Unified Pathway Research Articles

"Blade of Polarized Water Molecule" Is the Key to Hydrolase Catalysis Regulation.

Hydrolysis catalyzed by aspartic proteases is a crucial reaction in many biological processes. However, anchoring water molecules and unifying multiple catalytic pathways remain significant challenges. Consequently, molecular design often compromises by focusing on enhancing substrate specificity. Using our self-developed polarizable point charge (PPC) force field, we determined the significant role of polarization in the hydrolase of pepsin for the first time. To be stably anchored in the active site, the water should be intensely polarized with a charge higher than -0.94e. Induced by this polarization, the pepsin was shown to support three general base/general acid pathways, with a preference for the gemdiol-intermediate-based pathway. Consequently, we proposed the "Blade of Polarized Water Molecule" model for rational enzyme design, highlighting that the polarization of both the attacking water and the attacked carbonyl is crucial for enhancing hydrolysis. Mutants D290Q and S172P showed activity enhancements of 191.23% and 324.70%, respectively. The improved polarization of water, carbonyl, and relevant nucleophilic attack distances in the mutants reaffirmed the crucial role of polarization in improving hydrolysis. This study provides a new perspective on hydrolase analysis and modification.

Patient-centred stoma care support: ileostomy patients.

Stoma patients require continuous support throughout their entire journey with a stoma. Although many Stoma Care Services across the UK offer patient follow-up pathways, there is not one unified pathway. Patients may not be prepared for life with a stoma because, depending on their stoma type, they will have specific needs, and if patients and healthcare professionals are not prepared to manage these stoma-specific needs, complications and hospital readmissions may occur, worsening patients' outcomes and quality of life. Ileostomy patients are known to be more likely to experience complications, including hospital readmissions, and therefore, special care should be taken when preparing these patients for life with a stoma. They should be informed and educated to prevent complications, and if this is not always possible, thye should at least be able to recognise and manage early signs and symptoms of complications. This will empower them to self-care and know when to seek medical attention.

High resolution landscape of ribosomal RNA processing and surveillance.

Ribosomal RNAs are processed in a complex pathway. We profiled rRNA processing intermediates in yeast at single-molecule and single-nucleotide levels with circularization, targeted amplification and deep sequencing (CircTA-seq), gaining significant mechanistic insights into rRNA processing and surveillance. The long form of the 5' end of 5.8S rRNA is converted to the short form and represents an intermediate of a unified processing pathway. The initial 3' end processing of 5.8S rRNA involves trimming by Rex1 and Rex2 and Trf4-mediated polyadenylation. The 3' end of 25S rRNA is formed by sequential digestion by four Rex proteins. Intermediates with an extended A1 site are generated during 5' degradation of aberrant 18S rRNA precursors. We determined precise polyadenylation profiles for pre-rRNAs and show that the degradation efficiency of polyadenylated 20S pre-rRNA critically depends on poly(A) lengths and degradation intermediates released from the exosome are often extensively re-polyadenylated.

Enediyne natural product biosynthesis unified by a diiodotetrayne intermediate.

Enediyne natural products are renowned for their potent cytotoxicities but the biosynthesis of their defining 1,5-diyne-3-ene core moiety remains largely enigmatic. Since the discovery of the enediyne polyketide synthase cassette in 2002, genome sequencing has revealed thousands of distinct enediyne biosynthetic gene clusters, each harboring the conserved enediyne polyketide synthase cassette. Here we report that (1) the products of this cassette are an iodoheptaene, a diiodotetrayne and two pentaynes; (2) the diiodotetrayne represents a common biosynthetic intermediate for all known enediynes; and (3) cryptic iodination can be exploited to increase enediyne titers. These findings establish a unified biosynthetic pathway for the enediynes, set the stage to further advance enediyne core biosynthesis and enable fundamental breakthroughs in chemistry, enzymology and translational applications of enediyne natural products.

