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Abo-Ogiala, Atef, Caroline Carsjens, Heike Diekmann, Payam Fayyaz, Cornelia Herrfurth, Ivo Feussner, and Andrea Polle. "Temperature-induced lipocalin (TIL) is translocated under salt stress and protects chloroplasts from ion toxicity." Journal of Plant Physiology 171, no. 3-4 (February 2014): 250–59. http://dx.doi.org/10.1016/j.jplph.2013.08.003.
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Miguel, María Carolina, Marisa Ethel Giménez, and Alberto Ramón Meder. "Neutrophil gelatinase-associated lipocalin (NGAL) Biomarcador de injuria renal aguda en perros / Neutrophil gelatinase-associated lipocalin (NGAL): Biomarker of acute kidney injury in dogs." Brazilian Journal of Animal and Environmental Research 4, no. 2 (June 14, 2021): 2490–503. http://dx.doi.org/10.34188/bjaerv4n2-076.
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La evaluación de la función renal se ha realizado históricamente en base a la medición de la tasa de filtración glomerular, la cual fue reemplazada, posteriormente, por la determinación de creatinina sérica debido a su amplia disponibilidad y accesibildad. Sin embargo, en los trabajos actuales, cuando es comparada con el aumento de los biomarcadores de injuria renal aguda (IRA), se observa que la elevación desde su valor basal ante una lesión renal repentina sucede horas más tarde que el Neutrophil Gelatinase-Associated Lipocalin (NGAL) urinario. El objetivo de la presente revisión sobre NGAL, y sus alcances como marcador biológico temprano de injuria renal aguda, es conocer su validación como tal y su aplicabilidad en la práctica médica de pacientes caninos, ya que la identificación temprana de un proceso que altere la funcionalidad y viabilidad de los riñones, permite intervenir de manera terapéutica y anticiparse a un grado irreversible de lesión renal. El método utilizado para la revisión fue consultar revistas científicas internacionales y, sobre estas, seleccionar trabajos de la última década que demuestran resultados estadísticamente significativos sobre el aumento de NGAL entre dos (2) y doce (12) horas antes que la creatinina. Si bien los datos aportados por los trabajos de investigación son claros, aún queda un camino por recorrer en relación a los tipos de injuria renal aguda y sus diferentes causas. NGAL, como biomarcador renal, presente un espectro favorable en cuanto a utilidad clínica, dada la accesibilidad a los métodos complementarios que permiten valorarlo en medicina veterinaria.
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Mogielnicka-Brzozowska, Marzena, Mariola Słowińska, Leyland Fraser, Paweł Wysocki, Rafał Strzeżek, Karolina Wasilewska, and Władysław Kordan. "Identification and characterization of non-phosphorylcholine-binding and phosphorylcholine-binding proteins of canine seminal plasma." Annals of Animal Science 17, no. 3 (July 26, 2017): 771–86. http://dx.doi.org/10.1515/aoas-2017-0005.
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Abstract Seminal plasma (SP) proteins participate in the process of fertilization by binding to the sperm membrane, particularly to the phosphorylcholine-containing lipids. This study aimed to identify and characterize non-phosphorylcholine-binding and phosphorylcholine-binding proteins (nPch- BPs and PchBPs, respectively) of canine SP. The nPchBPs and PchBPs were isolated from canine SP by affinity chromatography. Electrophoretic studies revealed that the nPchBPs and PchBPs occurred in their native state as high-molecular-weight aggregates. Immunofluorescent staining showed preferential binding of nPchBPs to the sperm acrosome membrane, whereas PchBPs coating was uniformly distributed on the sperm post-acrosomal membrane, mid-piece and tail regions. Analysis with mass spectrometry confirmed that canine prostate specific esterase (CPSE) is a component of the nPchBPs and PchBPs, which is implicated in key mechanisms of protein-coating on the sperm plasma membrane surface. In addition, proteins of known binding properties such as prostaglandin-H2 D-isomerase and lipocalin-like 1 protein, identified in canine SP, might have a specific role in the fertilization-associated processes.
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Mizurini, Daniella M., Ivo M. B. Francischetti, John F. Andersen, and Robson Q. Monteiro. "Nitrophorin 2, a factor IX(a)-directed anticoagulant, inhibits arterial thrombosis without impairing haemostasis." Thrombosis and Haemostasis 104, no. 12 (2010): 1116–23. http://dx.doi.org/10.1160/th10-03-0186.
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SummaryNitrophorin 2 (NP2) is a 20 kDa lipocalin identified in the salivary gland of the blood sucking insect, Rhodnius prolixus. It functions as a potent inhibitor of the intrinsic pathway of coagulation upon binding to factor IX (FIX) or FIXa. Herein we have investigated the in vivo antithrombotic properties of NP2. Surface plasmon resonance assays demonstrated that NP2 binds to rat FIX and FIXa with high affinities (KD = 43 and 47 nM, respectively), and prolongs the aPTT without affecting the PT. In order to evaluate NP2 antithrombotic effects in vivo two distinct models of thrombosis in rats were carried out. In the rose Bengal/laser induced injury model of arterial thrombosis, NP2 increased the carotid artery occlusion time by ≈35 and ≈155%, at doses of 8 and 80 μg/kg, respectively. NP2 also inhibited thrombus formation in an arterio-venous shunt model, showing ≈60% reduction at 400 μg/kg (i.v. administration). The antithrombotic effect lasted for up to 48 hours after a single i.v. dose. Notably, effective doses of NP2 did not increase the blood loss as evaluated by tail-transection model. In conclusion, NP2 is a potent and long-lasting inhibitor of arterial thrombosis with minor effects on haemostasis. It might be regarded as a potential agent for the treatment of human cardiovascular diseases.
