Miracle-dz/UniFAR-FTUs huggingface dataset:
Cathepsin B as a potential cystatin M/E target in the mouse hair follicle

Deficiency of the cysteine protease inhibitor cystatin M/E (Cst6) in mice leads to disturbed epidermal cornification, impaired barrier function, and neonatal lethality. We report the rescue of the lethal skin phenotype of ichq (Cst6‐deficient; Cst6−/−) mice by transgenic, epidermis‐specific, reexpression of Cst6 under control of the human involucrin (INV) promoter. Rescued Tg(INV‐Cst6)Cst6ichq/ichq mice survive the neonatal phase, but display severe eye pathology and alopecia after 4 mo. We observed keratitis and squamous metaplasia of the corneal epithelium, comparable to Cst6−/− Ctsl+/− mice, as we have reported in other studies. We found the INV promoter to be active in the hair follicle infundibulum; however, we did not observe Cst6 protein expression in the lower regions of the hair follicle in Tg(INV‐Cst6)Cst6ichq/ichq mice. This result suggests that unrestricted activity of proteases is involved in disturbance of hair follicle biology, eventually leading to baldness. Using quenched activity‐based probes, we identified mouse cathepsin B (CtsB), which is expressed in the lower regions of the hair follicle, as an additional target of mouse Cst6. These data suggest that Cst6 is necessary to control CtsB activity in hair follicle morphogenesis and highlight Cst6‐controlled proteolytic pathways as targets for preventing hair loss. —Oortveld, M. A. W. , van Vlijmen‐Willems, I. M. J. J., Kersten, F. F. J., Cheng, T. , Verdoes, M. , van Erp, P. E. J., Verbeek, S. , Reinheckel, T. , Hendriks, W. J. A. J., Schalkwijk, J., Zeeuwen, P. L. J. M. Cathepsin B as a potential cystatin M/E target in the mouse hair follicle. FASEB J. 31, 4286–4294 (2017). www.fasebj. org
Proprotein convertase Furin1 expression in the Drosophila fat body is essential for a normal antimicrobial peptide response and bacterial host defense

Invading pathogens provoke robust innate immune responses in Dipteran insects, such as Drosophila melanogaster. In a systemic bacterial infection, a humoral response is induced in the fatbody. Gram‐positive bacteria trigger the Toll signaling pathway, whereas gram‐negative bacterial infections are signaled via the immune deficiency (IMD) pathway. We show here that the RNA interference‐mediated silencing of Furin1—a member of the proprotein convertase enzyme family—specifically in the fat body, results in a reduction in the expression of antimicrobial peptides. This, in turn, compromises the survival of adult fruit flies in systemic infections that are caused by both gram‐positive and ‐negative bacteria. Furin1 plays a nonredundant role in the regulation of immune responses, as silencing of Furin2, the other member of the enzyme family, had no effect on survival or the expression of antimicrobial peptides upon a systemic infection. Furin1 does not directly affect the Toll or IMD signaling pathways, but the reduced expression of Furin1 up‐regulates stress response factors in the fat body. We also demonstrate that Furin1 is a negative regulator of the Janus kinase/signal transducer and activator of transcription signaling pathway, which is implicated in stress responses in the fly. In summary, our data identify Furin1 as a novel regulator of humoral immunity and cellular stress responses in Drosophila. —Aittomäki, S. , Valanne, S. , Lehtinen, T. , Matikainen, S. , Nyman, T. A. , Rämet, M. , Pesu, M. Proprotein convertase Furin1 expression in the Drosophila fat body is essential for a normal antimicrobial peptide response and bacterial host defense. FASEB J. 31, 4770–4782 (2017). www.fasebj. org
E‐selectin‐mediated rolling facilitates pancreatic cancer cell adhesion to hyaluronic acid

