Photoactive protochlorophyllide-enzyme complexes reconstituted with PORA, PORB and PORC proteins of A. thaliana : fluorescence and catalytic properties

Photoactive Pchlide-POR-NADPH complexes were reconstituted using protochlorophyllide (Pchlide) and recombinant light-dependent protochlorophyllide oxidoreductase (POR) proteins, His₆-PORA, His₆-PORB and His₆-PORC, from Arabidopsis thaliana. We did not observe any differences in the kinetics of the protochlorophyllide photoreduction at room temperature among the PORA, PORB and PORC proteins. In contrast, the PORC protein showed lower yield of Chlide formation than PORA and PORB when preincubated in the dark for 30 min and then illuminated for a short time. The most significant observation was that reconstituted Pchlide-POR-NADPH complexes showed fluorescence maxima at 77 K similar to those observed for highly aggregated Pchlide-POR-NADPH complexes in prolamellar bodies (PLBs) in vivo. Homology models of PORA, PORB and PORC of Arabidopsis thaliana were developed to compare predicted structures of POR isoforms. There were only slight structural differences, mainly in the organisation of helices and loops, but not in the shape of whole molecules. This is the first comparative analysis of all POR isoforms functioning at different stages of A. thaliana development

Assessment of nutritional status disorders, general fatness and fat distribution in women and men aged 20–30

Wprowadzenie i cel pracy. W ostatnich latach wzrasta zainteresowanie tematyką zaburzeń stanu odżywienia organizmu. Celem głównym pracy była ocena częstości występowania dwóch skrajnych stanów odżywienia organizmu: niedożywienia białkowo-energetycznego (niedoborowej masy ciała) oraz stanu przeżywienia – nadmiaru (nadwaga i otyłość) u młodych kobiet i mężczyzn. W celach szczegółowych uwzględniono ocenę składu tkankowego ciała oraz dystrybucji tkanki tłuszczowej. Materiał i metody. Badaniami objęto łącznie 528 osób (373 kobiety i 155 mężczyzn) w wieku od 20. do 30. roku życia (22,2 ± 3,0 lat). Wykonano podstawowe pomiary antropometryczne: masy ciała (kg), wysokości ciała (cm), obwodów talii i bioder (cm). Na podstawie bezpośrednich pomiarów obliczono: wskaźnik masy ciała BMI (Body Mass Index), wskaźnik dystrybucji tkanki tłuszczowej WHR (Waist to Hip Ratio) oraz WC (Waist), wskaźnik BAI, obliczono procentową zawartość tkanki tłuszczowej w ciele oraz zastosowano metodę bioelektrycznej impedancji. Wyniki. W zbadanej grupie zaobserwowano na podstawie wskaźnika BMI (łącznie u 28,4% zbadanych) występowanie skrajnych stanów odżywienia charakterystycznych dla wychudzenia organizmu, związanego z niedożywieniem białkowo-energetycznym, oraz stany świadczące o przeżywieniu, czyli otyłości, w tym również otyłości II i III stopnia. W toku analiz stwierdzono przeważający (83,3%) odsetek osób o prawidłowej zawartości tkanki tłuszczowej. Odnotowano jednak wśród zbadanych kobiet i mężczyzn przypadki zbyt niskiej zawartości tłuszczu w ciele, jak również zawartości zbyt wysokiej, przekraczającej normę. Otyłość brzuszna (wisceralna) występowała u 40,7% do 48,7% zbadanych osób ogółem, w zależności od zastosowanego wskaźnika. Wśród kobiet za dominujący typ dystrybucji tkanki tłuszczowej (u 66,2%) uznano otłuszczenie pośladkowo-udowe (gynoidalne). Wśród mężczyzn dominował typ androidalny rozmieszczenia tkanki tłuszczowej w ciele. Natomiast na podstawie wyłącznie obwodu talii stwierdzono częstsze o 8,4% występowanie typu brzusznego wśród kobiet w porównaniu do zbadanych mężczyzn. Wnioski. Zaburzenia stanu odżywienia w postaci niedożywienia oraz przeżywienia organizmu występują zarówno wśród kobiet, jak i mężczyzn. Warto rozważyć działania profilaktyczne w kierunku zapobiegania czynnikom warunkującym ten niekorzystny dla zdrowia stan organizmu.Introduction and objective. In recent years, an increase has been observed in the scope of problems concerning nutritional status disorders. The primary goal of the study was the assessment of the frequency of occurrence of two extreme nutritional states: protein-energy malnutrition (deficiency of body mass) and over-eating excess state (overweight and obesity) in young men and women. For specific purposes, assessment of the body tissue composition and fat distribution was undertaken. Material and methods. The studies included a total of 528 people (373 women and 155 men), aged from 20–30 (22.2 ± 3.0) Basic anthropometric measurements were performed: body weight (kg), body height (cm), waist and hips circumferences (cm). Based on direct measurements, body mass index BMI, waist to hip ratio WHE and Waist Ratio WC, BAI index, the percentage content of the fatty tissue in the body was calculated, and the bioelectric impedance method was used. Results. In the examined group (in 28.4%), based on the BMI, the occurrence was observed of extreme nutritional states characteristic for body emaciation connected with protein-energy malnutrition and states evidencing over-nutrition, i.e. obesity, including obesity degree II and III. In the course of analyses, the highest percentage of individual with a normal content of fatty tissue was found (83.3%). However, among the studied men and women, cases were noted of a too low content of fatty tissue in the body, as well as too high – exceeding the standard. Abdominal obesity (visceral) occurred in 40.7 – 48.7% of the total population examined, according to the indicator applied. According to gender, the dominant type of fatty tissue distribution among women (in 66.2%) was gynoidal fatness. Among men dominated the androidal type of fatty tissue distribution. Considering exclusively waist circumference, the more frequent occurrence of the abdominal type by 8.4% was found among women compared men. Conclusions. Disorders of the nutritional state in the form of malnutrition and over-nutrition of the body occur both among men and women. Preventive measures are worth considering towards the prevention of factors conditioning this adverse state of the body

