivate Pa at doses and conditions relevant to implementation in indoor clinical settings. We assessed the effectiveness of UV-C (265 nm) at up to seven doses on the decay of nebulized Pa aerosols (clonal Pa strain) under a range of experimental conditions. Experiments were done in a 400 L rotating sampling drum. A six-stage Andersen cascade impactor was used to collect aerosols inside the drum and the particle size distribution was characterized by an optical particle counter. UV-C effectiveness was characterized relative to control tests (no UV-C) of the natural decay of Pa. We performed 112 tests in total across all experimental conditions. The addition of UV-C significantly increased the inactivation of Pa compared with natural decay alone at all but one of the UV-C doses assessed. UV-C doses from 246–1968 µW s/cm2 had an estimated effectiveness of approximately 50–90% for airborne Pa. The effectiveness of doses ≥984 µW s/cm2 were not significantly different from each other (p-values: 0.365 to ~1), consistent with a flattening of effectiveness at higher doses. Modelling showed that delivering the highest dose associated with significant improvement in effectiveness (984 µW s/cm2) to the upper air of three clinical rooms would lead to lower room doses from 37–49% of the 8 h occupational limit. Our results suggest that UV-C can expedite the inactivation of nebulized airborne Pa under controlled conditions, at levels that can be delivered safely in occupied settings. These findings need corroboration, but UV-C may have potential applications in locations where people with CF congregate, coupled with other indoor and administrative infection control measures.

Semi-Arid-Habitat-Adapted Plant-Growth-Promoting Rhizobacteria Allows Efficient Wheat Growth Promotion

Plant-growth-promoting rhizobacteria (PGPR) introduced into agricultural ecosystems positively affect agricultural production and constitute an ecological method for sustainable agriculture. The present study demonstrated the effects of two PGPR, Pantoea agglomerans strain Pa and Bacillus thuringiensis strain B25, on seed germination, on the plant growth of two durum wheat varieties, Bousselam and Boutaleb, and on the frequency of the cultivable beneficial bacterial community. The bacterial strains were used as seed primers (individually or in consortia) by coating them with carboxymethyl cellulose (CMC 1%). The effect of PGPR was negligible on germinative ability but improved seed vigor in the Boutaleb variety after inoculation with the Pa strain alone or in combination with the B25 strain. The results showed that the germination capacity depends on the wheat variety. It seemed to be better in the Bousselam variety. Analysis of the results of morphological plant parameters in sterile compost after 75 days under controlled conditions (16 h light, 26/16 °C day/night) showed a significant improvement in plant growth in both wheat varieties with the Pa strain alone or in combination. Chlorophyll (a, and total), carotenoid, and total soluble sugars were significantly increased, while proline and MDA were strongly reduced by inoculation of the Bousselam variety. Bacterial survival of the Pa and B25 strains in the rhizosphere of sterile compost was appreciable (105–107 CFU/g) for both the Pa and B25 strains. Only the Pa strain was endophytic and able to colonize roots. Contrary to sterile compost, the different inoculation treatments in natural soil (after 114 days) significantly improved all morphological parameters and chlorophyll pigments in both wheat varieties. The reduction of proline contents at the leaf level was observed with Pa, mainly in Bousselam. Bacterial densities of the rhizosphere and endophyte cultivable communities did not differ significantly. However, the number of cultivable beneficial bacteria isolated from roots and rhizosphere with multiple PGP traits was significantly increased. Bacterial survival of CMC-coated seed inoculum was appreciable and remained constant, especially for the Pa strain, during 21 months at room temperature. Based on these results, the PGPR used after seed priming would be a feasible and sustainable strategy to improve soil fertility and promote growth of durum wheat in stressful and non-stressful environments.

