Proteomic analysis of serum and sputum analytes distinguishes controlled and poorly controlled asthmatics.

BACKGROUND: A major goal of asthma therapy is to achieve disease control, with maintenance of lung function, reduced need for rescue medication, and prevention of exacerbation. Despite current standard of care, up to 70% of patients with asthma remain poorly controlled. Analysis of serum and sputum biomarkers could offer insights into parameters associated with poor asthma control. OBJECTIVE: To identify signatures as determinants of asthma disease control, we performed proteomics using Olink proximity extension analysis. METHODS: Up to 3 longitudinal serum samples were collected from 23 controlled and 25 poorly controlled asthmatics. Nine of the controlled and 8 of the poorly controlled subjects also provided 2 longitudinal sputum samples. The study included an additional cohort of 9 subjects whose serum was collected within 48 hours of asthma exacerbation. Two separate pre-defined Proseek Multiplex panels (INF and CVDIII) were run to quantify 181 separate protein analytes in serum and sputum. RESULTS: Panels consisting of 9 markers in serum (CCL19, CCL25, CDCP1, CCL11, FGF21, FGF23, Flt3L, IL-10Rß, IL-6) and 16 markers in sputum (tPA, KLK6, RETN, ADA, MMP9, Chit1, GRN, PGLYRP1, MPO, HGF, PRTN3, DNER, PI3, Chi3L1, AZU1, and OPG) distinguished controlled and poorly controlled asthmatics. The sputum analytes were consistent with a pattern of neutrophil activation associated with poor asthma control. The serum analyte profile of the exacerbation cohort resembled that of the controlled group rather than that of the poorly controlled asthmatics, possibly reflecting a therapeutic response to systemic corticosteroids. CONCLUSIONS AND CLINICAL RELEVANCE: Proteomic profiles in serum and sputum distinguished controlled and poorly controlled asthmatics, and were maintained over time. Findings support a link between sputum neutrophil markers and loss of asthma control.

Measured and perceived indices of fluid balance in professional athletes. The use and impact of hydration assessment strategies.

BACKGROUND: To determine athletes perceived and measured indices of fluid balance during training and the influence of hydration strategy use on these parameters. METHODS: Thirty-three professional rugby union players completed a 120 minute training session in hot conditions (35°C, 40% relative humidity). Pre-training hydration status, sweat loss, fluid intake and changes in body mass (BM) were obtained. The use of hydration assessment techniques and players perceptions of fluid intake and sweat loss were obtained via a questionnaire. RESULTS: The majority of players (78%) used urine colour to determine pre-training hydration status but the use of hydration assessment techniques did not influence pre-training hydration status (1.025 ± 0.005 vs. 1.023 ± 0.013 g.ml-1, P = .811). Players underestimated sweat loss (73 ± 17%) to a greater extent than fluid intake (37 ± 28%) which resulted in players perceiving they were in positive fluid balance (0.5 ± 0.8% BM) rather than the measured negative fluid balance (-1.0 ± 0.7% BM). Forty-eight percent of players used hydration monitoring strategies during exercise but no player used changes in BM to help guide fluid replacement. CONCLUSION: Players have difficulty perceiving fluid intake and sweat loss during training. However, the use of hydration monitoring techniques did not affect fluid balance before or during training.

Alterations in motor activity, sleep, and biochemistry in a cycling manic-depressive patient.

Biochemical and electrophysiological factors were studied longitudinally in a rapidly cycling manic-depressive patient. Slow changes in mood, motor activity, sleep, and urinary norepinephrine levels during the course of each depressed and manic episode are reported, as well as rapid alterations in many variables at the time of mood switch. Urinary concentrations of norepinephrine and its metabolite, 3-methoxy-4-hydroxyphenyl glycol (MHPG) were significantly lower in depression than in mania; norepinephrine but not MHPG excretion increased prior to the switch. We postulate that the slow behavioral and biological changes preceding switches in this patient are an important manifestation of the cyclic process in manic-depressive illness.

Progressive effects of cocaine on behavior and central amine metabolism in rhesus monkeys: relationship to kindling and psychosis.

Chronic administration of the same dose of cocaine to rhesus monkeys for up to 6 months was associated with progressive alterations in pathological behavior and increased susceptibility to seizures. Monkeys initially displaying prominent hyperactive stereotypic responses for up to 2 months began to demonstrate increasing amounts of inhibitory behavior, consisting of catalepsy, motor inhibition, and abnormal visual tracking and staring. Four of 13 animals developed increasing intensities of lingual-buccal dyskinesias after 10 weeks of chronic cocaine. Animals initially showing no convulsions to a given dose of cocaine eventually developed convulsions to the same dose, and then displayed an increased frequency of convulsions following subsequent injections. Levels of the dopamine metabolite, homovanillic acid (HVA), in the cisternal cerebrospinal fluid were significantly elevated during both excitatory stereotypic and inhibitory syndromes; a similar trend was observed for HVA after probenecid administration. Only the probenecid-induced accumulations of the serotonin metabolite 5-hydroxyindoleacetic acid, following acute cocaine administration, were significantly elevated. The progressive increases in convulsions, dyskinesias, and the inhibitory syndrome did not appear related to alterations in peak levels of cocaine in plasma or CSF, and a pharmacological kindling model is suggested as an alternate explanation of the data. The study extends the current models of stimulant-induced psychoses by highlighting the progressive alterations in behavior and neurological sequelae and in suggesting that this progressive mechanism may also be important in the development of psychosis in man.

