SWATH-MS identification of CXCL7, LBP, TGFß1 and PDGFRß as novel biomarkers in human systemic mastocytosis.

Mastocytosis is a rare myeloproliferative disease, characterised by accumulation of neoplastic mast cells in one or several organs. It presents as cutaneous or systemic. Patients with advanced systemic mastocytosis have a median survival of 3.5 years. The aetiology of mastocytosis is poorly understood, patients present with a broad spectrum of varying clinical symptoms that lack specificity to point clearly to a definitive diagnosis. Discovery of novel blood borne biomarkers would provide a tractable method for rapid identification of mastocytosis and its sub-types. Moving towards this goal, we carried out a clinical biomarker study on blood from twenty individuals (systemic mastocytosis: n = 12, controls: n = 8), which were subjected to global proteome investigation using the novel technology SWATH-MS. This identified several putative biomarkers for systemic mastocytosis. Orthogonal validation of these putative biomarkers was achieved using ELISAs. Utilising this workflow, we identified and validated CXCL7, LBP, TGFß1 and PDGF receptor-ß as novel biomarkers for systemic mastocytosis. We demonstrate that CXCL7 correlates with neutrophil count offering a new insight into the increased prevalence of anaphylaxis in mastocytosis patients. Additionally, demonstrating the utility of SWATH-MS for the discovery of novel biomarkers in the systemic mastocytosis diagnostic sphere.

blaIMP-27 on transferable plasmids in Proteus mirabilis and Providencia rettgeri.

OBJECTIVES: A carbapenem-resistant Providencia rettgeri (PR1) isolate was recovered from a wound infection in Missouri, USA. This isolate possessed an EDTA-inhibitable carbapenemase that was unidentified using the Xpert CARBA-R assay. Our objective was to elucidate the molecular determinant of carbapenem resistance in this isolate. We then sought to test the transmissibility of blaIMP-27 loci in clinical P. rettgeri and Proteus mirabilis isolates. METHODS: In October 2016 the novel ambler Class B carbapenemase blaIMP-27, was reported in two different Proteus mirabilis (PM185 and PM187) isolates. Broth mating assays for transfer of carbapenemase activity were performed for the three clinical isolates with recipient sodium azide-resistant Escherichia coli J53. Antibiotic susceptibility testing and phenotypic carbapenemase activity testing were performed on the clinical isolates, J53 and transconjugants using the Kirby-Bauer disc diffusion method according to CLSI guidelines. Plasmid DNA from PM187, PR1 and their transconjugants were used as input for Nextera Illumina sequencing libraries and sequenced on a NextSeq platform. RESULTS: PR1 was resistant to both imipenem and meropenem. PM187 and PR1 could transfer resistance to E. coli through plasmid conjugation (pPM187 and pPR1). pPM187 had a virB/virD4 type IV secretion system whereas pPR1 had a traB/traD type IV secretion system. CONCLUSION: Two of three blaIMP-27-bearing clinical isolates tested could conjugate resistance into E. coli. The resulting transconjugants became positive for phenotypic carbapenemase production but did not pass clinical resistance breakpoints. blaIMP-27 can be transmitted on different plasmid replicon types that rely on distinct classes of type IV secretion system for horizontal transfer.

Self-presentation in everyday interactions: effects of target familiarity and gender composition.

This study examined people's self-presentation motives in unstructured, everyday social interaction as a function of participants' gender similarity to, and general familiarity with, the targets of their self-presentations. Participants maintained a variant of the Rochester Interaction Record for 1 week. For every interaction that lasted 10 min or more, they rated the degree to which they wanted to make each of 4 impressions (likable, competent, ethical, and attractive), how much they thought about the impressions others in the interaction formed of them, and how nervous they felt in the interaction. In general, participants' self-presentational motives were lower in interactions with highly familiar people of their own sex than they were either in interactions with less familiar people of their sex or in interactions with people of the other sex regardless of familiarity. When participants' interactions with only their 3 most familiar interactants were examined, self-presentational concerns decreased with familiarity in same-sex interactions but increased with familiarity in cross-sex interactions.

An in vitro study of the effect endodontic access preparation has upon the retention of porcelain fused to metal crowns of maxillary central incisors.

