Mortality among patients hospitalized for COVID‐19 has declined over the course of the pandemic. Mortality trends specifically in solid organ transplant recipients (SOTR) are unknown. Using data from a multicenter registry of SOTR hospitalized for COVID‐19, we compared 28‐day mortality between early 2020 (March 1, 2020–June 19, 2020) and late 2020 (June 20, 2020–December 31, 2020). Multivariable logistic regression was used to assess comorbidity‐adjusted mortality. Time period of diagnosis was available for 1435/1616 (88.8%) SOTR and 971/1435 (67.7%) were hospitalized: 571/753 (75.8%) in early 2020 and 402/682 (58.9%) in late 2020 ( p < .001). Crude 28‐day mortality decreased between the early and late periods (112/571 [19.6%] vs. 55/402 [13.7%]) and remained lower in the late period even after adjusting for baseline comorbidities (aOR 0.67, 95% CI 0.46–0.98, p = .016). Between the early and late periods, the use of corticosteroids (≥6 mg dexamethasone/day) and remdesivir increased (62/571 [10.9%] vs. 243/402 [61.5%], p < .001 and 50/571 [8.8%] vs. 213/402 [52.2%], p < .001, respectively), and the use of hydroxychloroquine and IL‐6/IL‐6 receptor inhibitor decreased (329/571 [60.0%] vs. 4/492 [1.0%], p < .001 and 73/571 [12.8%] vs. 5/402 [1.2%], p < .001, respectively). Mortality among SOTR hospitalized for COVID‐19 declined between early and late 2020, consistent with trends reported in the general population. The mechanism(s) underlying improved survival require further study.

Precise and accurate quantification of protein expression levels in a complex biological setting is challenging. Here, we describe a method for absolute quantitation of endogenous proteins in cell lysates using an automated capillary immunoassay system (the size-based Simple Western system, ProteinSimple, CA). The method was able to accurately measure the absolute amounts of target proteins at picogram or sub-picogram levels per nanogram of cell lysates. The measurements were independent of the cell matrix or the cell lysis buffer and were not affected by different antibody affinities for their specific epitopes. We then applied this method to quantitate absolute levels of expression of protein kinase C (PKC) isoforms in LNCaP and U937 cells, two cell lines used extensively for probing the downstream biological responses to PKC targeted ligands. Our absolute quantitation confirmed the predominance of PKCδ in both cells, supporting the important functional role of this PKC isoform in these cell lines. The method described here provides an approach to accurately quantitate levels of protein expression and correlate protein level with function. In addition to enhanced accuracy relative to conventional western analysis, it circumvents the distortions inherent in comparison of signal intensities from different antibodies with different affinities.

Lung transplant recipients (LTR) with coronavirus disease 2019 (COVID‐19) may have higher mortality than non‐lung solid organ transplant recipients (SOTR), but direct comparisons are limited. Risk factors for mortality specifically in LTR have not been explored. We performed a multicenter cohort study of adult SOTR with COVID‐19 to compare mortality by 28 days between hospitalized LTR and non‐lung SOTR. Multivariable logistic regression models were used to assess comorbidity‐adjusted mortality among LTR vs. non‐lung SOTR and to determine risk factors for death in LTR. Of 1,616 SOTR with COVID‐19, 1,081 (66%) were hospitalized including 120/159 (75%) LTR and 961/1457 (66%) non‐lung SOTR (p = .02). Mortality was higher among LTR compared to non‐lung SOTR (24% vs. 16%, respectively, p = .032), and lung transplant was independently associated with death after adjusting for age and comorbidities (aOR 1.7, 95% CI 1.0–2.6, p = .04). Among LTR, chronic lung allograft dysfunction (aOR 3.3, 95% CI 1.0–11.3, p = .05) was the only independent risk factor for mortality and age >65 years, heart failure and obesity were not independently associated with death. Among SOTR hospitalized for COVID‐19, LTR had higher mortality than non‐lung SOTR. In LTR, chronic allograft dysfunction was independently associated with mortality.

