Drug-induced trafficking of p-glycoprotein in human brain capillary endothelial cells as demonstrated by exposure to mitomycin C.

P-glycoprotein (Pgp; ABCB1/MDR1) is a major efflux transporter at the blood-brain barrier (BBB), restricting the penetration of various compounds. In other tissues, trafficking of Pgp from subcellular stores to the cell surface has been demonstrated and may constitute a rapid way of the cell to respond to toxic compounds by functional membrane insertion of the transporter. It is not known whether drug-induced Pgp trafficking also occurs in brain capillary endothelial cells that form the BBB. In this study, trafficking of Pgp was investigated in human brain capillary endothelial cells (hCMEC/D3) that were stably transfected with a doxycycline-inducible MDR1-EGFP fusion plasmid. In the presence of doxycycline, these cells exhibited a 15-fold increase in Pgp-EGFP fusion protein expression, which was associated with an increased efflux of the Pgp substrate rhodamine 123 (Rho123). The chemotherapeutic agent mitomycin C (MMC) was used to study drug-induced trafficking of Pgp. Confocal fluorescence microscopy of single hCMEC/D3-MDR1-EGFP cells revealed that Pgp redistribution from intracellular pools to the cell surface occurred within 2 h of MMC exposure. Pgp-EGFP exhibited a punctuate pattern at the cell surface compatible with concentrated regions of the fusion protein in membrane microdomains, i.e., lipid rafts, which was confirmed by Western blot analysis of biotinylated cell surface proteins in Lubrol-resistant membranes. MMC exposure also increased the functionality of Pgp as assessed in three functional assays with Pgp substrates (Rho123, eFluxx-ID Gold, calcein-AM). However, this increase occurred with some delay after the increased Pgp expression and coincided with the release of Pgp from the Lubrol-resistant membrane complexes. Disrupting rafts by depleting the membrane of cholesterol increased the functionality of Pgp. Our data present the first direct evidence of drug-induced Pgp trafficking at the human BBB and indicate that Pgp has to be released from lipid rafts to gain its full functionality

Age-dependent and sex-dependent disparity in mortality in patients with adrenal incidentalomas and autonomous cortisol secretion: an international, retrospective, cohort study

Background: The association between cortisol secretion and mortality in patients with adrenal incidentalomas is controversial. We aimed to assess all-cause mortality, prevalence of comorbidities, and occurrence of cardiovascular events in uniformly stratified patients with adrenal incidentalomas and cortisol autonomy (defined as non-suppressible serum cortisol on dexamethasone suppression testing). Methods: We conducted an international, retrospective, cohort study (NAPACA Outcome) at 30 centres in 16 countries. Eligible patients were aged 18 years or older with an adrenal incidentaloma (diameter ≥1 cm) detected between Jan 1, 1996, and Dec 31, 2015, and availability of a 1 mg dexamethasone suppression test result from the time of the initial diagnosis. Patients with clinically apparent hormone excess, active malignancy, or follow-up of less than 36 months were excluded. Patients were stratified according to the 0800–0900 h serum cortisol values after an overnight 1 mg dexamethasone suppression test; less than 50 nmol/L was classed as non-functioning adenoma, 50–138 nmol/L as possible autonomous cortisol secretion, and greater than 138 nmol/L as autonomous cortisol secretion. The primary endpoint was all-cause mortality. Secondary endpoints were the prevalence of cardiometabolic comorbidities, cardiovascular events, and cause-specific mortality. The primary and secondary endpoints were assessed in all study participants. Findings: Of 4374 potentially eligible patients, 3656 (2089 [57·1%] with non-functioning adenoma, 1320 [36·1%] with possible autonomous cortisol secretion, and 247 [6·8%] with autonomous cortisol secretion) were included in the study cohort for mortality analysis (2350 [64·3%] women and 1306 [35·7%] men; median age 61 years [IQR 53–68]; median follow-up 7·0 years [IQR 4·7–10·2]). During follow-up, 352 (9·6%) patients died. All-cause mortality (adjusted for age, sex, comorbidities, and previous cardiovascular events) was significantly increased in patients with possible autonomous cortisol secretion (HR 1·52, 95% CI 1·19–1·94) and autonomous cortisol secretion (1·77, 1·20–2·62) compared with patients with non-functioning adenoma. In women younger than 65 years, autonomous cortisol secretion was associated with higher all-cause mortality than non-functioning adenoma (HR 4·39, 95% CI 1·93–9·96), although this was not observed in men. Cardiometabolic comorbidities were significantly less frequent with non-functioning adenoma than with possible autonomous cortisol secretion and autonomous cortisol secretion (hypertension occurred in 1186 [58·6%] of 2024 patients with non-functioning adenoma, 944 [74·0%] of 1275 with possible autonomous cortisol secretion, and 179 [75·2%] of 238 with autonomous cortisol secretion; dyslipidaemia occurred in 724 [36·2%] of 1999 patients, 547 [43·8%] of 1250, and 123 [51·9%] of 237; and any diabetes occurred in 365 [18·2%] of 2002, 288 [23·0%] of 1250, and 62 [26·7%] of 232; all p values <0·001). Interpretation: Cortisol autonomy is associated with increased all-cause mortality, particularly in women younger than 65 years. However, until results from randomised interventional trials are available, a conservative therapeutic approach seems to be justified in most patients with adrenal incidentaloma. Funding: Deutsche Forschungsgemeinschaft, Associazione Italiana per la Ricerca sul Cancro, Università di Torino

