Translational control in the spinal cord regulates gene expression and pain hypersensitivity in the chronic phase of neuropathic pain.

Sensitization of spinal nociceptive circuits plays a crucial role in neuropathic pain. This sensitization depends on new gene expression that is primarily regulated via transcriptional and translational control mechanisms. The relative roles of these mechanisms in regulating gene expression in the clinically relevant chronic phase of neuropathic pain are not well understood. Here, we show that changes in gene expression in the spinal cord during the chronic phase of neuropathic pain are substantially regulated at the translational level. Downregulating spinal translation at the chronic phase alleviated pain hypersensitivity. Cell-type-specific profiling revealed that spinal inhibitory neurons exhibited greater changes in translation after peripheral nerve injury compared to excitatory neurons. Notably, increasing translation selectively in all inhibitory neurons or parvalbumin-positive (PV+) interneurons, but not excitatory neurons, promoted mechanical pain hypersensitivity. Furthermore, increasing translation in PV+ neurons decreased their intrinsic excitability and spiking activity, whereas reducing translation in spinal PV+ neurons prevented the nerve injury-induced decrease in excitability. Thus, translational control mechanisms in the spinal cord, particularly in inhibitory neurons, play a role in mediating neuropathic pain hypersensitivity.

Functional analysis of the AUG initiator codon context reveals novel conserved sequences that disfavor mRNA translation in eukaryotes.

mRNA translation is a fundamental process for life. Selection of the translation initiation site (TIS) is crucial, as it establishes the correct open reading frame for mRNA decoding. Studies in vertebrate mRNAs discovered that a purine at -3 and a G at +4 (where A of the AUG initiator codon is numbered + 1), promote TIS recognition. However, the TIS context in other eukaryotes has been poorly experimentally analyzed. We analyzed in vitro the influence of the -3, -2, -1 and + 4 positions of the TIS context in rabbit, Drosophila, wheat, and yeast. We observed that -3A conferred the best translational efficiency across these species. However, we found variability at the + 4 position for optimal translation. In addition, the Kozak motif that was defined from mammalian cells was only weakly predictive for wheat and essentially non-predictive for yeast. We discovered eight conserved sequences that significantly disfavored translation. Due to the big differences in translational efficiency observed among weak TIS context sequences, we define a novel category that we termed 'barren AUG context sequences (BACS)', which represent sequences disfavoring translation. Analysis of mRNA-ribosomal complexes structures provided insights into the function of BACS. The gene ontology of the BACS-containing mRNAs is presented.

Impact of eIF2α phosphorylation on the translational landscape of mouse embryonic stem cells.

The integrated stress response (ISR) is critical for cell survival under stress. In response to diverse environmental cues, eIF2α becomes phosphorylated, engendering a dramatic change in mRNA translation. The activation of ISR plays a pivotal role in the early embryogenesis, but the eIF2-dependent translational landscape in pluripotent embryonic stem cells (ESCs) is largely unexplored. We employ a multi-omics approach consisting of ribosome profiling, proteomics, and metabolomics in wild-type (eIF2α+/+) and phosphorylation-deficient mutant eIF2α (eIF2αA/A) mouse ESCs (mESCs) to investigate phosphorylated (p)-eIF2α-dependent translational control of naive pluripotency. We show a transient increase in p-eIF2α in the naive epiblast layer of E4.5 embryos. Absence of eIF2α phosphorylation engenders an exit from naive pluripotency following 2i (two chemical inhibitors of MEK1/2 and GSK3α/ß) withdrawal. p-eIF2α controls translation of mRNAs encoding proteins that govern pluripotency, chromatin organization, and glutathione synthesis. Thus, p-eIF2α acts as a key regulator of the naive pluripotency gene regulatory network.

4E-BP1-dependent translation in nociceptors controls mechanical hypersensitivity via TRIM32/type I interferon signaling.