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Oleaga, Ana, Beatriz Soriano, Carlos Llorens, and Ricardo Pérez-Sánchez. "Sialotranscriptomics of the argasid tick Ornithodoros moubata along the trophogonic cycle." PLOS Neglected Tropical Diseases 15, no. 2 (February 5, 2021): e0009105. http://dx.doi.org/10.1371/journal.pntd.0009105.
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The argasid tick Ornithodoros moubata is the main vector of human relapsing fever (HRF) and African swine fever (ASF) in Africa. Salivary proteins are part of the host-tick interface and play vital roles in the tick feeding process and the host infection by tick-borne pathogens; they represent interesting targets for immune interventions aimed at tick control. The present work describes the transcriptome profile of salivary glands of O. moubata and assesses the gene expression dynamics along the trophogonic cycle using Illumina sequencing. De novo transcriptome assembling resulted in 71,194 transcript clusters and 41,011 annotated transcripts, which represent 57.6% of the annotation success. Most salivary gene expression takes place during the first 7 days after feeding (6,287 upregulated transcripts), while a minority of genes (203 upregulated transcripts) are differentially expressed between 7 and 14 days after feeding. The functional protein groups more abundantly overrepresented after blood feeding were lipocalins, proteases (especially metalloproteases), protease inhibitors including the Kunitz/BPTI-family, proteins with phospholipase A2 activity, acid tail proteins, basic tail proteins, vitellogenins, the 7DB family and proteins involved in tick immunity and defence. The complexity and functional redundancy observed in the sialotranscriptome of O. moubata are comparable to those of the sialomes of other argasid and ixodid ticks. This transcriptome provides a valuable reference database for ongoing proteomics studies of the salivary glands and saliva of O. moubata aimed at confirming and expanding previous data on the O. moubata sialoproteome.
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Vicentini, F., L. Griffin, C. Keenan, J. Cavin, K. Nieves, S. A. Hirota, and K. A. Sharkey. "A37 ENTERIC MICROBIOTA CONTRIBUTE TO BEHAVIORAL ALTERATIONS OBSERVED IN MICE WITH COLITIS." Journal of the Canadian Association of Gastroenterology 3, Supplement_1 (February 2020): 44–45. http://dx.doi.org/10.1093/jcag/gwz047.036.
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Abstract Background The enteric microbiota has been recognized as an essential regulator of both gut and brain physiology, a complex interaction generally termed the microbiota-gut-brain axis. Disturbances to gastrointestinal physiology lead to alterations in the composition of the enteric microbiota, whereas dysbiosis can also contribute to pathophysiology. Inflammatory bowel diseases (IBD) are chronic, relapsing inflammatory conditions of the gastrointestinal tract, associated with microbial dysbiosis. Interestingly, IBD patients exhibit an increased incidence of mental illness (i.e. anxiety and depression), often termed “sickness behavior”, even during the remitting phase of their disease. Nevertheless, it is unclear if alterations in the enteric microbiota associated with IBD are responsible for the observed modification in brain function and behavior. Here, we hypothesized that sickness behavior is driven by alterations in microbial composition, which occur in the context of intestinal inflammation. Aims We sought to determine whether transfer of the microbiota from colitic mice, exhibiting sickness behaviour, into healthy counterparts would induce behavioral changes. Methods Male mice (C57Bl/6J; 8 weeks old) were used in all experiments. Colitis was induced by administration of 2.5% dextran sodium sulfate (DSS) in the drinking water for 5 days. Colonic inflammation was assessed by measuring fecal lipocalin-2 and the expression of pro-inflammatory mediators via qPCR. Cecal matter from donor mice (control or DSS treated) were collected for fecal microbiota transplant (FMT). FMT was performed via oral gavage in antibiotic-treated recipient mice. Gut bacteria were evaluated by 16S rRNA sequencing in cecal samples. Anxiety- and depression-like behavior were assessed by elevated plus maze and tail suspension test, respectively. Results DSS-treated mice exhibited clinical disease, reflected by body weight loss, increased fecal lipocalin-2 and elevated colonic pro-inflammatory cytokine transcripts. An increase in anxiety-like behavior was observed in mice with colitis, although no alterations in depression-like behavior were detected. Colitic mice exhibited a unique microbial community. Transferring cecal material from colitic mice into recipient, antibiotic-treated mice, recapitulated alterations in behavior seen in colitic donors, as shown by increased anxiety-like behavior and unexpectedly, increased depression-like behavior. These behavioral changes occurred in the absence of colonic or brain inflammation in the recipient mice, but were associated with changes in stress-related gene expression (i.e. Crh). Conclusions Colitis-associated sickness behavior can be transmitted to antibiotic-treated recipient mice via FMT, which occurs in the absence of overt intestinal or brain inflammation. Funding Agencies CIHRNational Council for Scientific and Technological Development (CNPq-Brazil)
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Mary, Sheon, Philipp Boder, Giacomo Rossitto, Lesley Graham, Kayley Scott, Arun Flynn, David Kipgen, Delyth Graham, and Christian Delles. "Salt loading decreases urinary excretion and increases intracellular accumulation of uromodulin in stroke-prone spontaneously hypertensive rats." Clinical Science 135, no. 24 (December 2021): 2749–61. http://dx.doi.org/10.1042/cs20211017.
