Grain Boundary Driven Plateau-Rayleigh Instability in Multilayer Nanocrystalline Thin Film: A Phase-field Study

Thermal stability of nanocrystalline multilayer thin film is of paramount importance as the applications often involve high temperature. Here we report on the layer instability phenomenon in binary polycrystalline thin film initiating from the grain boundary migrations at higher temperatures using phase-field simulations. Effect of layer thickness, bilayer spacing and the absence of grain boundary are also investigated along with the grain boundary mobility of individual phases on the layer stability. Layer instability in the polycrystalline film is shown to arise from the grain boundary grooving which originates spontaneously from the presence of grain boundaries. Our results show that the growth of the perturbation generated from the differential curvature follows Plateau-Rayleigh instability criterion. Increase in layer thickness, lower bilayer thickness as well as lower grain boundary mobility improve layer stability. Phase-field simulations show similar microstructural evolution as has been observed in our Zirconium (Zr)/Zirconium Nitride (ZrN) system experimentally. Detail analysis performed in this work to understand the mechanisms of layer instability leads us to predict measures which will improve the thermal stability of multilayer nanocrystalline thin film

Proteomic Analysis of Benzo[a]pyrene-Mediated Bladder Toxicity

The major objective of this study was to evaluate whether the environmental contaminant benzo[a]pyrene B[a]P, one of the most important polycyclic aromatic hydrocarbons (PAH), is capable of mediating DNA damage in urinary bladder epithelial cells and hence potentially bladder carcinogenesis. To pursue this goal, effects of B[a]P on urinary bladder epithelial cells were investigated by applying a proteomic approach with the purpose of identifying proteins and pathways involved in B[a]P toxicity. First, the ability of bladder epithelial cells for B[a]P uptake and metabolism was determined. Secondly, a proteome map of primary porcine urinary bladder epithelial cells (PUBEC), the cell model used in the majority of the studies, was established as basis for comparative investigations. In the same model, investigations on time- and concentration-dependent expression changes of proteins after B[a]P exposure followed. The proteins were separated by using 2D gel electrophoresis and identified by MALDI-TOF-MS analysis in these studies. Finally, to elucidate mechanisms by which B[a]P mediates its toxicity, signaling pathways were studied in RT4 cells by using Blue Native PAGE analysis. Besides offering some insights into B[a]P-mediated toxic effects, the studies also point towards the possibility of bladder cancer development induced by B[a]P exposure. B[a]P is a ubiquitous environmental pollutant formed during the combustion of fossil fuels, grilling, barbecuing, and smoking of food. Although much information is available on the carcinogenic properties of B[a]P, the mechanism by which this chemical is taken up by cells is still not known. In Chapter 3 of this thesis, attempts were made to investigate the dynamics of B[a]P uptake, subcellular distribution, and metabolism in PUBEC. It was found that exposure to 0.5 μM B[a]P led to an increase in intracellular concentration of B[a]P in bladder epithelial cells in a time-dependent manner but without approaching saturation. Also, a marked difference in B[a]P uptake was observed among various PUBEC pools used for the studies. Subcellular partitioning studies of B[a]P by using confocal microscopy revealed that a significant amount of B[a]P accumulated in the cell membrane of PUBEC, while only a slight but significant increase in B[a]P fluorescence intensity was observed in the cytosol and nucleus. Quantification of B[a]P uptake by bladder epithelial cells by spectrofluorometric and gas chromatographic-mass spectrometric analysis yielded intracellular concentrations ranging from 7.28 μM to 35.07 μM in cells exposed to 0.5 μM B[a]P and from 29.9 μM to 406.64 μM in cells exposed to 10 μM B[a]P. The formation of 3-OH-B[a]P in all of the B[a]P-exposed PUBEC determined by GC-MS analysis demonstrated for the first time that oxygenated B[a]P metabolites are actually formed in this