Endodontic therapy often requires cutting access preparations through existing restorations. The purpose of this study was to determine whether endodontic access preparation affected the retention of porcelain fused to metal crowns of maxillary central incisors. Retentive forces required to displace porcelain fused to metal crowns of maxillary central incisors were measured using an Instron constant displacement rate testing machine. Using a paired t test analysis, a significant difference was found between crown retention before and after endodontic access preparation.

An in vitro study of the effect endodontic access preparation and amalgam restoration have upon incisor crown retention.

Endodontic access preparation leads to a significant reduction in crown retention. This study sought to determine whether this retention can be regained after access. Eighteen extracted human maxillary incisors had PFM crowns fabricated. Crowns were cemented, retention measured, recemented, access preparations cut, and retention once again measured. Then, crowns were recemented and accesses restored with dental amalgam and new retention measured. This format allowed each crown to be used as a control of itself. For crowns cemented with zinc phosphate cement and restored with amalgam, a 126% increase over original retention was measured. For crowns cemented with polycarboxylate cement and restored with amalgam, a 237% increase over original retention was measured. For crowns cemented with polycarboxylate cement and restored with amalgam, a 237% increase over original retention was measured. These results suggest that recementing crowns secondary to endodontic access and restoring the access with amalgam regains and even surpasses the original retention.

Medication use and health care contacts among symptomatic children with asthma.

OBJECTIVE: Asthma morbidity and mortality continue to increase despite the availability of improved therapies. Little is known about the degree to which children with asthma use medications and health care services during symptomatic periods. This study documents prospectively the use of medications and health care contacts among children with active asthma symptoms. METHODS: Children age 6--19 years from 11 primary care settings in upstate New York were eligible for this study if they had 3 or more asthma-related medical visits during the prior year. We collected extensive information on asthma symptoms, medication use, and contacts with health care providers from biweekly phone interviews and daily diaries during a 3-month period. Symptoms were evaluated as the average number of symptomatic days per week. We tabulated the proportion of children using anti-inflammatory medications and having health care contacts according to the frequency of their symptoms during this 3-month period. Chi-square and regression analyses were used. RESULTS: One hundred sixty-five children participated (67% White, 24% Black, 9% Other). Sixty-five percent of the children in this sample had an average of more than 2 symptomatic days per week or more than 2 symptomatic nights per month during the 3-month study period and thus had mild persistent to severe asthma. Among these children, 25% received prednisone, and 46% reported the use of an inhaled maintenance medication during the monitoring period. Ten percent of children in this sample experienced an average of 6 or more symptomatic days per week during the study period. Among these highly symptomatic children, only 19% received prednisone, and 56% used a maintenance medication. Further, the proportion of children having contact with a health care provider during this 3-month period was 50% or less, even among the children experiencing the most frequent asthma symptoms. There were no differences in the proportion of children with health care contacts, prednisone use, or maintenance anti-inflammatory use among different gender or race categories or with different insurance types or places of residence. CONCLUSIONS: Even among children experiencing almost daily asthma symptoms, inadequate anti-inflammatory therapy is common, and few contacts with health care providers occur. These children are silently suffering at home and likely are experiencing preventable morbidity.

An asthma management program for urban minority children.

Morbidity and mortality continue to increase for children with asthma. Minority children have disproportionately higher rates of adverse outcomes on almost all disease measures. An asthma management program for urban minority children was developed with research-based intervention strategies and insights gained from the child and family perspectives on illness and health care delivery. The goal of the intervention program was to deliver care that was culturally sensitive, focused on decreasing barriers to appropriate self-management, and committed to promoting partnerships among children, families, the health care system, and the broader community.

Illness narratives of children with asthma.

A child's own story of the illness experience is a necessary starting point for intervention strategies and outcomes research. A descriptive study of perceptions of asthma among 28 children (6-18 years old) with moderate or severe asthma was part of a larger study of symptom perception of asthma in childhood. The study sample included 36% Caucasian, 46% African American, and 18% other ethnic groups, encompassing a range of socioeconomic groups. Open-ended, semi-structured interviews were used to elicit children's perceptions of the impact of asthma on their lives, things that bothered them about having asthma, and worries they experienced. Visual analogue scales were used to assess several values. Themes that emerged in data analysis included (a) "I can't," (b) restrictions, (c) symptoms and treatments, (d) death, and (e) adaptation. Overall, asthma had a profound impact on study children, and certain themes were more predominant among subgroups of race and age; notably, the death theme was more prominent among African-American children.