Comorbid functional dyspepsia reflects IL-33–mediated airway neuronal dysfunction in asthma

BackgroundNeuronal dysfunction is implicated in the pathophysiology of asthma and functional dyspepsia (FD). However, the relationship between these diseases remains unclear. ObjectiveThis study aimed to clarify the clinical implications of comorbid FD in asthma and to explore the unified pathway between asthma and FD by focusing on airway neuronal dysfunction. MethodsClinical indices and biomarkers, including capsaicin cough sensitivity (C-CS), were compared between patients with asthma with and without FD. C-CS was determined based on the capsaicin concentration that induced at least two (C2) or five coughs (C5). Additionally, the associations of airway inflammation with airway innervation and gastrointestinal motility were evaluated in mouse models of type 2 airway inflammation. ResultsPatients with asthma with FD had worse asthma control and cough severity and lower C2 and C5 thresholds than those without FD. The severity of FD symptoms was negatively correlated with C2 and C5 thresholds. FD and poor asthma control were predictors of heightened C-CS (defined as C5 of ≤ 2.44 μM) in asthma. A mouse model of papain-induced airway inflammation developed airway hyperinnervation and gastrointestinal dysmotility, and both pathologies were ameliorated by an anti-interleukin (IL)-33 antibody. Moreover, papain-induced gastrointestinal dysmotility was mitigated by silencing the airway sensory neurons using QX-314, a sodium channel blocker. Furthermore, sputum IL-33 levels were significantly elevated in patients with asthma with FD or heightened C-CS compared with those in their counterparts. ConclusionFD is significantly associated with airway neuronal dysfunction in asthma. IL-33-mediated airway neuronal dysfunction may contribute to the interaction between asthma and FD.

From Hematopoietic Stem Cells to Platelets: Unifying Differentiation Pathways Identified by Lineage Tracing Mouse Models.

Platelets are the terminal progeny of megakaryocytes, primarily produced in the bone marrow, and play critical roles in blood homeostasis, clotting, and wound healing. Traditionally, megakaryocytes and platelets are thought to arise from multipotent hematopoietic stem cells (HSCs) via multiple discrete progenitor populations with successive, lineage-restricting differentiation steps. However, this view has recently been challenged by studies suggesting that (1) some HSC clones are biased and/or restricted to the platelet lineage, (2) not all platelet generation follows the "canonical" megakaryocytic differentiation path of hematopoiesis, and (3) platelet output is the default program of steady-state hematopoiesis. Here, we specifically investigate the evidence that in vivo lineage tracing studies provide for the route(s) of platelet generation and investigate the involvement of various intermediate progenitor cell populations. We further identify the challenges that need to be overcome that are required to determine the presence, role, and kinetics of these possible alternate pathways.

Challenges and solutions to system-wide use of precision oncology as the standard of care paradigm.

The personalised oncology paradigm remains challenging to deliver despite technological advances in genomics-based identification of actionable variants combined with the increasing focus of drug development on these specific targets. To ensure we continue to build concerted momentum to improve outcomes across all cancer types, financial, technological and operational barriers need to be addressed. For example, complete integration and certification of the 'molecular tumour board' into 'standard of care' ensures a unified clinical decision pathway that both counteracts fragmentation and is the cornerstone of evidence-based delivery inside and outside of a research setting. Generally, integrated delivery has been restricted to specific (common) cancer types either within major cancer centres or small regional networks. Here, we focus on solutions in real-world integration of genomics, pathology, surgery, oncological treatments, data from clinical source systems and analysis of whole-body imaging as digital data that can facilitate cost-effectiveness analysis, clinical trial recruitment, and outcome assessment. This urgent imperative for cancer also extends across the early diagnosis and adjuvant treatment interventions, individualised cancer vaccines, immune cell therapies, personalised synthetic lethal therapeutics and cancer screening and prevention. Oncology care systems worldwide require proactive step-changes in solutions that include inter-operative digital working that can solve patient centred challenges to ensure inclusive, quality, sustainable, fair and cost-effective adoption and efficient delivery. Here we highlight workforce, technical, clinical, regulatory and economic challenges that prevent the implementation of precision oncology at scale, and offer a systematic roadmap of integrated solutions for standard of care based on minimal essential digital tools. These include unified decision support tools, quality control, data flows within an ethical and legal data framework, training and certification, monitoring and feedback. Bridging the technical, operational, regulatory and economic gaps demands the joint actions from public and industry stakeholders across national and global boundaries.

Dose-dependent regulation of kidney mitochondrial function by angiotensin II.

