Adaptation of sea turtles to climate warming: Will phenological responses be sufficient to counteract changes in reproductive output?

Sea turtles are vulnerable to climate change since their reproductive output is influenced by incubating temperatures, with warmer temperatures causing lower hatching success and increased feminization of embryos. Their ability to cope with projected increases in ambient temperatures will depend on their capacity to adapt to shifts in climatic regimes. Here, we assessed the extent to which phenological shifts could mitigate impacts from increases in ambient temperatures (from 1.5 to 3°C in air temperatures and from 1.4 to 2.3°C in sea surface temperatures by 2100 at our sites) on four species of sea turtles, under a "middle of the road" scenario (SSP2-4.5). Sand temperatures at sea turtle nesting sites are projected to increase from 0.58 to 4.17°C by 2100 and expected shifts in nesting of 26-43 days earlier will not be sufficient to maintain current incubation temperatures at 7 (29%) of our sites, hatching success rates at 10 (42%) of our sites, with current trends in hatchling sex ratio being able to be maintained at half of the sites. We also calculated the phenological shifts that would be required (both backward for an earlier shift in nesting and forward for a later shift) to keep up with present-day incubation temperatures, hatching success rates, and sex ratios. The required shifts backward in nesting for incubation temperatures ranged from -20 to -191 days, whereas the required shifts forward ranged from +54 to +180 days. However, for half of the sites, no matter the shift the median incubation temperature will always be warmer than the 75th percentile of current ranges. Given that phenological shifts will not be able to ameliorate predicted changes in temperature, hatching success and sex ratio at most sites, turtles may need to use other adaptive responses and/or there is the need to enhance sea turtle resilience to climate warming.

Performance Comparison of Different Flat Plate Solar Collectors by Means of the Entropy Generation Rate Using Computational Fluid Dynamics.

In this work, a numerical analysis of three different flat plate solar collectors was conducted using their entropy generation rates. Specifically, the Computational Fluid Dynamics (CFD) technique was used to compare the detailed performance of conventional and zigzag tube geometries of flat plate solar collectors (FPCs) in terms of their entropy generation rates. The effects of fluid viscosity, heat transfer, and heat loss of the flat plate solar collectors were considered for the local and global entropy generation rate analyses. Variations on the inlet volumetric flow rate of the FPCs from 1.0 to 9.0 L/min were simulated under the average solar radiation for one year in the state of Guanajuato, Mexico. The results illustrate and discuss the temperatures, pressures, and global entropy generation rates for volumetric flow variations. The velocity, temperature, and pressure distributions and the maps of the local entropy generation rates inside the collectors are presented and analyzed for the case with a flow rate of 3.0 L/min. These results demonstrate that the zigzag geometries achieved higher outlet temperatures and greater entropy generation rates than the conventional geometry for all the volumetric flow rates considered.

Identification of Somatic Proteins in Haemonchus Contortus Infective Larvae (L3) and Adults.

Haemonchus contortus is considered the most pathogenic nematode in sheep production systems based on grazing. Comparing infective larvae (L3) with adult parasites can lead to the identification of proteins that play an important role in parasite-host interactions. In this study, we report a list of H. contortus somatic proteins and made a comparative analysis of somatic proteins of L3 and adult worms. L3 and adult parasites were subjected to protein extraction and subsequently to peptide fractionation. Peptides were analysed by mass spectrometry and LC-MS/MS data analysis. Data analysis and search on SEQUEST and MASCOT against H. contortus from the WormBase ParaSite database resulted in the identification of 775 unique peptide sequences corresponding to 227 proteins at 1 % FDR. From these, 18 proteins were specific to L3 and 63 to adult parasites. The gene ontology (GO) enrichment analysis of the proteins specific to L3 and adult worms to gain insight into cellular components, molecular functions and biological processes that affect the parasite-host interaction showed some differences between the two parasite stages. The list of proteins found provides a database to identify target proteins that could be useful as biomarkers of the infection or in the generation of anthelmintic drugs that inhibit proteins essential for the establishment of the infection and the survival of adult parasites. They can also serve as new candidates for vaccine research.