Symptom reporting in childhood asthma: a comparison of assessment methods.

BACKGROUND: One barrier to receiving adequate asthma care is inaccurate estimations of symptom severity. AIMS: To interview parents of children with asthma in order to: (1) describe the range of reported illness severity using three unstructured methods of assessment; (2) determine which assessment method is least likely to result in a "critical error" that could adversely influence the child's care; and (3) determine whether the likelihood of making a "critical error" varies by sociodemographic characteristics. METHODS: A total of 228 parents of children with asthma participated. Clinical status was evaluated using structured questions reflecting National Asthma Education and Prevention Panel (NAEPP) criteria. Unstructured assessments of severity were determined using a visual analogue scale (VAS), a categorical assessment of severity, and a Likert scale assessment of asthma control. A "critical error" was defined as a parent report of symptoms in the lower 50th centile for each method of assessment for children with moderate-severe persistent symptoms by NAEPP criteria. RESULTS: Children with higher severity according to NAEPP criteria were rated on each unstructured assessment as more symptomatic compared to those with less severe symptoms. However, among the children with moderate-severe persistent symptoms, many parents made a critical error and rated children in the lower 50th centile using the VAS (41%), the categorical assessment (45%), and the control assessment (67%). The likelihood of parents making a critical error did not vary by sociodemographic characteristics. CONCLUSIONS: All of the unstructured assessment methods tested yielded underestimations of severity that could adversely influence treatment decisions. Specific symptom questions are needed for accurate severity assessments.

The action of lecithin monolayers on mosquitoes I. General observations.

Monolayers of pure egg-lecithin at maximum equilibrium pressure were tested against eight species of mosquito in the laboratory, Anopheline larvae were prevented from surfacing and died in water of low dissolved oxygen content, but culicine larvae survived. The pupae of all species except Culex p. fatigans were killed. Effects on floating eggs, pupal-adult eclosion and adult behaviour are also reported.

Symptom perception and evaluation in childhood asthma.

BACKGROUND: Inaccuracies in symptom perception may contribute to morbidity and mortality in childhood asthma. OBJECTIVE: To systematically examine the accuracy of symptom perception on the part of children with asthma and their parents, as well as their interpretation and evaluation of the symptoms. METHOD: Twenty-eight patient/parent pairs from suburban and underserved urban pediatric populations participated in a 5-week protocol tracking subjective assessments of asthma severity (visual analog scales) and peak expiratory flow rates. Relationships between perceptual accuracy and demographic and disease factors were investigated. RESULTS: Adolescents were more accurate than school-aged children, more accurate children had better morbidity outcomes, and African American parents were more accurate than Caucasian parents. Socioeconomic status did not affect accuracy. Both children and parents missed early symptoms and waited too long prior to intervening in an exacerbation. CONCLUSIONS: There are multiple opportunities for error in symptom perception and evaluation. Identification of the source of error is critical to the effective utilization of education on self-management.

The biosynthesis of rubber: Incorporation of isopentenyl pyrophosphate into purified rubber particles by a soluble latex serum enzyme.

1. The rubber particles in Hevea brasiliensis latex have been partially purified by ;washing' with buffer solution, and separated into active fractions of different particle size. 2. The enzyme responsible for incorporating isopentenyl pyrophosphate into rubber is distributed between the surface of the rubber particles and the aqueous serum phase of the latex. The enzyme at the surface can be removed or inactivated if the rubber particles are washed sufficiently with buffer solution. Enzyme in the serum phase can be concentrated by fractional precipitation with ammonium sulphate. 3. To incorporate isopentenyl pyrophosphate into rubber in vitro, active rubber particles are required as well as enzyme and soluble cofactors. The activity of the rubber particles per unit surface area increases with diminishing particle size.

Circadian modification of drug-induced teratogenesis in rat fetuses.