Activation of the mechanistic target of rapamycin complex 1 (mTORC1) contributes to the development of chronic pain. However, the specific mechanisms by which mTORC1 causes hypersensitivity remain elusive. The eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) is a key mTORC1 downstream effector that represses translation initiation. Here, we show that nociceptor-specific deletion of 4E-BP1, mimicking activation of mTORC1-dependent translation, is sufficient to cause mechanical hypersensitivity. Using translating ribosome affinity purification in nociceptors lacking 4E-BP1, we identified a pronounced translational up-regulation of tripartite motif-containing protein 32 (TRIM32), an E3 ubiquitin ligase that promotes interferon signaling. Down-regulation of TRIM32 in nociceptors or blocking type I interferon signaling reversed the mechanical hypersensitivity in mice lacking 4E-BP1. Furthermore, nociceptor-specific ablation of TRIM32 alleviated mechanical hypersensitivity caused by tissue inflammation. These results show that mTORC1 in nociceptors promotes hypersensitivity via 4E-BP1-dependent up-regulation of TRIM32/interferon signaling and identify TRIM32 as a therapeutic target in inflammatory pain.

C8ORF88: A Novel eIF4E-Binding Protein.

Translation initiation in eukaryotes is regulated at several steps, one of which involves the availability of the cap binding protein to participate in cap-dependent protein synthesis. Binding of eIF4E to translational repressors (eIF4E-binding proteins [4E-BPs]) suppresses translation and is used by cells to link extra- and intracellular cues to protein synthetic rates. The best studied of these interactions involves repression of translation by 4E-BP1 upon inhibition of the PI3K/mTOR signaling pathway. Herein, we characterize a novel 4E-BP, C8ORF88, whose expression is predominantly restricted to early spermatids. C8ORF88:eIF4E interaction is dependent on the canonical eIF4E binding motif (4E-BM) present in other 4E-BPs. Whereas 4E-BP1:eIF4E interaction is dependent on the phosphorylation of 4E-BP1, these sites are not conserved in C8ORF88 indicating a different mode of regulation.

The Association between Statins Intake and Risk of Post Stroke Pneumonia: A Systematic Review and Meta-analysis.

AIM: This research aimed to examine the relationship between the intake of statins and the risk of post-stroke pneumonia in a systematic review and meta-analysis study. METHODS: An extensive search of published articles on March 21st , 2023, was done in several databases, like Web of Science (ISI), PubMed, Cochrane Library, Embase, Scopus, and Google Scholar. The Newcastle Ottawa Scale (NOS) checklist was employed to evaluate the quality of observational studies. Statistical tests (Chi-square test and I2 ) and graphical techniques (Forest plot) were used to determine whether heterogeneity existed in the meta-analysis studies. Funnel plots and Begg and Egger's tests were used to assess the publication bias. RESULTS: Seven studies (5 cohort and 2 case-control studies) were retrieved to examine the association between statins and post-stroke pneumonia. The sample size of the studies compiled in the meta-analysis was obtained to be 68,966 participants. Meta-analysis demonstrated that the overall odds of post-stroke pneumonia in the statin group was equal to 0.87 (95% CI: 0.67 - 1.13; p-value 0.458). Subgroup analysis indicated that the odds of post-stroke pneumonia in the statin group was equal to 0.93 (95% CI: 0.73-1.18; p-value=0.558) in the cohort studies, and equal to 0.92 (95% CI: 0.37-2.26; p-value=0.857) in the case-control studies. The examination of the association between the intake of statins and post-stroke pneumonia showed no evidence of publication bias (Begg's test, p-value = 0.368; Eggers test, p-value = 0.282). CONCLUSION: In this study, no relationship has been observed between receiving statins and the risk of post-stroke pneumonia.

A Systematic Review and Meta-analysis on the Role of Statins in the Prevention of Mortality Following Pancreatic Cancer.

