Inhaled antibiotics for the treatment of chronic Pseudomonas aeruginosa infection in cystic fibrosis patients: challenges to treatment adherence and strategies to improve outcomes

Réka Bodnár,1,2 Ágnes Mészáros,2 Máté Oláh,2 Tamás Ágh3 1Department of Pediatric Pulmonology, Heim Pál Children’s Hospital, Budapest, Hungary; 2University Pharmacy Department of Pharmacy Administration, Semmelweis University, Budapest, Hungary; 3Syreon Research Institute, Budapest, Hungary Background: Inhaled antibiotics (ABs) are recommended for use in the therapy of chronic Pseudomonas aeruginosa infection in patients with cystic fibrosis (CF). The aim of this systematic literature review was to identify level of adherence to inhaled ABs and to determine predictors and consequences of nonadherence in CF. Methods: A systematic literature search of English-language articles was conducted in April 2015 using Medline and Embase. No publication date limit was applied. The literature screening was conducted by two independent reviewers. All of the included studies were assessed for quality. Results: The search yielded 193 publications, of which ten met the inclusion criteria and underwent data extraction. Seven studies focused on inhaled tobramycin, one on inhaled colistimethate, one on inhaled levofloxacin, and one on inhaled aztreonam lysine. Medication adherence to inhaled ABs was analyzed by pharmacy refill history, daily phone diary, parent and child self-reports, vials counting, or electronic monitoring. In randomized controlled trials (n=3), proportion of adherent patients (>75%–80% of required doses taken) ranged from 86% to 97%; in prospective cohort studies (n=3), adherence rates ranged between 36% and 92%, and in retrospective studies (n=4) it ranged between 60% and 70%. The adherence to inhaled ABs in CF was found to be associated with the complexity of treatment, time of drug administration, age of patients, treatment burden (adverse events, taste), and patient satisfaction. Conclusion: The high diversity of adherence data was because of the different study designs (randomized controlled trials vs real-world studies) and the lack of a commonly accepted consensus on the definition of adherence in the reviewed articles. Routine adherence monitoring during CF care, discussing the possible reasons of suboptimal adherence with the patient, and changing treatment regimens on the basis of patient burden can individualize CF therapy for patients and may improve the level of adherence. Keywords: cystic fibrosis, antibiotics, adherence, compliance, Pseudomonas aeruginos

On compatible linear connections of two-dimensional generalized Berwald manifolds: a classical approach

summary:In the paper we characterize the two-dimensional generalized Berwald manifolds in terms of the classical setting of Finsler surfaces (Berwald frame, main scalar etc.). As an application we prove that if a Landsberg surface is a generalized Berwald manifold then it must be a Berwald manifold. Especially, we reproduce Wagner's original result in honor of the 75th anniversary of publishing his pioneering work about generalized Berwald manifolds

Tumorgenezis és a szomatosztatin hatásmechanizmusának vizsgálata humán hipofízis adenomákban = Tumorigenesis and the effects of somatostatin in human pituitary adenomas

Protein western array módszerrel vizsgáltuk ép és tumoros hypophysis mintákból a fehérjeexpressziót. Humán hypophysis szövetben korábban nem ismert 116 fehérjét detektáltunk, továbbá a hypophysis tumogenezisében feltételezhetően szerepet játszó fehérjéket. A Raf/MEK/ERK és a P13K/Akt/mTOR kaszkádok kulcsfontosságú jelátviteli utak -egymással interakcióban- a sejtnövekedés és tumorgenezis szabályozásában. Hypophysis adenomák vizsgálata során mindkét útvonal kezdeti kaszkádjának felülregulációjára következtethettünk. Az IGF II mRNA binding protein 3 (IMP3) expressziója általában korrelál különböző tumorok agresszivitásával. Vizsgálatunk alapján az IMP 3 mind normális, mind neoplasiás (adenoma, carcinoma) hypophysisben hasonló mértékben expresszálódik. A familiáris izololált hypophysisadenoma (FIPA) autoszomális dominánsan öröklődő megbetegedés. A családok 25%-ában mutatható ki a tumor szuppresszor AIP gén mutációja, amely eltérő fenotípussal jár. A hazánkban eddig identifikált FIPA családok részletes klinikai, genetikai vizsgálatait végeztük el. Nem tisztázott, hogy a hypophysis carcinomák de novo képződnek vagy adenomából alakulnak ki. Egy fiatal hypophysis daganatos nőbeteg 14 éves kórtörténete, a daganat (végül ACTH termelő hypophysis carcinoma) radiológiai, műtéti, morfológiai, hisztokémiai leletei alapján megállapítottuk, hogy a tumor kezdettől fogva agresszív volt, azonban de novo carcinoma jelenléte nem volt igazolható. | Using protein western array to analyse the protein expression of normal and adenomatous pituitary samples disclosed both novel proteins (116) and putative protein candidates for pituitary tumorigenesis. Raf/MEK/ERK and P13K/Akt/mTOR cascades are key signalling pathways interacting with each other to regulate cell growth and tumorigenesis. Our results indicate that in pituitary adenomas both pathways are up-regulated in the initial cascade of the pathways. The IGF II mRNA binding protein 3 (IMP3) is correlated with increased tumor agressiveness. Our results indicate that IMP3 is expressed both in normal and in nepolastic (adenoma, carcinoma) pituitary gland tissues without significant differences in expression levels. Familial isolated pituitary adenoma (FIPA) syndrome is an autosomal dominant disease. In 25% of families with this syndrome, the tumor suppressor AIP gene mutation was identified, these families have distinct phenotype. The FIPA families, identified so far in Hungary, were analysed in detail. It is unclear whether pituitary carcinomas arise de novo or evolve from adenomas. Studying the 14y clinical course, radiographic and operative findings, morphologic/immunohistochemical features of the pituitary tumor (finally an ACTH secreting carcinoma) of a young patient, all suggested that it was an aggressive tumor from the outset, but de novo carcinoma could not be confirme