Tumor cell extravasation is a multistep process preceded by cell rolling and arrest on the vessel wall via the formation of specific receptor–ligand bonds. The strength, availability, and number of receptor–ligand bonds regulate the rate by which tumor cells tether, roll, and adhere to vascular walls. Although the mechanics of selectin‐mediated rolling have been extensively studied, little is known regarding how tumor cell rolling on selectins facilitates adhesion to a distinct substrate‐bound protein with different kinetic properties. By using multicomponent protein patterning and a microfluidic system, we evaluated how E‐selectin‐dependent rolling modulates hyaluronic acid (HA) adhesion as a function of fluid shear, contact time, and the spacing between E‐selectin and HA regions patterned on the substrate. We show that tumor cells rolling on E‐selectin were ~40‐fold more likely to bind to HA than nonrolling cells in shear flow. Furthermore, E‐selectin‐dependent rolling promotes adhesion to HA by both physically slowing cells and enabling them to position proximal to the surface, thereby increasing the on rate of adhesion. A better understanding of tumor cell adhesion under physiologic shear would lead to the development of new diagnostic assays and pave the way to clinical approaches aimed ultimately to halt metastasis. —Shea, D. J., Li, Y. W. , Stebe, K. J., Konstantopoulos, K. E‐selectin‐mediated rolling facilitates pancreatic cancer cell adhesion to hyaluronic acid. FASEB J. 31, 5078–5086 (2017). www.fasebj. org
Specificity of anti‐saliva immune response in mice repeatedly bitten by Phlebotomus sergenti

Sand flies are bloodsucking insects transmitting parasites of genus Leishmania, the causative agents of diseases in humans and dogs. Experimental hosts repeatedly exposed to sand fly saliva can control Leishmania infection. Cell‐mediated anti‐saliva immune response is most likely responsible for this protective effect; however, there is no study so far concerning its antigenic specificity towards different sand fly vectors. In this study, splenocytes from BALB/c mice repeatedly exposed to the bites of Phlebotomus sergenti were challenged ex vivo with salivary gland homogenates from three different sand fly vectors –P. sergenti, P. papatasi, or P. arabicus. Mice bitten by P. sergenti had higher proliferative response to homologous antigen than splenocytes from naive mice. Splenocytes from P. sergenti bitten mice as well as anti‐P. sergenti antibodies partially cross‐reacted with P. papatasi saliva. In contrast, no cross‐reactivity was found with P. arabicus saliva. Our data indicate that both arms of the immune system, cellular and humoral, react in a species‐specific manner. Therefore, the presence of antibodies against salivary components of a certain species indicates the specificity of cell‐mediated immune response as well. The data suggest that unique transmission‐blocking vaccine would be required for each vector –Leishmania combination.
Specific phage‐displayed peptides discriminate different forms of neurocysticercosis by antibody detection in the serum samples

Neurocysticercosis (NC), caused by Taenia solium metacestode, infects the central nervous system and is a devastating parasitic infection. Diagnosis is based on symptoms, imaging, serology and epidemiology. Current markers present variable sensitivity and specificity, frequent cross‐reactions and are not able to discriminate NC clinical forms. The aim of this study was to select mimotopes of T. solium metacestode antigens that may be used in NC immunodiagnosis, specifically to discriminate between active and inactive forms. A random peptide phage display library was screened against IgY from chickens immunized with total saline extract from T. solium metacestodes and validated against 110 serum samples, classified into active NC (18), inactive NC (22), cross‐reactive parasitic diseases (40) and healthy controls (30). We have successfully selected seven peptides with significant immunoreactivity to IgG of NC patients, with sensitivity ranging from 95·5% to 100% to detect the inactive form and specificity varied from 85·7% to 94·3%. One phage‐displayed peptide (Cc48) can be directly used as biomarker to distinguish inactive from active forms with an accuracy of 95·7%, and this novel mimotope may also be used as an auxiliary tool to neuroimaging tests and treatment follow‐up.
Acetylcholinesterase-catalyzed hydrolysis allows ultrasensitive detection of pathogens with the naked eye.

Major State Basic Research Development Program of China (973 Program) [2013CB733802, 2014CB744503]; National Science Foundation of China (NSFC) [81101101, 51273165, 81201086, 81201190, 81371596]; Key Project of the Chinese Ministry of Education [212149]; Intramural Research Program (IRP) of the National Institute of Biomedical Imaging and Bioengineering (NIBIB); National Institutes of Health (NIH); NIH-NIBIB/NIST NRC
Ophiopogonin D alleviates high‐fat diet‐induced metabolic syndrome and changes the structure of gut microbiota in mice