BACKGROUND: Pancreatic cancer (PC) is a type of cancer with a high incidence and case-fatality rate. OBJECTIVE: This study aimed to evaluate the role of statins in preventing mortality following PC based on scientific evidence with systematic review and meta-analysis method. METHODS: This meta-analysis considered studies published from 1980 till the end of 2022 in ISI Web of Science, Scopus, PubMed, Cochrane, Science Direct, Google Scholar, and Embase databases. Funnel diagrams and Begg's and Egger's tests were used to assess the publication bias. RESULTS: In general, this meta-analysis has included 19 studies (13 cohort studies, 4 case-control, and 2 randomized clinical trials (RCTs)) and a total of 100,888 patients with PC. The risk of mortality of PC in statin users in total was 0.86 (95% CI: 0.80 - 0.92, P-value <0.001); in the case-control studies, it was equal to 0.53 (0.34-0.83); in the cohort studies, it was equal to 0.87 (0.82-0.92, P-value <0.001); in RCTs, it was equal to 1.19 (0.99-1.42, P-value <0.001); in studies with good quality score category, it was equal to 0.92 (0.86-0.99, P-value <0.001), and in articles of the moderate quality score category, it was equal to 0.73 (0.64-0.84, P-value <0.001). The results of statistical tests indicated the existence of publication bias (Begg's test (P-value = 0.002) and Egger's test (P-value = 0.004)). CONCLUSION: Statins reduce the risk of mortality in patients with PC. However, no significant relation has been observed in RCTs. Therefore, it is necessary to be cautious in interpreting the results.

C-Reactive Protein and D-dimer as Prognostic Markers for Clinical Outcomes in Patients with Mild Traumatic Brain Injury: A Cross-Sectional Study.

Objective: To investigate the use of prognostic markers such as C-reactive protein (CRP) and D-dimer for clinical outcomes in patients with mild traumatic brain injury (TBI). Methods: This cross-sectional study was conducted on patients with mild head trauma who were admitted to the Emergency Department of Imam Khomeini Hospital (Sari, Iran). Data were collected from 2018 to 2019. Age, sex, the time of injury hospitalization, length of hospitalization, length of unconsciousness, blood pressure, heart rate, respiratory rate, and concomitant symptoms were all recorded using a pre-designed checklist. The patient's Glasgow Coma Scale (GCS), CRP, and D-dimer were also measured. Moreover, all patients underwent CT scan. Results: This study included 74 patients with TBI. The mean age of the participants was 36.92±3.54. The mean CRP and D-dimer values were 5.69±0.77 and 0.58±0.11 in these patients, respectively. At the cut-off point of 11.50 for CRP, the sensitivity and specificity to detect the pathological lesions in CT scan were 75% and 95.50%, respectively (p<0.001). Additionally, with a D-dimer cut-off point of 0.90, the sensitivity and specificity for diagnosing pathological lesions in CT scan were 100% and 98.50%, respectively (p<0.001). Conclusion: In general, the CRP and D-dimer levels of patients with mild TBI (GCS≥13) can be assessed to protect against CT-induced radiation exposure and subsequent disorders; if they do not exhibit clinical signs to increase the risk of adverse brain damage, such as reduced level of consciousness, drowsiness, and prolonged periods of unconsciousness.

Medication Errors among Iranian Intensive Care Nurses: A Systematic Review.

