Enhancing validity, reliability and participation in self-reported health outcome measurement for children and young people: a systematic review of recall period, response scale format, and administration modality.

INTRODUCTION: Self-report is the gold standard for measuring children's health-related outcomes. Design of such measures is complex and challenging. This review aims to systematically appraise the evidence on recall period, response scale format, mode of administration and approaches needed to enable children and young people < 19 years to participate in valid and reliable self-reporting of their health outcomes. METHOD: PsycInfo, Medline, CINAHL and Embase were searched from 1 January 1990 to 15 March 2020, and citation searching undertaken in Scopus. Articles were included if they were primary research or case reports of ≥ 3 participants reporting the following: recall period, response scale selection, administration modality. Quality was assessed using QualSyst, and results synthesised narratively. This review was conducted and reported according to PRISMA guidelines. RESULTS: 81 of 13,215 retrieved articles met the inclusion criteria. Children < 5 years old cannot validly and reliably self-report health outcomes. Face scales demonstrate better psychometric properties than visual analogue or Likert scales. Computerised and paper scales generally show equivalent construct validity. Children prefer computerised measures. Children ≤ 7 years old think dichotomously so need two response options. Those > 8 years old can reliably use a 3-point scale. CONCLUSION: The results of this review have both clinical and research implications. They can be used to inform appropriate choice of PROM for use with CYP in the clinical setting. We also give eight recommendations for future development of self-reported outcome measures for children and young people.

Plagiarism and data falsification are the most common reasons for retracted publications in obstetrics and gynaecology.

OBJECTIVE: To determine the number of retracted articles and to examine the reasons for retraction within the obstetrics and gynaecology literature. DESIGN: Retrospective review of the PubMed database. SETTING: N/A. POPULATION: Obstetrics and gynaecology articles published from indexation until June 2018. METHODS: Articles were identified using keywords for retracted articles in obstetrics and gynaecology. Descriptive statistics were performed. MAIN OUTCOME MEASURES: Incidence of article retraction, the reasons given for retraction, and article demographics. RESULTS: A total of 176 articles were identified with a median time to retraction of 2 years; over three-quarters were retracted within the last decade (n = 136; 77.3%). The median journal impact factor was 2.5 (range of 0.26-52.67). Subspecialties with the highest number of retractions were gynaecological oncology (n = 76; 43.2%), gynaecology (n = 36; 20.5%), and obstetrics (n = 31; 17.6%). Among 176 first authors, 18 authors (10.2%) had two or more retracted articles. Clinical research articles accounted for approximately one-half of the retracted articles (n = 87; 49.4%). Among the clinical studies, 10.2% (n = 18) were randomised control trials, 16.5% (n = 29) were prospective trials and 13.1% (n = 23) were retrospective studies. Plagiarism (n = 40; 22.7%) and data falsification (n = 37; 21.0%) were the most common reasons given for retraction. CONCLUSION: Article retraction within the obstetrics and gynaecology literature is increasing. The most frequently cited reasons for article retractions were plagiarism, errors in data, fabricated results, article duplication, and compromised peer review. Consequences of article retractions to patient care and the scientific community can be significant. The thorough screening of manuscripts prior to publication should be prioritised. TWEETABLE ABSTRACT: The rate of retraction in obstetrics and gynaecology is increasing; the most common reason for retraction is plagiarism.

Effects of personal air pollution exposure on asthma symptoms, lung function and airway inflammation.

BACKGROUND: There is evidence that air pollution increases the risk of asthma hospitalizations and healthcare utilization, but the effects on day-to-day asthma control are not fully understood. OBJECTIVE: We undertook a prospective single-centre panel study to test the hypothesis that personal air pollution exposure is associated with asthma symptoms, lung function and airway inflammation. METHODS: Thirty-two patients with a clinical diagnosis of asthma were provided with a personal air pollution monitor (Cairclip NO2 /O3 ) which was kept on or around their person throughout the 12-week follow-up period. Ambient levels of NO2 and particulate matter were modelled based upon satellite imaging data. Directly measured ozone, NO2 and particulate matter levels were obtained from a monitoring station in central Leicester. Participants made daily electronic records of asthma symptoms, peak expiratory flow and exhaled nitric oxide. Spirometry and asthma symptom questionnaires were completed at fortnightly study visits. Data were analysed using linear mixed effects models and cross-correlation. RESULTS: Cairclip exposure data were of good quality with clear evidence of diurnal variability and a missing data rate of approximately 20%. We were unable to detect consistent relationships between personal air pollution exposure and clinical outcomes in the group as a whole. In an exploratory subgroup analysis, total oxidant exposure was associated with increased daytime symptoms in women but not men. CONCLUSIONS AND CLINICAL RELEVANCE: We did not find compelling evidence that air pollution exposure impacts on day-to-day clinical control in an unselected asthma population, but further studies are required in larger populations with higher exposure levels. Women may be more susceptible than men to the effects of air pollution, an observation which requires confirmation in future studies.