Pharmacological activity of sea cucumber Cucumaria frontosa derived amylase enzyme against blood stream pathogen

At presently availability of resistance pathogen to numerous drugs is a common feature of hospital-acquired Pseudomonas aeruginosa strains. So, the present study collect the 80 blood sample from blood cancer patient in Revathi Medical Hospital, Tirupur City for isolating the PA strain from blood sample and confirmed the bacterial strain using standard test method followed by drug susceptibility test for identification of resistant’s. In this study the maximum resistant was recorded in antibiotics Vancomycin 99% and maximum sensitive was observed in antibiotic Levofloxacin 99%. The blood stream infection causing PA plasmid extracted in 19 carcinogenic isolates which was showed more than 50% resistance selected for plasmid isolation. Due to the ability of resistant mechanism present in BSI pathogen the identification of new resources for novel antibiotics used urgently need to carry out a specific task to deal the blood stream infection. Moreover the enzyme especially the amylase have been isolated and purified from marine sea cucumber associated microorganism and the antimicrobial assay was done by using well techniques. The maximum zone of inhibition at 50, 100 and 150µl (, 25mm, 26mm and 16mm, 22mm, 23mm) of amylase extract was observed in the PAVG43,60 isolates. So hence this motto of this research proved the sea cucumber associated extracted amylase enzyme play a major role in the diagnosis of blood stream infection in cancer patients.
 Keywords: Blood Stream Infection, Pseudomonas aeruginosa, Sea Cucumber, Amylase

Aedes Albopictus and Cache Valley virus: a new threat for virus transmission in New York State

ABSTRACT We report surveillance results of Cache Valley virus (CVV; Peribunyaviridae, Orthobunyavirus) from 2017 to 2020 in New York State (NYS). Infection rates were calculated using the maximum likelihood estimation (MLE) method by year, region, and mosquito species. The highest infection rates were identified among Anopheles spp. mosquitoes and we detected the virus in Aedes albopictus for the first time in NYS. Based on our previous Anopheles quadrimaculatus vector competence results for nine CVV strains, we selected among them three stains for further characterization. These include two CVV reassortants (PA and 15041084) and one CVV lineage 2 strain (Hu-2011). We analyzed full genomes, compared in vitro growth kinetics and assessed vector competence of Aedes albopictus. Sequence analysis of the two reassortant strains (PA and 15041084) revealed 0.3%, 0.4%, and 0.3% divergence; and 1, 10, and 6 amino acid differences for the S, M, and L segments, respectively. We additionally found that the PA strain was attenuated in vertebrate (Vero) and mosquito (C6/36) cell culture. Furthemore, Ae. albopictus mosquitoes are competent vectors for CVV Hu-2011 (16.7–62.1% transmission rates) and CVV 15041084 (27.3–48.0% transmission rates), but not for the human reassortant (PA) isolate, which did not disseminate from the mosquito midgut. Together, our results demonstrate significant phenotypic variability among strains and highlight the capacity for Ae. albopictus to act as a vector of CVV.

MUC1 ectodomain is a flagellin-targeting decoy receptor and biomarker operative during Pseudomonas aeruginosa lung infection

We previously reported that flagellin-expressing Pseudomonas aeruginosa (Pa) provokes NEU1 sialidase-mediated MUC1 ectodomain (MUC1-ED) desialylation and MUC1-ED shedding from murine lungs in vivo. Here, we asked whether Pa in the lungs of patients with ventilator-associated pneumonia might also increase MUC1-ED shedding. The levels of MUC1-ED and Pa-expressed flagellin were dramatically elevated in bronchoalveolar lavage fluid (BALF) harvested from Pa-infected patients, and each flagellin level, in turn, predicted MUC1-ED shedding in the same patient. Desialylated MUC1-ED was only detected in BALF of Pa-infected patients. Clinical Pa strains increased MUC1-ED shedding from cultured human alveolar epithelia, and FlaA and FlaB flagellin-expressing strains provoked comparable levels of MUC1-ED shedding. A flagellin-deficient isogenic mutant generated dramatically reduced MUC1-ED shedding compared with the flagellin-expressing wild-type strain, and purified FlaA and FlaB recapitulated the effect of intact bacteria. Pa:MUC1-ED complexes were detected in the supernatants of alveolar epithelia exposed to wild-type Pa, but not to the flagellin-deficient Pa strain. Finally, human recombinant MUC1-ED dose-dependently disrupted multiple flagellin-driven processes, including Pa motility, Pa biofilm formation, and Pa adhesion to human alveolar epithelia, while enhancing human neutrophil-mediated Pa phagocytosis. Therefore, shed desialylated MUC1-ED functions as a novel flagellin-targeting, Pa-responsive decoy receptor that participates in the host response to Pa at the airway epithelial surface.