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Adult transplant hepatologists face challenges in providing care to young adults who received liver transplants during childhood. Because prior studies have focused mainly on pediatric providers, understanding these issues from the perspective of the adult hepatologist may provide novel insights and identify key barriers to care in this population. We conducted a national survey of adult transplant hepatologists to assess factors that may affect the transition of recipients from pediatric care to adult care. More than half of transplant hepatologists from all United Network for Organ Sharing regions (236/408 or 57.8%) completed the survey, and they reported that only 46.1% of patients arrived at their first adult clinic with both adequate knowledge of their condition and a parent/guardian. Moreover, 32.4% reported having no transition strategy, and only 15.5% reported having a formal transition program. The respondents reported that the greatest barriers to optimal care were patients' poor adherence and their limited knowledge and management of their condition. Those who reported participating in a formal transition program were less likely to report an inability of patients to discuss the impact of their condition on their overall daily life, fitness, and sexuality as a barrier to transition (odds ratio 5 0.40, 95% confidence interval 5 0.16-1.00). Our survey suggests that a formalized transition process is uncommon in adult transplant hepatology clinics and that improving patient knowledge, understanding specific components of effective transition programs, and incorporating input from adult providers in designing such programs may improve this process. Liver Transpl 21:213-223, 2015. V C 2014 AASLD.

Developing proteomic biomarkers is valuable for evaluating therapeutic effects of drugs and generating better treatment strategies. However, conventional protein analysis is often challenging due to inadequate sample size of clinical specimens, lack of assay reproducibility, accuracy and sensitivity. A novel capillary isoelectricfocusing (IEF) immunoassay system (NanoPro) was used to study the dynamic phosphorylation status of signaling molecules in non-small cell lung cancer (NSCLC) cells treated with EGFR tyrosine kinase and MEK inhibitors. NanoPro showed the same dynamic ERK phosphorylation as western blotting with good assay reproducibility using 1,000 times less protein. The IEF separation in NanoPro system enables multiple protein phosphorylation isoforms to be resolved and detected simultaneously. With NanoPro, we identified a specific on-target MEK response pattern to MEK inhibitor PD325901, which was not detectable by western blotting. We also revealed a MEK2 signal that may be associated with NSCLC cell sensitivity to EGFR inhibitor erlotinib, and distinguish erlotinib-sensitive from -intrinsic as well as -acquired resistant cells. Moreover, NanoPro could differentiate human ERK1 isoforms from the mouse isoforms based on their pI differences and demonstrated that erlotinib effectively inhibited ERK phosphorylation in targeted human xenograft cancer cells but not in surrounding mouse stromal cells. With 8 ug of tumor aspirates, we precisely quantified the response of 18 signaling molecules to erlotinib and MEK1 inhibitor treatments in a NSCLC patient. NanoPro’s higher sensitivity, better resolution of protein phosphorylation status and reduced tissue requirement warrant NanoPro’s investigation for future drug development and evaluation of drug effects of targeted therapies.

The bryostatins are a group of twenty macrolides isolated by Pettit and coworkers from the marine organism Bugula neritina. Bryostatin 1, the flagship member of the family, has been the subject of intense chemical and biological investigations due to its remarkably diverse biological activities, including promising indications as therapy for cancer, Alzheimer’s disease, and HIV. Other bryostatins, however, have attracted far less attention, most probably due to their relatively low natural abundance and associated scarcity of supply. Among all macrolides in this family, bryostatin 7 is biologically the most potent PKC (protein kinase C) ligand (in terms of binding affinity) and also the first bryostatin to be synthesized in the laboratory. Nonetheless, almost no biological studies have been carried out on this agent. We describe herein the total synthesis of bryostatin 7 based on our pyran annulation technology, which allows for the first detailed biological characterizations of bryostatin 7 with side-by-side comparisons to bryostatin 1. The results suggest that the more easily synthesized and less lipophilic bryostatin 7 may be an effective surrogate for bryostatin 1.

The 6MWT is a simple test of physical functioning but may be difficult to apply in LT recipients. The 6MWT performance improved following LT but was lower than expected, suggesting a low level of fitness up to 1 year following LT.