Intrarenal hypoxia has been suggested a unifying pathway to chronic kidney disease (CKD) and increased mitochondria leak respiration, which increases mitochondrial oxygen usage and is one important mechanism contributing to the development of the hypoxia. Previous studies indicate that angiotensin II (Ang II) effects on mitochondria function could be dose dependent. We investigated how moderate and high levels of Ang II affect kidney mitochondria function and pathways of leak respiration. C57 black 6 mice were treated with either vehicle or Ang II in low dose (400 ng/kg/min) or high dose (1,000 ng/kg/min) for 4 weeks. The function of kidney cortex mitochondria was measured by high-resolution respirometry. Ang II effects on gene expression in kidney tissue were measured by quantitative real-time PCR. Thiobarbituric acids reactive substances were determined as a marker of oxidative stress, and urinary protein excretion was measured as a maker of kidney injury. Low-dose Ang II induced overall mitochondria respiration, without compromising capacity of ATP production. Mitochondrial leak respiration was increased, and levels of oxidative stress were unchanged. However, high-dose Ang II decreased overall mitochondria respiration and reduced mitochondrial capacity for ATP production. Mitochondrial leak respiration was decreased, and oxidative stress increased in kidney tissue. Furthermore, gene expression of mediators that stimulate vasoconstriction and ROS production was increased, while components of counteracting pathways were decreased. In conclusion, Ang II dose-dependently affects mitochondrial function and leak respiration. Thus, Ang II has the potential to directly affect cellular metabolism during conditions of altered Ang II signaling.

Characterization of monoclonal gammopathy of undetermined significance progression to multiple myeloma through meta-analysis of GEO data

The etiology of monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM) is still obscure as are the processes that enable the progression of MGUS to MM. Understanding the unique vs. shared transcriptomes can potentially elucidate why individuals develop one or the other. Furthermore, highlighting key pathways and genes involved in the pathogenesis of MM or the development of MGUS to MM may allow the discovery of novel drug targets and therapies.We employed STARGEO platform to perform three separate meta-analysis to compare MGUS and MM transcriptomes. For these analyses we tagged (1) 101 MGUS patient plasma cells from bone marrow samples and 64 plasma cells from healthy controls (2) 383 MM patient CD138+ cells from bone marrow and the 101 MGUS samples in the first analysis as controls (3) 517 MM patient peripheral blood samples and 97 peripheral blood samples from healthy controls. We then utilized Ingenuity Pathway Analysis (IPA) to analyze the unique genomic signatures within and across these samples.Our study identified genes that may have unique roles in MGUS (GADD45RA and COMMD3), but also newly identified signaling pathways (EIF2, JAK/STAT, and MYC) and gene activity (NRG3, RBFOX2, and PARP15) in MGUS that have previously been shown to be involved in MM suggesting a spectrum of molecular overlap. On the other hand, genes such as DUSP4, RN14, LAMP5, differentially upregulated in MM, may be seen as tipping the scales from benignity to malignancy and could serve as drug targets or novel biomarkers for risk of progression. Furthermore, our analysis of MM identified newly associated gene/pathway activity such as inhibition of Wnt-signaling and defective B cell development. Finally, IPA analysis, suggests the multifactorial, oncogenic qualities of IFNγ signaling in MM may be a unifying pathway for these diverse mechanisms and prompts the need for further studies.

Inhibition of Cell Motility by Cell-Penetrating Dynamic Covalent Cascade Exchangers: Integrins Participate in Thiol-Mediated Uptake.

Integrins are cell surface proteins responsible for cell motility. Inspired by the rich disulfide exchange chemistry of integrins, we show here the inhibition of cell migration by cascade exchangers (CAXs), which also enable and inhibit cell penetration by thiol-mediated uptake. Fast-moving CAXs such as reversible Michael acceptor dimers, dithiabismepanes, and bioinspired epidithiodiketopiperazines are best, much better than Ellman's reagent. The implication that integrins participate in thiol-mediated uptake is confirmed by reduced uptake in integrin-knockdown cells. Although thiol-mediated uptake is increasingly emerging as a unifying pathway to bring matter into cells, its molecular basis is essentially unknown. These results identify the integrin superfamily as experimentally validated general cellular partners in the dynamic covalent exchange cascades that are likely to account for thiol-mediated uptake. The patterns identified testify to the complexity of the dynamic covalent networks involved. This work also provides chemistry tools to explore cell motility and expands the drug discovery potential of CAXs from antiviral toward antithrombotic and antitumor perspectives.