Background: Medication Error (ME) is a major patient safety concern in Intensive Care Units (ICUs). Critical care nurses play a crucial role in the safe administration of medication. This study was conducted to comprehensively review the literature concerning the prevalence of ME and associated factors and outcomes in Iranian ICU nurses. Materials and Methods: An extensive search of the literature was carried in international databases including PubMed, Web of Science, Scopus, and Google Scholar, as well as Persian databases such as Magiran and Scientific Information Database (SID) using ME-related keywords and the Persian equivalent of these keywords, from the first article written in this field to artcles published on March 30, 2021. The appraisal tool (AXIS tool) was used to assess the quality of the included studies. Results: Fifteen studies were included in this systematic review. The prevalence of MEs made by ICU nurses was 53.34%. The most common types of MEs were wrong infusion rate (14.12%), unauthorized medication (11.76%), and wrong time (8.49%) errors, respectively. MEs occurred more frequently in morning work shifts (44.44%). MEs happened more frequently for heparin, vancomycin, ranitidine, and amikacin. The most important influential factor in the occurrence of MEs in ICUs was management and human factors. Conclusions: The prevalence of MEs made by Iranian ICU nurses is high. Therefore, nurse managers and policymakers should develop appropriate strategies, including training programs, to reduce the occurrence of MEs made by nurses in ICUs.

The mRNA translation initiation factor eIF4G1 controls mitochondrial oxidative phosphorylation, axonal morphogenesis, and memory.

mRNA translation initiation plays a critical role in learning and memory. The eIF4F complex, composed of the cap-binding protein eIF4E, ATP-dependent RNA helicase eIF4A, and scaffolding protein eIF4G, is a pivotal factor in the mRNA translation initiation process. eIF4G1, the major paralogue of the three eIF4G family members, is indispensable for development, but its function in learning and memory is unknown. To study the role of eIF4G1 in cognition, we used an eIF4G1 haploinsufficient (eIF4G1-1D) mouse model. The axonal arborization of eIF4G1-1D primary hippocampal neurons was significantly disrupted, and the mice displayed impairment in hippocampus-dependent learning and memory. Translatome analysis showed that the translation of mRNAs encoding proteins of the mitochondrial oxidative phosphorylation (OXPHOS) system was decreased in the eIF4G1-1D brain, and OXPHOS was decreased in eIF4G1-silenced cells. Thus, eIF4G1-mediated mRNA translation is crucial for optimal cognitive function, which is dependent on OXPHOS and neuronal morphogenesis.

The Role of Vitamin D in Carpal Tunnel Syndrome Risk and Supplementation Outcomes: A Systematic Review.

BACKGROUND: Carpal tunnel syndrome (CTS) is a debilitating neuropathy that accompanies pain and other physical limitations and disrupts the normal functioning of the victims' lives. OBJECTIVE: We aimed to investigate Vitamin D's preventive and therapeutic effects on the occurrence and remission of CTS symptoms. METHODS: In this systematic review the PRISMA statement has been designed primarily. An extensive search was undertaken in various databases, including PubMed, Cochrane library, Web of Science, EMBASE, and Scopus. After considering the inclusion and exclusion criteria of the study, finally, 19 articles were retrieved. The raw data were extracted and entered into an Excel form, and the study outcomes were investigated. RESULTS: The main symptoms and tests, including functional score, nerve conduction, and pain, were improved after Vitamin D supplementation in CTS patients. However, they revealed worse scores in people with low Vitamin D levels. In addition, the scores of mentioned indices were worsened in people with lower serum Vitamin D levels. Nevertheless, some studies did not find a significant relationship between low serum 25(OH)D and more significant pain scores in CTS patients. In addition, Vitamin D inserts its effects on CTS by regulating cell proliferation, nerve growth factor, suppression of oxidative stress and inflammatory cytokines, and improvement in cartilage and microvascular damage. CONCLUSION: Vitamin D supplementation can improve the symptoms in CTS patients, and low serum 25(OH)D can aggravate the symptoms of the disease and could be a risk factor for its occurrence. However, more observational studies and clinical trials are needed.

The ribosome quality control factor Asc1 determines the fate of HSP70 mRNA on and off the ribosome.