Increased FcγRIIB dominance contributes to the emergence of resistance to therapeutic antibodies in chronic lymphocytic leukaemia patients.

Resistance to therapeutic antibodies in chronic lymphocytic leukaemia (CLL) is common. In this study, we show that therapeutic antibodies against CD62L (CD62L-Ab) or CD20 (obinutuzumab) were able to induce antibody-dependent cell-mediated cytotoxicity (ADCC) and phagocytosis (ADP) in primary cultures of CLL cells. CLL cells derived from patients with active disease requiring treatment displayed resistance to these antibodies, whereas patients with stable disease were sensitive. Using enrichment strategies and transcriptomic analyses, we show that antibody-dependent tumour cell killing was FcγR-dependent and mediated by macrophages. Moreover, we show that resistance cannot be attributed to total numbers or established subtypes of monocytes/macrophages, or the efficiency with which they bind an immune complex. Rather, ADCC/ADP resistance was due to reduced signalling activity through the activating FcγRs resulting in the transfer of dominance to the inhibitory FcγRIIb within macrophages. Most significantly, we show that resistance is an actionable event that could be reversed using inhibitors of FcγRIIb signalling in primary cultures of CLL cells that were previously insensitive to obinutuzumab or CD62L-Ab.

Beyond expectations: the physiological basis of sensory enhancement of satiety.

BACKGROUND/OBJECTIVES: Consumption of high-energy beverages has been implicated as a risk factor for weight gain, yet why nutrients ingested as beverages fail to generate adequate satiety remains unclear. In general, consumers do not expect drinks to be satiating, but drinks generate greater satiety when their sensory characteristics imply they may be filling. These findings challenge traditional bottom-up models of how gut-based satiety signals modify behaviour to suggest that beliefs at the point of ingestion modify gut-based satiety signalling. SUBJECTS/METHODS: Healthy volunteers (n=23) consumed four different beverages, combining an overt sensory manipulation (thin, low sensory (LS) or thicker and more creamy, enhanced sensory (ES)) and covert nutrient manipulation (low energy (LE), 78 kcal; high energy (HE), 267 kcal) on different days. Effects on satiety were assessed through rated appetite and levels of glucose, insulin, pancreatic polypeptide (PP) and cholesystokinin (CCK) recorded periodically over 90 min, and through intake at an ad libitum test lunch. RESULTS: Intake at the test lunch and rated appetite were both altered by both the sensory and nutrient manipulations, with lowest intake and greatest suppression of hunger post-drink in the ESHE condition. Insulin increased more after HE than LE drinks, and after ES than LS drinks, whereas PP levels were higher after ES than LS versions. CCK levels only increased after the ESHE drink. CONCLUSIONS: These data confirm acute sensitivity of satiety after consuming a drink both to the sensory characteristics and nutrient content of the drink, and suggest that this may be, at least in part, due to top-down modulation of release of satiety-related gut hormones.

Effects of ambient air temperature, humidity and rainfall on annual survival of adult little penguins Eudyptula minor in southeastern Australia.