On designing of diaphragm walls

W pracy przedstawiono proces weryfikacji i porównania obliczeń otrzymanych z programu bazującego na metodzie elementów skończonych z wynikami analitycznymi dla analizowanej ściany szczelinowej z przyporą ziemną. Dodatkowo autorzy dokonali krótkiego przeglądu literatury z zakresu ścian szczelinowych.The paper presents the process of verification and comparison of calculations received from the program based on the finite element method with analytical results for the analyzed diaphragm wall with an earth buttress. In addition, the authors have prepared a short review of literature in the field of diaphragm walls

Impact of starch mixture composition on properties of gluten-free bread

Celem podjętych badań było opracowanie składu mieszanek skrobiowych, przeznaczonych do wypieku chleba bezglutenowego, umożliwiających: uzyskanie poprawy właściwości fizycznych ciasta, podwyższenie jakości chleba i osiągnięcie korzystnego bilansu białka i błonnika pokarmowego. Podstawę mieszanek bezglutenowych stanowiły: skrobia pszenna, kukurydziana, ziemniaczana oraz mąka kukurydziana. Na podstawie charakterystyki amylograficznej badanych surowców opracowano skład mieszanki o lepkości 470 j.B., cechującej się dużą stabilnością. Mieszankę wzbogacano w błonnik pokarmowy różnego pochodzenia oraz białko sojowe. W chlebach doświadczalnych uzyskano zarówno bardziej miękką skórkę, jak i miękisz niż w pieczywie kontrolnym. W chlebach tych stwierdzono mniejsze ubytki wilgoci i wolniejszą utratę elastyczności miękiszu podczas prób przechowalniczych. Spowolnienie procesu starzenia się pieczywa, wyprodukowanego z opracowanych mieszanek, było wyraźnie związane z użyciem błonnika w recepturze ciasta. Udało się jednocześnie uzyskać zwiększenie wartości żywieniowej chleba bezglutenowego, gdyż zawarta w nim ilość błonnika pokarmowego odpowiada wartościom charakterystycznym dla tradycyjnego chleba razowego. Modyfikacje recepturowe spowodowały jednocześnie zmniejszenie wartości energetycznej uzyskiwanych chlebów, proporcjonalne do zastosowanej dawki błonnika. Chleby bezglutenowe otrzymane według proponowanych receptur charakteryzowały się równocześnie wydatnie zwiększoną zawartością białka, odpowiadającą poziomowi typowemu dla wyrobów pszennych. Ocena punktowa wykazała, że są to produkty w pełni akceptowane przez konsumentów. Uzyskały one znacznie wyższy stopień pożądalności konsumenckiej (89–93 %) niż chleb wyprodukowany z gotowej mieszanki handlowej (71 %).The objective of the study was to develop a composition of starch mixtures intended for baking gluten-free bread, which could contribute to the improvement of both the physical properties of the dough and the quality of the final bread, and to obtaining a favourable balance of protein and dietary fibre. Gluten-free mixtures were based on wheat, maize, and potato starches, and on maize flour. On the basis of amylographic profile of the raw materials studied, a composition of the mixture was developed that showed a viscosity of 470 B.U. and a high stability. The mixture was supplemented with dietary fibre of different origin and with soy protein. The experimental breads were characterized by both the softer crust and crumb compared to the control bread. They also showed the lower water depletion and the slower rate of decrease in the crumb elasticity during their storage. The inhibition of staling process of the bread made with the addition of the mixtures developed was clearly connected with adding dietary fibre to the dough recipe. Simultaneously, it was possible to win an increase in the nutritional value of gluten-free bread since the content of dietary fibre in the dough corresponded to the values characteristic for the conventional wholemeal bread. Modifications in the dough recipe also resulted in lowering the energetic value of the bread baked and were proportional to the quantity of fibre added. Furthermore, the gluten-free breads obtained according to the recipes suggested were characterised by the considerably raised protein content matching the level typical for wheat bakery products. The assessment method by points proved that the proposed breads were fully accepted by consumers. They received a much higher degree of the consumer acceptability (89–93 %) than the bread prepared using a ready-to-use mixture available on the market (71 %)

Function of dietary fibre in forming functional properties of gluten free dough and bread