Lipase B from Candida antarctica Immobilized on Epoxy-functionalized Hollow Silica Microspheres: Efficient Biocatalysts for Enantiomer Selective Acylation of Alcohols and Amines

Hollow silica microspheres with promising physical properties (MAT540TM) as support for enzyme immobilization and biocatalyst were investigated in this study. The amine-functionalized MAT540TM was activated by six bisepoxides inclosing different spacers and used as epoxy-functionalized carrier for immobilization of lipase B from Candida antarctica (CaLB). The novel, covalently fixed CaLB biocatalysts were compared in kinetic resolution (KR) of racemic 1-phenyethanol rac-1 and five racemic amines rac-3a-e using shaken flasks and continuous-flow packed-bed microreactors. Mechanic stability, re-usability and the effect of temperature (0–90 °C) on productivity and enantiomer selectivity of the covalently immobilized CaLB were investigated. The best performing CaLB biocatalyst showed good mechanic stability after 24 h operation time in continuous-flow mode at 60 °C and provided in KRs of racemic 1-phenyethanol rac-1 with vinyl acetate and of five racemic amines with isopropyl 2-ethoxyacetate as acylating agent the non-reacted (S)-alcohol [(S)-1] or (S)-amines [(S)-3a-e] and the forming (R)-ester [(R)-2] or (R)-amide [(R)-4a-e] in good yields with high enantiomeric excess (ee > 99 %, for all)

Expression and purification of recombinant phenylalanine ammonia-lyase from Petroselinum crispum

In the present study the molecular cloning, expression and purification of recombinant PcPAL, with a cleavable N-terminal His-tag is described. The PcPAL gene was cloned into pET-19b vector and transformed to different E.coli host cells. The optimization of expression and purification processes provided recombinant protein with high purity in its native, tetrameric fold with a yield of 7-8 mg protein / 1 L culture. The activity of the recombinant protein was tested towards its natural substrate L-Phe, the KM, and kcat values suggesting excellent catalytic properties of the recombinant enzyme

Immobilization engineering – How to design advanced sol-gel systems for biocatalysis?

An immobilization engineering approach using bioinformatics and experimental design tools was applied to improve the sol–gel enzyme entrapment methodology. This strategy was used for the immobilization of lipase B from Candida antarctica (CaLB), a versatile enzyme widely used even on the industrial scale. The optimized entrapment of CaLB in sol–gel matrices is reported by the response-surface methodology enabling efficient process development. The immobilized CaLBs characterized by functional efficiency and enhanced recovery provided economical and green options for flow chemistry. Various ternary mixtures of sol–gel precursors allowed the creation of tailored entrapment matrices best suited for the enzyme and its targeted substrate. The sol–gel-entrapped forms of CaLB were excellent biocatalysts in the kinetic resolutions of secondary alcohols and secondary amines with aromatic or aliphatic substituents both in batch and continuous-flow biotransformations

Characterisation of bioenergetic pathways and related regulators by multiple assays in human tumour cells

Background: Alterations in cellular metabolism are considered as hallmarks of cancers, however, to recognize these alterations and understand their mechanisms appropriate techniques are required. Our hypothesis was to determine whether dominant bioenergetic mechanism may be estimated by comparing the substrate utilisation with different methods to detect the labelled carbon incorporation and their application in tumour cells. Methods: To define the bioenergetic pathways different metabolic tests were applied: (a) measuring CO2 production from [1-14C]-glucose and [1-14C]-acetate; (b) studying the effect of glucose and acetate on adenylate energy charge; (c) analysing glycolytic and TCA cycle metabolites and the number of incorporated 13C atoms after [U-13C]-glucose/[2-13C]-acetate labelling. Based on [1-14C]-substrate oxidation two selected cell lines out of seven were analysed in details, in which the highest difference was detected at their substrate utilization. To elucidate the relevance of metabolic characterisation the expression of certain regulatory factors, bioenergetic enzymes, mammalian target of rapamycin (mTOR) complexes (C1/C2) and related targets as important elements at the crossroad of cellular signalling network were also investigated. Results: Both [U-13C]-glucose and [1-14C]-substrate labelling indicated high glycolytic capacity of tumour cells. However, the ratio of certain 13C-labelled metabolites showed detailed metabolic differences in the two selected cell lines in further characterisation. The detected differences of GAPDH, β-F1-ATP-ase expression and adenylate energy charge in HT-1080 and ZR-75.1 tumour cells also confirmed the altered metabolism. Moreover, the highly limited labelling of citrate by [2-13C]-acetate-representing a novel functional test in malignant cells-confirmed the defect of TCA cycle of HT-1080 in contrast to ZR-75.1 cells. Noteworthy, the impaired TCA cycle in HT-1080 cells were associated with high mTORC1 activity, negligible protein level and activity of mTORC2, high expression of interleukin-1β, interleukin-6 and heme oxygenase-1 which may contribute to the compensatory mechanism of TCA deficiency. Conclusions: The applied methods of energy substrate utilisation and other measurements represent simple assay system using 13C-acetate and glucose to recognize dominant bioenergetic pathways in tumour cells. These may offer a possibility to characterise metabolic subtypes of human tumours and provide guidelines to find biomarkers for prediction and development of new metabolism related targets in personalized therapy. © 2016 Jeney et al