Improving VTE prophylaxis in ward and ICU surgical urology patients: a Six-Sigma DMAIC methodology improvement project

PurposeThis study intends to improve the quality of venous thromboembolism (VTE) prophylaxis practices including proper VTE risk assessment and the appropriate prophylaxis measures for surgical urology patients.Design/methodology/approachThe authors applied the Six-Sigma define, measure, analyze, improve and control (DMAIC) improvement methodology in a pre–post interventional study that involved all adult patients above 18 years old indicated and scheduled for urology surgical interventions including endoscopic urological surgeries in a urology specialized 60-bed hospital. The pre-intervention sample included all patients meeting the inclusion criteria over a period of six months. Post-intervention sample included all patients meeting the inclusion criteria over a period of six months. The improvement areas included both the VTE risk assessment as well as the VTE prophylaxis prescription.FindingsDMAIC methodology has achieved a substantial sustained improvement in surgical urology VTE prophylaxis practices with an average of 70% on both levels; VTE risk assessment practices and VTE prophylaxis prescribing practices were statistically significant. The post-intervention results also showed a statistically controlled process with no special cause variations. Based on the study results, the Six-Sigma DMAIC methodology can be considered of high value when applied in healthcare clinical practice improvement projects.Research limitations/implicationsThe project study includes some pitfalls that can be addressed as follows: 1. The lack of VTE rate incidence tracking. This limitation can be partly refuted when the authors conduct a literature review and explore that the VTE prophylaxis effectiveness had been proven with sufficient evidence to an extent that pushed several scientific societies to develop their own guidelines to support VTE prophylaxis. (Algattas et al., 2018). 2. Another limitation of this study can be that it handled only surgical patients and more specifically surgical urology patients. Of course, VTE prophylaxis is a crucial life-threatening problem not only for the surgical admitted patients but also for all the medical admitted patients either in hospital wards or ICUs. However, the prediction that surgical patients especially surgical urology patients are more prone to VTE development risk as they have -in several cases-two or three main additive risk factors which are age, procedure duration and malignancy in elderly men. (Tikkinen et al., 2014). So, the authors consider the study project to be a prototype that hopefully can be utilized for future study projects that will manage both other surgical specialty patients and medical patients on the national level and can track accurately and effectively report the VTE incidence rates.Practical implicationsSeveral recommendations can be extracted from the research project that is summarized in the following points: Paying focused attention to continuous healthcare quality improvement initiatives and projects as a main approach for healthcare improvement especially for the public health-related problems. This might be achieved through periodic region-specific or specialty-specific focus groups from which public health problems could be addressed and prioritized to be considered as a part of country healthcare campaigns regarding cost-utility and feasibility studies. The adoption of a system thinking approach in dealing with the improvement strategies; all efforts and resources are to be employed to achieve a common objective. This includes the generation of a national-wide electronic health information system that can aid in healthcare resource allocation and direct the healthcare efforts towards the most important, high-priority public health problems. Electronic national-wide health record is really an effort, and resources consuming activity, but actually, it's worth exerting efforts, and its valuable outcomes may be seen several years later. 3. Development of unified national specialized VTE prophylaxis pathways to standardize the patient-specific VTE prophylaxis plans. Standardization of healthcare pathways enables healthcare professionals to follow an evidence-based practice which will be reflected on the improvement of healthcare quality level, cost-effectiveness enhancement, and timely patient care on all levels especially in high critical areas like ER and ICU. 4. Incorporation of VTE prophylaxis costs in the universal health insurance diagnosis-related group (DRG) insurance packages and service pricing. Universal health insurance is a nationwide strategy that is aiming to cover all Egypt residents by the year 2030. Universal health insurance is being following the DRG reimbursement policy that is thought to control all the healthcare-associated costs so, the VTE prophylaxis costs shall be added as the main cost item to encourage all healthcare facilities to follow an evidence-based VTE prophylaxis pathway taking into consideration the high-risk patient categories who will definitely represent a high-cost burden on the long run if they suffer a VTE event.Originality/valueDMAIC improvement methodology applications in healthcare are still relatively limited, especially on the clinical level. The study can be considered one of a kind in Egypt dealing with a comprehensive DMAIC methodology application on the clinical level.