cell model. These results indicate that bladder epithelial cells are capable of a strong accumulation and metabolic activation of B[a]P and suggest that B[a]P may act as a bladder cancer-inducing chemical. Also, the differences in B[a]P uptake by the various PUBEC pools is an explanation for the inter-individual variation in PAH toxicity as observed in humans. Urinary bladder epithelial cells (also known as transitional epithelial cells) are the innermost cells of the bladder which are involved in accommodating the fluctuation of liquid volume in this organ and also help to protect it against caustic/toxic effects of urine. These cells are also the first ones to come in contact with urinary toxicants and thus account for 90 % of bladder cancers known as transitional cell carcinomas. As a prerequisite for proteomic studies, the first reference proteome and phosphoproteome maps of porcine bladder epithelia cells were generated by applying 2DE gel electrophoresis. This is discussed in Chapter 4. A total of 120 selected protein spots were identified by MALDI-TOF-MS analysis, among which 31 phosphoproteins were enriched by using a method based on the precipitation with lanthanum ions (La3+). All identified proteins were bioinformatically annotated according to their physiochemical characteristic, subcellular location, and function. Most of the proteins were distributed in an area of pI 4-10 and a molecular mass range between 20 kDa and 100 kDa. The 2DE map with the complete range of expressed proteins, especially with information about phosphoproteins, provides a valuable resource for comparative proteomic analysis of normal and pathological conditions affecting the bladder function. The studies described in Chapter 5 of the thesis deal with a series of events leading from DNA damage to apoptosis that were investigated by using a proteomic approach. 2DE gel electrophoresis mapped the differences between cells exposed to 0.5 μM B[a]P and control cells. Twenty-five differentially expressed proteins involved in DNA repair, mitochondrial dysfunction, and apoptosis were identified by MALDI-TOF-MS analysis. A concentration-dependent increase in DNA damage was observed after an exposure period of 24 h. The expression of VDAC2, CTSD, HSP27, and HSP70 indicated towards the intrinsic apoptotic mitochondrial pathway, although the analysis of mitochondrial dysfunction pointed towards an alternate pathway of cell death: The mitochondrial membrane potential (MMP), although disturbed during the initial exposure period, was nearly retained after 24 h of B[a]P treatment. In conclusion, the studies indicated DNA damage caused by B[a]P at low concentrations during an exposure period of 24 h and also shed light on a possible apoptotic mechanism induced by DNA damage. Studies on protein-protein interactions involved in B[a]P toxicity are described in Chapter 6. A comparative analysis of proteomic complexes involving the two AhR ligands B[a]P and TCDD was carried out by using 2D BN/SDS-PAGE. For the enrichment of the protein complexes, a subcellular fractionation of unexposed cells and cells exposed to B[a]P and TCDD was carried out. BN/SDS-PAGE of these fractions revealed an effective separation of protein species and complexes of various origins, including mitochondria, plasma membrane, and intracellular compartments. The major differences in the protein maps obtained from samples of control cells and cells exposed to B[a]P and TCDD, respectively, concerned the expression of many calcium- and iron-containing proteins. On the basis of these findings, the intracellular calcium content of cells exposed to TCDD and B[a]P was evaluated, revealing an increase only after 24 h of exposure but with no transient elevation. The cells exposed to TCDD also showed an alteration in the labile iron pool (LIP) of the cells, but no such changes were observed in B[a]P-exposed cells. The increase in the LIP was strongly inhibited by the calmodulin (CaM) antagonist W-7 (10 μM). These results point towards a possible interaction between the iron and calcium signaling of the cells. The analysis of nitric oxide generation by using the Griess assay revealed a substantial increase in NO content of both B[a]P- and TCDD-exposed cells. Also in these cells, the basal NO generation was inhibited