Structure-Based Design, Docking and Binding Free Energy Calculations of A366 Derivatives as Spindlin1 Inhibitors.

The chromatin reader protein Spindlin1 plays an important role in epigenetic regulation, through which it has been linked to several types of malignant tumors. In the current work, we report on the development of novel analogs of the previously published lead inhibitor A366. In an effort to improve the activity and explore the structure-activity relationship (SAR), a series of 21 derivatives was synthesized, tested in vitro, and investigated by means of molecular modeling tools. Docking studies and molecular dynamics (MD) simulations were performed to analyze and rationalize the structural differences responsible for the Spindlin1 activity. The analysis of MD simulations shed light on the important interactions. Our study highlighted the main structural features that are required for Spindlin1 inhibitory activity, which include a positively charged pyrrolidine moiety embedded into the aromatic cage connected via a propyloxy linker to the 2-aminoindole core. Of the latter, the amidine group anchor the compounds into the pocket through salt bridge interactions with Asp184. Different protocols were tested to identify a fast in silico method that could help to discriminate between active and inactive compounds within the A366 series. Rescoring the docking poses with MM-GBSA calculations was successful in this regard. Because A366 is known to be a G9a inhibitor, the most active developed Spindlin1 inhibitors were also tested over G9a and GLP to verify the selectivity profile of the A366 analogs. This resulted in the discovery of diverse selective compounds, among which 1s and 1t showed Spindlin1 activity in the nanomolar range and selectivity over G9a and GLP. Finally, future design hypotheses were suggested based on our findings.

Urea-Substituted Tetramethylcyclopentadienyl Ligands for Supramolecularly Accelerated RhIII -Catalyzed ortho-C-H Olefination of Benzoic Acid Derivatives.

The design and synthesis of air-stable and conveniently crystallizable RhIII -cyclopentadienyl catalysts substituted with a urea moiety, which are able to accelerate the C-H olefination of benzoic acid derivatives, is reported. Through kinetic studies and NMR titration experiments, the catalysts' substrate recognition ability mediated by hydrogen bonding was identified to be the reason for this effect. Introduction of pyridone-phosphine ligands capable of forming additional H-bond interactions increased the catalytic performance even further. By unveiling a proportionality between reaction rate and relative complex formation enthalpy the hypothesis of a supramolecular catalyst preformation was supported. Its application to a variety of substrates proved the catalyst system's advantages, generally increasing the yields when compared to the results obtained with widely used [RhCp*Cl2 ]2 .

Facilitating Volunteer Community Engagement Service.

BACKGROUND: Community engagement is an emerging form of pedagogy in nursing education that requires students and faculty to go beyond the traditional classroom setting to generate meaningful community-based experiences. Service-learning and volunteering are strategies nurse educators use for community service work. There is a gap in specific guidance for the faculty role in facilitating community-engaged volunteer service. METHOD: A case study describes collaboration between two faculty members and a community RN to develop volunteer service weekend trips requested by undergraduate nursing students. They shared responsibilities for planning and offering the trips through a co-facilitator role based on community engagement principles. RESULTS: The trip facilitators' collaboration resulted in a positive and productive community volunteer service experience for undergraduate nursing students. CONCLUSION: The community-engaged pedagogy provided a framework for building capacity between an academic and a community organization for student volunteer service. [J Nurs Educ. 2020;59(3):166-168.].

Do trauma courses change practice? A qualitative review of 20 courses in East, Central and Southern Africa.

BACKGROUND: Trauma courses have been shown to improve clinical knowledge and patient outcomes. However, little is known about the individual drivers of change in practice amongst course participants in their home clinic environment. METHODS: Front-line healthcare workers participated in a two-day Primary Trauma Care (PTC) course. Immediately after the course participants completed an evaluation survey on intended change in the management of trauma patients. Six months after the course, participants completed a survey on actual changes that had occurred. RESULTS: A total of 451 participants were sampled, with 321 responding at 6 months, from 40 courses across East, Central and Southern Africa. The most commonly reported intended change was the adoption of an ABCDE/systematic approach (53%). Six months after the course, 92.7% of respondents reported that they had made changes in their management, with adoption of an ABCDE/systematic approach (50.0%) remaining most common. 77% of participants reported an improvement in departmental trauma management, 26% reported an increase in staffing, 29% an increase in equipment and 68% of participants had gone on to train other healthcare workers in PTC. CONCLUSION: The findings suggest that PTC courses not only improve individual management of trauma patients but also but is also associated with beneficial effects for participants' host institutions with regards to staffing, equipment and training.