C-Jun N-terminal Kinase Promotes Stress Granule Assembly and Neurodegeneration in C9orf72-mediated ALS and FTD.

Stress granules are the RNA/protein condensates assembled in the cells under stress. They play a critical role in the pathogenesis of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). However, how stress granule assembly is regulated and related to ALS/FTD pathomechanism is incompletely understood. Mutation in the C9orf72 gene is the most common cause of familial ALS and FTD. C9orf72 mutation causes the formation of toxic dipeptide repeats. Here we show, the two most toxic dipeptide repeats i.e. poly(GR) and poly(PR) activate JNK via the ER-stress response protein IRE1 using fly and cellular models. Further, we show activated JNK promotes stress granule assembly in cells by promoting the transcription of one of the key stress granule proteins i.e. G3BP1 by inducing histone 3 phosphorylation. Consistent with these findings, JNK or IRE1 inhibition reduced stress granule formation, histone 3 phosphorylation, G3BP1 mRNA and protein levels, and neurotoxicity in cells overexpressing poly(GR) and poly(PR) or neurons derived from male and female C9ALS/FTD patient-induced pluripotent stem cells (iPSCs). Our findings connect ER stress, JNK activation, and stress granule assembly in a unified pathway contributing to C9ALS/FTD neurodegeneration.Significance StatementJNK is a part of the mitogen-activated protein kinase pathway which is the central node for the integration of multiple stress signals. Cells are under constant stress in neurodegenerative diseases and how these cells respond to stress signals is a critical factor in determining their survival or death. Previous studies have shown JNK as a major contributor to cellular apoptosis. Here, we show the role of JNK in stress granule assembly. We identify toxic dipeptide repeats produced in ALS/FTD conditions activate JNK. The activated JNK in the nucleus can induce histone modifications which increase G3BP1 expression thus promoting stress granule assembly and neurodegeneration.

Unsupervised voxel-based segmentation reveals landscape of bacterial ribosome large subunit early assembly.

Ribosome biogenesis is a complex but efficient process in rapidly growing bacteria, where assemble a functional 70S ribosome takes ∼2 min, involving participation of 3 rRNAs, 50 r-proteins and dozens of assembly factors. In vitro reconstitution using different subsets of large subunit (50S, LSU) proteins with rRNAs, pioneered by Nierhaus lab, resulted in the Nierhaus assembly map, embodying the cooperativity and dependency for binding of LSU r-proteins to 23S rRNA. Critically absent from the Nierhaus map is the underlying folding of the rRNA that creates the binding sites for the r-proteins. In addition, the relationship of the observed cooperativity in vitro to the co-transcriptional assembly in cells remains to be determined. Pre-50S intermediates accumulate at low temperature in ∆deaD, a DEAD-box helicase implicated in 50S assembly. We solved 21 pre-50S density maps from intermediate-containing fractions using cryo-EM. In the newly solved maps, we discovered the earliest intermediate ever reported, consisting of domain I at the 5’-end of 23S rRNA. To probe the mechanism behind the maps during assembly, we developed a novel density map segmentation and dependency analysis method. Ten cooperative assembly blocks were identified from segmentation of the maps, and these were organized into a block dependency map. This is the first time the dependencies on folding of rRNA helices and ribosomal protein binding could be integrated into a complete assembly map. In addition, we showed how the exit tunnel is folded on the solvent side, serving as a scaffold for 50S maturation. Using this new segmentation analysis method, we revisited previously reported bL17-depletion and ∆srmB datasets. Most remarkably, the other two datasets are also consistent with the block dependency, implying a unified early assembly pathway and flexible maturation landscape in early 50S biogenesis.

Psoriasis associated with asthma and allergic rhinitis: a US-based cross-sectional study using the All of US Research Program.

