Methionine enkephalin (MENK) protected macrophages from ferroptosis by downregulating HMOX1 and ferritin.

OBJECTIVE: The aim of this work was to investigate the immunological effect of MENK by analyzing the protein spectrum and bioinformatics of macrophage RAW264.7, and to explore the relationship between macrophage and ferroptosis. RESULT: We employed proteomic analysis to identify differentially expressed proteins (DEPs) between macrophages and macrophages intervened by MENK. A total of 208 DEPs were identified. Among these, 96 proteins had upregulated expression and 112 proteins had downregulated expression. Proteomic analysis revealed a significant enrichment of DEPs associated with iron metabolism. The identification of hub genes was conducted using KEGG pathway diagrams and protein-protein interaction (PPI) analysis. The hub genes identified in this study include HMOX1 and Ferritin (FTH and FTL). A correlation was established between HMOX1, FTH, and FTL in the GO and KEGG databases. The results of PCR, WB and immunofluorescence showed that MENK downregulated the level of HMOX1 and FTH. CONCLUSION: MENK had the potential to become an adjuvant chemotherapy drug by regulating iron metabolism in macrophages, reducing levels of HMOX1 and ferritin. We proposed an innovative research direction on the therapeutic potential of MENK, focusing on the relationship between ferroptosis and macrophage activity.

Proteomics analysis of methionine enkephalin upregulated macrophages against infection by the influenza-A virus.

Macrophages have a vital role in phagocytosis and antiviral effect against invading influenza viruses. Previously, we found that methionine enkephalin (MENK) inhibited influenza virus infection by upregulating the "antiviral state" of macrophages. To investigate the immunoregulatory mechanism of action of MENK on macrophages, we employed proteomic analysis to identify differentially expressed proteins (DEPs) between macrophages infected with the influenza-A virus and cells infected with the influenza-A virus after pretreatment with MENK. A total of 215 DEPs were identified: 164 proteins had upregulated expression and 51 proteins had downregulated expression. Proteomics analysis showed that DEPs were highly enriched in "cytokine-cytokine receptor interaction", "phagosome", and "complement and coagulation cascades pathway". Proteomics analysis revealed that MENK could be an immune modulator or prophylactic for the prevention and treatment of influenza. MENK promoted the polarization of M1 macrophages, activated inflammatory responses, and enhanced phagocytosis and killing function by upregulating opsonizing receptors.

The Role of Mechanical Thrombectomy for Acute Massive Pulmonary Embolism in a Patient With Unilateral Lung Transplant and Atrial Septal Defect.

PURPOSE: The risk of thromboembolic disease is high in patients with lung transplantation and is associated with significant morbidity and mortality with single healthy transplanted lung. We present a case involving successful endovascular management of life-threatening acute massive pulmonary embolism (PE) in a patient with single lung transplant and atrial septal defect (ASD). CASE REPORT: A 65-year-old man with a history of interstitial lung disease status post single left orthotopic lung transplant in 2012 presented with acute massive PE and clot burden in the pulmonary arteries of the transplanted left lung. Severe right heart dysfunction, hemodynamic instability, and requirement for vasopressors persisted post systemic thrombolytic therapy. As a result, the patient underwent successful endovascular mechanical thrombectomy with immediate improvement in oxygen saturation and hemodynamic status. The procedure was performed without adverse outcomes or paradoxical embolization despite the presence of ASD. The right heart dysfunction resolved, the patient was extubated the next day, and was discharged to home 2 days post procedure. CONCLUSIONS: Endovascular mechanical thrombectomy was safely used to treat acute massive PE in a single transplanted lung in the presence of ASD. CLINICAL IMPACT: Endovascular mechanical thrombectomy could be safely utilized to treat patients with lung transplant and acute massive or submassive pulmonary embolism. However, safely of mechanical thrombectomy should be determined in case-based scenarios and based on time interval from transplantation to when the thrombectomy is required.

Using 3D-Printed Mesh-Like Brain Cortex with Deep Structures for Planning Intracranial EEG Electrode Placement.