when the cells were blocked with the CaM antagonist W-7. The results led to the conclusion that alterations in calcium and iron homeostasis upon exposure to TCDD and B[a]P is linked by NO that is produced by CaM-activated nitric oxide synthase (NOS). The NO thus produced by interacting with the iron centers of IRPAs modulated the activity of TfR1 and FTH1 which in turn changed the LIP of the cells and hence the toxicity. Although some new mechanistic insights into the mechanisms of B[a]P- and TCDD-induced toxicity were provided by these studies, further investigations are still required for the validation of these initial results.Die wesentliche Zielsetzung der in der vorliegenden Arbeit dargestellten Untersuchungen war die Klärung der Frage, ob Benzo[a]pyren (B[a]P), die Leitsubstanz der polyzyklischen aromatischen Kohlenwasserstoffe (PAH) und eine der bedeutendsten umweltrelevanten Gefahrstoffe, in Urothelzellen DNA-Schäden und damit potenziell Harnblasenkarzinome verursachen kann. Nach Einschätzung der International Agency for Research on Cancer (IARC) liegen entsprechende Indizien aus epidemiologischen Studien vor. Nach zunächst durchgeführten Untersuchungen zur Toxikokinetik von B[a]P in primären Schweineurothelzellen, dem überwiegend eingesetzten Zellmodell, lag der Schwerpunkt der weiteren Studien in der Proteom-basierten Analyse von B[a]P-induzierten Effekten, die möglicherweise in einen kanzerogenen Prozess münden. Basierend auf den Ergebnissen der Proteomanalysen wurden Hypothesen entwickelt, die in weiteren Untersuchungen mit konventionellen toxikologischen Methoden überprüft wurden. Zum besseren Verständnis der in Schweineurothelzellen unter physiologischen Bedingungen exprimierten Proteine wurden zunächst Proteomkarten angefertigt. Im selben Modell wurden anschließend Veränderungen des Proteoms nach Exposition gegen B[a]P analysiert. Hierzu wurden die Proteine aus Gesamtzelllysaten mit der zweidimensionalen Gelelektrophorese getrennt und nach tryptischem Verdau mittels MALDI-TOF-MS-Analyse ihrer Peptid-Fingerprintspektren identifiziert. In Zellen einer Harnblasen¬karzinom-Zelllinie (RT4) wurden Veränderungen von Proteinkomplexen bzw. Protein-Protein-Interaktionen nach B[a]P-Exposition mit Hilfe der Blue Native-PAGE untersucht. Insgesamt wurden durch die vorliegende Arbeit vertiefte Einblicke in die toxischen Wirkungen von B[a]P in Urothelzellen erhalten, darunter auch in solche, die auf einen B[a]P-induzierten kanzerogenen Effekt hinweisen. B[a]P kommt ubiquitär in der Umwelt vor. Es entsteht bei der unvollständigen Verbrennung fossiler Brennstoffe, ist im Zigarettenrauch vorhanden und bildet sich beim Braten, Grillen und Räuchern von Nahrungsmitteln. Die genotoxische Wirkung von B[a]P ist relativ gut untersucht, hingegen ist nur wenig über die zelluläre Aufnahme dieser Substanz bekannt. Nach einleitenden Bemerkungen im ersten Kapitel dieser Arbeit und der Darstellung der für die Studien verwendeten Materialien und Methoden im zweiten Kapitel wurde im dritten Kapitel die zeitabhängige Aufnahme von B[a]P in Schweineurothelzellen, die intrazelluläre Verteilung und der Metabolismus untersucht. Die 24-stündige Exposition gegen 0,5 µM B[a]P führte zu einem kontinuierlichen Anstieg der intrazellulären B[a]P-Konzentration ohne eine Sättigung zu erreichen, wobei ein Unterschied in der Kinetik zwischen den einzelnen Zellpools zu beobachten war. Die Analyse der subzellulären B[a]P-Verteilung mit einem konfokalen Laserscan-Mikroskop zeigte, dass B[a]P überwiegend in zellulären Membranen und nur zu einem kleinen Teil im Zytosol und im Zellkern akkumulierte. Die spektralfluorimetrische und gaschromatographisch-massenspektrometrische Quantifizierung ergab, dass nach Exposition gegen 0,5 µM B[a]P zwischen 7,28 µM und 35,07 µM B[a]P und nach einer Exposition gegen 10 µM zwischen 29,9 µM und 406,64 µM B[a]P aufgenommen wurde. Außerdem wurde erstmalig in Urothelzellen die Bildung eines oxidativen B[a]P-Metaboliten, die von 3-OH-B[a]P, mittels GC-MS nachgewiesen. Diese Ergebnisse zeigen, dass B[a]P intrazellulär akkumuliert und metabolisch aktiviert wird, was essentielle Voraussetzungen für eine kanzerogene Wirkung sind. Außerdem scheinen Schweineurothelzellen - analog zu humanen Urothelzellen - über eine unterschiedliche Aufnahmekapazität und