Psoriasis is a common chronic inflammatory disease with multiple known comorbidities. Increasing evidence suggests some mechanistic overlap in the immunopathogenesis of psoriasis and some cases of asthma and allergic rhinitis (AR), but the potential association between psoriasis and asthma and AR has not been thoroughly investigated. The study aimed to investigate the association between psoriasis and asthma and AR. We used data from the NIH All of US Research Program, a nationwide longitudinal cohort of US adults, collected from 2018 to present. The source population comprised a demographically and socioeconomically diverse cohort of over 300,000 Americans. We used multivariable logistic regression models to examine the association between psoriasis and asthma and AR, after adjusting for sociodemographic variables, body mass index, and smoking status. In total, 235,551 participants (mean [SD] age, 54.7 [16.6] years; 59.3% female), including 5165 individuals with psoriasis and 230,386 individuals without psoriasis, were included in our analysis. Participants with psoriasis had significantly higher prevalence of asthma (26.1% vs. 12.9%; P < 0.001) and AR (31.8% vs. 13.4%; P < 0.001) compared to participants without psoriasis. Psoriasis was significantly associated with both asthma [adjusted odds ratio (aOR) 2.22; 95% confidence interval (CI) 2.08-2.37] and AR (aOR, 2.57; 95% CI 2.42-2.73). In subgroup analyses, associations remained stable in multivariable analyses after stratification by age, sex, and income. Psoriasis is associated with both asthma and AR in our sample of US adults. Further research is needed to explore potentially unifying inflammatory pathways among psoriasis, asthma, and AR.

Comparative molecular analysis of two HFpEF/diastolic dysfunction models – in search of a unifying disease mechanism

Despite progress in the molecular analyses of Heart Failure with preserved Ejection Fraction (HFpEF), it remains unclear if the syndrome represents many diseases that manifest themselves with similar phenotypes, or if there is an underlying unifying pathway responsible for the disease etiology.

A Unified Primary Care Approach for the Management of Insomnia and Confusion Using the Clinical Pathway

Background: Delirium and confusion are risk factors for falls and its related injuries. Therefore, we designed a unified clinical pathway to reduce the risk of delirium among hospitalized patients. Methods: A unified approach of pharmacotherapy for managing insomnia and confusion, which is subdivided based on the patients’ age (&lt;70 versus ≥70 years) and presence of diabetes mellitus, was implemented in Nagoya Memorial Hospital, Japan. Risk factors for delirium were assessed via a multidisciplinary approach. For older high-risk patients, suvorexant or trazodone was prophylactically prescribed to promote sleep. If delirium occurred, either quetiapine or perospirone was administered prior to the usage of risperidone or haloperidol. The amounts of prescribed sleep inducers and antipsychotic agents were examined. The application rate of the clinical pathway and incidence of falls were compared before and after its introduction. Results: The application of a unified approach for the management of insomnia and confusion significantly decreased the prescribed amounts of benzodiazepines and may be associated with the reduced incidence of falls among hospitalized patients. Conclusions: A clinical pathway assisted the selection of sleep inducers and antipsychotic agents. This is useful for managing inpatients safely by preventing delirium and subsequent falls.

Nucleation and crystallization mechanism of heavy hydrocarbons in natural gas under flow field

The understanding of the crystallization mechanism of heavy hydrocarbons in natural gas directly determines the modification of nucleation theory and the development of new liquefaction processes. However, the mechanism of heavy hydrocarbon crystallization in the flow field is still not well understood. In this paper, molecular dynamics (MD) simulations were adopted to investigate heavy hydrocarbon crystallization under shear flow, and the nucleation pathway related to crystallization kinetics was explored. The results show that shear rates less than 1 × 108 s−1 promote the rapid growth of heavy hydrocarbon crystals with high final crystallinity, whereas higher shear rates cause crystals to grow slowly ending up with lower crystallinity. Compared with quiescent nucleation, the presence of a shear flow promotes the heavy hydrocarbon stretching and consequently accelerates the kinetic process of phase transition. In addition, molecular orientation fluctuations arising from the flow field are accompanied by the elastic deformation of solid nucleus, increasing the nucleation free energy barrier. Therefore, the nucleation rate of heavy hydrocarbon liquids peaks at the optimal shear rate 1 × 107 s−1. Moreover, a unified pathway for heavy hydrocarbon nucleation under shear flow was proposed.