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Abstract Uromodulin (UMOD) is the most abundant renal protein secreted into urine by the thick ascending limb (TAL) epithelial cells of the loop of Henle. Genetic studies have demonstrated an association between UMOD risk variants and hypertension. We aimed to dissect the role of dietary salt in renal UMOD excretion in normotension and chronic hypertension. Normotensive Wistar–Kyoto rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) (n=8/sex/strain) were maintained on 1% NaCl for 3 weeks. A subset of salt-loaded SHRSP was treated with nifedipine. Salt-loading in SHRSP increased blood pressure (ΔSBP 35 ± 5 mmHg, P<0.0001) and kidney injury markers such as kidney injury marker-1 (KIM-1; fold change, FC 3.4; P=0.003), neutrophil gelatinase-associated lipocalin (NGAL; FC, 2.0; P=0.012) and proteinuria. After salt-loading there was a reduction in urinary UMOD excretion in WKY and SHRSP by 26 and 55% respectively, compared with baseline. Nifedipine treatment reduced blood pressure (BP) in SHRSP, however, did not prevent salt-induced reduction in urinary UMOD excretion. In all experiments, changes in urinary UMOD excretion were dissociated from kidney UMOD protein and mRNA levels. Colocalization and ex-vivo studies showed that salt-loading increased intracellular UMOD retention in both WKY and SHRSP. Our study provides novel insights into the interplay among salt, UMOD, and BP. The role of UMOD as a cardiovascular risk marker deserves mechanistic reappraisal and further investigations based on our findings.
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Luo, Renfei, Kevin Yang, Fei Wang, Chuanming Xu, and Tianxin Yang. "(Pro)renin receptor decoy peptide PRO20 protects against adriamycin-induced nephropathy by targeting the intrarenal renin-angiotensin system." American Journal of Physiology-Renal Physiology 319, no. 5 (November 1, 2020): F930—F940. http://dx.doi.org/10.1152/ajprenal.00279.2020.
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Adriamycin (ADR) administration in susceptible rodents such as the BALB/c mouse strain produces injury to the glomerulus mimicking human chronic kidney disease due to primary focal segmental glomerulosclerosis. The goal of the present study was to use this model to investigate antiproteinuric actions of the (pro)renin receptor decoy inhibitor PRO20. BALB/c mice were pretreated for 1 day with PRO20 at 500 μg·kg−1·day−1 via an osmotic minipump followed by a single injection of vehicle or ADR (10 mg/kg) via the tail vein. Albuminuria and renal function were analyzed at the fourth week post-ADR administration. ADR-treated mice exhibited severe proteinuria, hypoalbuminemia and hyperlipidemia, glomerulosclerosis, podocyte loss, tubulointerstitial fibrosis, and oxidative stress, accompanied by elevated urinary neutrophil gelatinase-associated lipocalin and kidney injury molecule-1, all of which were significantly attenuated by PRO20. Urinary and renal renin activity and angiotensin II were elevated by ADR and suppressed by PRO20. In parallel, urinary and renal H2O2 levels and renal NADPH oxidase 4 (Nox4) and transient receptor potential channel C6 (TRPC6) expression in response to ADR were all similarly suppressed. Taken together, the results of the present study provide the first evidence that PRO20 can protect against podocyte damage and interstitial fibrosis in ADR nephropathy by preventing activation of the intrarenal renin-angiotensin system and upregulation of Nox4 and TRPC6 expression. PRO20 may have a potential application in the treatment of ADR nephropathy.
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Li, Yuanyuan, Weiwei Xia, Mengying Wu, Jie Yin, Qian Wang, Shuzhen Li, Aihua Zhang, Songming Huang, Yue Zhang, and Zhanjun Jia. "Activation of GSDMD contributes to acute kidney injury induced by cisplatin." American Journal of Physiology-Renal Physiology 318, no. 1 (January 1, 2020): F96—F106. http://dx.doi.org/10.1152/ajprenal.00351.2019.