Assessment of nutritional status disorders, general fatness and fat distribution in women and men aged 20-30

Wprowadzenie i cel pracy. W ostatnich latach wzrasta zainteresowanie tematyką zaburzeń stanu odżywienia organizmu. Celem głównym pracy była ocena częstości występowania dwóch skrajnych stanów odżywienia organizmu: niedożywienia białkowo-energetycznego (niedoborowej masy ciała) oraz stanu przeżywienia – nadmiaru (nadwaga i otyłość) u młodych kobiet i mężczyzn. W celach szczegółowych uwzględniono ocenę składu tkankowego ciała oraz dystrybucji tkanki tłuszczowej. Materiał i metody. Badaniami objęto łącznie 528 osób (373 kobiety i 155 mężczyzn) w wieku od 20. do 30. roku życia (22,2 ± 3,0 lat). Wykonano podstawowe pomiary antropometryczne: masy ciała (kg), wysokości ciała (cm), obwodów talii i bioder (cm). Na podstawie bezpośrednich pomiarów obliczono: wskaźnik masy ciała BMI (Body Mass Index), wskaźnik dystrybucji tkanki tłuszczowej WHR (Waist to Hip Ratio) oraz WC (Waist), wskaźnik BAI, obliczono procentową zawartość tkanki tłuszczowej w ciele oraz zastosowano metodę bioelektrycznej impedancji. Wyniki. W zbadanej grupie zaobserwowano na podstawie wskaźnika BMI (łącznie u 28,4% zbadanych) występowanie skrajnych stanów odżywienia charakterystycznych dla wychudzenia organizmu, związanego z niedożywieniem białkowo-energetycznym, oraz stany świadczące o przeżywieniu, czyli otyłości, w tym również otyłości II i III stopnia. W toku analiz stwierdzono przeważający (83,3%) odsetek osób o prawidłowej zawartości tkanki tłuszczowej. Odnotowano jednak wśród zbadanych kobiet i mężczyzn przypadki zbyt niskiej zawartości tłuszczu w ciele, jak również zawartości zbyt wysokiej, przekraczającej normę. Otyłość brzuszna (wisceralna) występowała u 40,7% do 48,7% zbadanych osób ogółem, w zależności od zastosowanego wskaźnika. Wśród kobiet za dominujący typ dystrybucji tkanki tłuszczowej (u 66,2%) uznano otłuszczenie pośladkowo-udowe (gynoidalne). Wśród mężczyzn dominował typ androidalny rozmieszczenia tkanki tłuszczowej w ciele. Natomiast na podstawie wyłącznie obwodu talii stwierdzono częstsze o 8,4% występowanie typu brzusznego wśród kobiet w porównaniu do zbadanych mężczyzn. Wnioski. Zaburzenia stanu odżywienia w postaci niedożywienia oraz przeżywienia organizmu występują zarówno wśród kobiet, jak i mężczyzn. Warto rozważyć działania profilaktyczne w kierunku zapobiegania czynnikom warunkującym ten niekorzystny dla zdrowia stan organizmu.Introduction and objective. In recent years, an increase has been observed in the scope of problems concerning nutritional status disorders. The primary goal of the study was the assessment of the frequency of occurrence of two extreme nutritional states: protein-energy malnutrition (deficiency of body mass) and over-eating excess state (overweight and obesity) in young men and women. For specific purposes, assessment of the body tissue composition and fat distribution was undertaken. Material and methods. The studies included a total of 528 people (373 women and 155 men), aged from 20–30 (22.2 ± 3.0) Basic anthropometric measurements were performed: body weight (kg), body height (cm), waist and hips circumferences (cm). Based on direct measurements, body mass index BMI, waist to hip ratio WHE and Waist Ratio WC, BAI index, the percentage content of the fatty tissue in the body was calculated, and the bioelectric impedance method was used. Results. In the examined group (in 28.4%), based on the BMI, the occurrence was observed of extreme nutritional states characteristic for body emaciation connected with protein-energy malnutrition and states evidencing over-nutrition, i.e. obesity, including obesity degree II and III. In the course of analyses, the highest percentage of individual with a normal content of fatty tissue was found (83.3%). However, among the studied men and women, cases were noted of a too low content of fatty tissue in the body, as well as too high – exceeding the standard. Abdominal obesity (visceral) occurred in 40.7 – 48.7% of the total population examined, according to the indicator applied. According to gender, the dominant type of fatty tissue distribution among women (in 66.2%) was gynoidal fatness. Among men dominated the androidal type of fatty tissue distribution. Considering exclusively waist circumference, the more frequent occurrence of the abdominal type by 8.4% was found among women compared men. Conclusions. Disorders of the nutritional state in the form of malnutrition and over-nutrition of the body occur both among men and women. Preventive measures are worth considering towards the prevention of factors conditioning this adverse state of the body