Ausstattung an metabolisierenden Enzymen zu verfügen, die zu interindividuellen Unterschieden in der Toxizität führen können. Die innere Schicht der Harnblasen wird von epithelialen Zellen ausgekleidet, die sich nicht nur den unterschiedlichen Füllungszuständen der Blase anpassen müssen, sondern auch das darunter liegende Gewebe vor der toxischen Wirkung des Urins schützen sollen. In 90 % aller Fälle gehen Harnblasenkarzinome von diesen Zellen aus. Um vergleichende proteomische Studien an diesen Zellen durchführen zu können, wurden erstmalig Protein- und Phosphoproteinkarten von Schweineurothelzellen nach zweidimensionaler gelelektrophoretischer Proteintrennung erstellt. Wie im vierten Kapitel beschrieben, wurden insgesamt 120 Proteine identifiziert. Darunter waren 31 Phosphoproteine, die durch eine Präzipita¬tion mit Lanthan¬ionen angereichert und schließlich bestimmt werden konnten. Alle Proteine wurden mittels Datenbanken spezifischen Funktionen, physikochemischen Eigenschaften und subzellulären Lokalisationen zugeordnet. Mit Hilfe dieser Protein- und Phosphoproteinreferenzkarten können bei vergleichenden Proteomstudien pathophysiologische Zustände leichter erfasst werden. Als Ergebnis weiterer proteomischer Studien wird im fünften Kapitel eine Folge von B[a]P-induzierten Ereignissen beschrieben, die über eine Schädigung der DNA zu Apoptose führen. Schweineurothelzellen wurden für 24 Stunden gegen 0,5 µM B[a]P exponiert und die Proteine im Zelllysat anschließend zweidimensional getrennt. 25 unterschiedlich regulierte Proteine wurden identifiziert, die in der DNA Reparatur, der mitochondrialen Funktion und der Apoptose involviert sind. Ergänzende toxikologische Untersuchungen (TUNEL-Methode, Comet-Assay) unterstützten diese Ergebnisse, da sowohl die Anzahl apoptotischer Zellen als auch das Ausmaß der DNA-Schädigung im gleichen Expositionszeitraum konzentrationsabhängig zunahm. Die Regulation von VDAC2, CTSD, HSP27 und HSP70 weist auf eine Aktivierung des intrinsischen mitochondrialen Apoptosewegs hin. Allerdings stützte die Messung des mitochondrialen Membranpotenzials diese Hypothese nicht. Zwar war das Membranpotential nach kurzzeitiger Exposition gestört, erholte sich aber annähernd vollständig nach 24 Stunden. Möglicherweise sind hier alternative apoptotische Mechanismen von Bedeutung. Insgesamt zeigten diese Untersuchungen, dass B[a]P in einer niedrigen Konzentration nach 24 Stunden die DNA schädigt und zu Apoptose führt. Außerdem wurden zahlreiche Proteine identifiziert, die an diesen Prozessen beteiligt sind. Veränderungen in der Protein-Protein-Interaktion nach B[a]P-Exposition werden im sechsten Kapitel beschrieben. Sie ergaben sich aus einer mittels 2D BN/SDS-PAGE durchgeführten vergleichenden Analyse von Proteinkomplexen nach Exposition von RT4-Zellen gegen B[a]P und TCDD, zwei Liganden des Arylhydrocarbon-Rezeptors (AhR). Zur Anreicherung der Proteinkomplexe wurden die Zellen subzellulär fraktioniert. Als größter Unterschied zwischen den beiden Liganden erwies sich die unterschiedliche Regulation von Eisen- und Calcium-haltigen Proteinen. Beide Liganden erhöhten den intrazellulären Calciumgehalt nach 24 Stunden. Zusätzlich erhöhte sich nur in den TCDD-exponierten Zellen der intrazelluläre labile Eisenpool, dessen Anstieg durch den Calmodulin-Antagonisten W-7 inhibiert wurde. Dieser Effekt deutete auf einen Zusammenhang zwischen der intrazellulären Eisen- und Calciumhomöostase hin, möglicherweise über die NO-Synthetase. Unterstützt wurde diese Hypothese durch den Nachweis eines NO-Anstiegs mittels Griess-Reaktion in den gegen beide Liganden exponierten Zellen. Wieder wurde der Anstieg durch den Calmodulin-Antagonisten W-7 inhibiert. Demnach scheint sowohl eine Exposition gegen TCDD als auch gegen B[a]P über Calmodulin die NO-Synthetase zu aktivieren, was wiederum zur erhöhten NO-Produktion führt. NO bindet an das Eisenzentrum im eisen-regulierenden Protein A (IRPA) und reguliert dadurch die Aktivität von TfR1 und FTH1, die ihrerseits den labilen Eisenpool beeinflussen und über diesen Mechanismus einen Teil der Toxizität der Liganden vermitteln. Weitere Studien sind erforderlich, um die in diesen Untersuchungen gewonnenen Ergebnisse, die bisher nicht bekannte toxische Wirkungsmechanismen von TCDD und B[a]P darstellen, zu validieren