A systematic review of worldwide data on tinea capitis: analysis of the last 20 years.

Dermatophyte infections are the most common fungal infections in humans; among them, tinea capitis (TC) - the most contagious fungal infection - is caused by anthropophilic, zoophilic and geophilic dermatophytes. The purpose of this systematic review was to determine the different aetiological variants involved in TC and the overall epidemiology of the causes of this infection in the last two decades. We searched the MEDLINE (PubMed) and Embase databases for articles published from July 2000 to August 2019 using the following search terms: 'Tinea capitis', 'Africa', 'America', 'Asia', 'Europe', 'Oceania', and the names of the countries on each continent. The flow of information through the different phases in this systematic review was depicted using a PRISMA flow diagram, which mapped the number of records identified, included and excluded, and the reasons for exclusion. Our findings indicate that the frequency of different aetiologic agents of TC in the reported studies varied globally, from 0.4-87.7% in Africa, 0.2-74.0% in North America, 0.0-91.2% in Eastern Asia, 0.0-69.0% in Eastern Europe and 2.9-86.4% in Oceania. Microsporum canis is the most frequent reported zoophilic agent worldwide, while Trichophyton violaceum and Trichophyton tonsurans are the predominant anthropophilic agents. Over time, the frequency of these latter fungal infections has increased globally, and these fungi have become the major species globally. Anthropophilic transmission - the most prevalent type of transmission - could be explained by two factors: (i) the socioeconomic status of affected countries and population groups with associated risk factors and (ii) movement of populations importing new causes of infection to areas where they had not been encountered previously. We observed that intercontinental migration and travel; globalization; environmental, climatic and ecological changes; and accelerated evolution of health technologies may influence the observed epidemiological changes and, consequently, contributed to the variations in the global status of TC.

Attenuation mechanisms and key parameters to enhance treatment performance in vegetation filters: A review.

In times when environmental concerns are on the rise and the search of ways to reduce waste generation and to create a circular economy is booming, Nature Based Solutions (NBSs) play a very important role. Vegetation Filters (VFs) are a type of Land Application System (LAS) in which wastewater is used to irrigate a forestry plantation to treat the water and produce biomass. VFs show multiple benefits that render this technology a suitable solution for wastewater treatment, especially for scattered populations or isolated buildings that lack of connection to sewer systems. This review aims to provide a comprehensive state of the art of VF implementation, highlighting the do's and don'ts for a successful performance focusing on those factors that are essential to water treatment. Results show that VFs have a great treatment capacity when all involving factors are considered, and their efficiency tends to increase with time, as the VF develops and "gets older". Indeed, the presence of fine-textured soils, the selection of a proper vegetation species, the use of pre-treated wastewater and a water balance-based irrigation schedule alternating wetting and -drying cycles are all factors that help to achieve the best performance. However, it is necessary to design and follow a simple but rigorous operation and maintenance schedule to avoid system failure, which could lead to NO3-N leaching towards groundwater.

Outcomes of an enhanced recovery after radical cystectomy program in a prospective multicenter study: compliance and key components for success.

OBJECTIVE: To investigate the effect of an Enhanced Recovery After Surgery (ERAS) program on complications and length of stay (LOS) after radical cystectomy (RC) and to assess if the number and type of components of ERAS play a key role on the decrease of surgical morbidity. MATERIALS AND METHODS: We analyzed the data of 277 patients prospectively recruited in 11 hospitals undergoing RC initially managed according to local practice (Group I) and later within an ERAS program (Group II). Two main outcomes were defined: 90-day complications rate and LOS. As secondary variables we studied 90-day mortality, 30-day readmission and transfusion rate. RESULTS: Patients in Group II had a higher use of ERAS measures (98.6%) than those in Group I (78.2%) (p < 0.05). Patients in Groups I and II experienced similar complications (70.5% vs. 66%, p = 0.42). LOS was not different between Groups I and II (12.5 and 14 days, respectively, p = 0.59). The risk of having any complication decreases for patients having more than 15 ERAS measures adopted [RR = 0.815; 95% confidence interval (CI) 0.667-0.996; p = 0.045]. Avoidance of transfusion and nasogastric tube, prevention of ileus, early ambulation and a fast uptake of a regular diet are independently associated with the absence of complications. CONCLUSIONS: Complications and LOS after RC were not modified by the introduction of an ERAS program. We hypothesize that at least 15 measures should be applied to maximize the benefit of ERAS.