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Cisplatin is one of the most effective antitumor agents, but its clinical use is highly limited by its severe side effects, especially nephrotoxicity. Recently, the active form of gasdermin D (GSDMD), termed GSDMD-N, was identified to mediate pyroptotic inflammatory cell death in several diseases. However, the role of the GSDMD-N fragment in cisplatin-induced acute kidney injury (AKI) remains unclear. In the present study, we found that pyroptosis was induced by cisplatin in both mouse kidney tissues and renal tubular epithelial cells, accompanied by increased expression of the GSDMD-N fragment. In GSDMD knockout mice with cisplatin-induced AKI, we found that cisplatin-induced loss of renal function, renal tubular injury, and inflammation was significantly attenuated compared with wild-type mice. Furthermore, the GSDMD-N fragment was overexpressed by an established rapid plasmid tail vein injection approach to evaluate the role of this cleaved form of GSDMD in AKI. As expected, mice with GSDMD-N fragment overexpression in the kidney were more susceptible to cisplatin-induced AKI than control mice, as evidenced by further elevated serum levels of blood urea nitrogen and creatinine, aggravated renal pathology, increased expression of neutrophil gelatinase-associated lipocalin and kidney injury molecule-1, and enhanced renal inflammatory cytokine secretion, which indicates a pathogenic role of GSDMD-N in cisplatin-induced AKI by triggering cell pyroptosis. Similar results were also observed in renal tubular epithelial cells overexpressing the GSDMD-N fragment. Thus these findings suggested that the activation of GSDMD contributes to cisplatin-induced AKI, possibly through triggering pyroptosis.
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Stepanenko, Olga V., Denis O. Roginskii, Olesya V. Stepanenko, Irina M. Kuznetsova, Vladimir N. Uversky, and Konstantin K. Turoverov. "Structure and stability of recombinant bovine odorant-binding protein: III. Peculiarities of the wild type bOBP unfolding in crowded milieu." PeerJ 4 (April 18, 2016): e1642. http://dx.doi.org/10.7717/peerj.1642.
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Contrary to the majority of the members of the lipocalin family, which are stable monomers with the specific OBP fold (a β-barrel consisting of a 8-stranded anti-parallel β-sheet followed by a short α-helical segment, a ninth β-strand, and a disordered C-terminal tail) and a conserved disulfide bond, bovine odorant-binding protein (bOBP) does not have such a disulfide bond and forms a domain-swapped dimer that involves crossing the α-helical region from each monomer over the β-barrel of the other monomer. Furthermore, although natural bOBP isolated from bovine tissues exists as a stable domain-swapped dimer, recombinant bOBP has decreased dimerization potential and therefore exists as a mixture of monomeric and dimeric variants. In this article, we investigated the effect model crowding agents of similar chemical nature but different molecular mass on conformational stability of the recombinant bOBP. These experiments were conducted in order to shed light on the potential influence of model crowded environment on the unfolding-refolding equilibrium. To this end, we looked at the influence of PEG-600, PEG-4000, and PEG-12000 in concentrations of 80, 150, and 300 mg/mL on the equilibrium unfolding and refolding transitions induced in the recombinant bOBP by guanidine hydrochloride. We are showing here that the effect of crowding agents on the structure and conformational stability of the recombinant bOBP depends on the size of the crowder, with the smaller crowding agents being more effective in the stabilization of the bOBP native dimeric state against the guanidine hydrochloride denaturing action. This effect of the crowding agents is concentration dependent, with the high concentrations of the agents being more effective.
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Knipp, Markus, Hideaki Ogata, Giancarlo Soavi, Giulio Cerullo, Alessandro Allegri, Stefania Abbruzzetti, Stefano Bruno, Cristiano Viappiani, Axel Bidon-Chanal, and F. Javier Luque. "Structure and dynamics of the membrane attaching nitric oxide transporter nitrophorin 7." F1000Research 4 (February 13, 2015): 45. http://dx.doi.org/10.12688/f1000research.6060.1.
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Nitrophorins represent a unique class of heme proteins that are able to perform the delicate transportation and release of the free-radical gaseous messenger nitric oxide (NO) in a pH-triggered manner. Besides its ability to bind to phospholipid membranes, the N-terminus contains an additional Leu-Pro-Gly stretch, which is a unique sequence trait, and the heme cavity is significantly altered with respect to other nitrophorins. These distinctive features encouraged us to solve the X-ray crystallographic structures of NP7 at low and high pH and bound with different heme ligands (nitric oxide, histamine, imidazole). The overall fold of the lipocalin motif is well preserved in the different X-ray structures and resembles the fold of other nitrophorins. However, a chain-like arrangement in the crystal lattice due to a number of head-to-tail electrostatic stabilizing interactions is found in NP7. Furthermore, the X-ray structures also reveal ligand-dependent changes in the orientation of the heme, as well as in specific interactions between the A-B and G-H loops, which are considered to be relevant for the biological function of nitrophorins. Fast and ultrafast laser triggered ligand rebinding experiments demonstrate the pH-dependent ligand migration within the cavities and the exit route. Finally, the topological distribution of pockets located around the heme as well as from inner cavities present at the rear of the protein provides a distinctive feature in NP7, so that while a loop gated exit mechanism to the solvent has been proposed for most nitrophorins, a more complex mechanism that involves several interconnected gas hosting cavities is proposed for NP7.
