Oxygen is toxic across all three domains of life. Yet, the underlying molecular mechanisms remain largely unknown. Here, we systematically investigate the major cellular pathways affected by excess ...molecular oxygen. We find that hyperoxia destabilizes a specific subset of Fe-S cluster (ISC)-containing proteins, resulting in impaired diphthamide synthesis, purine metabolism, nucleotide excision repair, and electron transport chain (ETC) function. Our findings translate to primary human lung cells and a mouse model of pulmonary oxygen toxicity. We demonstrate that the ETC is the most vulnerable to damage, resulting in decreased mitochondrial oxygen consumption. This leads to further tissue hyperoxia and cyclic damage of the additional ISC-containing pathways. In support of this model, primary ETC dysfunction in the Ndufs4 KO mouse model causes lung tissue hyperoxia and dramatically increases sensitivity to hyperoxia-mediated ISC damage. This work has important implications for hyperoxia pathologies, including bronchopulmonary dysplasia, ischemia-reperfusion injury, aging, and mitochondrial disorders.
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•Mechanisms of oxygen toxicity identified using genome-wide CRISPR-Cas9 screen•Hyperoxia degrades specific Fe-S cluster-containing proteins in vitro and in vivo•Hyperoxic ETC degradation decreases tissue oxygen consumption, causing further hyperoxia•Primary genetic ETC dysfunction increases oxygen sensitivity of same pathways
Baik et al. systematically investigate the major cellular pathways affected by excess molecular oxygen in vitro and in vivo. They find that hyperoxia impairs diphthamide synthesis, de novo purine biosynthesis, nucleotide excision repair, and ETC bioenergetics due to the degradation of specific labile Fe-S cluster-containing proteins. Additionally, they prove a new model of cyclic oxygen toxicity.
Allogeneic hematopoietic cell transplantation (HCT) after high-dose conditioning regimens imposes prohibitively high risks of morbidity and mortality for patients with high-risk acute myeloid ...leukemia (AML) who are older or have comorbid conditions. Here, we examined outcomes after nonmyeloablative allogeneic HCT in such patients.
Two hundred seventy-four patients (median age, 60 years) with de novo or secondary AML underwent allogeneic HCT from related (n = 118) or unrelated donors (n = 156) after conditioning with 2 Gy of total-body irradiation (TBI) with or without fludarabine. A calcineurin inhibitor and mycophenolate mofetil were used for postgrafting immunosuppression.
With a median follow-up of 38 months in surviving patients, the estimated overall survival at 5 years was 33%. The estimated 5-year relapse/progression and nonrelapse mortality rates were 42% and 26%, respectively. The cumulative incidences of grades 2, 3, and 4 acute graft-versus-host disease (GVHD) were 38%, 9%, and 5%, respectively. The cumulative incidence of chronic GVHD at 5 years was 44%. Patients in first and second complete remission had better survival rates than patients with more advanced disease (37% and 34% v 18%, respectively). Patients with HLA-matched related or unrelated donors had similar survivals. Unfavorable cytogenetic risk status was associated with increased relapse and subsequent mortality. Chronic GVHD was associated with lower relapse risk.
Allogeneic HCT from related or unrelated donors after conditioning with low-dose TBI and fludarabine, relying almost exclusively on graft-versus-leukemia effects, can result in long-term remissions in older or medically infirm patients with AML.
Exosomes can serve as delivery vehicles for advanced therapeutics. The components necessary and sufficient to support exosomal delivery have not been established. Here we connect biochemical ...composition and activity of exosomes to optimize exosome-mediated delivery of small interfering RNAs (siRNAs). This information is used to create effective artificial exosomes. We show that serum-deprived mesenchymal stem cells produce exosomes up to 22-fold more effective at delivering siRNAs to neurons than exosomes derived from control cells. Proteinase treatment of exosomes stops siRNA transfer, indicating that surface proteins on exosomes are involved in trafficking. Proteomic and lipidomic analyses show that exosomes derived in serum-deprived conditions are enriched in six protein pathways and one lipid class, dilysocardiolipin. Inspired by these findings, we engineer an "artificial exosome," in which the incorporation of one lipid (dilysocardiolipin) and three proteins (Rab7, Desmoplakin, and AHSG) into conventional neutral liposomes produces vesicles that mimic cargo delivering activity of natural exosomes.