Insulin resistance in non-diabetes patients with spondyloarthritis.

Objectives: Insulin resistance (IR) constitutes a major underlying abnormality driving cardiovascular disease in the general population and has been linked to inflammatory diseases. In this study, we aimed to determine the prevalence of IR in patients with spondyloarthritis (SpA) and whether IR can be explained by disease-related features in such cases. Method: The study included 577 subjects: 306 patients diagnosed with SpA according to Assessment of SpondyloArthritis international Society criteria and 271 controls. Insulin and C-peptide serum levels, IR and ß-cell function (%B) indices by homoeostatic model assessment (HOMA2), and lipid profiles were assessed in patients and controls. A multivariable regression analysis was performed to evaluate the differences in IR indices between patients and controls and to determine how IR is associated with disease-related characteristics in SpA patients. Results: HOMA2-%B and HOMA2-IR scores, both calculated with insulin or C-peptide, had significantly higher values in SpA patients compared to controls in multivariable analysis adjusted for age, gender, traditional IR-related factors, and glucocorticoid intake. Disease activity, functional status, and metrological SpA indices were positively related to IR, but only in univariable analysis. Disease duration and positivity for human leucocyte antigen-B27 were independently associated with a higher HOMA2-%B after multivariable analysis. Conclusion: Patients with SpA have an increased IR compared to controls. SpA disease-related data are independently associated with ß-cell dysfunction.

Development and field testing of low-cost, quantal microbial assays with volunteer reporting as scalable means of drinking water safety estimation.

AIMS: To evaluate a low-cost water quality test for at-scale drinking water safety estimation in rural India. METHODS AND RESULTS: Within a longitudinal study to characterize variability in household drinking water safety in rural Maharashtra, we piloted a low-cost presence-absence (LCPA) microbial test designed to be used by volunteer residents in rural areas. In comparing the LCPA results with standard laboratory methods for enumeration of Escherichia coli, we found that LCPA tests using modified mTec media were highly sensitive in detecting drinking water of moderate risk (88% of tests were positive at E. coli counts of 11-100 CFU per 100 ml) and high risk (96% of tests were positive at E. coli counts of 101 + CFU per 100 ml). The LCPA tests demonstrated low specificity for E. coli specifically, due to concurrent detection of Klebsiella: 38% of LCPA tests were positive even when E. coli was not detected in a 100 ml sample by membrane filtration, suggesting the test would be conservative in risk estimation. We also found that 47% of participants in rural villages in India were willing to conduct tests and return results after a brief training, with 45% of active participants sending their water testing results via short message service. CONCLUSIONS: Given their low cost (~US$0.50 as piloted) and open-source format, such tests may provide a compelling alternative to standard methods for rapid water quality assessments, especially in resource-limited settings. SIGNIFICANCE AND IMPACT OF THE STUDY: The lack of availability of water quality data constrains efforts to monitor, evaluate and improve the safety of water and sanitation infrastructure in underserved settings. Current water testing methods are not scalable because of laboratory and cost constraints. Our findings indicate the LCPA or similar low-cost microbial tests could be useful in rapid water safety estimation, including via crowdsourcing.

Activity and expression of Candida glabrata vacuolar proteases in autophagy-like conditions.