Plasma C4d+ Endothelial Microvesicles Increase in Acute Antibody-Mediated Rejection.

BACKGROUND: Antibody-mediated rejection (AMR) is a major cause of kidney allograft loss. Currently, AMR diagnosis relies on biopsy which is an invasive procedure. A noninvasive biomarker of acute AMR could lead to early diagnosis and treatment of this condition and improve allograft outcome. Microvesicles are membrane-bound vesicles released from the cell surface after injury. We hypothesized that because AMR is associated with allograft endothelial injury and C4d deposition, plasma microvesicles positive for endothelial (CD144) marker and C4d are increased in this condition. METHODS: We studied microvesicle concentration in the plasma of 95 kidney transplant patients with allograft dysfunction and compared with 23 healthy volunteers. Biopsy diagnosis and scoring was performed using Banff classification. RESULTS: In the 28 subjects with AMR, the density of C4d+/CD144+ microvesicles was on average 11-fold (P = 0.002) higher than transplant recipients with no AMR and 24-fold (P = 0.008) than healthy volunteers. Densities of C4d+ and C4d+/annexin V+ (C4d+/AVB+) microvesicles were also increased in AMR patients compared with no AMR and healthy subjects. C4d+/AVB+ microvesicles correlated with AMR biopsy severity. Nine patients with acute AMR that received treatment showed a mean 72% decrease (P = 0.01) in C4d+/CD144+ microvesicle concentration compared with pretreatment values. CONCLUSIONS: Quantification of plasma C4d+ microvesicles provides information about presence of AMR, its severity and response to treatment in transplant patients.

Development of a Coordinatively Unsaturated Chiral Indenylruthenium Catalyst.

An unprecedented coordinatively unsaturated chiral indenylruthenium complex 12 was designed and synthesized to provide additional coordination sites for Ru-catalyzed asymmetric transformations. In an attempt to catalyze an asymmetric enyne cycloisomerization reaction of 1,6-enyne, significant amounts of hydroxycyclization were observed. Up to 84:16 er of the hydroxycyclization product was obtained in 2-MeTHF. This chiral indenylruthenium catalyst could also perform an asymmetric redox isomerization/C-H insertion reaction in up to 90:10 er.

Midterm Survivorship of the Lefèvre Constrained Liner: A Consecutive Multisurgeon Series of 166 Cases.

BACKGROUND: Constrained liners are used as part of a salvage procedure to provide stability for patients at high risk for dislocation after a total hip arthroplasty. However, no recent studies exist highlighting their effectiveness and/or limitations. METHOD: This prospective review included 166 consecutive hip arthroplasties, either primary (27%) or revision (73%), with a unique design of a constrained liner: Lefèvre retentive cup. There were 113 females (69%), and the average age at index surgery was 75.9 years (range, 35-94). The mean follow-up was 6.2 years (range, 0.3-11). RESULTS: Twenty patients had a reoperation; 10 for infection (4 acute and 6 chronic joint infection) and 10 for cup failure (5 fixation failure, 3 aseptic loosening, and 2 dislocation). Ten-year survivals for cup revision were 89% (CI, 83-94) and 92% (CI, 89-97) for all revision and revision for noninfectious reasons, respectively. When solely evaluating for dislocation, the survival at 10 years was 99% (CI, 97-100). Considering primary and revision cases, 10-year survivals cup revision for aseptic reasons were 92.4% (CI, 84-100) and 92.5% (CI, 87-98), respectively. CONCLUSIONS: The Lefèvre retentive cup demonstrated excellent 10-year's survivorship. With the rate of aseptic loosening around 2% and a dislocation rate around 1%, the cup is as effective as other available devices and is therefore a cost-effective tool to reduce the risk of dislocation in at-risk patients undergoing hip arthroplasty.