We reported encouraging early results of allogeneic hematopoietic cell transplantation (HCT) after nonmyeloablative conditioning in 64 patients who had advanced chronic lymphocytic leukemia (CLL). ...Here, we have extended the follow-up to a median of 5 years and have included data on an additional 18 patients.
Eighty-two patients, age 42 to 72 years, who had fludarabine-refractory CLL were conditioned with 2 Gy total-body irradiation alone or combined with fludarabine followed by HCT from related (n = 52) or unrelated (n = 30) donors.
Complete remission (CR) and partial remission were achieved in 55% and 15% of patients, respectively. Higher CR rates were noted after unrelated HCT (67% v 48%). The 5-year incidences of nonrelapse mortality (NRM), progression/relapse, overall survival, and progression-free survival were 23%, 38%, 50%, and 39%, respectively. Among 25 patients initially reported in CR, 8% relapsed and 8% died as a result of NRM, whereas 84% have remained alive and in CR. Among 14 responding patients who were tested and who had molecular eradication of their disease, two died as a result of NRM, two relapsed, and 10 have remained negative. At 5 years, 76% of living patients were entirely well, whereas 24% continued to receive immunosuppression for chronic graft-versus-host disease; the median performance status in each group was 100% and 90%, respectively. Lymphadenopathy > or = 5 cm, but not cytogenetic abnormalities at HCT, predicted relapse. In a risk-stratification model, patients who had lymphadenopathy less than 5 cm and no comorbidities had a 5-year OS of 71%.
Nonmyeloablative HCT resulted in a median survival of 5 years for patients who had fludarabine-refractory CLL with sustained remissions and in the continued resolution of chronic graft-versus-host disease in surviving patients.
LPCAT3 is an integral membrane acyltransferase in the Lands cycle responsible for generating C20:4 phospholipids and has been implicated in key biological processes such as intestinal lipid ...absorption, lipoprotein assembly, and ferroptosis. Small-molecule inhibitors of LPCAT3 have not yet been described and would offer complementary tools to genetic models of LPCAT3 loss, which causes neonatal lethality in mice. Here, we report the discovery by high-throughput screening of a class of potent, selective, and cell-active inhibitors of LPCAT3. We provide evidence that these compounds inhibit LPCAT3 in a biphasic manner, possibly reflecting differential activity at each subunit of the LPCAT3 homodimer. LPCAT3 inhibitors cause rapid rewiring of polyunsaturated phospholipids in human cells that mirrors the changes observed in LPCAT3-null cells. Notably, these changes include not only the suppression of C20:4 phospholipids but also corresponding increases in C22:4 phospholipids, providing a potential mechanistic explanation for the partial but incomplete protection from ferroptosis observed in cells with pharmacological or genetic disruption of LPCAT3.
Today, there is a huge effort to develop cancer immunotherapeutics capable of combating cancer cells as well as the biological environment in which they can grow, adapt, and survive. For such ...treatments to benefit more patients, there is a great need to dissect the complex interplays between tumor cells and the host's immune system. Monitoring mechanisms of resistance to immunotherapeutics can delineate the evolution of key players capable of driving an efficacious antitumor immune response. In doing so, simultaneous and systematic interrogation of multiple biomarkers beyond single biomarker approaches needs to be undertaken. Zooming into cell-to-cell interactions using technological advancements with unprecedented cellular resolution such as single-cell spatial transcriptomics, advanced tissue histology approaches, and new molecular immune profiling tools promises to provide a unique level of molecular granularity of the tumor environment and may support better decision-making during drug development. This review will focus on how such technological tools are applied in clinical settings, to inform the underlying tumor-immune biology of patients and offer a deeper understanding of cancer immune responsiveness to immuno-oncology treatments.