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Knipp, Markus, Hideaki Ogata, Giancarlo Soavi, Giulio Cerullo, Alessandro Allegri, Stefania Abbruzzetti, Stefano Bruno, Cristiano Viappiani, Axel Bidon-Chanal, and F. Javier Luque. "Structure and dynamics of the membrane attaching nitric oxide transporter nitrophorin 7." F1000Research 4 (August 18, 2015): 45. http://dx.doi.org/10.12688/f1000research.6060.2.
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Nitrophorins represent a unique class of heme proteins that are able to perform the delicate transportation and release of the free-radical gaseous messenger nitric oxide (NO) in a pH-triggered manner. Besides its ability to bind to phospholipid membranes, the N-terminus of NP7, a member of the NO transporter nitrophorin family, contains an additional Leu-Pro-Gly stretch, which is a unique sequence trait, and the heme cavity is significantly altered with respect to other nitrophorins. These distinctive features encouraged us to solve the X-ray crystallographic structures of NP7 at low and high pH and bound with different heme ligands (nitric oxide, histamine, imidazole). The overall fold of the lipocalin motif is well preserved in the different X-ray structures and resembles the fold of other nitrophorins. However, a chain-like arrangement in the crystal lattice due to a number of head-to-tail electrostatic stabilizing interactions is found in NP7. Furthermore, the X-ray structures also reveal ligand-dependent changes in the orientation of the heme, as well as in specific interactions between the A-B and G-H loops, which are considered to be relevant for the biological function of nitrophorins. Fast and ultrafast laser triggered ligand rebinding experiments demonstrate the pH-dependent ligand migration within the cavities and the exit route. Finally, the topological distribution of pockets located around the heme as well as from inner cavities present at the rear of the protein provides a distinctive feature in NP7, so that while a loop gated exit mechanism to the solvent has been proposed for most nitrophorins, a more complex mechanism that involves several interconnected gas hosting cavities is proposed for NP7.
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Bota, Daniela A., David E. Piccioni, Christopher M. Duma, Renato V. LaRoca, Santosh Kesari, Mehrdad Abedi, Robert D. Aiken, Aleksandra J. Poole, Gabriel I. Nistor, and Robert O. Dillman. "Abstract CT571: Plasma proteomic markers prognostic or predictive for survival of patients with newly diagnosed glioblastoma who were treated in a phase II trial with standard care and the addition of the novel patient-specific dendritic cell vaccine AV-GBM-1." Cancer Research 82, no. 12_Supplement (June 15, 2022): CT571. http://dx.doi.org/10.1158/1538-7445.am2022-ct571.
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Abstract Standard aggressive therapy for primary newly diagnosed glioblastoma (GBM), which includes surgical resection followed by concurrent radiation therapy and temozolomide chemotherapy (RT/TMZ) and then maintenance TMZ, is associated with poor survival rates. Adding treatment with AV-GBM-1, a personal vaccine consisting of autologous dendritic cells (DC) pulsed with autologous tumor antigens (ATA) may improve survival. AV-GBM-1 is produced by incubating ATA with autologous DC. ATA are from a lysate of irradiated autologous GBM cells that were self-renewing in serum-free medium with factors that favor survival and proliferation of tumor initiating cells, i.e., stem cells and early progenitor cells. After recovery from RT/TMZ, GBM patients were injected subcutaneously with AV-GBM-1 admixed in granulocyte-macrophage colony-stimulating factor (GM-CSF) at weeks 1, 2, 3, 8, 12, 16, 20, and 24. This study examined changes in plasma proteomics before and after injections of AV-GBM-1. 57 patients were treated during November 2018 to October 2020. Median progression-free and overall survival were 10.3 and 16.0 months respectively. Plasma samples obtained at baseline (week-0), and just prior to the third injection (week-2) were cryopreserved and subsequently analyzed for 448 proteomic markers using quantitative, multiplex enzyme-linked immunosorbent assays (Raybiotech, Inc., Norcross, GA.). 54 patients had a baseline week-0 sample prior to the 1st vaccination; 50 had a week-2 sample just prior to the 3rd dose; 49 had paired samples for both time-points. The averages of paired samples were measured for all patients and for cohorts defined by median survival that were compared by 2-tail T-Tests. After two weekly injections there were significant (p<0.01) increases in thymus and activation-regulated chemokine (TARC, CCL17), chemerin, lipocalin-2 and angiopoietin, and decreases in thrombospondin-5, angiotensinogen, and beta-fibroblast growth factor. The changes in TARC are due to GM-CSF. 25 patients survived more than 15 months and 24 less than 15 months. Prognostic baseline markers associated with longer survival were higher CD14 (p=0.009), higher tissue-inhibitor of metalloproteinases (TIMP-1), and higher insulin growth factor binding proteins (IGFBP) 1 and 2 (p<0.02). The only markers that appeared predictive for survival (p<0.05) based on changes from baseline to week-2 were increased lipocalin-2, associated with longer survival (p=0.0369), and decreased brain-derived neurotrophic factor (BDNF) associated with shorter survival (p=0.0186). Because of the large number of analyses, these results are considered hypothesis generating and not definitive. Citation Format: Daniela A. Bota, David E. Piccioni, Christopher M. Duma, Renato V. LaRoca, Santosh Kesari, Mehrdad Abedi, Robert D. Aiken, Aleksandra J. Poole, Gabriel I. Nistor, Robert O. Dillman. Plasma proteomic markers prognostic or predictive for survival of patients with newly diagnosed glioblastoma who were treated in a phase II trial with standard care and the addition of the novel patient-specific dendritic cell vaccine AV-GBM-1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT571.