Candida glabrata is an emerging opportunistic pathogen that has intrinsic resistance to azoles. During infection or while living as a commensal, it encounters nutritional stresses such as deficiency of carbon or nitrogen sources. Herein, we investigate the expression and activity of PrA, Ape1, Ape3 and CpY vacuolar proteases during these stressful nutrimental conditions. Our findings demonstrate a differential activity profile depending on the addition or lack of carbon, nitrogen or both. Of the four proteases tested, PrA and Ape3 showed a higher activity in the absence of nitrogen. Steady-state RNA levels for all the proteases were also differentially expressed although not always correlated with its activity, suggesting multiple levels of regulation. Microscopy observations of C. glabrata cells subjected to the different conditions showed an increase in the vacuolar volume. Moreover, the presence of ATG8-PE and an increased expression of ATG8 were observed in the yeast under the tested conditions suggesting that C. glabrata is in autophagy stage. Taken together, our results showed that PrA, Ape1, Ape3 and CpY have varying activities and expression depending on whether nitrogen or carbon is added to the media, and that these vacuolar proteases might have a role in the autophagy process.

The dual role of DksA protein in the regulation of Escherichia coli pArgX promoter.

Gene expression regulation by the stringent response effector, ppGpp, is facilitated by DksA protein; however DksA and ppGpp can play independent roles in transcription. In Escherichia coli, the pArgX promoter which initiates the transcription of four tRNA genes was shown to be inhibited by ppGpp. Our studies on the role of DksA in pArgX regulation revealed that it can stimulate transcription by increasing the binding of RNA polymerase to the promoter and the productive transcription complex formation. However, when DksA is present together with ppGpp a severe down-regulation of promoter activity is observed. Our results indicate that DksA facilitates the effects of ppGpp to drive formation of inactive dead-end complexes formed by RNA polymerase at the ArgX promoter. In vivo, ppGpp-mediated regulation of pArgX transcription is dependent on DksA activity. The potential mechanisms of opposing pArgX regulation by ppGpp and DksA are discussed. pArgX is the first reported example of the promoter stimulated by DksA and inhibited by ppGpp in vitro when an overall inhibition occurs in the presence of both regulators. A dual role is thus proposed for DksA in the regulation of the pArgX promoter activity.

Decreased expression of ectonucleotidase E-NPP1 in leukocytes from subjects with severe asthma exacerbation.

Several studies suggest that ATP and related nucleotides play a role in the pathophysiology of asthma. However, the functionality of ectonucleotidases in this disease has been scantly investigated. We studied total ectonucleotidase activity in leukocytes from patients suffering from asthma exacerbation and explored the expression of E-NTPDase 1, 2, 3, and 8, and E-NPP1, 2, and 3, in their polymorphonuclear cells by immunofluorescence and qPCR. Leukocytes from patients with mild or moderate asthma exacerbation had similar ectonucleotidase activity than leukocytes from healthy subjects, while in patients with severe asthma exacerbation, this activity was lower. Of the ectonucleotidases studied, only E-NPP1 displayed diminished immunofluorescence and a significant decrease in its mRNA expression, both in patients with severe asthma exacerbation. This reduced E-NPP1 expression could be responsible for increased amounts of ATP or other nucleotides, capable of worsening asthma exacerbation, and warranting further investigation.

[Pilot tests using molecular diagnostic assay cervicovaginal infection during pregnancy]. / Ensayo piloto del uso de pruebas moleculares para el diagnóstico de infecciones cervicovaginales en pacientes embarazadas.