Cells survive harsh environmental conditions by potently upregulating molecular chaperones such as heat shock proteins (HSPs), particularly the inducible members of the HSP70 family. The life cycle of HSP70 mRNA in the cytoplasm is unique-it is translated during stress when most cellular mRNA translation is repressed and rapidly degraded upon recovery. Contrary to its 5' untranslated region's role in maximizing translation, we discovered that the HSP70 coding sequence (CDS) suppresses its translation via the ribosome quality control (RQC) mechanism. The CDS of the most inducible Saccharomyces cerevisiae HSP70 gene, SSA4, is uniquely enriched with low-frequency codons that promote ribosome stalling during heat stress. Stalled ribosomes are recognized by the RQC components Asc1p and Hel2p and two novel RQC components, the ribosomal proteins Rps28Ap and Rps19Bp. Surprisingly, RQC does not signal SSA4 mRNA degradation via No-Go-Decay. Instead, Asc1p destabilizes SSA4 mRNA during recovery from heat stress by a mechanism independent of ribosome binding and SSA4 codon optimality. Therefore, Asc1p operates in two pathways that converge to regulate the SSA4 mRNA life cycle during stress and recovery. Our research identifies Asc1p as a critical regulator of the stress response and RQC as the mechanism tuning HSP70 synthesis.

The relationship between causes of suicidal attempts in Iran and individual and social variables: a retrospective study.

OBJECTIVE: Determine the prevalence of suicide attempts and the relationships between the different causes of attempts with sociodemographic and clinical characteristics among individuals in Iran. METHODS: A retrospective review of data about suicide attempts from poisoning care centers in Babol city between 2017 and 2021. Multinomial regression analysis (with mental illness being the reference variable) was used to determine the factors associated with the different causes of suicide attempts (addiction, romantic relationship problems, and economic problems). RESULTS: The overall prevalence of completed suicide in the population sampled was 10.8% (95% confidence interval 9.5-12.1) (244/2,263). Relative to mental disorder, given that other variables in the model are held constant the following were associated with suicide attempts. A previous history of suicide attempts was associated with increasing the relative risk ratio of attempting suicide while having no positive history of smoking was associated with reducing the relative risk ratio of a suicidal attempt. However, the use of multiple drugs to attempt suicide was associated with an increased relative risk ratio of attempting suicide with romantic relationship problems and addiction as causes of suicide attempts. The first year of data collection (2017) and the female gender were both associated with an increased relative risk ratio of having a suicide attempt due to romantic relationships and economic problems. A family history of suicide was associated with an increased relative risk ratio of suicide attempts due to romantic relationship problems. However, using Pesticides-aluminum phosphide and detergent and javel water to attempt reduced the relative risk ratio of attempting due to romantic relationship problems. Age, self-employment, middle income, and married were associated with an increased relative risk ratio of suicide attempts among individuals due to addiction. However, staying longer at the emergency department was associated with a reduced relative risk ratio of having had a suicide attempt due to addiction. CONCLUSIONS: This study highlights the interplay between romantic hardships, addiction, economic hardships as reasons for suicide attempts and various sociable variables in a population in Northern Iran. The most associated reason for suicide attempts was romantic relationship hardships. Therefore, interventions such as sessions on conflict resolution, boundary setting, and management of grieving would greatly benefit this society and reduce the rate of suicide, especially among individuals with a history of suicide attempts.

Attitudes of Emergency Nurses and Patients' Family Members Towards the Presence of Family Members During Cardiopulmonary Resuscitation; a Cross-sectional Study.