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Kao, Ting-Hsien, Yi-Jen Peng, Hsi-Kai Tsou, Donald M. Salter, and Herng-Sheng Lee. "Nerve growth factor promotes expression of novel genes in intervertebral disc cells that regulate tissue degradation." Journal of Neurosurgery: Spine 21, no. 4 (October 2014): 653–61. http://dx.doi.org/10.3171/2014.6.spine13756.
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Object Increased neurotrophin activity in degenerative intervertebral discs (IVDs) is one potential cause of chronic low-back pain (LBP). The aim of the study was to assess if nerve growth factor (NGF) might alter gene expression of IVD cells and contribute to disc degeneration by enhancing expression or activity of factors that cause breakdown of IVD matrix. Methods Rat-tail IVD cells were stimulated by NGF and subjected to microarray analysis. Real-time polymerase chain reaction, Western blotting, and immunocytochemistry of rat and human IVD cells and tissues treated with NGF in vitro in the absence or presence of the NGF inhibitor Ro 08-2750 were used to confirm findings of the microarray studies. Phosphorylation of mitogen-activated protein kinase (MAPK) was used to identify cell signaling pathways involved in NGF stimulation in the absence or presence of Ro 08-2750. Results Microarray analysis demonstrated increased expression of chitinase 3-like 1 (Chi3l1), lipocalin 2 (Lcn2), and matrix metalloproteinase–3 (Mmp3) following NGF stimulation of rat IVD cells in vitro. Increased gene expression was confirmed by real-time polymerase chain reaction with a relative increase in the Mmp/Timp ratio. Increased expression of Chi3l1, Lcn2, and Mmp3 following NGF stimulation was also demonstrated in rat cells and human tissue in vitro. Effects of NGF on protein expression were blocked by an NGF inhibitor and appear to function through the extracellular-regulation kinase 1/2 (ERK1/2) MAPK pathway. Conclusions Nerve growth factor has potential effects on matrix turnover activity and influences the catabolic/anabolic balance of IVD cells in an adverse way that may potentiate IVD degeneration. Anti-NGF treatment might be beneficial to ameliorate progressive tissue breakdown in IVD degeneration and may lead to pain relief.
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Couto, Sheila M. F., Douglas I. Machado, Carolina Conde, Vinicius C. Silva, Adriana A. Souza, Karina B. Peres, Beatriz A. Brandi, and Maria De Fátima F. Vattimo. "Physical Training Is a Potential Modifier of Risk for Contrast-Induced Acute Kidney Injury in Diabetes Mellitus." BioMed Research International 2020 (November 17, 2020): 1–6. http://dx.doi.org/10.1155/2020/1830934.
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Background. Iodinated contrast (IC) is a leading cause of hospital-based acute kidney injury (AKI). Contrast-induced acute kidney injury (CI-AKI) is a decline in renal function due to iodinated contrast administration and occurs more frequently in individuals with increasingly common risk factors, such as diabetes mellitus (DM). Physical training (PT) can have renoprotective effects on CI-AKI in diabetic nephropathy. The aim of this study was to evaluate the injury in kidneys of diabetic rats submitted to treatment with IC, evaluating the impact of PT on hemodynamics and renal function in addition to oxidative profile in diabetic rats submitted to IC-AKI. Materials and Methods. Adult male Wistar rats are randomized into four groups: citrate ( n = 7 ): control group, citrate buffer (streptozotocin-STZ vehicle), intravenous tail (iv), single dose; DM ( n = 7 ): STZ, 60 mg/kg, iv, single dose; DM+IC ( n = 7 ): DM rats treated with IC (sodium meglumine ioxithalamate, 6 mL/kg, intraperitoneal (ip), single dose); DM+IC+PT ( n = 7 ): DM rats treated with IC as mentioned and submitted to physical training. Renal function parameters (inulin clearance, neutrophil gelatinase-associated lipocalin (NGAL), serum creatinine, and urinary albumin), hemodynamics (renal blood flow and renal vascular resistance), and oxidative profile (urinary peroxides, urinary TBARS, urinary nitric oxide, and renal tissue thiols) were evaluated. Results. It was possible to observe a decrease in inulin clearance, renal blood flow, and thiols in renal tissue accompanied by an increase in urinary flow, serum creatinine, urinary albumin, renal vascular resistance, urinary peroxides, urinary nitrate, and TBARS in the DM group compared to the citrate group. The DM+IC group showed a reduction in inulin clearance, and the renal dysfunction was also seen by the increased NGAL. Renal hemodynamics and oxidative profile compared were also worsened in the DM group. PT improved renal function by increasing renal blood flow and thiol levels in renal tissue and reduced renal vascular resistance, metabolites of reactive oxygen, nitrogen species, and lipid peroxidation in the DM+IC+PT group compared to DM+IC. Conclusions. Our results confirmed that DM induction increases renal vulnerability to the toxicity of IC and an association between DM with IC predisposes to severe AKI with reduced renal function alongside with renal hemodynamic alterations and oxidative mechanism of injury. The PT showed a renoprotective effect in DM animals subjected to damage with IC by modulating renal hemodynamics and oxidative profile, confirming a potential to modify the risk of CI-AKI when diabetes mellitus is present.