Celem pracy było określenie wpływu suplementacji chleba bezglutenowego preparatami błonnikowymi różnego pochodzenia, wprowadzanymi w postaci past, na właściwości reologiczne ciasta bezglutenowego oraz objętość i porowatość miękiszu chleba. Receptury na próbne wypieki chleba bezglutenowego opracowano tak, aby zwiększyć zawartość błonnika pokarmowego i białka do wartości typowych dla chleba pszenno-żytniego i żytniego. Podstawową mieszankę receptury (80 %) stanowiły skrobie: pszenna, kukurydziana, ziemniaczana oraz mąka kukurydziana. Pozostałe 20 % stanowiły mieszanki preparatów błonnikowych, mąki sojowej i gumy guar. Jako preparaty błonnikowe zastosowano błonnik sojowy lub łubinowy, pektynę i inulinę. Preparaty błonnikowe (15 %) mieszano z mąką sojową (4 %) i gumą guar (1 %), a następnie poddawano hydratacji do konsystencji pasty i w takiej postaci dodawano do ciasta. Pomiarów reologicznych past i ciast dokonano za pomocą analizatora tekstury TA.XT plus. Ciasta poddano fermentacji z udziałem drożdży piekarskich w ciągu 40 min. Chleb wypiekano w temperaturze 200 ºC w ciągu 40 min. Pomiar objętości chleba bezglutenowego wykonano zgodnie z zaleceniami odnośnych norm. Pasty różniły się znacznie pod względem twardości w zależności od źródła pochodzenia błonnika. Najmniejszą twardość wykazywała pasta zawierająca błonnik łubinowy z dodatkiem inuliny. Stwierdzono, że dodatek inuliny do błonnika łubinowego w znacznym stopniu wpłynął na właściwości reologiczne mieszaniny. Pomiary przylepności i adhezji potwierdziły korzystny wpływ błonnika łubinowego na właściwości reologiczne ciast. Najniżej oceniono ciasto z udziałem błonnika sojowego i inuliny. Najkorzystniej na objętość chleba bezglutenowego wpłynęła suplementacja pastą sporządzoną z udziałem błonnika łubinowego.The objective of the study was to assess the impact of supplementing the gluten free bread with fibre preparations of different origin, added in the form of pastes, on the gluten free dough and on the volume and porosity of bread crumb. To make samples of gluten free bread, special recipes were developed. Their specific composition had to increase the content of proteins and dietary fibres to the values appearing typical for wheat-rye and rye bread. The basic mixture of the recipe (80 %) consisted of wheat, maize, and potato starches, and of maize flour. The rest (20 %) were mixtures of the fibre preparations, soya flour, and guar gum. The soya or lupine fibre, pectin, and inulin were used as the dietary fibre preparations. The fibre preparations (15 %) were mixed with soya flour (4 %) and guar gum (1 %) and, then, hydrated to obtain the consistency of paste. In this form, they were added to the gluten free dough. The rheological measurements of the pastes and dough were performed using a TA.XT plus texture analyser system. The doughs were fermented by applying bakery yeast, during a period of 40 min. The bread was baked at 200 °C during a 40 min period. The gluten free bread volume was measured according to the recommended bakery standards. The pastes significantly differed in their hardness values depending on the original source of dietary fibre. The paste containing lupine fibre and the inulin added showed the lowest hardness. It was found that the addition of inulin into lupine fibre significantly impacted the rheological properties of the mixture. The stickiness and adhesion values measured confirmed the advantageous impact of the lupine fibre on the rheological properties of gluten free doughs. The dough supplemented with the soya fibre and inulin was assessed as the worst. The most advantageous effect on the gluten free bread volume was achieved when the dough was supplemented with a paste containing the lupine fibre

Technical and economic evaluating the efficiency of agricultural machinery maintenance service

Metodą zbierania informacji na podstawie uprzednio przygotowanych kwestionariuszy o strukturze zamkniętej i jawnym celu, oraz odpowiednich zestawień i porównań ww. informacji, przeprowadzono badania, na podstawie których oceniono stan zużycia korozyjnego oraz sytuację w zakresie konserwacji oraz obsługi i napraw bieżących maszyn rolniczych. Na podstawie wyników uzyskanych w trakcie badań populacji 970 maszyn, przeprowadzonych w 56 gospodarstwach oceniono, że możliwe jest dokonywanie oceny efektywności obsługi konserwacyjnej maszyn rolniczych przez wyznaczenie zależności pomiędzy technicznie uzasadnionymi nakładami ponoszonymi na konserwację, a stopniem stanu zużycia korozyjnego oraz nakładami ponoszonymi na obsługę i naprawę bieżącą ww. maszyn.Investigations were carried out to state the corrosive wear of agricultural machines as well as to evaluate the situation dealing with their maintenance, service and running repairs. The method of information gathering was based on previously elaborated questionnaires of closed structure and evident purpose, as well as on adequate lists and comparisons of completed data. The results obtained at testing the population of 970 machines on 56 farms, showed the possibility of evaluating the maintenance service efficiency for the farm machines through determination of the dependencies between technically justified inputs on the maintenance and a degree of corrosive wear and running repairs of the farm machines under study

An application of critically refracted longitudinal ultrasonic subsurface waves (LCR) to industrial nondestructive testing of thin walled pipe shaped elements of shock absorbers, axle shafts and transmission shafts

The paper describes the application of critically refracted longitudinal ultrasonic subsurface waves (LCR, known also as lateral, creeping or head waves) to nondestructive flaw detection in friction welding or MAIB welded thin walled pipe shaped parts. The theory, a practical laboratory and industrial applications are presented