Gut dysbiosis is believed to play a critical role in the pathogenesis of metabolic diseases, including obesity. Ophiopogonin D (OP‐D), a steroidal glycoside and an active component extracted from the root of the plant Ophiopogon japonicus (also known as radix ophiopogonis), can regulate multiple physiologic processes. Here we show that OP‐D administration reduces body weight, hyperglycemia, hyperlipidemia, and insulin resistance in male mice fed a high‐fat diet (HFD). Pyrosequencing of the V4 regions of 16S rRNA genes in mouse feces revealed a deviation of the gut microbiota in response to OP‐D treatment. In particular, the decreased Firmicutes‐to‐Bacteroidetes ratios and endotoxin‐bearing Proteobacteria levels indicated that OP‐D reversed HFD‐induced gut dysbiosis. More importantly, the effects of OP‐D on modulation of obesity and microbiota were transferable via horizontal feces transfer from OP‐D‐treated mice to HFD‐fed mice. Taken together, our results suggest that OP‐D may be used as a prebiotic agent to treat obesity‐associated gut dysbiosis and metabolic syndrome. —Chen, S. , Li, X. , Liu, L. , Liu, C. , Han, X. Ophiopogonin D alleviates high‐fat diet‐induced metabolic syndrome and changes the structure of gut microbiota in mice. FASEB J. 32,1139‐1153 (2018). www.fasebj. org
The Transcription of FOXO Genes Is Stimulated by FOXO3 and Repressed by Growth Factors*

FOXO (Forkhead box O) transcription factors induce cell growth arrest and apoptosis, which can be prevented by FOXO phosphorylation by AKT in response to growth factors such as platelet-derived growth factors (PDGF) and insulin-like growth factor I (IGF-I). In addition to this well characterized post-translational modification, we showed that FOXO1, FOXO3, and FOXO4 were also regulated at the transcriptional level. PDGF, fibroblast growth factors (FGF), and IGF-I repressed the expression of FOXO genes in human fibroblasts. This process was sensitive to phosphatidylinositol 3-kinase inhibition by LY294002. FOXO1-specific shRNA decreased FOXO1 mRNA expression and enhanced fibroblast proliferation, mimicking the effects of growth factors. Conversely, ectopic FOXO3 activation blocked the proliferation of fibroblasts and induced the expression of FOXO1, FOXO4, and p27-KIP1. Using luciferase reporter assays and chromatin immunoprecipitations, we identified a conserved FOXO-binding site in the promoter of the FOXO1 gene, which was required for regulation by PDGF, and mediated the up-regulation of FOXO1 by itself and by FOXO3. Altogether, our results suggest that the expression of FOXO1 and FOXO4 genes is stimulated by FOXO3 and possibly by other FOXO factors in a positive feedback loop, which is disrupted by growth factors.
Inhibitor of serine peptidase 2 enhances Leishmania major survival in the skin through control of monocytes and monocyte-derived cells

Leishmania major is the causative agent of the neglected tropical disease, cutaneous leishmaniasis. In the mouse, protective immunity to Leishmania is associated with inflammatory responses. Here, we assess the dynamics of the inflammatory responses at the lesion site during experimental long‐term, low‐dose intradermal infection of the ear, employing noninvasive imaging and genetically modified L. major. Significant infiltrates of neutrophils and monocytes occurred at 1‐4 d and 2‐4 wk, whereas dermal macrophage and dendritic cell (DC) numbers were only slightly elevated in the first days. Quantitative whole‐body bioluminescence imaging of myeloperoxidase activity and the quantification of parasite loads indicated that the Leishmania virulence factor, inhibitor of serine peptidase 2 (ISP2), is required to modulate phagocyte activation and is important for parasite survival at the infection site. ISP2 played a role in the control of monocyte, monocyte‐derived macrophage, and monocyte‐derived DC (moDC) influx, and was required to reduce iNOS expression in monocytes, monocyte‐derived cells, and dermal DCs; the expression of CD80 in moDCs; and levels of IFN‐γ in situ. Our findings indicate that the increased survival of L. major in the dermis during acute infection is associated with the down‐regulation of inflammatory monocytes and monocyte‐derived cells via ISP2. —Goundry, A. , Romano, A. , Lima, A. P. C. A. , Mottram, J. C. , Myburgh, E. Inhibitor of serine peptidase 2 enhances Leishmania major survival in the skin through control of monocytes and monocyte‐derived cells. FASEB J. 32,1315‐1327 (2018). www.fasebj. org
Critical determinants of combined sprint and endurance performance: an integrative analysis from muscle fiber to the human body