Microfluidic Obstacle Arrays Induce Large Reversible Shape Change in Red Blood Cells.

Red blood cell (RBC) shape change under static and dynamic shear stress has been a source of interest for at least 50 years. High-speed time-lapse microscopy was used to observe the rate of deformation and relaxation when RBCs are subjected to periodic shear stress and deformation forces as they pass through an obstacle. We show that red blood cells are reversibly deformed and take on characteristic shapes not previously seen in physiological buffers when the maximum shear stress was between 2.2 and 25 Pa (strain rate 2200 to 25,000 s−1). We quantify the rates of RBC deformation and recovery using Kaplan–Meier survival analysis. The time to deformation decreased from 320 to 23 milliseconds with increasing flow rates, but the distance traveled before deformation changed little. Shape recovery, a measure of degree of deformation, takes tens of milliseconds at the lowest flow rates and reached saturation at 2.4 s at a shear stress of 11.2 Pa indicating a maximum degree of deformation was reached. The rates and types of deformation have relevance in red blood cell disorders and in blood cell behavior in microfluidic devices.

Chemical and microbiological stability of diluted ceftazidime in three different solutions under three storage temperatures over a 28day period.

Ceftazidime (CAZ) solutions are being used based on anecdotal reports for otitis externa complicated by multidrug-resistant Pseudomonas aeruginosa (MDR PA). The chemical and microbiological stability of these proposed compounded solutions have not been evaluated, and likely are affected by the diluent and storage duration or temperature. Compounded CAZ solutions would show variable degradation dependent on diluent, time and temperature. The antimicrobial activity of the solutions would reflect changes in concentration and not alterations to the chemical compound. Ceftazidime was compounded with 100mL 0.9% sodium chloride (NA+CAZ), 118mL Triz-EDTAAqueous flush (TE+CAZ) and 125mL Douxo Micellar Solution (MI+CAZ). Aliquots of the solutions were stored at 25°C, 4°C and -20°C for 28 days. High-performance liquid chromatography was used to analyse CAZ recovery from compounded solutions at weekly intervals. A modified broth dilution technique was utilised to assess minimum inhibitory concentration (MIC) to monitor antimicrobial activity against a reference PA strain. Temperature, duration of storage and diluent each had independent effects on the chemical stability of CAZ. CAZ concentrations decreased over time as well as with increased temperature. NA+CAZ solutions exhibited the least degradation compared to the other solutions. The MIC for PA was most consistent for NA+CAZ solutions regardless of storage temperature and duration of storage. Chemical and microbiological stability of compounded CAZ solutions varied by diluent, storage temperature and duration of storage. Dilution in NA resulted in the lowest variation in stability over 28 days when stored at refrigerated or frozen temperatures compared to other diluents.

The effect of subcutaneous local spraying of Pseudomonas aeruginosa preparation to reduce postoperative drainage time in patients with breast cancer.

To evaluate the efficacy of subcutaneous local spraying of Pseudomonas aeruginosa (P. aeruginosa) preparation (PAP) to reduce the postoperative drainage time in patients with breast cancer (BC). The PAP was prepared from an inactivated P. aeruginosa-mannose sensitive hemagglutinin (PA-MSHA) strain, a genetically engineered heat-inactivated PA strain with, mannose-sensitive binding activity, which can induce tumor cell apoptosis. This study is its retrospective nature, a total of 65 BC patients (PAP group) who underwent subcutaneous local spraying of PAP in the operation area at West China Hospital of Sichuan University between June 2019 and October 2019, and 32 BC patients without treatment (non-PAP group, without intraoperative subcutaneous local spraying of PAP in the operation area) were enrolled. Daily drainage volume, drainage time, presence of seroma after extubation, and treatment complications (such as fever, incision infection, and flap necrosis) were recorded. No significant differences in age, height, weight, age of initial menstruation, menopause, body mass index (BMI), tumor size, lymph node metastasis, or treatment complications were found between the two groups (P>0.05). Drainage volume and drainage time in the PAP group were significantly lower than those in the non-PAP group (P<0.001). Additionally, the incidence of seroma after extubation was significantly lower in the PAP group compared to the non-PAP group (P=0.009). Subcutaneous local spraying of PAP on the operation area may be helpful to reduce drainage volume, shorten drainage time, and prevent the occurrence of seroma after BC surgery.

