bluuebunny/crossref_metadata_2025 huggingface dataset:
Perihippocampal failure after hippocampal-avoidance whole-brain radiotherapy in cancer patients with brain metastases
Authors: Shieh Li-Tsun , Lee Sung-Wei , Chen Chia-Chun , Ho Yi-Chia , Wang Yu-Wen , Ho Sheng-Yow
Published: April 2022

Perihippocampal failure is a rare clinical scenario in brain metastatic cancer patients following hippocampal-avoidance (HA) whole-brain radiotherapy (HA-WBRT). The clinical features have not been fully identified because clinical data on intracranial failure after HA-WBRT are limited. It is thus necessary to accumulate clinical data.

We retrospectively analyzed cancer patients with brain metastases who were diagnosed between January 2014 and September 2020 at a regional referral hospital. The medical records of patients who underwent HA-WBRT were reviewed. The clinical features of intracranial recurrence were described. Dosimetry parameters were compared in terms of deviation from the recommended protocol of the Radiation Therapy Oncology Report 0933.

Twenty-four eligible patients with brain metastases who underwent HA-WBRT were identified; 13 (54%) were male. Seventeen patients (71%) had lung cancer, 6 (25%) had breast cancer, and 1 (4%) had liver cancer. The median overall survival was 12 months. Three patients developed intracranial failure during clinical follow-up, and 2 relapsed with intracranial failure in the perihippocampal region at 13 and 22 months, respectively. The perihippocampal failure rate was about 8%. One patient with small cell lung cancer received HA-prophylactic cranial irradiation; the minimum and maximum doses to the hippocampi were 6.8 and 10.7 Gy, respectively. Another patient with brain metastases from lung adenocarcinoma received HA-WBRT; the minimum and maximum doses to the hippocampi were 5.4 and 10.6 Gy, respectively.

We reported unusual cases of intracranial failure in the perihippocampal region following HA-WBRT. Perihippocampal failure could be attributed to an under-dose of radiation partially or be resulted from aggressiveness of cancer per se. Further research on this topic is encouraged.
Strategies to Understand What Matters to Advanced Cancer Patients in Advance Care Planning: A Qualitative Study Using the Lifeline Interview Method
Authors: Takenouchi Sayaka , Chikada Ai , Mori Masanori , Tamura Keiko , Nin Kazuko
Published: April 2022

Little is known about how health care providers should conduct advance care planning to identify the values of East Asian patients who have serious illnesses. This study aims to explore whether and how patients from an East Asian culture and with advanced cancer express their values and priorities when nurses utilize the lifeline interview method to enable patients to reflect on their life trajectories and if it can bridge advance care planning discussions. Data obtained from individual, semistructured interviews of 11 patients with advanced lung cancer were analyzed using qualitative content analysis. Seven main themes were identified: (1) treatment and essential elements to maintaining everyday life, (2) beliefs regarding and support for being “myself”, (3) emotional ups and downs in response to physical condition, (4) competency to cope, (5) what I want to do “now” to achieve my goals, (6) goals that bring hope for life, and (7) wishes and preferences for end-of-life decision-making. Results suggest that the lifeline interview method is an excellent means for nurses and other health care professionals to elicit patients' values and priorities. Moreover, it bridges advance care planning discussions to reflect on what matters to patients in future palliative care.
SMARCB1-deficient and SMARCA4-deficient Malignant Brain Tumors With Complex Copy Number Alterations and TP53 Mutations May Represent the First Clinical Manifestation of Li-Fraumeni Syndrome
Authors: Hasselblatt Martin , Thomas Christian , Federico Aniello , Nemes Karolina , Johann Pascal D. , Bison Brigitte , Bens Susanne , Dahlum Sonja , Kordes Uwe , Redlich Antje , Lessel Lienhard , Pajtler Kristian W. , Mawrin Christian , Schüller Ulrich , Nolte Kay , Kramm Christof M. , Hinz Felix , Sahm Felix , Giannini Caterina , Penkert Judith , Kratz Christian P. , Pfister Stefan M. , Siebert Reiner , Paulus Werner , Kool Marcel , Frühwald Michael C.
Published: April 2022

Atypical teratoid/rhabdoid tumor (AT/RT) is a malignant central nervous system tumor predominantly affecting infants. Mutations of SMARCB1 or (rarely) SMARCA4 causing loss of nuclear SMARCB1 or SMARCA4 protein expression are characteristic features, but further recurrent genetic alterations are lacking. Most AT/RTs occur de novo, but secondary AT/RTs arising from other central nervous system tumors have been reported. Malignant gliomas, IDH wild-type, arising in patients with Li-Fraumeni syndrome typically show somatic mutations of TP53 as well as complex copy number alterations, but little is known about the loss of SMARCB1 or SMARCA4 protein expression in this context. Here, we report 2 children in whom malignant supratentorial brain tumors with SMARCB1 deficiency, complex copy number alterations, and somatic TP53 mutations lead to the discovery of pathogenic/likely pathogenic TP53 variants in the germline. Screening of the molecularneuropathology.org dataset for cases with similar genetic and epigenetic alterations yielded another case with SMARCA4 deficiency in a young adult with Li-Fraumeni syndrome. In conclusion, SMARCB1-deficient or SMARCA4-deficient malignant brain tumors with complex copy number alterations and somatic TP53 mutations in children and young adults may represent the first clinical manifestation of Li-Fraumeni syndrome and should prompt genetic counseling and investigation for TP53 germline status.
“The More We Know, the More We're Able to Help”: Participatory Development of an Evaluation Framework for Community Health Worker Programs
Authors: Sherman Mya , Covert Hannah H. , Lichtveld Maureen Y.
Published: April 2022

Context:

It is critical to evaluate community health worker (CHW) programs to maximize effectiveness. However, there is little consensus, and structure, for how to evaluate such programs.

