A comparative study of the structural and physicochemical properties of the major proteins from Camelina sativa (L.) Crantz and Brassica napus L.

Camelina sativa (L.) Crantz is a new industrial oilseed crop suitable for the Canadian prairies and which shows potential benefits for the Canadian bio-economy. This study was carried out to identify the major proteins of the oil-free residue (meal) of C. sativa (camelina) while investigating their structural and physicochemical properties. Canola (Brassica napus L.) was used as the control in the study. Camelina seeds were treated with Viscozyme® (0.1 mL/g) to remove mucilage. The mucilage free meal contained 51.3% protein (dwb, %N×6.25) which was greater than in canola. Both camelina and canola meals shared similar profiles for polypeptides and amino acids. At acidic pHs, canola meal had higher soluble protein content than did camelina meal, but the opposite was observed when the pH moved toward alkaline. A pH of 4.5 identified as the apparent isoelectric point (pI) of the protein from these two meals, which is presumably a cruciferin-napin complex. The 11S and 2S proteins of both seed types were isolated and purified using liquid chromatography. The purified 11S protein from camelina and canola contained predominantly cruciferin with minor contamination with non-targeted storage proteins. Of the non-cruciferin contamination of camelina, vicilin (7S) found to be abundant. The purified 2S protein from camelina contained napin and a noticeable amount of late embryogenesis abundant (LEA) protein, whereas non-napin contamination was minor in canola. In camelina, cruciferin, napin and vicilin expressed from eleven, four and six genes, respectively, were identified. The oil body proteins were also isolated and several isoforms of oleosin were found in camelina and canola, whereas putative isoforms of caleosin and steroleosin were found only in camelina. The structural and physicochemical properties of cruciferin and napin were studied in response to changing pH and temperature. The predominance of beta-structure and α-helix content in the 2˚ structure of cruciferin and napin, respectively, was confirmed for both camelina and canola. Cruciferin from camelina and canola exhibited acid-induced structural unfolding at the 3˚structure level. Cruciferin was not completely unfolded and assumed an intermediate state, plausibly a molten globule. Napin structure was not as sensitive as cruciferin to changing medium pH or an increase in temperature. Cruciferin exhibited high thermal stability (>80˚C) at neutral and alkaline pH, whereas the opposite was observed at acidic pH. Results showed that the cruciferin and napin responded differently to changing pH and temperature. Therefore, conditions of oil extraction and protein recovery from meal may affect these two storage proteins differently

VALUE ADDED FRACTIONATION OF CANARYSEED (Phalaris canariensis L.) STORAGE PROTEINS

The overall goal of this research was to develop an aqueous-based protein fractionation process from canaryseed (Phalaris canariensis L.), an important specialty crop for Canada, for value addition in the food industry. Evaluation of the microstructural features of canaryseed showed that it is composed of a bran, germ (embryo) and a starch-rich (starchy) endosperm, which is a typical characteristic of a cereal grain. The germ covers ~8-12% of the total endosperm area and it is highly concentrated with oil. Except for the germ, oil is distributed in the starchy endosperm and accounts for most of the oil in the whole seed. Presence of oil distributed in the starchy endosperm would be challenging to apply aqueous-based processing techniques for protein fractionation. The whole (full bran) canaryseed flour was high in oil, ash, fiber, phytic acid and phenolic content than that of white (low bran) flour prepared from roller-milling whereas, they contained 21.3% and 21.4% of protein concentration, respectively, which is not significantly different (p>0.05). Therefore, white flour could be a purer starting material for protein fractionation than whole flour. However, whole-flour showed better oil and water holding, emulsifying capacities, and digestibility in terms of in vitro protein digestibility corrected amino acid score (IV-PDCAAS) compared to that of white-flour whereas, emulsion and foaming stability, and foaming capacity were similar. Both hexane and ethanol were tested as de-oiling solvents for canaryseed, and it was found that linoleic acid and phosphatidylcholine were the major fatty acid and phospholipid extracted by these two solvents. Neither hexane nor ethanol denatured the canaryseed protein (peak denaturation temperature of 107-108°C) and did not cause any negative impact on their techno-functionality and digestibility. A laboratory-scale-enzyme-assisted-aqueous process was developed to prepare a protein concentrate (>70% protein purity) using yellow-canaryseed-white flour. The same process was successfully adapted for brown canaryseed. However, lower protein purity can be expected depending on the protein content in the white flour if the protein content in the seed is lower due to genetic and environment factors. The aqueous treatment applied for de-oiling reduced the initial oil content (6%) in yellow-seed flour to 1.2%. The enzymatic treatment applied afterward degrade the starch in the flour improving the protein content from 21% to 74.8%. It also increased the oil content from 1.2% to 12.1%, which was higher than anticipated oil content in the final product. Same trend was observed for brown canaryseed processing. The protein concentrates from yellow and brown canaryseed prepared using this aqueous-based method showed a least gelation concentration at 16% (w/w), which is higher than commercial soy protein concentrate (CSPC, 13%), but lower than the commercial vital wheat gluten (CVWG, 19%). At the least gelation concentration, the protein gels from both yellow and brown canaryseed showed significantly lower (p<0.05) strength than that of CSPC even with the presence of mono- and divalent salts. On the other hand, yellow and brown canaryseed protein concentrates showed comparable bread-dough improving properties to CVWG at lower inclusion levels (1-3%). Noticeable differences of gelation properties between yellow and brown canaryseed was not observed. During the scaling up of the lab process, some modifications to the original aqueous process was performed to address issues related to upstream decanter separation. The modified process reduced the oil content to <1% and degraded ~85% starch to prepare the final protein product. However, proteins were lost into the waste stream due to the modifications, subsequently lowering the protein recovery into the final product. Further investigations on process modifications and optimization are required to prevent protein losses and improve protein recovery. In summary, this research was able to develop an enzyme-assisted aqueous process for canaryseed protein fractionation for value addition despite having a major fraction of oil distributed within the starchy endosperm. This process used canaryseed white flour obtained using roller milling as the starting material for protein fractionation, which is purer in chemical composition compared to whole-seed flour. The developed method can be used for both yellow and brown canaryseed to fractionate protein. Canaryseed protein has high thermal stability and the use of hexane and ethanol as de-oiling solvent does not denature the protein and affect negatively for its techno-functional and nutritional properties. The fractionated protein did not display uniquely improved techno-functional properties compared to commercial soy protein or vital wheat gluten. However, canaryseed protein showed comparable bread-dough-forming properties to that of vital wheat gluten. The enzyme-assisted aqueous process was scaled up using pilot-scale equipment and was able to successfully de-oil white canaryseed flour. However, some process modifications were required to address the issues encountered during scaling up which caused lower protein recovery and purity in the final fractionated product. Further investigations and modifications are required to improve and optimize the scaled-up process for protein recovery