P117 Association between Pseudomonas aeruginosa (Pa) strain genotype and viable bacterial community profiles in sputum

P117 Association between Pseudomonas aeruginosa (Pa) strain genotype and viable bacterial community profiles in sputum

Cystic Fibrosis Patients Infected With Epidemic Pseudomonas aeruginosa Strains Have Unique Microbial Communities.

Pseudomonas aeruginosa is the archetypal cystic fibrosis (CF) pathogen. However, the clinical course experienced by infected individuals varies markedly. Understanding these differences is imperative if further improvements in outcomes are to be achieved. Multiple studies have found that patients infected with epidemic P. aeruginosa (ePA) strains may have a worse clinical prognosis than those infected with unique, non-clonal strains. Additionally, the traditionally uncultured CF lung bacterial community (i.e., CF microbiome) may further influence the outcome. We sought to identify if these two important variables, not identified through routine culture, associate and together may contribute to disease pathogenesis. Patients were classified as being infected with Prairie Epidemic ePA (PES) or a non-clonal strain, unique PA strains (uPA), through a retrospective assessment of a comprehensive strain biobank using a combination of PFGE and PES-specific PCR. Patients were matched to age, sex, time-period controls and sputum samples from equivalent time periods were identified from a sputum biobank. Bacterial 16S rRNA gene profiling and Pseudomonas qPCR was used to characterize the respiratory microbiome. We identified 31 patients infected with PES and matched them with uPA controls. Patients infected with PES at baseline have lower microbial diversity (P = 0.02) and higher P. aeruginosa relative abundance (P < 0.005). Microbial community structure was found to cluster by PA strain type, although it was not the main determinant of community structure as additional factors were also found to be drivers of CF community structure. Communities from PES infected individuals were enriched with Pseudomonas, Streptococcus and Prevotella OTUs. The disproportionate disease experienced by ePA infected CF patients may be mediated through a combination of pathogen-pathogen factors as opposed to strictly enhanced virulence of infecting P. aeruginosa strains.

Detection of 5 Kinds of Genes Related to Plasmid-Mediated Quinolone Resistance in Four Species of Nonfermenting Bacteria with 2 Drug Resistant Phenotypes.

Objective This study aimed to detect 5 kinds of genes related to plasmid-mediated quinolone resistance in four species of nonfermenting bacteria with 2 drug resistance phenotypes (multidrug resistance and pandrug resistance), which were Acinetobacter baumannii (Ab), Pseudomonas aeruginosa (Pa), Stenotrophomonas maltophilia (Sm), and Elizabethkingia meningoseptica (Em). Methods The Phoenix NMIC/ID-109 panel and API 20NE panel were applied to 19 isolated strains, including 6 Ab strains (2 strains with multidrug resistance and 4 strains with pandrug resistance), 6 Pa strains (3 strains with multidrug resistance and 3 strains with pandrug resistance), 4 Sm strains (2 strains with multidrug resistance and 2 strains with pandrug resistance), and 3 Cm strains (2 strains with multidrug resistance and 1 strain with pandrug resistance). After strain identification and drug susceptibility test, PCR was applied to detect 5 genes related to plasmid-mediated quinolone resistance. The genes detected were quinolone resistance A (qnrA), aminoglycoside acetyltransferase ciprofloxacin resistance variant, acc(6′)-Ib-cr, and 3 integrons (intI1, intI2, and intI3). The amplified products were analyzed by 1% agarose gel electrophoresis and sequenced. Sequence alignment was carried out using the bioinformatics technique. Results Of 19 strains tested, 8 strains carried acc(6′)-Ib-cr and 6 of them were of pandrug resistance phenotype (3 Ab strains, 2 Pa strains, and 1 Sm strain). The carrying rate of acc(6′)-Ib-cr was 60.0% for strains of pandrug resistance (6/10). Two strains were of multidrug resistance (1 Ab strain and 1 Pa strain), and the carrying rate of acc(6′)-Ib-cr was 22.0% (2/9). The carrying rate was significantly different between strains of multidrug resistance and pandrug resistance (P < 0.05). The class 1 integron was detected in 11 strains, among which 6 strains were of pandrug resistance (3 Ab strains, 2 Pa strains, and 1 Sm strain). The carrying rate of the class 1 integron was 60.0% (6/10). Five strains were of multidrug resistance (3 Pa strains, 1 Ab strain, and 1 Em strain), and the carrying rate was 55.6% (5/9). The carrying rate of the class 1 integron was not significantly different between strains of multidrug resistance and pandrug resistance (P > 0.05). Both acc(6′)-Ib-cr and intI1 were detected in 6 strains, which were negative for qnrA, intI2, and intI3. Conclusion Quinolone resistance of isolated strains was related to acc(6′)-Ib-cr and intI1 but not to qnrA, intI2, or intI3. The carrying rate of acc(6′)-Ib-cr among the strains of pandrug resistance was much higher than that among the strains of multidrug resistance. But, the strains of two drug resistant phenotypes were not significantly different in the carrying rate of intI1. The detection rates of the two genes were high and similar in Ab and Pa strains. 1 Em strain carried the class 1 integron.