Objective:

The purpose of this study was to develop a flexible framework for evaluating components of CHW programs in community and clinical settings.

Design:

Semistructured interviews were conducted with CHWs and their supervisors to determine evaluation categories for the framework. A survey was then administered to pilot the framework at federally qualified health centers and community-based organizations with existing CHW programs.

Setting:

Eighteen federally qualified health centers and community-based organizations in Louisiana, Mississippi, Alabama, and Florida.

Participants:

Participants consisted of 23 CHWs and 19 CHW supervisors.

Results:

Interview participants directly informed the development of an evaluation framework for CHW programs. The framework consists of 7 evaluation categories: client relations, intraorganizational relations, interorganizational relations, capacity development, program effectiveness, cost-efficiency, and sustainability. Survey respondents specified best practices for program implementation for each evaluation category. Recommendations for CHW program evaluation include tailoring evaluation efforts and data collection methods to program context, using mixed-methods approaches for collecting evaluation data, and streamlining evaluation efforts with an organization's existing evaluation systems.

Conclusions:

The evaluation framework is a flexible and practical model for collecting information needed for monitoring and evaluating CHW programs. By relying on practitioners' perspectives, this study contributes to an evidence base for implementing and evaluating CHW programs.
Developing a National Trauma Research Action Plan: Results from the acute resuscitation, initial patient evaluation, imaging, and management research gap Delphi survey
Authors: Costantini Todd W. , Galante Joseph M. , Braverman Maxwell A. , Phuong Jim , Price Michelle A. , Cuschieri Joseph , Godat Laura N. , Holcomb John B. , Coimbra Raul , Bulger Eileen M. ,   NTRAP Acute Resuscitation Panel
Published: April 2022

BACKGROUND

Injury is the leading cause of death in patients aged 1 to 45 years and contributes to a significant public health burden for individuals of all ages. To achieve zero preventable deaths and disability after injury, the National Academies of Science, Engineering and Medicine called for the development of a National Trauma Research Action Plan to improve outcomes for military and civilian trauma patients. Because rapid resuscitation and prompt identification and treatment of injuries are critical in achieving optimal outcomes, a panel of experts was convened to generate high-priority research questions in the areas of acute resuscitation, initial evaluation, imaging, and definitive management on injury.

METHODS

Forty-three subject matter experts in trauma care and injury research were recruited to perform a gap analysis of current literature and prioritize unanswered research questions using a consensus-driven Delphi survey approach. Four Delphi rounds were conducted to generate research questions and prioritize them using a 9-point Likert scale. Research questions were stratified as low, medium, or high priority, with consensus defined as ≥60% of panelists agreeing on the priority category. Research questions were coded using a taxonomy of 118 research concepts that were standard across all National Trauma Research Action Plan panels.

RESULTS

There were 1,422 questions generated, of which 992 (69.8%) reached consensus. Of the questions reaching consensus, 327 (33.0%) were given high priority, 621 (62.6%) medium priority, and 44 (4.4%) low priority. Pharmaceutical intervention and fluid/blood product resuscitation were most frequently scored as high-priority intervention concepts. Research questions related to traumatic brain injury, vascular injury, pelvic fracture, and venous thromboembolism prophylaxis were highly prioritized.

CONCLUSION

This research gap analysis identified more than 300 high-priority research questions within the broad category of Acute Resuscitation, Initial Evaluation, Imaging, and Definitive Management. Research funding should be prioritized to address these high-priority topics in the future.
A comparison of management and outcomes following blunt versus penetrating pancreatic trauma: A secondary analysis from the Western Trauma Association Multicenter Trials Group on Pancreatic Injuries
Authors: Biffl Walter L. , Ball Chad G. , Moore Ernest E. , West Michaela , Russo Rachel M. , Balogh Zsolt , Kornblith Lucy , Callcut Rachael , Schaffer Kathryn B. , Castelo Matthew ,   the WTA Multicenter Trials Group on Pancreatic Injuries
Published: April 2022

BACKGROUND

The impact of injury mechanism on outcomes of pancreatic trauma has not been well studied, and current guidelines do not differentiate recommendations for blunt and penetrating injuries. The purpose of this study was to analyze interventions and outcomes as they relate to mechanism. We hypothesized that penetrating pancreatic trauma results in greater morbidity than blunt trauma because of more frequent operative exploration without imaging and thus more aggressive surgical management.

METHODS

Secondary analysis of a multicenter retrospective review of pancreatic injuries in patients 15 years and older from 2010 to 2018 was performed. Deaths within 24 hours of admission were excluded from analysis of the primary outcome, pancreas-related complications (PRCs). Data were analyzed by injury mechanism using various statistical tests where appropriate.

RESULTS

Thirty-three centers reported on 1,240 patients (44% penetrating). Penetrating trauma patients were twice as likely to undergo resection (45% vs. 23%) and suffer PRCs (39% vs. 20%). However, differences varied widely based on injury grade and management. There were fewer resections and more nonoperative management in blunt grades I to III injury. Pancreas-related complications occurred in 40% of high-grade injuries with no difference between mechanisms and in 40% of patients after resection, regardless of mechanism or injury grade. High-grade pancreatic injury (odds ratio [OR], 2.39; 95% confidence interval [CI], 1.55–3.67), penetrating injury (OR, 1.99; 95% CI, 1.31–3.05), and management in a low-volume center (i.e., five or fewer cases/year) (OR, 1.65; 95% CI, 1.16–2.35) were independent predictors of PRCs.