Magnetic-order induced effects in nanocrystalline NiO probed by Raman spectroscopy

The magnetic-order induced effects in nanocrystalline NiO are investigated through the phonons and magnons observed in the Raman spectra. The key observations are (i) an anisotropy of the first-order transverse and longitudinal optical phonons, with a splitting on the order of 5 meV and (ii) a marked size and excitation wavelength variation of the two-magnon peak, which varies linearly with a redshift of similar to 50 cm(-1) with a size reduction from 105 to 30 mm The magnon-related peaks, in contrast to the phonons, are suppressed for near-resonance laser excitations. The experimental results are interpreted in terms of the exchange interactions and strong electron-electron correlations. The magnetization measurements shows a crossover to ferromagnetism with large coercivities and magnetization with decreasing size, which is shown to be due to the thermoinduced contribution

IMPORTANCE OF NANOCARRIERS AND PROBIOTICS IN THE TREATMENT OF ULCERATIVE COLITIS

Ulcerative colitis (UC) is an inflammatory chronic disease primarily affecting the colonic mucosa; the extent and severity of colon involvement are variable. Ulcerative colitis is identified by mucus diarrhea, tenesmus, bowel distension, and anemia. 5-aminosalicylic acid drugs, steroids, and immune suppressants are used for the therapy of ulcerative colitis. The mainchallenges in the management of thediseaseare drug-related side-effects and local targeting. To overcome these challengesprobiotics and micro and Nanoparticulate systemauspiciousapproaches to overcome drug-related adverse side effects and local targeting.Upon ingestion, the probiotics can result in beneficial health effects. Probiotics and micro and nanoparticulate approaches for suitable targeting and overcome the drug-associated side effect. Probiotics are mainly used as gut modulators but are also nowadays explored for their use in ulcerative colitis.The current therapeutic goals are to achieve clinical remission along with mucosal healing, avoidance of complications such as side effects of the drug and to improve the quality of life. The use of probiotics to increase the health of the intestine and used to block or manage intestinal disorders. They may prevent the induction of inflammatory reactions. Probiotics must be inspected for efficacy in the prevention and management of a wide spectrum of gastrointestinal diseases, like antibiotic-associated diarrhea.Micro and Nanoparticulate drug delivery system has been achieving huge importance for targeting of the drug to colon locally at a controlled and sustained rate

Quality By Design: A Systematic Approach for the Analytical Method Validation

The scientific way to develop an easy and robust analytical technique for critical analysis is a QbD approach. QbD is a systematic approach to product or method development that begins with predefined objectives and uses science and risk management approaches to achieve product and method understanding and ultimately method control. The aim of the analytical QbD is to achieve quality in measurement. The main objective of this review to explain different steps involved in method development by the QbD approach for analytical method development and describes the implementation of QbD in analytical procedure validation. The advantages of applying QbD principles to analytical technique include discovering and minimizing the source of variability that may lead to poor method robustness and ensuring that the method meets its intended performance need throughout the product and method lifecycle. Keywords: Quality by design (QbD), Risk Analysis, Analytical method validatio

Factor analysis for udder and teat type traits in Sahiwal and Karan Fries cows

Selecting female cows for productivity based on udder and teat traits is essential in field due to lack of available records. The objective of the present study was to reduce the dimensionality of the 17 udder and teat traits and to analyse their impact on milk productivity. The data on 256 cattle comprising 133 Sahiwal and 123 Karan Fries cows were used in this study over the years 2017-2019 from Livestock Research Centre (LRC) of ICAR-National Dairy Research Institute, Karnal. The 17 udder and teat traits were fore udder attachment (FUA), rear udder width (RUW), rear udder height (RUH), udder balance (UB), udder depth (UD), udder length (UL), udder width (UW), udder circumference (UC), central ligament/udder cleft (CL), teat circumference (TC), fore teat length (FTL), rear teat length (RTL), distance between fore and rear teat (DFR), distance between left and right teat (DLR), shortest distance from floor to fore teat (SDFT), shortest distance from floor to rear teat (SDRT) and teat diameter (TD). In the factor analysis, first five latent factors accounted for 62.22% of total variance in udder and teat measurements for Sahiwal cows and 65.67% in Karan Fries (KF) cows, respectively. In Sahiwal cows F1 represented better udder support and wideness (wide udders, udders were supported by strong suspensory ligament), whereas in KF cows, first factor reflected better udder dimension and distance between teats (longer length, width of udder and well placed teats). Factor analysis could reduce the multicollinearity of the data. It can be concluded that inclusion of udder and teat measurements in selection index can be a reliable criteria for selecting cows for higher milk yield