COMMUNITY RAINWATER HARVESTING FINANCIAL PAYBACK ANALYSES - CASE STUDY IN MALAYSIA

Malaysian water demand is increasing at an alarming rate reaching 27 to 38% higher than the World Health Organisation recommended consumption limit of 165 liters per capita per day. Therefore, the Malaysian water shortage crisis is quite possible in future due to this water demand uptrend. The average annual rainfall of Malaysia is 2,400 mm but large portion of this fresh water resource becomes runoff and lost through our catchments. Urban flash flood is also becoming more frequent due to fast pace of urban development and anthropogenic induced runoff. Malaysia has experienced drought and flooding in different areas and therefore, it is crucial to study the feasibility of alternate water resources in Malaysia to manage and maintain the sustainability of urban township. This study reviewed a past rain water harvesting system (RWHS) case and conducted the financial payback analyses on its proposed system. If there were 177 rain days per year with at least 52 mm of rainfall event depth, the payback period of the proposed RWHS would be 5.8 years when the discount rate (i%) = 2% and 8.2 years if i% = 10%. The payback period became longer when the annual rain days dropped below 106 and 89 rain days per year. If the proposed RWHS only serve the community under this study, it will take 12 days to consume 800 m3 stored water, while any rainfall of consecutive days will not be harvested as the underground storage tank is in full capacity. The proposed RWHS must be filled up at least 38 times per year in order to break even with the proposed annual maintenance cost but will never be able to achieve any payback from its initial investment. Rain water harvesting and full utilisation is the only way to achieve high water cost savings and shorter payback period, and maximise urban excess runoff reduction.

Durum Wheat Stress Tolerance Induced by Endophyte Pantoea agglomerans with Genes Contributing to Plant Functions and Secondary Metabolite Arsenal

In the arid region Bou-Saâda at the South of Algeria, durum wheat Triticum durum L. cv Waha production is severely threatened by abiotic stresses, mainly drought and salinity. Plant growth-promoting rhizobacteria (PGPR) hold promising prospects towards sustainable and environmentally-friendly agriculture. Using habitat-adapted symbiosis strategy, the PGPR Pantoea agglomerans strain Pa was recovered from wheat roots sampled in Bou-Saâda, conferred alleviation of salt stress in durum wheat plants and allowed considerable growth in this unhostile environment. Strain Pa showed growth up to 35 °C temperature, 5–10 pH range, and up to 30% polyethylene glycol (PEG), as well as 1 M salt concentration tolerance. Pa strain displayed pertinent plant growth promotion (PGP) features (direct and indirect) such as hormone auxin biosynthesis, production of 1-aminocyclopropane-1-carboxylate (ACC) deaminase, and ammonia and phosphate solubilization. PGPR features were stable over wide salt concentrations (0–400 mM). Pa strain was also able to survive in seeds, in the non-sterile and sterile wheat rhizosphere, and was shown to have an endophytic life style. Phylogenomic analysis of strain Pa indicated that Pantoea genus suffers taxonomic imprecision which blurs species delimitation and may have impacted their practical use as biofertilizers. When applied to plants, strain Pa promoted considerable growth of wheat seedlings, high chlorophyll content, lower accumulation of proline, and favored K+ accumulation in the inoculated plants when compared to Na+ in control non-inoculated plants. Metabolomic profiling of strain Pa under one strain many compounds (OSMAC) conditions revealed a wide diversity of secondary metabolites (SM) with interesting salt stress alleviation and PGP activities. All these findings strongly promote the implementation of Pantoea agglomerans strain Pa as an efficient biofertilizer in wheat plants culture in arid and salinity-impacted regions.