CONCLUSION

Management of grades I to III, but not grades IV/V, pancreatic injuries varies based on mechanism. Penetrating injury is an independent risk factor for PRCs, but main pancreatic duct injury and resection are associated with high rates of PRCs regardless of the injury mechanism. Resection appears to offer better outcomes for grade IV/V injuries, and grade I and II injuries should be managed nonoperatively.

LEVEL OF EVIDENCE

Therapeutic/Care Management; Level III.
Hemodynamic events during en route critical care for patients with traumatic brain injury
Authors: Davis William T. , Medellin Kimberly L. , Cutright Julie E. , Arana Allyson A. , Strilka Richard , Sorensen Derek , Maddry Joseph K. , Savell Shelia C. , Ng Patrick C.
Published: April 2022

BACKGROUND

Exposure to stressors of flight may increase risk of secondary insults among critically injured combat casualties wounded with traumatic brain injury (TBI). The primary objective of this study was to describe the prevalence of hemodynamic events by phase of transport among patients with TBI transported by Critical Care Air Transport Teams (CCATT).

METHODS

We performed a secondary analysis of a retrospective cohort of 477 adults with moderate to severe TBI, who required transport by CCATT to Germany from multiple hospitals in the Middle East between January 2007 and May 2014. We abstracted clinical data from handwritten CCATT medical records. Hemodynamic events included systolic blood pressure <100 mm Hg and cerebral perfusion pressure <60 mm Hg. We calculated the proportion of patients experiencing hemodynamic events for each phase of flight.

RESULTS

We analyzed 404 subjects after exclusions for catastrophic brain injury (n = 39) and missing timestamps (n = 34). Subjects had high Injury Severity Scores (median, 29; interquartile range [IQR], 21–35) and a median flight time of 423 minutes (IQR, 392.5–442.5 minutes). The median of documented in-flight vital signs was 8 measurements (IQR, 6.5–8 measurements). Documented systolic blood pressure in-flight events occurred in 3% of subjects during ascent, 7.9% during early flight, 7.7% during late flight, and 2.2% during descent, with an overall in-flight prevalence of 13.9%. Among patients with intracranial pressure monitoring (n = 120), documented cerebral perfusion pressure events occurred in 5% of subjects during ascent, 23% during early flight, 17% during late flight, and 5.8% during descent, with an overall in-flight prevalence of 30.8%.

CONCLUSION

Documented hemodynamic events occurred during each phase of flight in severely injured combat casualties wounded with TBI, and episodic documentation likely underestimated the actual in-flight frequency of secondary insults.

LEVEL OF EVIDENCE

Prognostic and epidemiological; Level IV.
Postinjury platelet aggregation and venous thromboembolism
Authors: Matthay Zachary A. , Hellmann Zane J. , Nunez-Garcia Brenda , Fields Alexander T. , Cuschieri Joseph , Neal Matthew D. , Berger Jeffrey S. , Luttrell-Williams Elliot , Knudson M. Margaret , Cohen Mitchell J. , Callcut Rachael A. , Kornblith Lucy Z.
Published: April 2022

BACKGROUND

Posttraumatic venous thromboembolism (VTE) remains prevalent in severely injured patients despite chemoprophylaxis. Importantly, although platelets are central to thrombosis, they are not routinely targeted in prevention of posttraumatic VTE. Furthermore, platelets from injured patients show ex vivo evidence of increased activation yet impaired aggregation, consistent with functional exhaustion. However, the relationship of this platelet functional phenotype with development of posttraumatic VTE is unknown. We hypothesized that, following injury, impaired ex vivo platelet aggregation (PA) is associated with the development of posttraumatic VTE.

METHODS

We performed a secondary analysis of 133 severely injured patients from a prospective observational study investigating coagulation and inflammation (2011–2019). Platelet aggregation in response to stimulation with adenosine diphosphate (ADP), collagen, and thrombin was measured at presentation (preresuscitation) and 24 hours (postresuscitation). Viscoelastic clot strength and lysis were measured in parallel by thromboelastography. Multivariable regression examined relationships between PA at presentation, 24 hours, and the change (δ) in PA between presentation and 24 hours with development of VTE.

RESULTS

The 133 patients were severely injured (median Injury Severity Score, 25), and 14% developed VTE (all >48 hours after admission). At presentation, platelet count and PA were not significantly different between those with and without incident VTE. However, at 24 hours, those who subsequently developed VTE had significantly lower platelet counts (126 × 109/L vs. 164 × 109/L, p = 0.01) and lower PA in response to ADP (p < 0.05), collagen (p < 0.05), and thrombin (p = 0.06). Importantly, the magnitude of decrease in PA (δ) from presentation to 24 hours was independently associated with development of VTE (adjusted odds ratios per 10 aggregation unit decrease: δ-ADP, 1.31 [p = 0.03]; δ-collagen, 1.36 [p = 0.01]; δ-thrombin, 1.41 [p < 0.01]).

CONCLUSION

Severely injured patients with decreasing ex vivo measures of PA despite resuscitation have an increased risk of developing VTE. This may have implications for predicting development of VTE and for studying platelet targeted chemoprophylaxis regimens.