The hedgehog processing pathway is required for NSCLC growth and survival.

Considerable interest has been generated from the results of recent clinical trials using smoothened (SMO) antagonists to inhibit the growth of hedgehog (HH) signaling-dependent tumors. This interest is tempered by the discovery of SMO mutations mediating resistance, underscoring the rationale for developing therapeutic strategies that interrupt HH signaling at levels distinct from those inhibiting SMO function. Here, we demonstrate that HH-dependent non-small cell lung carcinoma (NSCLC) growth is sensitive to blockade of the HH pathway upstream of SMO, at the level of HH ligand processing. Individually, the use of different lentivirally delivered shRNA constructs targeting two functionally distinct HH-processing proteins, skinny hedgehog (SKN) or dispatched-1 (DISP-1), in NSCLC cell lines produced similar decreases in cell proliferation and increased cell death. Further, providing either an exogenous source of processed HH or a SMO agonist reverses these effects. The attenuation of HH processing, by knocking down either of these gene products, also abrogated tumor growth in mouse xenografts. Finally, we extended these findings to primary clinical specimens, showing that SKN is frequently overexpressed in NSCLC and that higher DISP-1 expression is associated with an unfavorable clinical outcome. Our results show a critical role for HH processing in HH-dependent tumors, identifies two potential druggable targets in the HH pathway, and suggest that similar therapeutic strategies could be explored to treat patients harboring HH ligand-dependent cancers.

Cadaveric elbow allografts. A six-year experience.

Transplantation of total elbow allografts has been employed as a salvage procedure in an attempt to provide patients with a useful, painless range of motion of the elbow. Patients who are candidates for this procedure include those with disabling elbow joint symptoms who refuse an arthrodesis or are not candidates for conventional total elbow replacement because of excessive bone loss or young age. Allografts must be subjected to rigid internal fixation. Rush rod fixation used early in this series was associated with a high incidence of nonunion. In this series, ten patients followed for one to six years were provided with a functional elbow. However, long-term results are still unknown. Although not recommended for routine use, this operation is viewed as a salvage procedure. The use of allografts in elbow reconstruction does not preclude subsequent reconstruction with another allograft or fusion. In patients with deficient bone stock, the allografts reestablish bone mass to permit an arthrodesis or reconstructive arthroplasty.

Distribution and metabolism of topically applied ethanolamine.

The distribution and metabolism of topical [14C]ethanolamine was studied in vivo, using athymic nude mice, human skin grafted onto athymic nude mice, and in vitro, using excised pig skin. Ethanolamine was the only radioactive phospholipid base detected in the human skin grafts, in the mouse skin, and in the pig skin. Ethanolamine that penetrated human skin grafts or mouse skin was extensively metabolized in the animal. The liver is a major site for metabolism of ethanolamine, containing over 24% of the applied radioactive dose. The kidneys, lungs, brain, and the heart contained 2.53, 0.55, 0.27, and 0.15% of the dose, respectively. The hepatic phospholipid bases, ethanolamine, choline, and serine, were all highly radioactive. Hepatic, human skin graft, and mouse skin proteins were also highly radioactive. Over 18% of the topical radioactive dose was oxidized to 14CO2 and 4.6% was excreted in the urine over 24 hr. Urea, glycine, serine, choline, and uric acid were the urinary metabolites of ethanolamine. In vitro data showed that a relatively small percentage of the applied dose was lost from the skin by evaporation and percutaneous penetration. The bulk of the dose remained in the epidermis. Radiometric and enzymatic assays suggest that ethanolamine enhances the hydrolysis of topically applied diisopropylfluorophosphate.

Differences in root longevity of some tree species.

Although the importance of root production and mortality to nutrient fluxes in ecosystems is widely recognized, the difficulties associated with root measurements have limited the availability of reliable data. We have used minirhizotrons and image analysis to measure root longevity of Prunus avium L., Picea sitchensis (Bong.) Carrière, Acer pseudoplatanus L. and Populus x canadensis cv. Beaupre directly in cohorts of roots. Major differences in the longevity of roots among species were identified. For example, 40% of Prunus avium roots but only 6% of Picea sitchensis roots survived for more than 14 days. Survival analysis of cohorts of roots of Prunus avium and Populus x canadensis revealed differences in the distribution of longevity among cohorts. Genetic, biotic and abiotic factors that may influence longevity are discussed.