Seabirds are subject to the influences of local climate variables during periods of land-based activities such as breeding and, for some species, moult; particularly if they undergo a catastrophic moult (complete simultaneous moult) as do penguins. We investigated potential relationships between adult penguin survival and land-based climate variables (ambient air temperature, humidity and rainfall) using 46 years of mark-recapture data of little penguins Eudyptula minor gathered at a breeding colony on Phillip Island in southeastern Australia. Our results showed that adult penguin survival had a stronger association with land-based climate variables during the moult period, when birds were unable to go to sea for up to 3 weeks, than during the breeding period, when birds could sacrifice breeding success in favour of survival. Annual adult survival probability was positively associated with humidity during moult and negatively associated with rainfall during moult. Prolonged heat during breeding and moult had a negative association with annual adult survival. Local climate projections suggest increasing days of high temperatures, fewer days of rainfall which will result in more droughts (and by implication, lower humidity) and more extreme rainfall events. All of these predicted climate changes are expected to have a negative impact on adult penguin survival.

A fish perspective: detecting flow features while moving using an artificial lateral line in steady and unsteady flow.

For underwater vehicles to successfully detect and navigate turbulent flows, sensing the fluid interactions that occur is required. Fish possess a unique sensory organ called the lateral line. Sensory units called neuromasts are distributed over their body, and provide fish with flow-related information. In this study, a three-dimensional fish-shaped head, instrumented with pressure sensors, was used to investigate the pressure signals for relevant hydrodynamic stimuli to an artificial lateral line system. Unsteady wakes were sensed with the objective to detect the edges of the hydrodynamic trail and then explore and characterize the periodicity of the vorticity. The investigated wakes (Kármán vortex streets) were formed behind a range of cylinder diameter sizes (2.5, 4.5 and 10 cm) and flow velocities (9.9, 19.6 and 26.1 cm s(-1)). Results highlight that moving in the flow is advantageous to characterize the flow environment when compared with static analysis. The pressure difference from foremost to side sensors in the frontal plane provides us a useful measure of transition from steady to unsteady flow. The vortex shedding frequency (VSF) and its magnitude can be used to differentiate the source size and flow speed. Moreover, the distribution of the sensing array vertically as well as the laterally allows the Kármán vortex paired vortices to be detected in the pressure signal as twice the VSF.

Pathology of runting in farmed saltwater crocodiles (Crocodylus porosus) in Australia.

Extremely poor growth of some individuals within a birth cohort (runting) is a significant problem in crocodile farming. We conducted a pathological investigation to determine if infectious disease is associated with runting in farmed saltwater crocodiles (Crocodylus porosus) and to look for evidence of other etiologies. In each of 2005 and 2007, 10 normal and 10 runt crocodiles, with an average age of 5.5 months and reared under identical conditions, were sampled. Laboratory testing included postmortem; histological examination of a wide variety of tissues (with quantitation of features that were noted subjectively to be different between groups); hematology; serum biochemistry (total protein, albumin, globulins, total calcium, phosphorus, and iron); bacterial culture of liver and spleen (2005 only); viral culture of liver, thymus, tonsil, and spleen using primary crocodile cell lines (2007 only); and serum corticosterone (2007 only). The only evidence of infectious disease was mild cutaneous poxvirus infection in 45% of normal and 40% of runt crocodiles and rare intestinal coccidia in 5% of normal and 15% of runt crocodiles. Bacterial and viral culture did not reveal significant differences between the 2 groups. However, runt crocodiles exhibited significant (P < .05) increases in adrenocortical cell cytoplasmic vacuolation and serum corticosterone, decreased production of bone (osteoporosis), and reduced lymphoid populations in the spleen, tonsil, and thymus. Runts also exhibited moderate anemia, hypoalbuminemia, and mild hypophosphatemia. Taken together, these findings suggest an association between runting and a chronic stress response (hyperactivity of the hypothalamic-pituitary-adrenal axis).

Impact of the Ottawa Influenza Decision Aid on healthcare personnel's influenza immunization decision: a randomized trial.