Endothelial Cell Biomechanical Responses are Dependent on Both Fluid Shear Stress and Tensile Strain.

The goal of this study was to investigate how concurrent shear stress and tensile strain affect endothelial cell biomechanical responses. Human coronary artery endothelial cells were exposed to concurrent pulsatile shear stress and cyclic tensile strain in a programmable shearing and stretching device. Three shear stress-tensile strain conditions were used: (1) pulsatile shear stress at 1Pa and cyclic tensile strain at 7%, simulating normal stress/strain conditions in a healthy coronary artery; (2) shear stress at 3.7Pa and tensile strain at 3%, simulating pathological stress/strain conditions near a stenosis; (3) shear stress at 0.7Pa and tensile strain at 5%, simulating pathological stress/strain conditions in a recirculation zone. Cell morphology was quantified using immunofluorescence microscopy. Cell surface PECAM-1 phosphorylation, ICAM-1 expression, ERK1/2 and NF-κB activation were measured using ELISA or Western blot. Simultaneous stimulation from pulsatile shear stress and cyclic tensile strain induced a significant increase in cell area, compared to that induced by shear stress or tensile strain alone. The combined stimulation caused significant increases in PECAM-1 phosphorylation. The combined stimulation also significantly enhanced EC surface ICAM-1 expression (compared to that under shear stress alone) and transcriptional factor NF-κB activation (compared to that under control conditions). Pulsatile shear stress and cyclic tensile strain could induce increased but not synergistic effect on endothelial cell morphology or activation. The combined mechanical stimulation can be relayed from cell membrane to nucleus. Therefore, to better understand how mechanical conditions affect endothelial cell mechanotransduction and cardiovascular disease development, both shear stress and tensile strain need to be considered.

Neuraminidase 1–mediated desialylation of the mucin 1 ectodomain releases a decoy receptor that protects against Pseudomonas aeruginosa lung infection

Pseudomonas aeruginosa (Pa) expresses an adhesin, flagellin, that engages the mucin 1 (MUC1) ectodomain (ED) expressed on airway epithelia, increasing association of MUC1-ED with neuraminidase 1 (NEU1) and MUC1-ED desialylation. The MUC1-ED desialylation unmasks both cryptic binding sites for Pa and a protease recognition site, permitting its proteolytic release as a hyperadhesive decoy receptor for Pa. We found here that intranasal administration of Pa strain K (PAK) to BALB/c mice increases MUC1-ED shedding into the bronchoalveolar compartment. MUC1-ED levels increased as early as 12 h, peaked at 24-48 h with a 7.8-fold increase, and decreased by 72 h. The a-type flagellin-expressing PAK strain and the b-type flagellin-expressing PAO1 strain stimulated comparable levels of MUC1-ED shedding. A flagellin-deficient PAK mutant provoked dramatically reduced MUC1-ED shedding compared with the WT strain, and purified flagellin recapitulated the WT effect. In lung tissues, Pa increased association of NEU1 and protective protein/cathepsin A with MUC1-ED in reciprocal co-immunoprecipitation assays and stimulated MUC1-ED desialylation. NEU1-selective sialidase inhibition protected against Pa-induced MUC1-ED desialylation and shedding. In Pa-challenged mice, MUC1-ED-enriched bronchoalveolar lavage fluid (BALF) inhibited flagellin binding and Pa adhesion to human airway epithelia by up to 44% and flagellin-driven motility by >30%. Finally, Pa co-administration with recombinant human MUC1-ED dramatically diminished lung and BALF bacterial burden, proinflammatory cytokine levels, and pulmonary leukostasis and increased 5-day survival from 0% to 75%. We conclude that Pa flagellin provokes NEU1-mediated airway shedding of MUC1-ED, which functions as a decoy receptor protecting against lethal Pa lung infection.