Background: The prevalence of cervicovaginal infections during pregnancy has been associated with adverse perinatal outcomes however, the actual approach used for diagnosis is not effective. The aim of this study was to compare the diagnosis of vaginal infections in pregnant women using clinical, molecular diagnostic and traditional microbiological culture in a pilot study, to determine the prevalence and association with the development of preterm labor. Materials and methods: We performed a nested cross-sectional study composed by 54 women in a cohort of pregnant women in Mexico City. Cervicovaginal infections were evaluated by clinical methods, microbiology culture and a commercially available molecular biology test. Results: Prevalence of cervicovaginal infections during pregnancy was estimated between 28% and 50% according to methodologies. Considering the clinical diagnosis of preterm labor as the gold standard, all diagnostic tests were poor as predictors of preterm labor. Conclusion: Traditional approaches to establish the significance of cervicovaginal infection in pregnancy are exhausted, so be sought new ways to understand this complex relationship. Meanwhile it is recommended to continue to use traditional methods to identify infections during pregnancy in both knowledge of new methods aimed at understanding these relationships are sophisticated.

Serum sCD26 for colorectal cancer screening in family-risk individuals: comparison with faecal immunochemical test.

BACKGROUND: The development of specific screening programs for individuals with a family history of colorectal cancer (CRC) is a priority. This study evaluates the diagnostic performance of serum soluble CD26 (sCD26) in family-risk individuals and compares this marker with the faecal immunochemical test for the detection of advanced neoplasia (AN) (CRC or advanced adenomas; AA). METHODS: Five hundred and sixteen asymptomatic individuals with at least one first-degree relative with CRC were included. Serum sCD26 was measured in all the individuals who also underwent a colonoscopy (53 AA and four cancer cases were found) and a faecal immunochemical test. RESULTS: Setting specificity to 90% and 95%, respectively, sCD26 showed a sensitivity of 39.6% and 28.3% for AA, and of 42.1% and 28.1% for AN. The combination of sCD26 and the faecal test detected AA and AN with a 52.8% and 56.1% sensitivity, corresponding to 93.5% specificity. CONCLUSIONS: The combination of serum sCD26 and the faecal blood test could result a valuable strategy for detecting AN in familial-risk CRC screening.

Cooperative adsorption behavior of phosphopeptides on TiO2 leads to biased enrichment, detection and quantification.

The adsorption behavior of phosphopeptides onto TiO2 surfaces was studied using the quartz crystal microbalance with dissipation monitoring (QCM-D) as the main experimental technique. The main focus is the characterization of the emergence of positive cooperativity under conditions where the peptides have a positively charged C-term. It is shown that when carrying no net charge, small water-soluble peptides as a rule develop positive cooperativity. The impact of the adsorption mechanism on the outcome of TiO2 based enrichment methods was investigated with the help of matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS). The data presented illustrate how the phosphopeptide profile in the enriched material may deviate from that in the native sample, as cooperative phosphopeptides are overrepresented in the former. Furthermore, commonly employed washing and elution solutions may facilitate preferential release of certain peptides, leading to further bias in the recovered sample. Taken together, the results of the present study demonstrate that thorough understanding of the mechanisms behind the adsorption of phosphopeptides on the enrichment material is necessary in order to develop reliable qualitative and quantitative methods for phosphoproteomics.

Variability in Damage Caused by the Mite Tetranychus urticae (Trombidiformes: Tetranychidae) Koch on Three Varieties of Strawberry.

The strawberry, Fragaria×ananassa Duchesne (Rosales: Rosaceae), is an important crop in Mexico. We evaluated the tolerance of three newly developed Mexican strawberry varieties (CP0615, CPLE-7, and CPJacona) to Tetranychus urticae Koch (Trombidiformes: Tetranychidae), the most important pest of strawberry. We evaluated the effect of three different initial mite densities on population growth, duration of each developmental stage and survival of T. urticae on the three strawberry varieties. We also compared the photosynthetic activity (Pn), sub-stomatal CO2 concentration (Ci), stomatal conductance (gs) and the area of leaf damaged in the three varieties. The largest final density of mites occurred on the variety CP0615, followed by the varieties CPLE-7 and CPJacona. There were no significant differences in the duration of T. urticae developmental stages amongst the varieties, except for larvae where the shortest duration was on variety CPLE-7. The proportion of eggs reaching the adult stage (survival) was significantly lower on the variety CPLE-7. The number and morphology of the trichomes did not play an important role in the outcomes, as they were similar in the three varieties. There were no significant differences in Pn, Ci, and gs values amongst the three varieties in the presence and absence of T. urticae. The area of leaf damaged in variety CPLE-7 was significantly smaller than for the other varieties. Based on these results, and with regard to spider mite tolerance, we believe that the variety CPLE-7 has the greatest potential for further development, and eventually, for use on a commercial scale in Mexico.