Pregnant Sprague-Dawley rats were injected with hydroxyurea (750 mg/kg) or physiological saline on the 12th day of gestation. Hydroxyurea and saline (control) treated groups were each composed of six subgroups injected at consecutive 4-h intervals (i.e., group 1 at 00(00), group 2 at 04(00),...). All females were injected at the same circadian phase as they were mated. The developmental age of all fetuses was 288 +/- 2 hrs at the time of injection. The fetuses were taken by caesarean section on the 20th or 21st day of gestation. Teratogenesis was greatest when hydroxyurea was administered in the light phase (light-dark 12:12 cycle). Deformity rates correlate with motor activity, mitotic rates and DNA synthesis.

Standards and practice guidelines as the foundation for clinical practice.

The structure and organization of health care delivery are in the midst of rapid change. Health care providers from a variety of disciplines are being challenged to define their practice and the expected patient outcomes resulting from their processes of care delivery. Standards and clinical practice guidelines are important tools for enhancing the quality of health care delivery and for documenting care. The article describes a process for developing standards and clinical practice guidelines and presents an organizational scheme for them. Based on recommendations from diverse national groups, a format for practice guidelines is presented, and a system for implementation and ongoing evaluation is recommended.

Alamethicin-induced conductances in lipid bilayers: I. Data analysis and simple steady-state model.

Conductance flucutations induced by low concentrations of alamethicin in planar lipid bilayer membranes have been examined with a novel computer-aided analysis. A precise test is applied to show that the system is Markovian (except for the zeroth state) and an estimate is given for the (small) number of transitions between nonadjacent states: the system closely conforms to a birth-and-death process. The complete set of rate parameters governing the steady-state distribution is derived, and it is shown that these parameters can be used to reconstruct exactly the experimental relative frequency distribution. It is also shown that the electrochemical free energies of the conductance states vary quadratically with state number for low-lying states; the free energies of activation of both the birth and death processes are linearly related to the free-energy differences between states but the transfer coefficient is close to unity. A detailed model based on the nucleation of a two-dimensional pore accounts for these observations provided both birth and death take place via an intermediate expansion of the pore lumen. This model requires two energy parameters, the "edge" and "bulk" energies of the pore, together with a trigger rate of the initial process which is a sufficient description for the steady-state behavior of this voltage-controlled system.

Comparison of two standard chemotherapy regimens for good-prognosis germ-cell tumours: a randomised trial. Australian and New Zealand Germ Cell Trial Group.

BACKGROUND: Most patients with metastatic germ-cell tumours are cured with chemotherapy. However, the optimum chemotherapy regimen is uncertain, and there is variation in international practice. We did a multicentre randomised trial to compare two standard chemotherapy regimens for men with good-prognosis germ-cell tumours. METHODS: Good prognosis was defined by modified Memorial Sloan-Kettering criteria. The first regimen (regimen A) was based on treatment recommendations from Indiana University and comprised three cycles of 20 mg/m2 cisplatin on days 1-5, 100 mg/m2 etoposide on days 1-5, and 30 kU bleomycin on days 1, 8, and 15, repeated every 21 days. The second regimen (regimen B) was based on the control regimen of a published randomised clinical trial and comprised four cycles of 100 mg/m2 cisplatin on day 1, 120 mg/m2 etoposide on days 1-3, and 30 kU bleomycin on day 1, repeated every 21 days. The primary outcome measure was overall survival. Analysis was by intention to treat. FINDINGS: 166 patients were randomised, 83 to each regimen. The trial was stopped when the second planned interim analysis met predefined stopping rules. The median follow-up was 33 months. Overall survival was substantially better with regimen A (three vs 13 deaths, hazard ratio 0.22 [95% CI 0.06-0.77], p=0.008). This difference was due to deaths from cancer (one vs nine), and not deaths from treatment (two vs two) and remained significant after adjustment for other prognostic factors (0.25 [0.07-0.88], p=0.03). INTERPRETATION: In men with good-prognosis germ-cell tumours, the regimen developed at Indiana University is superior to the alternative regimen studied in this trial. The lower total dose and dose-intensity of bleomycin and the lower dose-intensity of etoposide in regimen B could be responsible for the worse outcome.