Asma bronquial i medi ambient

L'asma bronquial és una malaltia que es caracteritza per la presentació d'episodis d'obstrucció bronquial secundaris a una hiperreactivitat de les vies aèries, els quals desapareixen de forma espontània o bé per l'acció del tractament broncodilatador. El medi ambient, i de forma més específica les característiques de l'aire que respirem, incideix en el desenvolupament de crisis d'agudització en la majoria de malalts asmàtics. No hi ha dubte dels efectes nocius dels increments de pol·lució atmosfèrica sobre la mortalitat i la morbilitat dels individus amb patologia crònica de tipus càrdio-respiratori. En aquest sentit, són nombrosos els estudis que demostren que les condicions meteorològiques i els nivells de contaminants tenen un paper en el nombre i la gravetat de les crisis d'agudització d'aquests malalts. ..

Impact of Lung Function Decline on Mortality in Lung Transplant Recipients: Long-Term Results From the L-CsA-i Study for the Prevention of Bronchiolitis Obliterans Syndrome

BackgroundChronic lung allograft dysfunction (CLAD) is defined by a progressive loss of FEV1 and is associated with premature mortality. The aim of this study was to investigate the direct association between FEV1 decline and risk of mortality in patients after lung transplantation (LTx). Methods10-year follow up data from lung transplant recipients participating in randomized placebo-controlled clinical trial investigating the role of liposomal Cyclosporine A for inhalation (L-CsA-i) in the prevention of bronchiolitis obliterans syndrome (NCT01334892) was used. The association between the course of FEV1 over time and the risk of mortality was assessed using joint modeling and Cox regression analysis. ResultsA total of 130 patients were included. Predictors of FEV1 decline were a higher absolute FEV1 at baseline and male sex. The joint model analysis indicated a significant association of change of FEV1 and risk of mortality (p < 0.001), with a predicted 3.4% increase in mortality risk for each 1% decline in FEV1. Significant predictors of a progressive phenotype were single LTx and treatment with placebo (as opposed to L-CsA-i). At the end of follow-up, 82 patients (63.1%) were still alive. Cox regression analyses for mortality identified only single LTx as a predictor of higher risk. ConclusionBased on our observation of a close association between FEV1 and mortality over a period of 10 years we suggest FEV1 as a valid predictor of mortality and a suitable surrogate endpoint in the investigation of early interventions

Efficacy and safety of the combination of reduced duration prophylaxis followed by immuno-guided prophylaxis to prevent cytomegalovirus disease in lung transplant recipients (CYTOCOR STUDY) : An open-label, randomised, non-inferiority clinical trial