Comparison of Petroleum Hydrocarbons Degradation by Klebsiella pneumoniae and Pseudomonas aeruginosa

The aim of this work was to develop bacterial communities to effectively degrade petroleum hydrocarbons (PHs). We investigated the biotic and abiotic contributors to differences in PHs degradation efficacy between two bacterial strains, Klebsiella pneumoniae (Kp) and Pseudomonas aeruginosa (Pa), screened out from the activated sludge of a petroleum refinery. We characterized the temporal variations in degradation efficacy for diesel and its five major constituents as a sole carbon source and identified more constituents they degraded. The growth characteristics, surface tension, hydrophobicity and emulsifiability of these two strains were measured. We further estimated the relationships between their degradation efficacy and all the biotic and abiotic factors. Results showed that the Pa strain had higher diesel degradation efficacy (58% on Day 14) and utilized more diesel constituents (86%) compared to Kp. Additionally, the growth of the Pa strain in diesel medium was faster than that of the Kp strain. The Pa strain had a lower surface tension and higher hydrophobicity and emulsifiability than Kp, while the surfactant produced by Pa was identified as rhamnolipids. Degradation of PHs was positively related to bacterial growth, hydrophobicity and emulsification but negatively related to surface tension. Overall, differences in degrading capacity for diesel constituents, relative growth rate, and biosurfactant production contributed to the variation in the PHs degradation efficacy of these two bacterial strains.

697. Pseudomonas aeruginosa PcrV and Psl, the Molecular Targets of Bispecific Monoclonal Antibody MEDI3902, Are Conserved Among Diverse Hospital Isolates Collected From an International Surveillance Study

Background Pseudomonas aeruginosa is a frequent cause of life-threatening infections in mechanically ventilated patients and is associated with high mortality rates. Bispecific monoclonal antibody MEDI3902 targeting Pa type-3-secretion system (PcrV) and the Psl exopolysaccharide is currently under phase 2b development for the prevention of pneumonia in mechanically ventilated subjects with Pa colonization in the lower respiratory tract. In this study, we sought to survey a vast collection of global Pa clinical isolates for presence of pcrV and psl loci and MEDI3902 epitope conservation to evaluate the magnitude of Pa strain coverage by MEDI3902.Methods913 Pa clinical isolates were collected from diverse patients and geographical locations in 2004–2014. Whole genome sequencing of the full collection was performed via MiSeq 2 × 250 runs (Illumina®). PcrV and Psl expression was detected by immunoblotting and ELISA, respectively. The crystal structure of anti-PcrV fab and PcrV fragment complex-crystals was solved at 2.8 Å resolution. MEDI3902 activity against representative isolates was tested in cytotoxicity and opsonophagocytosis assays and in a murine pneumonia model.ResultsWhole-genome sequencing revealed intact pcrV and psl genetic elements in 99% and 94% of isolates, respectively. We identified 46 variants of PcrV that were all bound by the anti-PcrV moiety of MEDI3902 and confirmed through crystal structure analysis that antibody-antigen contact residues were preserved in all variants. Similarly, anti-Psl binding was confirmed for selected isolates containing the complete Psl operon and strains lacking non-essential psl genes. Importantly, 99.9% of isolates contained the full complement of either genetic element. Consistent with these results, we observed potent MEDI3902 activity against diverse strain types, including strains that expressed only a single target.ConclusionOur results indicate PcrV and Psl are highly prevalent in recent clinical isolates from around the world, suggesting that MEDI3902 can mediate broad coverage against Pa.Disclosures D. E. Tabor, Astra Zeneca: employee, Salary.