Impact of Allele-Level HLA Mismatch on Outcomes in Recipients of Double Umbilical Cord Blood Transplantation.

The impact of allele-level HLA mismatch is uncertain in recipients of double umbilical cord blood (UCB) transplantation. We report a single-center retrospective study of the clinical effect of using allele-level HLA mismatch HLA-A, -B, -C, -DRB1, and -DQB1 of the 2 UCB units. We studied 342 patients with hematologic malignancy. Donor-recipient pairs were grouped according to the number of matched HLA alleles, with 32 matched at 9-10/10, 202 at 6-8/10, and 108 at 2-5/10 alleles. The incidence of hematopoietic recovery, acute and chronic graft-versus-host disease, and nonrelapse mortality and treatment failure was similar between groups. In an exploratory analysis of 174 patients with acute leukemia, after adjusting for length of first remission and cytogenetic risk group, a 2-5/10 HLA match was associated with lower risk of relapse and treatment failure. These data indicate that a high degree of allele-level HLA mismatch does not adversely affect transplant outcomes and may be associated with reduced relapse risk in patients with acute leukemia.

Acute dietary nitrate supplementation does not augment submaximal forearm exercise hyperemia in healthy young men.

Despite the popularity of dietary nitrate supplementation and the growing evidence base of its potential ergogenic and vascular health benefits, there is no direct information about its effects on exercising limb blood flow in humans. We hypothesized that acute dietary nitrate supplementation from beetroot juice would augment the increases in forearm blood flow, as well as the progressive dilation of the brachial artery, during graded handgrip exercise in healthy young men. In a randomized, double-blind, placebo-controlled crossover study, 12 young (22 ± 2 years) healthy men consumed a beetroot juice (140 mL Beet-It Sport, James White Juice Company) that provided 12.9 mmol (0.8 g) of nitrate or placebo (nitrate-depleted Beet-It Sport) on 2 study visits. At 3 h postconsumption, brachial artery diameter, flow, and blood velocity were measured (Doppler ultrasound) at rest and during 6 exercise intensities. Nitrate supplementation raised plasma nitrate (19.5-fold) and nitrite (1.6-fold) concentrations, and lowered resting arterial pulse wave velocity (PWV) versus placebo (all p < 0.05), indicating absorption, conversion, and a biological effect of this supplement. The supplement-associated lowering of PWV was also negatively correlated with plasma nitrite (r = -0.72, p = 0.0127). Despite these systemic effects, nitrate supplementation had no effect on brachial artery diameter, flow, or shear rates at rest (all p ≥ 0.28) or during any exercise workload (all p ≥ 0.18). These findings suggest that acute dietary nitrate supplementation favorably modifies arterial PWV, but does not augment blood flow or brachial artery vasodilation during nonfatiguing forearm exercise in healthy young men.

Anti-HLA antibodies in double umbilical cord blood transplantation.

Recent registry data suggest that host-versus-graft alloreactions mediated by anti-donor HLA antibodies in recipients of adult allogeneic hematopoietic stem cells or single-unit umbilical cord blood (UCB) contribute to the risk of graft failure. The present study evaluated the impact of anti-HLA antibodies on engraftment and unit predominance in 126 double-UCB (dUCB) recipients. Eighteen dUCB recipients were identified with at least 1 of 2 UCB units recognized by anti-HLA antibodies directed against donor-directed HLA-specific antibodies (DSAs). Overall, 9 of 12 patients who had DSAs against 1 of the 2 UCB units composing the graft and 5 of 6 patients who had DSAs against both units engrafted. The cumulative incidence of engraftment was similar in patients with and without DSAs (83% vs 78%). Thus, our data do not support a negative effect of anti-HLA antibodies on engraftment, at least in the setting of cyclosporine and mycophenolate mofetil and the conditioning regimens used at the University of Minnesota, and argue against routine screening for use in graft selection before dUCB transplantation. Further studies are needed to fully understand the value of anti-HLA antibody testing in dUCB graft selection and its impact on transplantation outcomes.