Surgical evaluation of medically refractory epilepsy frequently necessitates implantation of multiple intracranial electrodes for the identification of the seizure focus. Knowledge of the individual brain's surface anatomy and deep structures is crucial for planning the electrode implantation. We present a novel method of 3D printing a brain that allows for the simulation of placement of all types of intracranial electrodes. We used a DICOM dataset of a T1-weighted 3D-FSPGR brain MRI from one subject. The segmentation tools of Materialise Mimics 21.0 were used to remove the osseous anatomy from brain parenchyma. Materialise 3-matic 13.0 was then utilized in order to transform the cortex of the segmented brain parenchyma into a mesh-like surface. Using 3-matic tools, the model was modified to incorporate deep brain structures and create an opening in the medial aspect. The final model was then 3D printed as a cerebral hemisphere with nylon material using selective laser sintering technology. The final model was light and durable and reflected accurate details of the surface anatomy and some deep structures. Additionally, standard surgical depth electrodes could be passed through the model to reach deep structures without damaging the model. This novel 3D-printed brain model provides a unique combination of visualizing both the surface anatomy and deep structures through the mesh-like surface while allowing repeated needle insertions. This relatively low-cost technique can be implemented for interdisciplinary preprocedural planning in patients requiring intracranial EEG monitoring and for any intervention that requires needle insertion into a solid organ with unique anatomy and internal targets.

Mechanism for Burgess Shale-type preservation.

Exceptionally preserved fossil biotas of the Burgess Shale and a handful of other similar Cambrian deposits provide rare but critical insights into the early diversification of animals. The extraordinary preservation of labile tissues in these geographically widespread but temporally restricted soft-bodied fossil assemblages has remained enigmatic since Walcott's initial discovery in 1909. Here, we demonstrate the mechanism of Burgess Shale-type preservation using sedimentologic and geochemical data from the Chengjiang, Burgess Shale, and five other principal Burgess Shale-type deposits. Sulfur isotope evidence from sedimentary pyrites reveals that the exquisite fossilization of organic remains as carbonaceous compressions resulted from early inhibition of microbial activity in the sediments by means of oxidant deprivation. Low sulfate concentrations in the global ocean and low-oxygen bottom water conditions at the sites of deposition resulted in reduced oxidant availability. Subsequently, rapid entombment of fossils in fine-grained sediments and early sealing of sediments by pervasive carbonate cements at bed tops restricted oxidant flux into the sediments. A permeability barrier, provided by bed-capping cements that were emplaced at the seafloor, is a feature that is shared among Burgess Shale-type deposits, and resulted from the unusually high alkalinity of Cambrian oceans. Thus, Burgess Shale-type preservation of soft-bodied fossil assemblages worldwide was promoted by unique aspects of early Paleozoic seawater chemistry that strongly impacted sediment diagenesis, providing a fundamentally unique record of the immediate aftermath of the "Cambrian explosion."

Leukemia cells induce changes in human bone marrow stromal cells.

BACKGROUND: Bone marrow stromal cells (BMSCs) are multipotent cells that support angiogenesis, wound healing, and immunomodulation. In the hematopoietic niche, they nurture hematopoietic cells, leukemia, tumors and metastasis. BMSCs secrete of a wide range of cytokines, growth factors and matrix proteins which contribute to the pro-tumorigenic marrow microenvironment. The inflammatory cytokines IFN-γ and TNF-α change the BMSC secretome and we hypothesized that factors produced by tumors or leukemia would also affect the BMSC secretome and investigated the interaction of leukemia cells with BMSCs. METHODS: BMSCs from healthy subjects were co-cultured with three myeloid leukemia cell lines (TF-1, TF-1α and K562) using a trans-well system. Following co-culture, the BMSCs and leukemia cells were analyzed by global gene expression analysis and culture supernatants were analyzed for protein expression. As a control, CD34+ cells were also cocultured with BMSCs. RESULTS: Co-culture induced leukemia cell gene expression changes in stem cell pluripotency, TGF-ß signaling and carcinoma signaling pathways. BMSCs co-cultured with leukemia cells up-regulated a number of proinflammatory genes including IL-17 signaling-related genes and IL-8 and CCL2 levels were increased in co-culture supernatants. In contrast, purine metabolism, mTOR signaling and EIF2 signaling pathways genes were up-regulated in BMSCs co-cultured with CD34+ cells. CONCLUSIONS: BMSCs react to the presence of leukemia cells undergoing changes in the cytokine and chemokine secretion profiles. Thus, BMSCs and leukemia cells both contribute to the creation of a competitive niche more favorable for leukemia stem cells.

Life cycle evolution in the trilobites Balangia and Duyunaspis from the Cambrian Series 2 (Stage 4) of South China.

The evolution process can be reconstructed by tracking the changes in the dynamic characters of life cycles. A number of related trilobites from the Cambrian of South China provide additional information for the study of trilobite evolutionary patterns, which has been hampered by previous incomplete fossil record though. Here, Balangia and Duyunaspis represent related Cambrian oryctocephalid trilobites from South China, are comprehensively discussed over the ontogeny, and the results show that, from B. balangensis via D. duyunensis to D. jianheensis, their exoskeletal morphology shows a directional evolution. Based on the direction of evolutionary changes in the development of Balangia and Duyunaspis, we speculate that Duyunaspis likely evolved from Balangia instead of Balangia evolved from Duyunaspis, as was previously assumed. This inference is also supported by the phylogenetic tree. This research provides not only a better understanding of the mechanisms of evolution in trilobites, but also new insights for the relationship between developmental evolutionary changes and phylogeny in trilobites.