Introduction: Family presence during cardiopulmonary resuscitation (CPR) is one of the elements where family-centered care is practiced. This study aimed to investigate the attitudes of Iranian emergency nurses and patients' family members regarding the presence of family during CPR. Methods: In a cross-sectional study, 350 emergency nurses and 254 family members of patients admitted to the emergency department of an educational hospital in Iran were enrolled. Data were collected from May to November 2020 using convenience sampling and using a 27-item questionnaire of participants' attitudes towards family presence during CPR. Results: The mean attitude scores of nurses and family members of patients regarding family presence during CPR were 86.79 ± 7.50 and 92.48 ± 6.77, respectively (p < 0.001). The highest and lowest mean scores of nurses' attitude towards family presence during CPR were related to "CPR performance will be negatively influenced" and "Family members have the right to be present during CPR of their relatives/ Allows relatives to stay with the patient until the end/ Makes the patient less worried ", respectively. The highest and lowest mean scores of family members' attitude towards family presence during CPR were related to "Family members may interfere with CPR" and " May be beneficial to the relatives' grieving process ", respectively. Conclusion: Overall, the results of this study showed that the attitude of emergency nurses and patients' family members towards family presence during CPR was positive. Of course, the mean attitude score of nurses in this regard was significantly lower.

Audit of the functional preparedness of the selected military hospital in response to incidents and disasters: participatory action research.

INTRODUCTION: Since hospitals play an important role in dealing with disaster victims, this study was conducted to audit the functional preparedness of the selected military hospital in response to incidents and disasters. MATERIALS AND METHODS: This applied action research was conducted in all wards of a military hospital from September 2020 to September 2021. The functional preparedness of the hospital was assessed using a functional preparedness checklist containing 17 domains and the weaknesses of the hospital were identified. Then, during the hospital audit cycle, a plan was developed to improve work processes and the functional preparedness of different wards of the hospital in response to incidents and disasters using the FOCUS-PDCA model. The functional preparedness of the hospital was compared before and after the intervention and analyzed using SPSS22. RESULTS: The relative mean score of hospital preparedness in response to disasters was 508 out of 900 (56.44%) before the intervention, which was moderate. The relative mean score of the hospital preparedness in response to disasters was 561 (63.63%) after the intervention, which was good. The highest preparedness was related to risk assessment (85%) and the lowest preparedness was related to victims' dead bodies (44%). CONCLUSION: Considering the effect of action research on improving the hospital's functional preparedness in response to disasters, other healthcare facilities are encouraged to incorporate auditing into their work plans.

The amino acid sensor GCN2 controls red blood cell clearance and iron metabolism through regulation of liver macrophages.

GCN2 (general control nonderepressible 2) is a serine/threonine-protein kinase that controls messenger RNA translation in response to amino acid availability and ribosome stalling. Here, we show that GCN2 controls erythrocyte clearance and iron recycling during stress. Our data highlight the importance of liver macrophages as the primary cell type mediating these effects. During different stress conditions, such as hemolysis, amino acid deficiency or hypoxia, GCN2 knockout (GCN2-/-) mice displayed resistance to anemia compared with wild-type (GCN2+/+) mice. GCN2-/- liver macrophages exhibited defective erythrophagocytosis and lysosome maturation. Molecular analysis of GCN2-/- cells demonstrated that the ATF4-NRF2 pathway is a critical downstream mediator of GCN2 in regulating red blood cell clearance and iron recycling.

UBR4/POE facilitates secretory trafficking to maintain circadian clock synchrony.

Ubiquitin ligases control the degradation of core clock proteins to govern the speed and resetting properties of the circadian pacemaker. However, few studies have addressed their potential to regulate other cellular events within clock neurons beyond clock protein turnover. Here, we report that the ubiquitin ligase, UBR4/POE, strengthens the central pacemaker by facilitating neuropeptide trafficking in clock neurons and promoting network synchrony. Ubr4-deficient mice are resistant to jetlag, whereas poe knockdown flies are prone to arrhythmicity, behaviors reflective of the reduced axonal trafficking of circadian neuropeptides. At the cellular level, Ubr4 ablation impairs the export of secreted proteins from the Golgi apparatus by reducing the expression of Coronin 7, which is required for budding of Golgi-derived transport vesicles. In summary, UBR4/POE fulfills a conserved and unexpected role in the vesicular trafficking of neuropeptides, a function that has important implications for circadian clock synchrony and circuit-level signal processing.