LEVEL OF EVIDENCE

Prognostic/Epidemiological; Level III.
Rhizorhabdus phycosphaerae sp. nov., isolated from the phycosphere of Microcystis aeruginosa
Authors: Kim Minkyung , Bae Junshik , Park Woojun
Published: April 2022

A novel bacterial strain, designated MK52T, was isolated from the phycosphere of

Microcystis aeruginosa

. Strain MK52T is a Gram-stain-negative, pink-pigmented, rod-shaped, strictly aerobic bacterium. In 16S rRNA phylogenetic analysis, the MK52T strain was most closely related to

Rhizorhabdus wittichii

RW1T (98.66 %) and

Rhizorhabdus histidinilytica

UM2T (98.51 %). The genomic DNA G+C content of strain MK52T was calculated to be 65.5 mol%. The average nucleotide identity values of strain MK52T with

R. wittichii

RW1T and

R. histidinilytica

UM2T were 80.35 and 80.23 %, respectively, with digital DNA–DNA hybridization values of 23.6 and 22.9 %, respectively, and average amino acid identities of 75.59 and 75.79 %, respectively. The major isoprenoid quinone was Q-10, and the predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and sphingoglycolipid. Fatty acid methyl ether analysis showed that summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) was the main cellular fatty acid in strain MK52T. Strain MK52T cells grew at 21–34 °C (optimum, 30 °C), pH 5–8 (optimum, pH 7) and with 0–2 % (w/v) NaCl (optimum, 0.5 % NaCl). Rhizorhabdus phycosphaerae sp. nov. is proposed as a new species (=KCTC 72877T=DSM 111424T) based on its genotypic and phenotypic characteristics.
Estimating the risk of hospitalisation and death in England’s remaining unvaccinated population
Authors: Shingleton Joseph , Dyke Steven , Herrick Archie , Walker Brodie , Finnie Thomas
Published: April 2022

The roll-out of COVID-19 vaccines in the England has generally been very good – with over 80% of over 12s having received two doses of vaccine by the start of February 2022, and 67% having received a further booster dose. Despite this, there is a small section of the population who remain unvaccinated, either due to lack of access, hesitancy or resistant to vaccination. In this report we estimate that, during 2021, there were approximately 3,500 deaths in unvaccinated people, who could otherwise have reasonably been expected to receive a vaccination. Further, we show that if all of the remaining unvaccinated population in England were to become infected (or reinfected) with the Omicron variant of COVID-19, we would expect to see approximately 11,700 further deaths and 29,600 hospitalisations. These number could fall to 5,300 and 19,600 respectively, if all but the most vaccine resistant individuals become fully vaccinated.
Persistent cross-species SARS-CoV-2 variant infectivity predicted via comparative molecular dynamics simulation
Authors: Rajendran Madhusudan , Babbitt Gregory A.
Published: April 2022

Widespread human transmission of SARS-CoV-2 highlights the substantial public health, economic, and societal consequences of virus spillover from wildlife and also presents a repeated risk of reverse spillovers back to naïve wildlife populations. We employ comparative statistical analyses of a large set of short-term molecular dynamic (MD) simulations to investigate potential human to bat (Rhinolophus macrotis) cross-species infectivity allowed by the binding of SARS-CoV-2 receptor-binding domain (RBD) to angiotensin-converting enzyme 2 (ACE2) across the bat progenitor strain and emerging human strain variants of concern (VOC). We statistically compare the dampening of atom motion during binding across protein sites upon the formation of the RBD/ACE2 binding interface using bat vs. human target receptors (i.e. bACE2 and hACE2). We report that while the bat progenitor viral strain RaTG13 shows some pre-adaption to binding hACE2, it also exhibits stronger overall affinity to bACE2. However, while the early emergent human strains and later VOC’s exhibit robust binding to both hACE2 and bACE2, the delta and omicron variants exhibit evolutionary adaption of binding to hACE2. However, we conclude there is a still significant risk of mammalian cross-species infectivity of human VOC’s during upcoming waves of infection as COVID-19 transitions from a pandemic to endemic status.
The role of environmental calcium in the extreme acid tolerance of northern banjo frog (Limnodynastes terraereginae) larvae
Authors: Hird Coen , Franklin Craig E. , Cramp Rebecca L.
Published: April 2022

Many aquatically respiring animals inhabiting low pH waters can suffer acute inhibition of ion uptake and loss of branchial (gill) epithelial integrity, culminating in a fatal, rapid loss of body Na+. Environmental calcium levels ([Ca2+]e) are pivotal in maintaining branchial junction integrity, with supplemental Ca2+reversing the negative effects of low pH in some animals. Tolerance of some naturally acidic environments by aquatic animals is further complicated by low [Ca2+]e, yet many of these environments are surprisingly biodiverse. How these animals overcome the combined damaging actions of low pH and low environmental Ca2+remains unknown. Here, we examined the effects of [Ca2+]eon the response to low pH in larvae of the highly acid tolerant frogLimnodynastes terraereginae. Acute exposure to low pH water in the presence of low [Ca2+]eincreased net Na+efflux. Provision of additional [Ca2+]ereduced net Na+efflux, but the effect was saturable. Acclimation to both low and high [Ca2+]eimproved the resistance of larvae to Na+efflux at low pH. Inhibition of apical Ca2+uptake by ruthenium red resulted in an abrupt loss of tolerance to low pH in larvae acclimated to low pH water. Acclimation to acidic water increased branchial gene expression of the intracellular Ca2+transport protein calbindin, consistent with a role for increased transcellular Ca2+trafficking in the tolerance of acidic water. This study confirmed the physiological challenge of low [Ca2+]eon branchial integrity in acidic waters and highlighted a potential role for maintenance of transcellular Ca2+uptake in the acid tolerance ofL. terraereginae.