Independent Lung Ventilation-Experimental Studies on a 3D Printed Respiratory Tract Model

Independent lung ventilation (ILV) is a life-saving procedure in unilateral pulmonary pathologies. ILV is underused in clinical practice, mostly due to the technically demanding placement of a double lumen endotracheal tube (ETT). Moreover, the determination of ventilation parameters for each lung in vivo is limited. In recent years, the development of 3D printing techniques enabled the production of highly accurate physical models of anatomical structures used for in vitro research, considering the high risk of in vivo studies. The purpose of this study was to assess the influence of double-lumen ETT on the gas transport and mixing in the anatomically accurate 3D-printed model of the bronchial tree, with lung lobes of different compliances, using various ventilation modes. The bronchial tree was obtained from Respiratory Drug Delivery (RDD Online, Richmond, VA, USA), processed and printed by a dual extruder FFF 3D printer. The test system was also composed of left side double-lumen endotracheal tube, Siemens Test Lung 190 and anesthetic breathing bag (as lobes). Pressure and flow measurements were taken at the outlets of the secondary bronchus. The measured resistance increased six times in the presence of double-lumen ETT. Differences between the flow distribution to the less and more compliant lobe were more significant for the airways with double-lumen ETT. The ability to predict the actual flow distribution in model airways is necessary to conduct effective ILV in clinical conditions

Patterns of pleural pressure amplitude and respiratory rate changes during therapeutic thoracentesis

Abstract Background Although the impact of therapeutic thoracentesis on lung function and blood gases has been evaluated in several studies, some physiological aspects of pleural fluid withdrawal remain unknown. The aim of the study was to assess the changes in pleural pressure amplitude (Pplampl) during the respiratory cycle and respiratory rate (RR) in patients undergoing pleural fluid withdrawal. Methods The study included 23 patients with symptomatic pleural effusion. Baseline pleural pressure curves were registered with a digital electronic manometer. Then, the registrations were repeated after the withdrawal of consecutive portions of pleural fluid (200 ml up to 1000 ml and 100 ml above 1000 ml). In all patients the pleural pressure curves were analyzed in five points, at 0, 25%, 50%, 75% and 100% of the relative volume of pleural effusion withdrawn in particular patients. Results There were 11 and 12 patients with right sided and left sided pleural effusion, respectively (14 M, 9F, median age 68, range 46–85 years). The most common cause of pleural effusion were malignancies (20 pts., 87%). The median total volume of withdrawn pleural fluid was 1800 (IQR 1500–2400) ml. After termination of pleural fluid withdrawal Pplampl increased in 22/23 patients compared to baseline. The median Pplampl increased from 3.4 (2.4–5.9) cmH2O to 10.7 (8.1–15.6) cmH2O (p < 0.0001). Three patterns of Pplampl changes were identified. Although the patterns of RR changes were more diversified, a significant increase between RR at baseline and the last measurement point was found (p = 0.0097). Conclusions In conclusion, therapeutic thoracentesis is associated with significant changes in Pplampl during the respiratory cycle. In the vast majority of patients Pplampl increased steadily during pleural fluid withdrawal. There was also an increase in RR. The significance of these changes should be elucidated in further studies. Trial registration ClinicalTrial.gov, registration number: NCT02192138, registration date: July 1st, 2014

Relative activity of the POR proteins, defined as the increase in Chlide fluorescence intensity per time unit and per protein concentration (in mg), calculated for different Pchlide concentrations.

<p>Calculations were performed for series of fluorescence spectra measured at room temperature (<i>e</i>.<i>g</i>. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116990#pone.0116990.s002" target="_blank">S2 Fig.</a>). POR concentration in a single experiment was between 1 and 2.75 μM; NADPH concentration: 0.05 mM. Error bars are SD of two independent series of fluorescence spectra.</p

Representative 77 K fluorescence emission spectra of a reaction mixture containing Pchlide, NADPH and PORA or PORC.

<p>Spectra labelled as “dark” were measured after a 30-min incubation of the reaction mixture in darkness. After these measurements, the samples were thawed, illuminated, frozen again and used for fluorescence measurement (spectra labelled as “light”). See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116990#sec002" target="_blank">materials and methods</a> for the details. POR concentration: 6.3 ± 0.3 μM, Pchlide concentration: 1.3 μM, NADPH concentration: 0.2 mM. Pchlide:POR ratio = 0.21. Excitation wavelength: 440 nm.</p