BACKGROUND: Influenza immunization for healthcare personnel reduces frequency and severity of nosocomial influenza outbreaks and influenza-associated morbidity and mortality among patients. The Ottawa Influenza Decision Aid (OIDA) was developed to assist undecided healthcare workers in deciding whether or not to be immunized. AIM: To assess the impact of the OIDA, and to ascertain whether its use would increase the level of confidence in healthcare workers' influenza immunization decision and positively affect their intent to be immunized. METHODS: Single-centre, single-blind, parallel-group, randomized controlled trial. FINDINGS: Eight per cent (151 of 1886) of the unimmunized healthcare personnel were randomized. Of 107 eligible respondents, 48 were in the Ottawa Influenza Decision Aid (OIDA) group and 59 in the control group. A statistically significant (P = 0.020) greater improvement in confidence in immunization decision was observed in the OIDA group compared with the control group. Whereas the odds of changing intent to be immunized from 'no/unsure' to 'yes' was 2.4 times greater in the OIDA group, this result did not reach statistical significance after adjusting for intent to be immunized at baseline. The post-OIDA intent to be immunized in the OIDA and control groups compared to the pre-OIDA intent to be immunized showed that the OIDA had a significant effect on reducing uncertainty (P = 0.035). CONCLUSIONS: Using an accessible, balanced, understandable format for all healthcare personnel about their influenza immunization decision appears to have an impact on both healthcare personnel's confidence in their immunization decision and in their intent to be immunized.

Children's palliative care: a global concern.

Children's palliative care (CPC) is a specialty in itself, albeit closely related to adult palliative care (World Health Organization (WHO), 2002). However, although there are many children who require palliative care, in much of the world CPC has a poor profile and is inaccessible to those who need it (Downing et al, 2010; Knapp et al, 2011). The provision of high-quality palliative care for children is a global concern, with 27% of the world population being under the age of 15, rising to as many as 49% in countries such as Uganda (WHO, 2010). It has been estimated that as many as 7 million of these children around the world will need palliative care each year (Rushton et al, 2002), although the true figure is likely to be higher. The public health approach to palliative care is key to the development of CPC services, as is the development of models that integrate services into existing health structures.

Issues associated with the use of modified texture foods.

Use of modified texture foods (MTF) is common in the geriatric population. There is a potential for increased prevalence of use of MTF due in part to longer survival of persons with dementia, those who have suffered from a stroke, as well as other degenerative diseases that affect chewing and swallowing. Unfortunately, little clinical, nutritional and sensory research has been conducted on MTF to inform practice. This review highlights issues identified in the literature to date that influence nutritional and sensory quality and acceptability of these foods. Use of MTF is highly associated with undernutrition, however causality is difficult to demonstrate due to confounding factors such as the requirement for feeding assistance. Knowledge gaps and considerations that need to be taken into account when conducting research are identified.

Cloning and pharmacological characterization of the dog P2X7 receptor.

BACKGROUND AND PURPOSE: Human and rodent P2X7 receptors exhibit differences in their sensitivity to antagonists. In this study we have cloned and characterized the dog P2X7 receptor to determine if its antagonist sensitivity more closely resembles the human or rodent orthologues. EXPERIMENTAL APPROACH: A cDNA encoding the dog P2X7 receptor was isolated from a dog heart cDNA library, expressed in U-2 OS cells using the BacMam viral expression system and characterized in electrophysiological, ethidium accumulation and radioligand binding studies. Native P2X7 receptors were examined by measuring ATP-stimulated interleukin-1beta release in dog and human whole blood. KEY RESULTS: The dog P2X7 receptor was 595 amino acids long and exhibited high homology (>70%) to the human and rodent orthologues although it contained an additional threonine at position 284 and an amino acid deletion at position 538. ATP possessed low millimolar potency at dog P2X7 receptors. 2'-&3'-O-(4benzoylbenzoyl) ATP had slightly higher potency but was a partial agonist. Dog P2X7 receptors possessed relatively high affinity for a number of selective antagonists of the human P2X7 receptor although there were some differences in potency between the species. Compound affinities in human and dog blood exhibited a similar rank order of potency as observed in studies on the recombinant receptor although absolute potency was considerably lower. CONCLUSIONS AND IMPLICATIONS: Dog recombinant and native P2X7 receptors display a number of pharmacological similarities to the human P2X7 receptor. Thus, dog may be a suitable species for assessing target-related toxicity of antagonists intended for evaluation in the clinic.

Blood pressure variability and prevalence of hypertension using automated readings from multiple visits to a pharmacy-based community-wide programme.