Summary statement

Tolerance of naturally acidic, dilute, and soft waters by larvae of the frogLimnodynastes terraereginaeinvolves adaptations to the branchial calcium transport pathway which protects intercellular junctions against damage.
A Chitin-Binding Protein Ultra-highly expressed in the Outer Fold of Mantle is Related to Shell Colour in Pacific Oyster Crassostrea gigas
Authors: Li Mai , Tang Juyan , Huang Baoyu , Liu Yaqiong , Wei Lei , Han Yijing , Wang Xiaona , Sang Xiuxiu , Yuan Mengqiang , Fan Nini , Cai Shuai , Zheng Yanxin , Zhang Meiwei , Wang Xiaotong
Published: April 2022

Molluscs constitute the second largest phylum of animals in the world, and shell colour and stripes are one of their most important phenotypic characteristics. Studies on the mechanism of shell pigmentation help understand the evolutionary and ecological significance of shell colour and serve as the basis for shell colour breeding. In this study, a matched-pair design was used in comparing the black- and white-striped mantles of the same oyster. The result showed that the stripes of shell surface are corresponding to the stripes of the mantle edge. Transcriptomic analysis identified an uncharacterized protein gene (we named it as Crassostrea gigas chitin-binding protein, CgCBP) highly expressed in the black mantles. Three folds (inner fold, middle fold and outer fold) were found on the mantle edge of oyster, but only the outer fold has the same colour as the shell. Transcriptomic comparison indicated that the three folds of the mantle edge are functionally differentiated, and the CgCBP gene is ultra-highly expressed only in the outer fold. We obtained new black and white shell periostraca from the shell notching experiment and found their structural differences by scanning electron microscope. Chitin was successfully extracted and identified from the shell periostraca. Proteomic analysis revealed differences in protein composition between the two shell periostraca. Particularly, the black shell periostraca have more proteins related to melanin biosynthesis and chitin binding compared with the white ones, and melanin particles were observed in the black mantle edge using transmission electron microscope. Magnetic bead binding and Western blot experiments showed that the CgCBP protein can specifically bind to chitin and in vivo RNAi screening indicated that CgCBP knockdown can change the structure of the shell periostracum and reduce its pigmentation. All these results suggest that the outer fold of mantle may have more important roles in shell pigmentation, shell periostracum structure is functionally correlated with shell pigmentation, and the CgCBP gene ultra-highly expressed in the outer fold may influence shell pigmentation by affecting its periostracum structure.
Transcriptome profiling of the dorsomedial prefrontal cortex in suicide victims
Authors: Dóra Fanni , Renner Éva , Keller Dávid , Palkovits Miklós , Dobolyi Árpád
Published: April 2022

The default mode network (DMN) plays an outstanding role in psychiatric disorders. Still, gene expressional changes in its major component, the dorsomedial prefrontal cortex (DMPFC), have not been characterized. We used RNA-sequencing in postmortem DMPFC samples to investigate suicide victims compared to control subjects. Most of the data variance (79%) was associated with expression changes between suicide and control samples. 1400 genes differed using log2FC > ± 1 and adjustedp-value < 0.05 criteria between groups. Genes associated with depressive disorder, schizophrenia and impaired cognition were strongly overexpressed in top differentially expressed genes. Gene set enrichment and protein-protein interaction network analysis revealed that pathways related to cytokine receptor signaling were enriched in downregulated while glutamatergic synaptic signaling in upregulated genes in suicide individuals. A validated differentially expressed gene, which is known to be associated with mGluR5, was the N-terminal EF-hand calcium-binding proteins 2 (NECAB2).In situhybridization histochemistry and immunohistochemistry proved that NECAB2 is expressed in 2 different types of inhibitory neurons located in layers II-IV and VI, respectively. Our results imply extensive gene expressional alterations in the DMPFC related to suicidal behavior. Some of these genes may contribute to the altered mental state and behavior of suicide victims.
Peer presence elicits task-independent changes within and beyond the mentalizing network across children and adults
Authors: Tricoche Leslie , Pélisson Denis , Longo Léa , Koun Eric , Poisson Alice , Prado Jérôme , Meunier Martine
Published: April 2022

There is ample behavioral evidence that others’ mere presence can affect any behavior in human and non-human animals, generally facilitating the expression of mastered responses while impairing the acquisition of novel ones. Much less is known about i) how the brain orchestrates the modulation of such a wide array of behaviors by others’ presence and ii) when these neural underpinnings mature during development. To address these issues, fMRI data were collected in children and adults alternately observed and unobserved by a familiar peer. Subjects performed two tasks. One, numerosity comparison, depends on number-processing brain areas, the other, phonological comparison, on language-processing areas. Consistently with previous behavioral findings, peer observation facilitated both tasks, and children’s improvement was comparable to adults’. Regarding brain activation, we found virtually no evidence of observation-driven changes within the number- or language-related areas specific to each task. Rather, we observed the same task-independent changes for both numerosity and phonological comparisons. This unique neural signature encompassed a large brain network of domain-general areas involved in social cognition, especially mentalizing, attention, and reward. It was also largely shared by children and adults. The one exception was children’s right temporoparietal junction, which failed to show the observation- driven lesser deactivation seen in adults. These findings indicate that social facilitation of some human education-related skills is i) primarily orchestrated by domain-general brain networks, rather than by task-selective substrates, and ii) relatively mature early in the course of education, thus having a protracted impact on academic achievements that may have heretofore been underestimated.

Highlight

Basic math and reading skills were measured in children and adults.

Participants were alternately observed and unobserved by a familiar peer.