Body impedance analyzer and anthropometric indicators; predictors of metabolic syndrome.

Aim: Metabolic syndrome is one of the outcomes of a sedentary lifestyle in the modern world. In this study, we want to introduce the predictors of metabolic syndrome using anthropometric indices and Bio-Electrical Impedance Analysis (BIA) test values. Method: This cross-sectional study was performed on 2284 employees of Tehran University of Medical Sciences in different job categories. Metabolic syndrome was determined according to IDF criteria. Anthropometric dimensions, para-clinical tests, basic information were collected from the participants. Also, the body analysis of the participants was performed using a BIA method. Result: The prevalence of metabolic syndrome in this study was 23.2% based on IDF criteria, which was 21% and 26.6% in men and women, respectively. The most important factor among the components of IDF criteria was HDL deficiency. In this study, neck circumference, fat mass, visceral fat, muscle mass percentage and waist to height ratio were observed as predictors of metabolic syndrome. Conclusion: This study realized that there is association between fat mass, fat-free mass, visceral fat and muscle mass which all are some elements of body composition analysis and metabolic syndrome as a major health issue.

Convention on the rights of persons with disabilities: Qualitative exploration of barriers to the implementation of articles 25 (health) and 26 (rehabilitation) in Iran.

Background: The International Convention on the Rights of Persons with Disabilities (CRPD) is the most important International Document for recognizing the rights of persons with disabilities, including the right to health and rehabilitation. Islamic Republic Iran acceded to the Convention in 2008, but still has a long way to go to achieve its desired status and in line with the objectives of the convention. This study aimed to identify the barriers to the implementation of articles 25 and 26 of the CRPD in Iran. Methods: This study was performed using conventional content analysis. Twenty-one individuals were recruited by purposive sampling with maximum variation and were continued until saturation. Data were gathered through in-depth, semi-structured interviews from June 2018 to May 2019. MAXQDA version 10 was used for analyzing data. Results: The resulting data analysis yielded 860 initial or open codes. The concepts were categorized into 27 subcategories and 7 categories. Main categories were included: "Structure inefficiency", "lack of comprehensive rehabilitation program", "inadequate awareness", "neglected economy of people with disabilities", "weak access to services", "cultural challenges" and "disregard for new technologies". Conclusion: The findings showed that the executive structures in the country have a lot of problems with health and rehabilitation programs for people with disabilities. It seems understanding the barriers to implementation of articles 25 and 26 of the international CRPD empowers officials in the field and improve services by providing a better view of the disabled. Nevertheless, it is recommended for policymakers to consider rehabilitation as a main element of the health system.

Identification and characterization of hippuristanol-resistant mutants reveals eIF4A1 dependencies within mRNA 5' leader regions.

Hippuristanol (Hipp) is a natural product that selectively inhibits protein synthesis by targeting eukaryotic initiation factor (eIF) 4A, a DEAD-box RNA helicase required for ribosome recruitment to mRNA templates. Hipp binds to the carboxyl-terminal domain of eIF4A, locks it in a closed conformation, and inhibits its RNA binding. The dependencies of mRNAs for eIF4A during initiation is contingent on the degree of secondary structure within their 5' leader region. Interest in targeting eIF4A therapeutically in cancer and viral-infected settings stems from the dependencies that certain cellular (e.g. pro-oncogenic, pro-survival) and viral mRNAs show towards eIF4A. Using a CRISPR/Cas9-based variomics screen, we identify functional EIF4A1 Hipp-resistant alleles, which in turn allowed us to link the translation-inhibitory and cytotoxic properties of Hipp to eIF4A1 target engagement. Genome-wide translational profiling in the absence or presence of Hipp were undertaken and our validation studies provided insight into the structure-activity relationships of eIF4A-dependent mRNAs. We find that mRNA 5' leader length, overall secondary structure and cytosine content are defining features of Hipp-dependent mRNAs.