Optimizing physical performance is a major goal in current physiology. However, basic understanding of combining high sprint and endurance performance is currently lacking. This study identifies critical determinants of combined sprint and endurance performance using multiple regression analyses of physiologic determinants at different biologic levels. Cyclists, including 6 international sprint, 8 team pursuit, and 14 road cyclists, completed a Wingate test and 15‐km time trial to obtain sprint and endurance performance results, respectively. Performance was normalized to lean body mass2/3 to eliminate the influence of body size. Performance determinants were obtained from whole‐body oxygen consumption, blood sampling, knee‐extensor maximal force, muscle oxygenation, whole‐muscle morphology, and muscle fiber histochemistry of musculus vastus lateralis. Normalized sprint performance was explained by percentage of fast‐type fibers and muscle volume (R2 = 0.65; P < 0.001) and normalized endurance performance by performance oxygen consumption (Vo2), mean corpuscular hemoglobin concentration, and muscle oxygenation (R2 = 0.92; P < 0.001). Combined sprint and endurance performance was explained by gross efficiency, performance Vo2 and likely by muscle volume and fascicle length (P = 0.056; P = 0.059). High performance Vo2 related to a high oxidative capacity, high capillarization x myoglobin, and small physiologic cross‐sectional area (R2 = 0.67; P < 0.001). Results suggest that fascicle length and capillarization are important targets for training to optimize sprint and endurance performance simultaneously. — Van der Zwaard, S. , van derLaarse, W. J., Weide, G. , Bloemers, F. W. , Hofmijster, M. J., Levels, K. , Noordhof, D. A. , de Koning, J. J., de Ruiter, C. J., Jaspers, R. T. Critical determinants of combined sprint and endurance performance: an integrative analysis from muscle fiber to the human body. FASEB J. 32, 2110–2123 (2018). www.fasebj. org
LGR4 modulates breast cancer initiation, metastasis, and cancer stem cells

The fourth member of the leucine‐rich repeat‐containing GPCR family (LGR4, frequently referred to as GPR48) and its cognate ligands, R‐spondins (RSPOs) play crucial roles in the development of multiple organs as well as the survival of adult stem cells by activation of canonical Wnt signaling. Wnt/β‐catenin signaling acts to regulate breast cancer; however, the molecular mechanisms determining its spatiotemporal regulation are largely unknown. In this study, we identified LGR4 as a master controller of Wnt/β‐catenin signaling‐mediated breast cancer tumorigenesis, metastasis, and cancer stem cell (CSC) maintenance. LGR4 expression in breast tumors correlated with poor prognosis. Either Lgr4 haploinsufficiency or mammary‐specific deletion inhibited mouse mammary tumor virus (MMTV)‐PyMT‐ and MMTV‐Wnt1‐driven mammary tumorigenesis and metastasis. Moreover, LGR4 down‐regulation decreased in vitro migration and in vivo xenograft tumor growth and lung metastasis. Furthermore, Lgr4 deletion in MMTV‐Wnt1 tumor cells or knockdown in human breast cancer cells decreased the number of functional CSCs by ~90%. Canonical Wnt signaling was impaired in LGR4‐deficient breast cancer cells, and LGR4 knockdown resulted in increased E‐cadherin and decreased expression of N‐cadherin and snail transcription factor ‐2 (SNAI2) (also called SLUG), implicating LGR4 in regulation of epithelial–mesenchymal transition. Our findings support a crucial role of the Wnt signaling component LGR4 in breast cancer initiation, metastasis, and breast CSCs. —Yue, Z. , Yuan, Z. , Zeng, L. , Wang, Y. , Lai, L. , Li, J., Sun, P., Xue, X. , Qi, J., Yang, Z. , Zheng, Y. , Fang, Y. , Li, D. , Siwko, S. , Li, Y. , Luo, J., Liu, M. LGR4 modulates breast cancer initiation, metastasis, and cancer stem cells. FASEB J. 32, 2422–2437 (2018). www.fasebj. org
Biochemical and Structural Analysis of Bacterial O-antigen Chain Length Regulator Proteins Reveals a Conserved Quaternary Structure*

Lipopolysaccharide (LPS) is a major component of the Gram-negative outer membrane and is an important virulence determinant. The O-antigen polysaccharide of the LPS molecule provides protection from host defenses, and the length of O-antigen chains plays a pivotal role. In the Wzy-dependent O-antigen biosynthesis pathway, the integral inner membrane protein Wzz determines the O-antigen chain length. How these proteins function is currently unknown, but the hypothesis includes activities such as a “molecular ruler” or a “molecular stopwatch,” and other possibilities may exist. Wzz homologs are membrane proteins with two transmembrane helices that flank a large periplasmic domain. Recent x-ray crystallographic studies of the periplasmic portions of Wzz proteins found multiple oligomeric forms, with quaternary structures favoring the “molecular ruler” interpretation. Here, we have studied full-length Wzz proteins with the transmembrane portions embedded in lipid membranes. Using electron microscopy and image analysis we find a unique hexameric state rather than differing oligomeric forms. The data suggest that in vivo Wzz proteins determine O-antigen chain length via subtle structure-function relationships at the level of primary, secondary, or tertiary structure within the context of a hexameric complex.
The Kaposi's Sarcoma-associated Herpesvirus-encoded vIRF-3 Inhibits Cellular IRF-5*