Long-term assessment of cutaneous inflammation and treatment using optical resolution photoacoustic microscopy.

Cutaneous inflammation is an acute skin disease characterized by edema and vascular hyperplasia. Longitudinal monitoring of vasculature is crucial for studying the development of inflammation and evaluating the therapeutic efficacy of drugs. Optical-resolution photoacoustic microscopy (OR-PAM) is a hybrid imaging tool for non-invasive and label-free visualization of microcirculations with a capillary-scale spatial resolution. In this study, we assess the feasibility of OR-PAM for long-term monitoring of vascular changes in 12-O-Tetradecanoylphorbol-13-Acetate (TPA)-induced mouse models, as well as the corresponding treatment process. Quantitative vascular evaluation is conducted based on derived key parameters, including vessel length, branchpoint number, vessel area fraction, vessel diameter, fractal dimension, vessel tortuosity and ear thickness, which reveal that vascular morphological changes are highly dependent on the concentration of TPA and existence of therapeutic drugs. Furthermore, the results show the potential of OR-PAM in the clinical management of inflammation and as an effective tool to evaluate vascular responses to pharmacological interventions in vivo.

Leanchoiliidae reveals the ancestral organization of the stem euarthropod brain.

Fossils provide insights into how organs may have diversified over geological time.1 However, diversification already accomplished early in evolution can obscure ancestral events leading to it. For example, already by the mid-Cambrian period, euarthropods had condensed brains typifying modern mandibulate lineages.2 However, the demonstration that extant euarthropods and chordates share orthologous developmental control genes defining the segmental fore-, mid-, and hindbrain suggests that those character states were present even before the onset of the Cambrian.3 Fossilized nervous systems of stem Euarthropoda might, therefore, be expected to reveal ancestral segmental organization, from which divergent arrangements emerged. Here, we demonstrate unsurpassed preservation of cerebral tissue in Kaili leanchoiliids revealing near-identical arrangements of bilaterally symmetric ganglia identified as the proto-, deuto-, and tritocerebra disposed behind an asegmental frontal domain, the prosocerebrum, from which paired nerves extend to labral ganglia flanking the stomodeum. This organization corresponds to labral connections hallmarking extant euarthropod clades4 and to predicted transformations of presegmental ganglia serving raptorial preocular appendages of Radiodonta.5 Trace nervous system in the gilled lobopodian Kerygmachela kierkegaardi6 suggests an even deeper prosocerebral ancestry. An asegmental prosocerebrum resolves its location relative to the midline asegmental sclerite of the radiodontan head, which persists in stem Euarthropoda.7 Here, data from two Kaili Leanchoilia, with additional reference to Alalcomenaeus,8,9 demonstrate that Cambrian stem Euarthropoda confirm genomic and developmental studies10-15 claiming that the most frontal domain of the euarthropod brain is a unique evolutionary module distinct from, and ancestral to, the fore-, mid-, and hindbrain.

Synchronized moulting behaviour in trilobites from the Cambrian Series 2 of South China.

The study of moulting behaviour in the fossil record is relatively well known in arthropods and this is especially true for trilobites. Nevertheless, while studies focusing on the style of moulting in social and semi-social groups of modern animals (e.g. arthropods) are common, very few works investigate moulting adaptations in deep time. Here we report a trilobite assemblage from the Cambrian Series 2 "Tsinghsutung" Formation of South China. Around 850 specimens were used for this study from three different levels across one section near Balang (SE Guizhou Province, South China). These levels preserve numerous trilobite clusters in some cases containing around 400 individual specimens. Up to four species have been found in these clusters, but two species are more common. Trilobite clusters bear a high percentage of disarticulated specimens that we interpret as moults. Additionally, measurements of bioclast orientation and the dorsoventral attitude suggests very quiet water conditions followed by rapid burial events, prior to scavenger disturbance. Together, this indicates that the fossil assemblages were a result of a biological phenomenon rather than mechanical processes, allowing us to interpret the position of the fossil parts as different moulting configurations. Since the trilobite assemblage seems to be in situ, the large number of exuviae suggests a local place of migration. This was triggered by the need for group protection while moulting, which is suggestive of gregarious behaviour, possibly synchronized. These trilobites from the Cambrian Epoch 2, Age 4 constitute one of the earliest known gregarious community of trilobites and has important implications for understanding the ecology of this group during their emergence in the Cambrian.