Impact of chronic intermittent external compressions on forearm blood flow capacity in humans.

During dynamic exercise, the vasculature embedded within skeletal muscle intermittently collapses due to increased intramuscular pressure (IMP). The aim of this study was to ascertain whether oscillations in IMP during muscle contractions independently contribute to exercise training-induced increases in blood flow capacity (BFC). Based on IMP measurements during handgrip exercise, we attempted to mimic the action of repeated vascular compressions by using external inflatable cuffs. Thus, 24 healthy young male subjects underwent a 4-week program (5 days/week, 1 h/day) of application of external compressions of the non-dominant forearm, while the dominant limb served as an internal control. To evaluate the impact of compression pressures of different magnitudes, subjects were randomly assigned to one of three groups: 50, 100 and 150 mmHg of external compression. Prior to the intervention and after 2 and 4 weeks of treatment, we measured peak forearm blood flow (PBF) (Doppler ultrasound) and calculated peak vascular conductance (PVC) following 10 min of forearm ischemia. In the 50 and 100 mmHg groups, application of intermittent compressions did not alter PBF in either control or intervention forearms. In the 150 mmHg group, there was a trend (P = 0.04) for greater increases in PBF from baseline after 4 weeks in the intervention forearm compared to the control forearm (delta PBF: 4.2 ± 2.5 vs. -2.1 ± 2.0 (ml(100 ml)(-1) min(-1)), in the intervention and control forearms, respectively), but the changes in PVC were not significant (P = 0.1). These findings suggest that repeated oscillations in IMP contribute minimally to exercise-induced increase in forearm BFC in healthy young humans.

High immunologic risk living donor kidney transplant using bortezomib in a novel induction regimen without acute antibody mediated rejection.

Desensitization therapies have been used with modest success in kidney transplantation. Some candidates, however, have such great breadth and depth of anti-HLA antibodies that they remain incompatible with potential donors. Bortezomib has been used without much success in desensitization regimens, but we hypothesized that its use during induction may be helpful in targeting antibody production by long-lived plasma cells. This report describes a high-risk positive crossmatch son-to-mother transplant that was performed after desensitization. The induction immunosuppression was supplemented with bortezomib. Pre- and post-transplant immunosuppression, antibody monitoring, biopsy data, and the clinical course are described in detail. Following transplant, the patient had excellent early graft function. Serial biopsies did not reveal acute antibody mediated rejection. Despite excellent graft function, the patient underwent withdrawal of care and died due to complications of calciphylaxis and deconditioning. This case details the first report of bortezomib used as part of induction therapy in solid organ transplant. Donor specific antibody production remained stable after transplant, with near complete abrogation of class I specificities. There were no bortezomib-related complications.

Mislabeled units of umbilical cord blood detected by a quality assurance program at the transplantation center.

We instituted procedures to check the identity of cord blood unit provided for transplantation by carrying out ABO and human leukocyte antigen (HLA) typing of the thawed units before transplantation. ABO typing is done using standard techniques. Rapid HLA class I serology is with monoclonal antibody trays (One Lambda Inc) using standard incubations. One mislabeled umbilical cord blood (UCB) unit was detected on the day of intended transplantation by repeat ABO typing of the thawed unit at our transplantation center. Because ABO typing will not detect all labeling errors, the rapid serologic class I HLA typing procedure was done on thawed units just before transplantation for all units without an attached segment. This procedure identified a second mislabeled unit. In a 6-year period, 2 of 871 (0.2%) cord blood units sent to us for transplantation were mislabeled and potentially would have been transplanted incorrectly. This error rate of 1 per 249 (0.4%) patients could have potentially devastating consequences.

Bortezomib for refractory antibody-mediated cardiac allograft rejection.

This experience demonstrates that a bortezomib-based regimen provided effective therapy for late, refractory AMR in an adult heart transplant recipient and was well tolerated. This remarkably positive experience despite the refractory nature of the AMR episode argues strongly for continued evaluation of bortezomib use in this patient population.