Blood pressure (BP) measurements taken outside the routine office context may be a useful adjunct strategy to monitor BP. Community-based BP data can also provide estimates of the prevalence of elevated BP. We compared multiple readings taken on different days in pharmacies using an automated BpTRU device during a cardiovascular health programme targeting community-dwelling older adults. Mean systolic (S) and diastolic (D) BP values were compared over time using repeated measures analysis of variance for all participants with at least three separate sets of readings (n=317). BP variability was then examined among four subgroups based on report of antihypertensive medication or no treatment, and normal or elevated SBP at the initial visit (< or >or=140, or 130 if diabetes reported). Prevalence of elevated BP was compared across visits. Overall, mean SBP decreased between visits 1 and 2 (140.4 vs 137.1 mm Hg; P<0.001). Among participants with normal SBP at the initial visit, SBP did not vary significantly, whether or not antihypertensive treatment was reported. Those with initially elevated SBP experienced a significant decrease between visits 1 and 2, also regardless of treatment status. Prevalence of elevated BP decreased from visits 1 to 2 (55.8 vs 48.9%; P=0.026) and from visits 1 to 3 (55.8 vs 42.9%; P<0.001). Analyses of BP data from a community-based programme using an accurate device showed that initial readings may inflate the population estimate of elevated BP. Findings suggest that more than one set of BP readings measured on different occasions are needed, particularly if the first set is elevated.

Negative and positive allosteric modulators of the P2X(7) receptor.

BACKGROUND AND PURPOSE: Antagonist effects at the P2X(7) receptor are complex with many behaving in a non-competitive manner. In this study, the effects of N-[2-({2-[(2-hydroxyethyl)amino]ethyl}amino)-5-quinolinyl]-2-tricyclo[3.3.1.1(3,7)]dec-1-ylacetamide (compound-17) and N (2)-(3,4-difluorophenyl)-N (1)-[2-methyl-5-(1-piperazinylmethyl)phenyl]glycinamide dihydrochloride (GW791343) on P2X(7) receptors were examined and their mechanism of action explored. EXPERIMENTAL APPROACH: Antagonist effects were studied by measuring agonist-stimulated ethidium accumulation in cells expressing human or rat recombinant P2X(7) receptors and in radioligand binding studies. KEY RESULTS: Compound-17 and GW791343 were non-competitive inhibitors of human P2X(7) receptors. Receptor protection studies using decavanadate and pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) showed that neither compound-17 nor GW791343 competitively interacted at the ATP binding site and so were probably negative allosteric modulators of the P2X(7) receptor. GW791343 prevented the slowly reversible blockade of the human P2X(7) receptor produced by compound-17 and inhibited [(3)H]-compound-17 binding to the P2X(7) receptor suggesting they may bind to similar or interacting sites. At rat P2X(7) receptors, compound-17 was a negative allosteric modulator but the predominant effect of GW791343 was to increase agonist responses. Antagonist interaction and radioligand binding studies revealed that GW791343 did not interact at the ATP binding site but did interact with the compound-17 binding site suggesting that GW791343 is a positive allosteric modulator of the rat P2X(7) receptor. CONCLUSIONS: Compound-17 was a negative allosteric modulator of human and rat P2X(7) receptors. GW791343 was a negative allosteric modulator of the human P2X(7) receptor but at the rat P2X(7) receptor its predominant effect was positive allosteric modulation. These compounds should provide valuable tools for mechanistic studies on P2X(7) receptors.

Seasonal temperature variation and the rate of donor deferral for low hematocrit in the American Red Cross.

BACKGROUND: Hematocrit (Hct) values in healthy adult populations exhibit seasonal variation, with the lowest values occurring in the summer. The extent to which environmental temperature contributes to the seasonal trend in deferral rates for unacceptable Hct in the American Red Cross was further analyzed. STUDY DESIGN AND METHODS: A centralized database of donations during 2002 to 2004, constituting 24.3 million donor presentations, was further characterized. Data on mean monthly temperature in the United States were obtained for the same period from a government agency. Multivariate regression analyses were performed to determine the relationship between Hct deferral rates among blood donors and environmental temperature and donor characteristics. RESULTS: Hct deferral rates were associated with mean monthly temperature in the United States (R(2) = 0.77). The relationship between the Hct deferral rate and environmental temperature was strongest in the region of the country with the highest seasonal variation in temperature, followed by regions with intermediate and low seasonal variation in temperature, respectively. The seasonal pattern in Hct deferral rates occurred in both sexes and across all age groups, with significantly higher Hct deferral rates occurring in June through August compared to other quarters (p < 0.0007). CONCLUSION: There is a significant seasonal pattern in Hct deferral rates that is associated with environmental temperature. The relationship between Hct deferral rates and temperature is strongest in areas of the country with greater temperature variability. The effect of seasonality on Hct deferrals should be taken into account for donor counseling, recruitment, and retention efforts.