East-West gradient in the incidence of inflammatory bowel disease in Europe: the ECCO-EpiCom inception cohort.

OBJECTIVE: The incidence of inflammatory bowel disease (IBD) is increasing in Eastern Europe. The reasons for these changes remain unknown. The aim of this study was to investigate whether an East-West gradient in the incidence of IBD in Europe exists. DESIGN: A prospective, uniformly diagnosed, population based inception cohort of IBD patients in 31 centres from 14 Western and eight Eastern European countries covering a total background population of approximately 10.1 million people was created. One-third of the centres had previous experience with inception cohorts. Patients were entered into a low cost, web based epidemiological database, making participation possible regardless of socioeconomic status and prior experience. RESULTS: 1515 patients aged 15 years or older were included, of whom 535 (35%) were diagnosed with Crohn's disease (CD), 813 (54%) with ulcerative colitis (UC) and 167 (11%) with IBD unclassified (IBDU). The overall incidence rate ratios in all Western European centres were 1.9 (95% CI 1.5 to 2.4) for CD and 2.1 (95% CI 1.8 to 2.6) for UC compared with Eastern European centres. The median crude annual incidence rates per 100,000 in 2010 for CD were 6.5 (range 0-10.7) in Western European centres and 3.1 (range 0.4-11.5) in Eastern European centres, for UC 10.8 (range 2.9-31.5) and 4.1 (range 2.4-10.3), respectively, and for IBDU 1.9 (range 0-39.4) and 0 (range 0-1.2), respectively. In Western Europe, 92% of CD, 78% of UC and 74% of IBDU patients had a colonoscopy performed as the diagnostic procedure compared with 90%, 100% and 96%, respectively, in Eastern Europe. 8% of CD and 1% of UC patients in both regions underwent surgery within the first 3 months of the onset of disease. 7% of CD patients and 3% of UC patients from Western Europe received biological treatment as rescue therapy. Of all European CD patients, 20% received only 5-aminosalicylates as induction therapy. CONCLUSIONS: An East-West gradient in IBD incidence exists in Europe. Among this inception cohort--including indolent and aggressive cases--international guidelines for diagnosis and initial treatment are not being followed uniformly by physicians.

Putrescine catabolism is a metabolic response to several stresses in Escherichia coli.

Genes whose products degrade arginine and ornithine, precursors of putrescine synthesis, are activated by either regulators of the nitrogen-regulated (Ntr) response or σ(S) -RNA polymerase. To determine if dual control regulates a complete putrescine catabolic pathway, we examined expression of patA and patD, which specify the first two enzymes of one putrescine catabolic pathway. Assays of PatA (putrescine transaminase) activity and ß-galactosidase from cells with patA-lacZ transcriptional and translational fusions indicate dual control of patA transcription and putrescine-stimulated patA translation. Similar assays for PatD indicate that patD transcription required σ(S) -RNA polymerase, and Nac, an Ntr regulator, enhanced the σ(S) -dependent transcription. Since Nac activation via σ(S) -RNA polymerase is without precedent, transcription with purified components was examined and the results confirmed this conclusion. This result indicates that the Ntr regulon can intrude into the σ(S) regulon. Strains lacking both polyamine catabolic pathways have defective responses to oxidative stress, high temperature and a sublethal concentration of an antibiotic. These defects and the σ(S) -dependent expression indicate that polyamine catabolism is a core metabolic response to stress.

Effect of oral anticoagulants on the outcome of faecal immunochemical test.