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Berterame, Nadia Maria, Stefano Bertagnoli, Vera Codazzi, Danilo Porro, and Paola Branduardi. "Temperature-induced lipocalin (TIL): a shield against stress-inducing environmental shocks in Saccharomyces cerevisiae." FEMS Yeast Research 17, no. 6 (July 29, 2017). http://dx.doi.org/10.1093/femsyr/fox056.
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Reck, Jose, Anelise Webster, Bruno Dall’Agnol, Ronel Pienaar, Minique H. de Castro, Jonathan Featherston, and Ben J. Mans. "Transcriptomic Analysis of Salivary Glands of Ornithodoros brasiliensis Aragão, 1923, the Agent of a Neotropical Tick-Toxicosis Syndrome in Humans." Frontiers in Physiology 12 (August 6, 2021). http://dx.doi.org/10.3389/fphys.2021.725635.
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Tick salivary glands produce and secrete a variety of compounds that modulate host responses and ensure a successful blood meal. Despite great progress made in the identification of ticks salivary compounds in recent years, there is still a paucity of information concerning salivary molecules of Neotropical argasid ticks. Among this group of ticks, considering the number of human cases of parasitism, including severe syndromes and hospitalization, Ornithodoros brasiliensis can be considered one of the major Neotropical argasid species with impact in public health. Here, we describe the transcriptome analysis of O. brasiliensis salivary glands (ObSG). The transcriptome yielded ~14,957 putative contigs. A total of 368 contigs were attributed to secreted proteins (SP), which represent approximately 2.5% of transcripts but ~53% expression coverage transcripts per million. Lipocalins are the major protein family among the most expressed SP, accounting for ~16% of the secretory transcripts and 51% of secretory protein abundance. The most expressed transcript is an ortholog of TSGP4 (tick salivary gland protein 4), a lipocalin first identified in Ornithodoros kalahariensis that functions as a leukotriene C4 scavenger. A total of 55 lipocalin transcripts were identified in ObSG. Other transcripts potentially involved in tick-host interaction included as: basic/acid tail secretory proteins (second most abundant expressed group), serine protease inhibitors (including Kunitz inhibitors), 5' nucleotidases (tick apyrases), phospholipase A2, 7 disulfide bond domain, cystatins, and tick antimicrobial peptides. Another abundant group of proteins in ObSG is metalloproteases. Analysis of these major protein groups suggests that several duplication events after speciation were responsible for the abundance of redundant compounds in tick salivary glands. A full mitochondrial genome could be assembled from the transcriptome data and confirmed the close genetic identity of the tick strain sampled in the current study, to a tick strain previously implicated in tick toxicoses. This study provides novel information on the molecular composition of ObSG, a Brazilian endemic tick associated with several human cases of parasitism. These results could be helpful in the understanding of clinical findings observed in bitten patients, and also, could provide more information on the evolution of Neotropical argasids.
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Kim, Ji Hwa, Sang Won Han, Seong Hwan Ahn, and Kyung-Yul Lee. "Abstract TP220: Neutrophil Gelatinase-associated Lipocalin in Patients With Acute Ischemic Stroke." Stroke 47, suppl_1 (February 2016). http://dx.doi.org/10.1161/str.47.suppl_1.tp220.
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Introduction: Neutrophil gelatinase-associated lipocalin (NGAL) appears to be a promising biomarker in diagnosis and prognosis in kidney and cardiovascular disease. However, the relationship between NGAL concentration and ischemic stroke is poorly understood. Hypothesis: We investigated plasma NGAL concentration in patients with acute ischemic stroke in relation to medical history, risk factors, hematologic and biochemical tests, stroke severity and subtype. Methods: Clinical data of 139 patients who admitted to a tertiary hospital with TIA (n=17) and ischemic stroke (n=122) were collected. We evaluated the following parameters: A) Medical history and risk factors of the patients; B) Hematologic and biochemical blood test at the time of admission; C) Stroke severity which was assessed using the National Institutes of Health Stroke Scale (NIHSS); D) Stroke subtype which was decided by the Trial of Org 10172 in Acute Stroke Treatment (TOAST) classification. Results: In our study (62% male, mean age 65 ± 14 years), median admission serum NGAL level was 75 ng/mL (interquartile range [IQR] 52-112 ng/mL). Serum NGAL level was elevated in 21 (15.1%) patients with the cutoff point of 150 ng/mL. High plasma NGAL concentrations were associated with age ( p = 0.018) and previous history of coronary artery occlusive disease ( p = 0.003). Mean NIHSS score in high NGAL group was higher than normal NGAL group (7.6 ±7.2 vs 4.4 ± 5.6, p = 0.021). We observed significant correlations between serum NGAL with C-reactive protein (spearman coefficient rho = 0.319, p < 0.001), erythrocyte sedimentation rate (r = 0.410, p < 0.001), creatinine (r = 0.237, p = 0.005), blood urea nitrogen (r = 0.353, p = 0.003), estimated glomerular filtration rate (r = -0.471, p < 0.001), fibrinogen (r = 0.327, p < 0.001) and homocysteine (r = 0.189, p = 0.032). There were no significant differences in mean serum NGAL level between TIA patients and stroke patients, and stroke subtypes by TOAST classification. Conclusions: Increased NGAL in acute ischemic stroke patients is associated with age, previous coronary artery occlusive disease history and stroke severity. Elevated plasma levels of NGAL appear to be correlated with some biomarkers associated with inflammation, coagulation and kidney dysfunction.