Introduction Prolonged use of antivirals to prevent the development of cytomegalovirus (CMV) disease in lung transplant patients has been shown to have significant side effects, for which alternatives are being sought to reduce their use. The monitoring of cell immunity against CMV could be an alternative as it has shown to be useful in identifying transplant patients at low risk of infection, who could benefit from shorter prophylaxis. The aim of the CYTOCOR study is to demonstrate that the combination of a reduced prophylaxis strategy with subsequent CMV-specific immunological monitoring would allow CMV infection to be controlled in lung transplant patients as effectively as the usual strategy (prophylaxis followed by pre-emptive therapy), while reducing the side effects of antivirals due to the shorter duration of prophylaxis. Methods and analysis Phase III randomised, open, multicentre, parallel, non-inferiority clinical trial to study the efficacy and safety of the combination of a prophylaxis strategy up to month +3 post-transplant followed by immuno-guided prophylaxis using the QuantiFERON-CMV technique up to month +12 post-transplant to prevent CMV disease in CMV-seropositive lung transplant recipients. This strategy will be compared with a combination of a usual prophylaxis strategy up to month +6 post-transplant followed by pre-emptive therapy up to month +12. To study the incidence of CMV disease, patients will be followed up to 18 months post-transplantation. A total of 150 patients are expected to be recruited for the study. Ethics and public dissemination The clinical trial has been approved by the Research Ethics Committees and authorised by the Spanish Agency of Medicines and Medical Devices (AEMPS). If the hypothesis of this clinical trial is verified, the dissemination of the results could change clinical practice by increasing knowledge about the safety and efficacy of discontinuing valganciclovir prophylaxis in lung transplant recipients. Trial registration number NCT03699254

Bronchopulmonary Penetration of Isavuconazole in Pulmonary Transplant Recipients (PBISA01): Protocol for a Phase IV Clinical Trial With a Single Treatment Arm

Background: Aspergillosis is the most frequently observed invasive fungal disease (IFD) in lung transplant recipients. Isavuconazole (ISA) has shown a better safety profile and noninferiority to voriconazole in the treatment of patients with IFD. Objective: The aim of this study is to describe the bronchopulmonary pharmacokinetic profile of oral ISA by analyzing the degree of penetration in the epithelial lining fluid and alveolar macrophages in patients receiving lung transplantation with a diagnosis of IFD. Methods: A total of 12 patients aged ≥18 years receiving a lung transplant with an IFD diagnosis and indication for ISA treatment and follow-up bronchoscopy will be included in the study. After 5 days of treatment with ISA and before the treatment is discontinued, the patients will be randomized (1:1:1:1) to perform the scheduled bronchoscopy at various times after the administration of ISA (2, 4, 8, and 12 hours). In total, 4 blood samples will be obtained per patient: at 72 hours after treatment initiation, on the day of the bronchoscopy, at the time of the bronchoalveolar lavage (simultaneously), and at 7 days after treatment initiation, to analyze tacrolimus and ISA plasma levels. ISA concentrations will be measured in plasma, epithelial lining fluid, and alveolar macrophages by a high-performance liquid chromatography/UV coupled to fluorescence method. Results: Enrollment for the PBISA01 trial began in October 2020 and was completed in October 2021. All samples will be analyzed once recruitment is complete, and the results are expected to be published in October 2022. Conclusions: There are no clinical studies that analyze the bronchopulmonary penetration of ISA. Bronchoalveolar lavage performed routinely in the follow-up of lung transplant recipients constitutes an opportunity to analyze the bronchopulmonary penetration of ISA. Trial registration: European Clinical Trials Register 2019-004240-30; www.clinicaltrialsregister.eu/ctr-search/trial/2019-004240-30/ES. International registered report identifier (irrid): DERR1-10.2196/37275.This work is supported funded by Pfizer (grant 54685521). Pfizer will have no role in the study’s design; the collection, management, analysis, and interpretation of data; writing of the report; and the decision to submit the report for publication.S

Bronchopulmonary penetration of isavuconazole in lung transplant recipients

Isavuconazole's (ISA) pharmacokinetics was studied among lung transplant recipients to evaluate its bronchopulmonary penetration. This study included 13 patients and showed mean serum concentrations of 3.30 (standard deviation [SD] 0.45), 5.12 (SD 1.36), and 6.31 (SD 0.95) at 2 h, 4 h, and 24 h respectively. Mean concentrations in the epithelial lining fluid were 0.969 (SD 0.895), 2.141 (SD 1.265), and 2.812 (SD 0.693) at the same time points. ISA is a drug with a tolerable safety profile that achieves adequate concentrations in the lung.This work was partially supported and funded by Pfizer (grant 54685521). Pfizer had no role in the study’s design; the collection, management, analysis, and interpretation of data; writing of the report; and the decision to submit the report for publicationS

Efficacy and safety of the combination of reduced duration prophylaxis followed by immuno-guided prophylaxis to prevent cytomegalovirus disease in lung transplant recipients (CYTOCOR STUDY): an open-label, randomised, non-inferiority clinical trial.