Itaconic acid and its applications for textile, pharma and agro-industrial purposes

Itaconic acid (IA) is a well-known bio-based monounsaturated organic acid (C5H6O4), with a white color and crystalline structure. It is widely used in the agro-based, plastics, textile, paint and pharmaceutical sectors, owing to its flexible structure, due to the presence of functional groups with covalent double bonds. IA is an alternative to the petrochemicals acrylic and methacrylic acids. Commercial manufacturing of IA using Aspergillus terreus is more economically effective and feasible, and the Department of Energy (DOE) of the United States added IA under the “top 12” organic chemicals in 2004. This review provides an overview on the synthesis of IA and improvement of its yield by mutagenesis and metabolic engineering of Aspergillus and other fungal strains, along with its wide applications for food, pharmaceutical and textile purposes

Quantification of human C1 esterase inhibitor protein using an automated turbidimetric immunoassay

BACKGROUND: Impaired levels or function of C1 inhibitor (C1-INH) results in angioedema due to increased bradykinin. It is important to distinguish between angioedema related to C1-INH deficiency and that caused by other mechanisms, as treatment options are different. In hereditary (HAE) and acquired (AAE) angioedema, C1-INH concentration is measured to aid patient diagnosis. Here, we describe an automated turbidimetric assay to measure C1-INH concentration on the Optilite® analyzer. METHODS: Linearity, precision, and interference were established over a range of C1-INH concentrations. The 95th percentile reference interval was generated from 120 healthy adult donors. To compare the Optilite C1-INH assay with a predicate assay used in a clinical laboratory, samples sent for C1-INH investigation were used. The predicate results were provided to allow comparison. RESULTS: The Optilite C1-INH assay was linear across the measuring range at the standard sample dilution. Intra and interassay variability was <6%. The 95th percentile adult reference interval for the assay was 0.21-0.38 g/L. There was a strong correlation between the Optilite concentrations and those generated with the predicate assay (R2 = 0.94, P < 0.0001, slope y = 0.83x). All patients with Type I HAE (n = 24) and AAE (n = 3) tested had concentrations below the measuring range in both assays, while all patients with unspecified angioedema (UAE), not diagnosed with HAE or AAE had values within the reference range. CONCLUSION: The Optilite assay allows the automated and precise quantification of C1-INH concentrations in patient samples. It could therefore be used as a tool to aid the investigation of patients with angioedema

British Lung Foundation/United Kingdom primary immunodeficiency network consensus statement on the definition, diagnosis, and management of granulomatous-lymphocytic interstitial lung disease in common variable immunodeficiency disorders

A proportion of people living with common variable immunodeficiency disorders develop granulomatous-lymphocytic interstitial lung disease (GLILD). We aimed to develop a consensus statement on the definition, diagnosis, and management of GLILD. All UK specialist centers were contacted and relevant physicians were invited to take part in a 3-round online Delphi process. Responses were graded as Strongly Agree, Tend to Agree, Neither Agree nor Disagree, Tend to Disagree, and Strongly Disagree, scored +1, +0.5, 0, −0.5, and −1, respectively. Agreement was defined as greater than or equal to 80% consensus. Scores are reported as mean ± SD. There was 100% agreement (score, 0.92 ± 0.19) for the following definition: “GLILD is a distinct clinico-radio-pathological ILD occurring in patients with [common variable immunodeficiency disorders], associated with a lymphocytic infiltrate and/or granuloma in the lung, and in whom other conditions have been considered and where possible excluded.” There was consensus that the workup of suspected GLILD requires chest computed tomography (CT) (0.98 ± 0.01), lung function tests (eg, gas transfer, 0.94 ± 0.17), bronchoscopy to exclude infection (0.63 ± 0.50), and lung biopsy (0.58 ± 0.40). There was no consensus on whether expectant management following optimization of immunoglobulin therapy was acceptable: 67% agreed, 25% disagreed, score 0.38 ± 0.59; 90% agreed that when treatment was required, first-line treatment should be with corticosteroids alone (score, 0.55 ± 0.51)