•Brain–behavior relationships probed with oddball and frequency-sweep paradigms.•Convergent behavioral and neural measures at group and individual levels.•Face individuation minimum at 50 ms and ...optimum at 170 ms stimulus duration.•Large inter-individual differences in neural and behavioral responses at mid-rates.•Neural peak latency at ~185 ms predicted individuals’ behavioral accuracy.
Establishing consistent relationships between neural activity and behavior is a challenge in human cognitive neuroscience research. We addressed this issue using variable time constraints in an oddball frequency-sweep design for visual discrimination of complex images (face exemplars). Sixteen participants viewed sequences of ascending presentation durations, from 25 to 333 ms (40–3 Hz stimulation rate) while their electroencephalogram (EEG) was recorded. Throughout each sequence, the same unfamiliar face picture was repeated with variable size and luminance changes while different unfamiliar facial identities appeared every 1 s (1 Hz). A neural face individuation response, tagged at 1 Hz and its unique harmonics, emerged over the occipito-temporal cortex at 50 ms stimulus duration (25–100 ms across individuals), with an optimal response reached at 170 ms stimulus duration. In a subsequent experiment, identity changes appeared non-periodically within fixed-frequency sequences while the same participants performed an explicit face individuation task. The behavioral face individuation response also emerged at 50 ms presentation time, and behavioral accuracy correlated with individual participants’ neural response amplitude in a weighted middle stimulus duration range (50–125 ms). Moreover, the latency of the neural response peaking between 180 and 200 ms correlated strongly with individuals’ behavioral accuracy in this middle duration range, as measured independently. These observations point to the minimal (50 ms) and optimal (170 ms) stimulus durations for human face individuation and provide novel evidence that inter-individual differences in the magnitude and latency of early, high-level neural responses are predictive of behavioral differences in performance at this function.
This study investigated several determinants of radiation safety culture among radiologic technologists to determine whether factors related to work shifts or workday length affect the perception of ...workplace radiation safety.
The secondary analysis used de-identified data from 425 radiologic technologists collected with the Radiation Actions and Dimensions of Radiation Safety (RADS) questionnaire, a 35-item survey with valid and reliable psychometric properties. Respondents included radiologic technologists working in radiography, computed tomography (CT), mammography, and hospital radiology administration. Descriptive statistics were used to report RADS survey item outcomes, and analysis of variance (ANOVA) tests with Games-Howell post hoc tests were conducted to analyze the hypotheses.
Mean differences in perception of teamwork across imaging stakeholders (
< .001) and leadership actions (
= .001) were found across shift-length groups. In addition, mean differences in perception of teamwork across imaging stakeholders (
= .007) were found across work-shift groups.
Longer shifts (≥ 12 hours) and night shifts are related to a diminished perception of the importance of radiation safety among radiologic technologists. The study showed a significant effect of these shift factors on the perception of teamwork and leadership actions concerning radiation safety.
These results underscore the importance of leadership actions and messaging, teamwork-building, and in-service training on radiation safety for technologists who frequently work long, after-hours shifts.
Crystal structure prediction (CSP) calculations can reduce risk and improve efficiency during drug development. Traditionally, CSP calculations use lattice energies computed through density ...functional theory. While this approach is often successful in predicting the low energy structures, it neglects the crucial role of thermal effects on polymorph stabilities. In the present study, we develop a robust and efficient protocol for predicting the relative stability of polymorphs at different temperatures. The protocol is executed on a highly parallel cloud computing infrastructure to produce results at time scales useful for drug development timelines. We demonstrate this protocol on molecule XXIII from the sixth crystal structure prediction blind test. Our results predict that Form D is the most stable experimentally observed polymorph at ambient temperature and Form C is the most stable at low temperature consistent with experiments also conducted in the present study.
Noonan syndrome, a developmental disorder characterized by congenital heart defects, reduced growth, facial dysmorphism and variable cognitive deficits, is caused by constitutional dysregulation of ...the RAS-MAPK signaling pathway. Here we report that germline NRAS mutations conferring enhanced stimulus-dependent MAPK activation account for some cases of this disorder. These findings provide evidence for an obligate dependency on proper NRAS function in human development and growth.
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