Behavior showed that children were as facilitated by peer observation as adults.

fMRI data showed task-independent, observation-driven changes in mentalizing, attention, and reward brain regions.

All adults’ neural changes were also found in children, except one located in right temporo- parietal junction.
Cryo-EM structures of Na+-pumping NADH-ubiquinone oxidoreductase from Vibrio cholerae
Authors: Kishikawa Jun-ichi , Ishikawa Moe , Masuya Takahiro , Murai Masatoshi , Kitazumi Yuki , Butler Nicole L. , Kato Takayuki , Barquera Blanca , Miyoshi Hideto
Published: April 2022

The Na+-pumping NADH-ubiquinone oxidoreductase (Na+-NQR) couples electron transfer from NADH to ubiquinone with Na+-pumping, generating an electrochemical Na+ gradient that is essential for energy-consuming reactions in bacteria. Since Na+-NQR is exclusively found in prokaryotes, it is a promising target for highly selective antibiotics. However, the molecular mechanism of inhibition is not well-understood for lack of the atomic structural information about an inhibitor-bound state. Here we present cryo-electron microscopy structures of Na+- NQR from Vibrio cholerae with or without a bound inhibitor at 2.5- to 3.1-Å resolution. The structures reveal the arrangement of all six redox cofactors including riboflavin, whose position has been under debate, and a newly assigned 2Fe-2SNqrD/E cluster located between the membrane embedded NqrD and NqrE subunits. A large part of the hydrophilic NqrF near the cytoplasmic membrane surface is barely visible in the density map, suggesting a high degree of flexibility. This flexibility may be responsible to reducing the long distance between the 2Fe- 2S centers in NqrF and NqrD/E, consistent with physiologically relevant electron transfer. Two different types of specific inhibitors (korormicin A and aurachin D-42) bind to the N-terminal region of NqrB, which is disordered in the absence of inhibitors. The current inhibitor-bound structures reasonably explain our previous biochemical findings obtained by different chemistry-based experiments. This study provides a definite foundation for understanding the function of Na+-NQR and the molecular mechanism of its specific inhibitors to support molecular design of new antibiotics targeting the enzyme.
Structures of the T cell potassium channel Kv1.3 with immunoglobulin modulators
Authors: Selvakumar Purushotham , Fernández-Mariño Ana I. , Khanra Nandish , He Changhao , Paquette Alice J. , Wang Bing , Huang Ruiqi , Smider Vaughn V. , Rice William J. , Swartz Kenton J. , Meyerson Joel R.
Published: April 2022

The Kv1.3 potassium channel is expressed abundantly on activated T cells and mediates the cellular immune response. This role has made the channel a target for therapeutic immunomodulation to block its activity and suppress T cell activation. We determined structures of human Kv1.3 alone, with a nanobody inhibitor, and with an antibody-toxin fusion blocker. Rather than block the channel directly, four copies of the nanobody bind the tetramer’s voltage sensing domains and the pore domain to induce an inactive pore conformation. In contrast, the antibody-toxin fusion docks its toxin domain at the extracellular mouth of the channel to insert a critical lysine into the pore. The lysine induces an active conformation of the pore yet blocks ion permeation. This study visualizes Kv1.3 pore dynamics, defines two distinct mechanisms to suppress Kv1.3 channel activity with exogenous inhibitors, and provides a framework to aid development of emerging T cell immunotherapies.
High-resolution transcriptomics of bovine purified protein derivative-stimulated peripheral blood from cattle infected withMycobacterium bovisacross an experimental time course
Authors: Correia Carolina N. , McHugo Gillian P. , Browne John A. , McLoughlin Kirsten E. , Nalpas Nicolas C. , Magee David A. , Whelan Adam O. , Villarreal-Ramos Bernardo , Vordermeier H. Martin , Gormley Eamonn , Gordon Stephen V. , MacHugh David E.
Published: April 2022

Objectives

Improved bovine tuberculosis (bTB) diagnostics with higher sensitivity and specificity are urgently required. A better understanding of the peripheral blood transcriptional response ofMycobacterium bovis-infected animals after bovine purified protein derivative (PPD-b) stimulation of whole blood—an important component of current bTB diagnostics—will provide new information for development of better diagnostics.

Methods

RNA sequencing (RNA-seq) was used to study the peripheral blood transcriptome after stimulation with PPD-b across four time points (-1 wk pre-infection, and +1 wk, +2 wk, and +10 wk post-infection) from a 14-weekM. bovisinfection time course experiment with ten age-matched Holstein-Friesian cattle.

Results

In vitro PPD-b stimulation of peripheral blood fromM. bovis-infected and non-infected cattle elicited a strong transcriptional response. Comparison of PPD-b stimulated, and unstimulated samples revealed higher expression of genes encoding cytokine receptors, transcription factors, and interferon-inducible proteins. Lower expression was seen for genes encoding proteins involved in antimicrobial activity, C-type lectin receptors, inhibition of signal transduction, and genes encoding metal ion transporters.