A central alarm system that gates multi-sensory innate threat cues to the amygdala.

SUMMARYPerception of threats is essential for survival. Previous findings suggest that parallel pathways independently relay innate threat signals from different sensory modalities to multiple brain areas, such as the midbrain and hypothalamus, for immediate avoidance. Yet little is known about whether and how multi-sensory innate threat cues are integrated and conveyed from each sensory modality to the amygdala, a critical brain area for threat perception and learning. Here, we report that neurons expressing calcitonin gene-related peptide (CGRP) in the parvocellular subparafascicular nucleus in the thalamus and external lateral parabrachial nucleus in the brainstem respond to multi-sensory threat cues from various sensory modalities and relay negative valence to the lateral and central amygdala, respectively. Both CGRP populations and their amygdala projections are required for multi-sensory threat perception and aversive memory formation. The identification of unified innate threat pathways may provide insights into developing therapeutic candidates for innate fear-related disorders.

Aβ42 oligomers trigger synaptic loss through CAMKK2-AMPK-dependent effectors coordinating mitochondrial fission and mitophagy

During the early stages of Alzheimer’s disease (AD) in both mouse models and human patients, soluble forms of Amyloid-β 1–42 oligomers (Aβ42o) trigger loss of excitatory synapses (synaptotoxicity) in cortical and hippocampal pyramidal neurons (PNs) prior to the formation of insoluble amyloid plaques. In a transgenic AD mouse model, we observed a spatially restricted structural remodeling of mitochondria in the apical tufts of CA1 PNs dendrites corresponding to the dendritic domain where the earliest synaptic loss is detected in vivo. We also observed AMPK over-activation as well as increased fragmentation and loss of mitochondrial biomass in Ngn2-induced neurons derived from a new APPSwe/Swe knockin human ES cell line. We demonstrate that Aβ42o-dependent over-activation of the CAMKK2-AMPK kinase dyad mediates synaptic loss through coordinated phosphorylation of MFF-dependent mitochondrial fission and ULK2-dependent mitophagy. Our results uncover a unifying stress-response pathway causally linking Aβ42o-dependent structural remodeling of dendritic mitochondria to synaptic loss.

Human–rat integrated microRNAs profiling identified a new neonatal cerebral hypoxic–ischemic pathway melatonin‐sensitive

Neonatal encephalopathy (NE) is a pathological condition affecting long‐term neurodevelopmental outcomes. Hypothermia is the only therapeutic option, but does not always improve outcomes; hence, researchers continue to hunt for pharmaceutical compounds. Melatonin treatment has benefitted neonates with hypoxic–ischemic (HI) brain injury. However, unlike animal models that enable the study of the brain and the pathophysiologic cascade, only blood is available from human subjects. Therefore, due to the unavailability of neonatal brain tissue, assumptions about the pathophysiology in pathways and cascades are made in human subjects with NE. We analyzed animal and human specimens to improve our understanding of the pathophysiology in human neonates. A neonate with NE who underwent hypothermia and enrolled in a melatonin pharmacokinetic study was compared to HI rats treated/untreated with melatonin. MicroRNA (miRNA) analyses provided profiles of the neonate's plasma, rat plasma, and rat brain cortexes. We compared these profiles through a bioinformatics tool, identifying Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways common to HI brain injury and melatonin treatment. After evaluating the resulting pathways and the literature, to validate the method, the key proteins expressed in HI brain injury were investigated using cerebral cortexes. The upregulated miRNAs in human neonate and rat plasma helped identify two KEGG pathways, glioma and long‐term potentiation, common to HI injury and melatonin treatment. A unified neonatal cerebral melatonin‐sensitive HI pathway was designed and validated by assessing the expression of protein kinase Cα (PKCα), phospho (p)‐Akt, and p‐ERK proteins in rat brain cortexes. PKCα increased in HI‐injured rats and further increased with melatonin. p‐Akt and p‐ERK returned phosphorylated to their basal level with melatonin treatment after HI injury. The bioinformatics analyses validated by key protein expression identified pathways common to HI brain injury and melatonin treatment. This approach helped complete pathways in neonates with NE by integrating information from animal models of HI brain injury.