IPSE, an abundant egg-secreted protein of the carcinogenic helminth Schistosoma haematobium, promotes proliferation of bladder cancer cells and angiogenesis.

BACKGROUND: Schistosoma haematobium, the helminth causing urogenital schistosomiasis, is a known bladder carcinogen. Despite the causal link between S. haematobium and bladder cancer, the underlying mechanisms are poorly understood. S. haematobium oviposition in the bladder is associated with angiogenesis and urothelial hyperplasia. These changes may be pre-carcinogenic events in the bladder. We hypothesized that the Interleukin-4-inducing principle of Schistosoma mansoni eggs (IPSE), an S. haematobium egg-secreted "infiltrin" protein that enters host cell nuclei to alter cellular activity, is sufficient to induce angiogenesis and urothelial hyperplasia. Methods: Mouse bladders injected with S. haematobium eggs were analyzed via microscopy for angiogenesis and urothelial hyperplasia. Endothelial and urothelial cell lines were incubated with recombinant IPSE protein or an IPSE mutant protein that lacks the native nuclear localization sequence (NLS-) and proliferation measured using CFSE staining and real-time monitoring of cell growth. IPSE's effects on urothelial cell cycle status was assayed through propidium iodide staining. Endothelial and urothelial cell uptake of fluorophore-labeled IPSE was measured. Findings: Injection of S. haematobium eggs into the bladder triggers angiogenesis, enhances leakiness of bladder blood vessels, and drives urothelial hyperplasia. Wild type IPSE, but not NLS-, increases proliferation of endothelial and urothelial cells and skews urothelial cells towards S phase. Finally, IPSE is internalized by both endothelial and urothelial cells. Interpretation: IPSE drives endothelial and urothelial proliferation, which may depend on internalization of the molecule. The urothelial effects of IPSE depend upon its NLS. Thus, IPSE is a candidate pro-carcinogenic molecule of S. haematobium. SUMMARY: Schistosoma haematobium acts as a bladder carcinogen through unclear mechanisms. The S. haematobium homolog of IPSE, a secreted schistosome egg immunomodulatory molecule, enhances angiogenesis and urothelial proliferation, hallmarks of pre-carcinogenesis, suggesting IPSE is a key pro-oncogenic molecule of S. haematobium.

A new high-resolution 3-D quantitative method for analysing small morphological features: an example using a Cambrian trilobite.

Taphonomic processes play an important role in the preservation of small morphological features such as granulation or pits. However, the assessment of these features may face the issue of the small size of the specimens and, sometimes, the destructiveness of these analyses, which makes impossible carrying them out in singular specimen, such as holotypes or lectotypes. This paper takes a new approach to analysing small-morphological features, by using an optical surface roughness (OSR) meter to create a high-resolution three-dimensional digital-elevation model (DEM). This non-destructive technique allows analysing quantitatively the DEM using geometric morphometric methods (GMM). We created a number of DEMs from three populations putatively belonging to the same species of trilobite (Oryctocephalus indicus) that present the same cranidial outline, but differ in the presence or absence of the second and third transglabellar furrows. Profile analysis of the DEMs demonstrate that all three populations show similar preservation variation in the glabellar furrows and lobes. The GMM shows that all populations exhibit the same range of variation. Differences in preservation are a consequence of different degrees of cementation and rates of dissolution. Fast cementation enhances the preservation of glabellar furrows and lobes, while fast dissolution hampers preservation of the same structures.

Interferon-γ and Tumor Necrosis Factor-α Polarize Bone Marrow Stromal Cells Uniformly to a Th1 Phenotype.

Activated T cells polarize mesenchymal stromal cells (MSCs) to a proinflammatory Th1 phenotype which likely has an important role in amplifying the immune response in the tumor microenvironment. We investigated the role of interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α), two factors produced by activated T cells, in MSC polarization. Gene expression and culture supernatant analysis showed that TNF-α and IFN-γ stimulated MSCs expressed distinct sets of proinflammatory factors. The combination of IFN-γ and TNF-α was synergistic and induced a transcriptome most similar to that found in MSCs stimulated with activated T cells and similar to that found in the inflamed tumor microenvironment; a Th1 phenotype with the expression of the immunosuppressive factors IL-4, IL-10, CD274/PD-L1 and indoleamine 2,3 dioxygenase (IDO). Single cell qRT-PCR analysis showed that the combination of IFN-γ and TNF-α polarized uniformly to this phenotype. The combination of IFN-γ and TNF-α results in the synergist uniform polarization of MSCs toward a primarily Th1 phenotype. The stimulation of MSCs by IFN-γ and TNF-α released from activated tumor infiltrating T cells is likely responsible for the production of many factors that characterize the tumor microenvironment.