Conclusions

A transcriptional signature associated with the peripheral blood response to PPD-b stimulation consisting of 170 genes was identified exclusively in the post-infection time points. Therefore, this represents a panel of potential biomarkers ofM. bovisinfection.
mTORC1 activity is essential for disease progression in chronic lymphocytic leukemia
Authors: Malik Natasha , Hay Jodie , Dunn Karen M. , Nakagawa Rinako , Sansom Owen J. , Michie Alison M.
Published: April 2022

The precise role of mechanistic target of rapamycin complex 1 (mTORC1) during chronic lymphocytic leukemia (CLL) pathogenesis remains to be elucidated. Targeted deletion of mTORC1 componentRaptorin adult mice reveals that mTORC1 function is essential for initiation and maintenance of CLL.Raptor-deficient bone marrow-derived PKCα-KR transduced haemopoietic progenitors failed to generate a CLL-like diseasein vitro, due to an inability to overcome the mTORC1-mediated block in B cell lineage commitment. Induction ofRaptor-deficiency in NSG mice transplanted withMx1-RaptorBM-derived PKCα-KR transduced cells after disease was established, revealed a reduced CLL-like disease load and a significant increase in survival in the mice. Interestingly in mice transplanted with an aggressive CLL-like disease, rapamycin treatment reduced disease burden more effectively than AZD2014 (dual mTORC1/2 inhibitor), indicating a skew towards mTORC1 sensitivity with more aggressive leukemic disease. Rapamycin efficiently targeted the translation elongation axis eEF2/eEF2K downstream of mTORC1, resulting in eEF2 inactivation through induction of eEF2T56phosphorylation. Rapamycin treatment of primary CLL cells halted proliferation, modulated eEF2K/eEF2 phosphorylation and inhibited MCL1 expression. Our studies demonstrate that mTORC1 plays an essential role in leukemia progressionin vitroandin vivoin our CLL mouse model, with evidence for increased rapamycin sensitivity in aggressive secondary CLL transplants. Furthermore, the suppression of translation elongation through inactivation of eEF2 may offer a novel therapeutic target for blocking CLL progression.
Tumor-derived biomarkers beyond antigen expression enhance efficacy of CD276/B7H3 antibody-drug conjugate in metastatic prostate cancer
Authors: Agarwal Supreet , Fang Lei , McGowen Kerry , Yin JuanJuan , Bowman Joel , Ku Anson T. , Alilin Aian Neil , Corey Eva , Roudier Martine , True Lawrence , Dumpit Ruthy , Coleman Ilsa , Lee John , Nelson Peter S. , Capaldo Brian J. , Mariani Aida , Hoover Clare , Senatorov Ilya S. , Beshiri Michael , Sowalsky Adam G. , Hurt Elaine M , Kelly Kathleen
Published: April 2022

Antibody-drug conjugates (ADCs) are promising targeted cancer therapy; however, patient selection based solely on target antigen expression without consideration for cytotoxic payload vulnerabilities has plateaued clinical benefits. Biomarkers to capture patients who might benefit from specific ADCs have not been systematically determined for any cancer. We present a comprehensive therapeutic and biomarker analysis of a B7H3-ADC with pyrrolobenzodiazepine(PBD) payload in 26 treatment-resistant, metastatic prostate cancer(mPC) models. B7H3 is a tumor-specific surface protein widely expressed in mPC, and PBD is a DNA cross-linking agent. B7H3 expression was necessary but not sufficient for B7H3-PBD-ADC responsiveness. RB1 deficiency and/or replication stress, characteristics of poor prognosis, conferred sensitivity and were associated with complete tumor regression in both neuroendocrine (NEPC) and androgen receptor positive(ARPC) prostate cancer models, even with low B7H3 levels. Non-ARPC models, which are currently lacking efficacious treatment, demonstrated the highest replication stress and were most sensitive to treatment. In RB1 wild-type ARPC tumors, SLFN11 expression or select DNA repair mutations in SLFN11 non-expressors governed response. Significantly, wild-type TP53 predicted non-responsiveness (7/8 models). Overall, biomarker-focused selection of models led to high efficacy of in vivo treatment. These data enable a paradigm shift to biomarker-driven trial designs for maximizing clinical benefit of ADC therapies.
NIPBL and WAPL balance cohesin activity to regulate chromatin folding and gene expression
Authors: Luppino Jennifer M. , Field Andrew , Nguyen Son C. , Park Daniel S. , Shah Parisha P. , Lan Yemin , Yunker Rebecca , Jain Rajan , Adelman Karen , Joyce Eric F.
Published: April 2022

The relationship between cohesin-mediated chromatin looping and gene expression remains unclear. We investigated the roles of NIPBL and WAPL, two regulators of cohesin activity, in chromatin folding and transcription in human cells. Consistent with their opposing roles in cohesin regulation, depletion of these factors showed opposite effects on levels of chromatin-bound cohesin and spatial insulation of neighboring domains. We find that NIPBL or WAPL depletion each alter the expression of ~2,000 genes, most of which are uniquely sensitive to either regulator. We find that each set of differentially expressed genes are enriched at chromatin loop anchors and clustered within the genome, suggesting there are genomic regions sensitive to either more or less cohesin. Remarkably, co-depletion of both regulators rescued chromatin misfolding and gene misexpression compared to either single knockdown. Taken together, we present a model in which the relative, rather than absolute, levels of NIPBL and WAPL are required to balance cohesin activity in chromatin folding to regulate transcription.
The mitogen-activated protein kinase kinase kinase, ILK5, regulates plant purinergic receptor-mediated, innate immunity
Authors: Kim Daewon , Chen Dongqin , Ahsan Nagib , Thelen Jay J. , Stacey Gary
Published: April 2022