Kaposi's sarcoma-associated herpesvirus encodes four genes with homology to the family of interferon regulatory factors (IRFs). At least one of these viral IRFs, vIRF-3, is expressed in latently Kaposi's sarcoma-associated herpesvirus-infected primary effusion lymphoma (PEL) cells and is essential for the survival of PEL cells. We now report that vIRF-3 interacts with cellular IRF-5, thereby inhibiting binding of IRF-5 to interferon-responsive promoter elements. Consequently, vIRF-3 blocked IRF-5-mediated promoter activation. A central double helix motif present in vIRF-3 was sufficient to abrogate both DNA binding and transcriptional transactivation by IRF-5. Upon DNA damage or activation of the interferon or Toll-like receptor pathways, cytoplasmic IRF-5 has been reported to be translocated to the nucleus, which results in induction of both p53-independent apoptosis and p21-mediated cell cycle arrest. We report here that IRF-5 is present in the nuclei of PEL cells without interferon stimulation. Silencing of vIRF-3 expression in PEL cells was accompanied by increased sensitivity to interferon-mediated apoptosis and up-regulation of IRF-5 target genes. In addition, vIRF-3 antagonized IRF-5-mediated activation of the p21 promoter. The data presented here indicate that vIRF-3 contributes to immune evasion and sustained proliferation of PEL cells by releasing IRF-5 from transcription complexes.
Mettl14 is required for mouse post-implantation development by facilitating epiblast maturation

N6-methyladenosine (m6A) is the most prevalent and reversible internal modification of mammalian messenger and noncoding RNAs mediated by specific m6A writer, reader, and eraser proteins. As an m6A writer, the METTL3–METTL14-WTAP complex dynamically regulates m6A modification and plays important roles in diverse biological processes. However, our knowledge about the complete functions of this RNA methyltransferase complex, the contributions of each component to the methylation and their impacts on different biological pathways, are still very limited. Here, by employing both in vivo and in vitro models, we report that METTL14 was indispensable for post-implantation embryonic development by facilitating the conversion from naïve to primed state of the epiblast. Depletion of Mettl14 lead to conspicuous embryonic growth retardation from E6.5 mainly as a result of resistance to differentiation, which further lead to embryonic lethality early in gestation. Our data highlight the critical function of METTL14, as an m6A modification regulator, in orchestrating early mouse embryogenesis.
Juvenile hormone–dependent Kazal‐type serine protease inhibitor Greglin safeguards insect vitellogenesis and egg production

In addition to preventing insect metamorphosis, juvenile hormone (JH) is known to stimulate aspects of insect reproduction. However, the molecular mechanisms of JH action in insect reproduction remain largely unknown. By reanalyzing the transcriptomic data from adults and other developmental stages of the migratory locust Locusta migratoria, we identified a gene coding for Kazal‐type protease inhibitor, previously named Greglin. Greglin is specifically expressed in adult females and most abundant in the fat body and ovaries. Interestingly, Greglin is among the top 3 of highly expressed genes in adult female locusts, after 2 vitellogenin (Vg) genes. Greglin is induced by JH and expressed at remarkably high levels in the vitellogenic stage. Knockdown of Greglin in adult female locusts results in accelerated degradation of serine protease substrate and significantly reduced levels of Greglin protein in hemolymph and ovaries. The consequent phenotypes include blocked oocyte maturation, arrested ovarian growth and shrunken follicular epithelium, as well as declines in egg number and hatchability. The data provide the first evidence, to our knowledge, that JH‐dependent Greglin is involved in locust vitellogenesis and oocyte maturation likely by protecting vitellogenesis and other forms of yolk precursors from proteolysis. The result offers new insights into the regulation of JH and function of protease inhibitors in insect vitellogenesis, oocyte maturation and fecundity. —Guo, W. , Wu, Z. , Yang, L. , Cai, Z. , Zhao, L. , Zhou, S. Juvenile hormone–dependent Kazal‐type serine protease inhibitor Greglin safeguards insect vitellogenesis and egg production. FASEB J. 33, 917–927 (2019). www.fasebj. org