Direct labelling of the human P2X7 receptor and identification of positive and negative cooperativity of binding.

BACKGROUND AND PURPOSE: The P2X(7) receptor exhibits complex pharmacological properties. In this study, binding of a [(3)H]-labelled P2X(7) receptor antagonist to human P2X(7) receptors has been examined to further understand ligand interactions with this receptor. EXPERIMENTAL APPROACH: The P2X(7) receptor antagonist, N-[2-({2-[(2-hydroxyethyl)amino]ethyl}amino)-5-quinolinyl]-2-tricyclo[3.3.1.1(3,7)]dec-1-ylacetamide (compound-17), was radiolabelled with tritium and binding studies were performed using membranes prepared from U-2 OS or HEK293 cells expressing human recombinant P2X(7) receptors. KEY RESULTS: Binding of [(3)H]-compound-17 was higher in membranes prepared from cells expressing P2X(7) receptors than from control cells and was inhibited by ATP suggesting labelled sites represented human P2X(7) receptors. Binding was reversible, saturable and modulated by P2X(7) receptor ligands (Brilliant Blue G, KN62, ATP, decavanadate). Furthermore, ATP potency was reduced in the presence of divalent cations or NaCl. Radioligand binding exhibited both positive and negative cooperativity. Positive cooperativity was evident from bell shaped Scatchard plots, reduction in radioligand dissociation rate by unlabelled compound-17 and enhancement of radioligand binding by KN62 and unlabelled compound-17. ATP and decavanadate inhibited binding in a negative cooperative manner as they enhanced radioligand dissociation. CONCLUSIONS: These data demonstrate that human P2X(7) receptors can be directly labelled and provide novel insights into receptor function. The positive cooperativity observed suggests that binding of compound-17 to one subunit in the P2X(7) receptor complex enhances subsequent binding to other P2X(7) subunits in the same complex. The negative cooperative effects of ATP suggest that ATP and compound-17 bind at separate, interacting, sites on the P2X(7) receptor.

Pharmacological profile of the alpha4beta2 nicotinic acetylcholine receptor partial agonist varenicline, an effective smoking cessation aid.

The preclinical pharmacology of the alpha4beta2 nicotinic acetylcholine receptor (nAChR) partial agonist varenicline, a novel smoking cessation agent is described. Varenicline binds with subnanomolar affinity only to alpha4beta2 nAChRs and in vitro functional patch clamp studies in HEK cells expressing nAChRs show that varenicline is a partial agonist with 45% of nicotine's maximal efficacy at alpha4beta2 nAChRs. In neurochemical models varenicline has significantly lower (40-60%) efficacy than nicotine in stimulating [(3)H]-dopamine release from rat brain slices in vitro and in increasing dopamine release from rat nucleus accumbens in vivo, while it is more potent than nicotine. In addition, when combined with nicotine, varenicline effectively attenuates the nicotine-induced dopamine release to the level of the effect of varenicline alone, consistent with partial agonism. Finally, varenicline reduces nicotine self-administration in rats and supports lower self-administration break points than nicotine. These data suggest that varenicline can reproduce to some extent the subjective effects of smoking by partially activating alpha4beta2 nAChRs, while preventing full activation of these receptors by nicotine. Based on these findings, varenicline was advanced into clinical development and recently shown to be an effective and safe aid for smoking cessation treatment.

Adenosine-evoked Na+ transport in human airway epithelial cells.