Mitogen-activated protein (MAP) kinase signaling cascades play important roles in the regulation of eukaryotic defense against various pathogens. Activation of the extracellular ATP (eATP) receptor P2K1 triggers MAP kinase 3 and 6 (MPK3/6) phosphorylation, which leads to elevated defense responses in Arabidopsis. However, the mechanism by which P2K1 activates the MAPK cascade is unclear. In this study, we identified Raf-like MAPKKK ILK5 as a downstream substrate of the P2K1 kinase. P2K1 phosphorylates ILK5 on serine 192. The interaction between P2K1 and ILK5 was confirmed both in vitro and in planta and their interaction was enhanced by ATP treatment. Similar to P2K1 expression, ILK5 expression levels were highly induced by treatment with ATP, flg22, Pseudomonas syringae pv. tomato DC3000, and various abiotic stresses, such as wounding. ILK5 interacts with both MKK4 and MKK5, but only MKK5 is phosphorylated by ILK5. Moreover, phosphorylation of MPK3/6 was significantly reduced upon ATP treatment in ilk5 mutant plants, relative to wild-type. The ilk5 mutant plants showed higher susceptibility to P. syringae pathogen infection relative to wild-type plants. Plants expressing only the mutant ILK5S192A protein, lacking kinase activity, did not activate the MAPK cascade upon ATP addition. Taken together, the results suggest that eATP activation of P2K1 results in transphosphorylation of the Raf-like MAPKKK ILK5, which subsequently triggers the MAPK cascade, culminating in activation of MAPK3 and 6 associated with an elevated innate immunity response.

Significance statement

Pathogens invasion and subsequent wound stress activates extracellular ATP-mediated purinergic signaling cascades, a danger associated molecular pattern (DAMP) signal, which induces phosphorylation of MAPKs. Previous studies revealed that the P2K1 purinergic receptor increases MPK3/6 phosphorylation in response to eATP signaling cascades in Arabidopsis. However, the specific mechanism by which this occurs remains unknown. Here, we describe the isolation and characterization of Raf-like MAPKKK ILK5 (Integrin-linked Kinase 5) as a downstream substrate of P2K1 kinase activity. Initiation of an eATP-dependent signaling pathway by phosphorylation of ILK5 with subsequent activation of MKK5, leading to activation of MPK3/6 and downstream events is crucial to the plant innate immunity response.
T cell Aryl Hydrocarbon Receptor Activity Tunes the Gut Microenvironment to Sustain Autoimmunity and Neuroinflammation
Authors: Merchak Andrea R. , Cahill Hannah J. , Brown Lucille C. , Brown Ryan M. , Rivet-Noor Courtney , Beiter Rebecca M. , Slogar Erica R. , Olgun Deniz G. , Gaultier Alban
Published: April 2022

Multiple sclerosis (MS) is a T cell driven autoimmune disease that attacks the myelin of the central nervous system and currently has no cure. MS etiology is linked to both the gut flora and external environmental factors but this connection is not well understood. One immune system regulator responsive to non-pathogenic external stimuli is the aryl hydrocarbon receptor (AHR). The AHR, which binds diverse molecules present in the environment in barrier tissues, is a therapeutic target for MS. However, AHR’s precise function in T lymphocytes, the orchestrators of MS, has not been described. Here we show that in a mouse model of MS, T cell specificAhrknockout leads to recovery driven by a decrease in T cell fitness. At the mechanistic level, we demonstrate that the absence of AHR changes the gut microenvironment composition to generate metabolites that impact T cell viability, such as bile salts and short chain fatty acids. Our study demonstrates a newly emerging role for AHR in mediating the interdependence between T lymphocytes and the microbiota, while simultaneously identifying new potential molecular targets for the treatment of MS and other autoimmune diseases.
Disruption of anterior temporal lobe reduces distortions in memory from category knowledge
Authors: Tompary Alexa , Xia Alice , Coslett H. Branch , Thompson-Schill Sharon L.
Published: April 2022

Memory retrieval does not provide a perfect recapitulation of past events, but instead an imperfect reconstruction of event-specific details and general knowledge. However, it remains unclear whether this reconstruction relies on mixtures of signals from different memory systems, including one supporting general knowledge. Here, we investigate whether the anterior temporal lobe (ATL) distorts new memories due to prior category knowledge. In this experiment (N=36), participants encoded and retrieved image-location associations. Most images’ locations were clustered according to their category, but some were in random locations. With this protocol, we previously demonstrated that randomly located images were retrieved closer to their category cluster relative to their encoded locations, suggesting an influence of category knowledge. We combined this procedure with transcranial magnetic stimulation (TMS) delivered to the left ATL before retrieval. We separately examined event-specific details (error) and category knowledge (bias) to identify distinct signals attributable to different memory systems. We found that TMS to ATL attenuated bias in location memory, but only for atypical category members. The magnitude of error was not impacted, suggesting that a memory’s fidelity can be decoupled from its distortion by category knowledge. This raises the intriguing possibility that retrieval is jointly supported by separable memory systems.
Sorting of secretory proteins at the trans-Golgi network by human TGN46
Authors: Lujan Pablo , Garcia-Cabau Carla , Wakana Yuichi , Lillo Javier Vera , Rodilla-Ramírez Carmen , Sugiura Hideaki , Malhotra Vivek , Salvatella Xavier , Garcia-Parajo Maria F. , Campelo Felix
Published: April 2022

Secretory proteins are sorted at the trans-Golgi network (TGN) for export into specific transport carriers. However, the molecular players involved in this fundamental process remain largely elusive. Here, we identified the human transmembrane protein TGN46 as a receptor for the export of secretory cargo protein PAUF in CARTS – a class of protein kinase D-dependent TGN-to-plasma membrane carriers. We show that TGN46 is necessary for cargo sorting and loading into nascent carriers at the TGN. By combining quantitative fluorescence microscopy and mutagenesis approaches, we further discovered that the lumenal domain of TGN46 encodes for its cargo sorting function. In summary, our results define a cellular function of TGN46 in sorting secretory proteins for export from the TGN.