INTRODUCTION: Prolonged use of antivirals to prevent the development of cytomegalovirus (CMV) disease in lung transplant patients has been shown to have significant side effects, for which alternatives are being sought to reduce their use. The monitoring of cell immunity against CMV could be an alternative as it has shown to be useful in identifying transplant patients at low risk of infection, who could benefit from shorter prophylaxis. The aim of the CYTOCOR study is to demonstrate that the combination of a reduced prophylaxis strategy with subsequent CMV-specific immunological monitoring would allow CMV infection to be controlled in lung transplant patients as effectively as the usual strategy (prophylaxis followed by pre-emptive therapy), while reducing the side effects of antivirals due to the shorter duration of prophylaxis. METHODS AND ANALYSIS: Phase III randomised, open, multicentre, parallel, non-inferiority clinical trial to study the efficacy and safety of the combination of a prophylaxis strategy up to month +3 post-transplant followed by immuno-guided prophylaxis using the QuantiFERON-CMV technique up to month +12 post-transplant to prevent CMV disease in CMV-seropositive lung transplant recipients. This strategy will be compared with a combination of a usual prophylaxis strategy up to month +6 post-transplant followed by pre-emptive therapy up to month +12. To study the incidence of CMV disease, patients will be followed up to 18 months post-transplantation. A total of 150 patients are expected to be recruited for the study. ETHICS AND PUBLIC DISSEMINATION: The clinical trial has been approved by the Research Ethics Committees and authorised by the Spanish Agency of Medicines and Medical Devices (AEMPS).If the hypothesis of this clinical trial is verified, the dissemination of the results could change clinical practice by increasing knowledge about the safety and efficacy of discontinuing valganciclovir prophylaxis in lung transplant recipient

Follow-up with Telemedicine in Early Discharge for COPD Exacerbations: Randomized Clinical Trial (TELEMEDCOPD-Trial)

The results reported by different studies on telemonitoring in patients with chronic obstructive pulmonary disease (COPD) have been contradictory, without showing clear benefits to date. The objective of this study was to ascertain whether an early discharge and home hospitalization telehealth program for patients with COPD exacerbation is as effective as and more efficient than a traditional early discharge and home hospitalization program. A prospective experimental non-inferiority study, randomized into two groups (telemedicine/control) was conducted. The telemedicine group underwent monitoring and was required to transmit data on vital constants and ECGs twice per day, with a subsequent telephone call and 2 home visits by healthcare staff (intermediate and at discharge). The control group received daily visits. The main variable was time until first exacerbation. The secondary variables were: number of exacerbations; use of healthcare resources; satisfaction; quality of life; anxiety-depression; and therapeutic adherence, measured at one and 6 months of hospital discharge. A total of 116 patients were randomized (58 to each group) without significant differences in baseline characteristics or time until first exacerbation, i.e. median 48 days (pp. 25-75:23-120) in the control group, and 47 days (pp. 25-75:19-102) in the intervention group; p = 0.52). A significant decrease in the number of visits was observed in the intervention versus the control group, 3.8 ± 1 vs 5.1 ± 2(p = 0.001), without significant differences in the number of exacerbations. In conclusion follow-up via a telemedicine program in early discharge after hospitalization is as effective as conventional home follow up, being the cost of either strategy not significantly different.This study was awarded a Health Research Fund (Fondo de Investigaciones Sanitarias/FIS) grant, dossier No. PI12/01161 of the Carlos III Institute of Health and the Ministry of Sciences, Innovation and Universities.S

Voriconazole and squamous cell carcinoma after lung transplantation: A multicenter study

This study evaluated the independent contribution of voriconazole to the development of squamous cell carcinoma (SCC) in lung transplant recipients, by attempting to account for important confounding factors, particularly immunosuppression. This international, multicenter, retrospective, cohort study included adult patients who underwent lung transplant during 2005-2008. Cox regression analysis was used to assess the effects of voriconazole and other azoles, analyzed as time-dependent variables, on the risk of developing biopsy-confirmed SCC. Nine hundred lung transplant recipients were included. Median follow-up time from transplant to end of follow-up was 3.51 years. In a Cox regression model, exposure to voriconazole alone (adjusted hazard ratio 2.39, 95% confidence interval 1.31-4.37) and exposure to voriconazole and other azole(s) (adjusted hazard ratio 3.45, 95% confidence interval 1.07-11.06) were associated with SCC compared with those unexposed after controlling for important confounders including immunosuppressants. Exposure to voriconazole was associated with increased risk of SCC of the skin in lung transplant recipients. Residual confounding could not be ruled out because of the use of proxy variables to control for some confounders. Benefits of voriconazole use when prescribed to lung transplant recipients should be carefully weighed versus the potential risk of SCC. This article is protected by copyright. All rights reserved