BACKGROUND AND PURPOSE: Absorptive epithelia express apical receptors that allow nucleotides to inhibit Na(+) transport but ATP unexpectedly stimulated this process in an absorptive cell line derived from human bronchiolar epithelium (H441 cells) whilst UTP consistently caused inhibition. We have therefore examined the pharmacological basis of this anomalous effect of ATP. EXPERIMENTAL APPROACH: H441 cells were grown on membranes and the short circuit current (I(SC)) measured in Ussing chambers. In some experiments, [Ca(2+)](i) was measured fluorimetrically using Fura -2. mRNAs for adenosine receptors were determined by the polymerase chain reaction (PCR). KEY RESULTS: Cross desensitization experiments showed that the inhibitory response to UTP was abolished by prior exposure to ATP whilst the stimulatory response to ATP persisted in UTP-pre-stimulated cells. Apical adenosine evoked an increase in I(SC) and this response resembled the stimulatory component of the response to ATP, and could be mimicked by adenosine receptor agonists. Pre-stimulation with adenosine abolished the stimulatory component of the response to ATP. mRNA encoding A(1), A(2A) and A(2B) receptor subtypes, but not the A(3) subtype, was detected in H441 cells and adenosine receptor antagonists could abolish the ATP-evoked stimulation of Na(+) absorption. CONCLUSIONS AND IMPLICATIONS: The ATP-induced stimulation of Na(+) absorption seems to be mediated via A(2A/B) receptors activated by adenosine produced from the extracellular hydrolysis of ATP. The present data thus provide the first description of adenosine-evoked Na(+) transport in airway epithelial cells and reveal a previously undocumented aspect of the control of this physiologically important ion transport process.

Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non-healing wounds.

BACKGROUND: Larvae of the greenbottle fly Lucilia sericata are used routinely for the clinical treatment of difficult necrotic and infected wounds. Degradation by proteinases contained in larval excretory/secretory (ES) products is thought to contribute to wound debridement by removal of dead tissue. However, proteinase activity may also affect host tissue remodelling processes. OBJECTIVES: To identify proteolytic enzymes derived from L. sericata ES products with activities against fibrin and extracellular matrix (ECM) components. METHODS: Larval proteinase activities were assayed in vitro using class-specific substrates and inhibitors. Their action against fibrin and ECM components was examined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. RESULTS: Three classes of proteolytic enzyme were detected in the secretions using fluorescein isothiocyanate-labelled casein as a model substrate. The predominant activity belonged to serine proteinases (pH optima 8-9) of two different subclasses (trypsin-like and chymotrypsin-like), with a weaker aspartyl proteinase (pH 5) and a metalloproteinase (pH 9) with exopeptidase characteristics also present. Using skin-relevant ECM components as substrates L. sericata ES products solubilized fibrin clots and degraded fibronectin, laminin and acid-solubilized collagen types I and III. Hydrolysis of ECM macromolecules was inhibited by preincubating ES products with phenylmethylsulphonyl fluoride but not 4-amidinophenylmethylsulphonyl fluoride, indicating that degradation was due to the 'chymotrypsin-like' serine proteinase. CONCLUSIONS: These data suggest that a combination of L. sericata ES proteinases involving chymotrypsin-like and trypsin-like activities could potentially influence wound healing events when maggots are introduced into necrotic and infected wounds, with the chymotrypsin-like activity involved in the remodelling of ECM components.

Ecological basis for fertility control in the house mouse (Mus domesticus) using immunocontraceptive vaccines.

Laboratory studies confirm the potential for fertility control in the house mouse Mus domesticus using mouse cytomegalovirus (MCMV) as a vector for an immunocontraceptive vaccine. This article presents an overview of key results from research in Australia on enclosed and field populations of mice and the associated epidemiology of MCMV. The virus is geographically widespread in Australia. It also persists in low population densities of mice, although if population densities are low for at least a year, transmission of the virus is sporadic until a population threshold of approximately 40 mice ha(-1) is reached. The serological prevalence of MCMV was high early in the breeding season of four field populations. Enclosure studies confirm that MCMV has minimal impact on the survival and breeding performance of mice and that it can be transmitted to most adults within 10-12 weeks. Other enclosure studies indicate that about two-thirds of females would need to be sterilized to provide effective control of the rate of growth of mouse populations. If this level is not maintained for 20-25 weeks after the commencement of breeding, the mouse population can compensate through increased recruitment per breeding female. The findings from this series of descriptive and manipulative population studies of mice support the contention that MCMV would be a good carrier for an immunocontraceptive vaccine required to sustain female sterility levels at or above 65%.