BACKGROUND: We aimed to evaluate whether oral anticoagulants (OACs) alter faecal immunochemical test (FIT) performance in average-risk colorectal cancer (CRC) screening. METHODS: Individuals aged 50-69 years were invited to receive one FIT sample (cutoff 75 ng ml(-1)) between November 2008 and June 2011. RESULTS: Faecal immunochemical test was positive in 9.3% (21 out of 224) of users of OAC and 6.2% (365 out of 5821) of non-users (P-trend=0.07). The positive predictive value (PPV) for advanced neoplasia (AN) in non-users was 50.4% vs 47.6% in users (odds ratio, 0.70; 95% CI, 0.3-1.8; P=0.5). The PPV for AN in OAC more antiplatelets (aspirin or clopidogrel) was 75% (odds ratio, 2; 95% CI, 0.4-10.8; P=0.4). CONCLUSIONS: Oral anticoagulant did not significantly modify the PPV for AN in this population-based colorectal screening program. The detection rate of advanced adenoma was higher in the combination OAC more antiplatelets.

SRBC/cavin-3 is a caveolin adapter protein that regulates caveolae function.

Caveolae are a major membrane domain common to most cells. One of the defining features of this domain is the protein caveolin. The exact function of caveolin, however, is not clear. One possible function is to attract adapter molecules to caveolae in a manner similar to how clathrin attracts molecules to coated pits. Here, we characterize a candidate adapter molecule called SRBC. SRBC binds PKCdelta and is a member of the STICK (substrates that interact with C-kinase) superfamily of PKC-binding proteins. We also show it co-immunoprecipitates with caveolin-1. A leucine zipper in SRBC is essential for both co-precipitation with caveolin and localization to caveolae. SRBC remains associated with caveolin when caveolae bud to form vesicles (cavicles) that travel on microtubules to different regions of the cell. In the absence of SRBC, intracellular cavicle traffic is markedly impaired. We conclude that SRBC (sdr-related gene product that binds to c-kinase) and two other family members [PTRF (Pol I and transcription release factor) and SDPR] function as caveolin adapter molecules that regulate caveolae function.

Inhibition of extracellular nucleotides hydrolysis intensifies the allergic bronchospasm. A novel protective role of ectonucleotidases.

BACKGROUND: Nucleotides released to the extracellular space stimulate purinergic receptors, and their effects are modulated by ectonucleotidases. The role of ATP in the allergic bronchospasm has been scantly studied. METHODS: We used several techniques (plethysmography, organ baths, confocal microscopy, RT-PCR, ATP measurement) to explore the role of nucleotides and ectonucleotidases in the allergic bronchospasm in guinea pigs. RESULTS: While allergenic challenge with a low-dose ovalbumin (OVA) only produced a small bronchospasm (~2-fold the basal lung resistance), previous inhibition of ectonucleotidases by ARL-67156 greatly intensified this response (~11-fold the basal lung resistance, with 44% mortality). Bronchoalveolar lavage fluid obtained during this bronchospasm contained increased ATP concentration. This potentiation was abolished by antagonism of purinergic receptors (suramin+RB2) or TXA2 receptor (SQ29548), or by intratracheal apyrase. In tracheal rings and lung parenchyma strips, OVA caused a concentration-dependent contraction. Suramin+RB2 or levamisole produced a significant rightward displacement of this response, and ARL-67156 did not modify it. Platelets stimulated with OVA released ATP. Confocal images of nonsensitized tracheas showed slight fluorescence for P2Y6 receptors in epithelium and none for P2Y4 . Sensitized animals showed strong fluorescence to both receptors and to alkaline phosphatase in the airway epithelium. This correlated with a large increment in mRNA for P2Y4 and P2Y6 receptors in sensitized animals. CONCLUSIONS: Nucleotides greatly potentiate the allergic bronchospasm when ectonucleotidases activity is diminished, and this effect is probably favored by the upregulation of P2Y4 and P2Y6 receptors in airway epithelium during sensitization. These results prompt for further research on these mechanisms in human asthma.