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Tarjus, Antoine, Soumaya El Moghrabi, Céline Latouche, Patrick Rossignol, Faiez Zannad, Nicolette Farman, Natalia López-Andrés, and Frédéric Jaisser. "Abstract 69: Role of Neutrophil Gelatinase Associated Lipocalin in Cardiovascular Remodeling Induced by Aldosterone." Hypertension 62, suppl_1 (September 2013). http://dx.doi.org/10.1161/hyp.62.suppl_1.a69.
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Neutrophil Gelatinase Associated Lipocalin (NGAL) is a circulating protein, member of the lipocalin family, which binds MMP9 and modulates its stability and activity. We have recently shown that NGAL is a primary target of aldosterone/mineralocorticoid receptor (MR) in endothelial cells, vascular smooth muscle cells and cardiomyocytes. We hypothesized that NGAL could be a mediator of aldosterone/MR profibrotic and proinflammatory effects in the cardiovascular system. Wild type (WT) and NGAL Knock Out (KO) mice were subjected to an uni-nephrectomy aldosterone salt challenge (NAS, 200μg/kg/day of aldo, 1% NaCl in tap water) for 4 weeks. Blood pressure (SBP) was measured by tail cuff method. Vascular reactivity was assessed by wire myograph. Cardiovascular fibrosis and inflammation were analyzed by RT-PCR, western blot, immunohistochemistry and ELISA. There was no difference in SBP between NGAL KO mice compared to WT mice in basal condition. With NAS challenge, SBP was increased only in WT (SBP; CT 107±3, CT NAS 133±5, KO 109±3, KO NAS 115±3 mmHg). Ex vivo aortic contraction induced by phenylephrine (Phe) and potassium chloride (KCl) were increased by NAS challenge in WT and KO (Emax Phe; CT 2.28±0.8, CT NAS 10.65±1.3, KO 1.45±0.3, KO NAS 9.32±0.9 mN). Responses to vasodilators (acetylcholine and sodium nitroprussiate) were altered in NAS condition in both groups. Quantification of pro collagen I N-terminal peptide (PINP) in plasma showed an increase of PINP due to NAS treatment in WT that was prevented by NGAL inactivation (CT 83±15, CT NAS 129±10, KO 70±19, KO NAS 59±12 μg/l). In myocardium, NAS treatment increased collagen type I and perivascular fibrosis in WT whereas KO were resistant to fibrosis (Collagen Volume Fraction; CT 19±3, CT NAS 28±2, KO 20±3, KO NAS 20±3 %). In aorta, collagen type I, vascular fibrosis and osteopontin were also increased by NAS in WT. These increases were prevented by NGAL inactivation (CVF; CT 24±4, CT NAS 34±5, KO 21±2, KO NAS 28±2 %). Our results show that NGAL plays a role in aldosterone/MR-mediated hypertension, vascular and cardiac peri-vascular fibrosis and inflammation, but not in vascular dysfunction.
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Trentelman, Jos J. A., Radek Sima, Nicolas Krezdorn, Julen Tomás-Cortázar, Diego Barriales, Katsuhisa Takumi, Joe M. Butler, et al. "A combined transcriptomic approach to identify candidates for an anti-tick vaccine blocking B. afzelii transmission." Scientific Reports 10, no. 1 (November 18, 2020). http://dx.doi.org/10.1038/s41598-020-76268-y.
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AbstractIxodes ricinus is the vector for Borrelia afzelii, the predominant cause of Lyme borreliosis in Europe, whereas Ixodes scapularis is the vector for Borrelia burgdorferi in the USA. Transcription of several I. scapularis genes changes in the presence of B. burgdorferi and contributes to successful infection. To what extend B. afzelii influences gene expression in I. ricinus salivary glands is largely unknown. Therefore, we measured expression of uninfected vs. infected tick salivary gland genes during tick feeding using Massive Analysis of cDNA Ends (MACE) and RNAseq, quantifying 26.179 unique transcripts. While tick feeding was the main differentiator, B. afzelii infection significantly affected expression of hundreds of transcripts, including 465 transcripts after 24 h of tick feeding. Validation of the top-20 B. afzelii-upregulated transcripts at 24 h of tick feeding in ten biological genetic distinct replicates showed that expression varied extensively. Three transcripts could be validated, a basic tail protein, a lipocalin and an ixodegrin, and might be involved in B. afzelii transmission. However, vaccination with recombinant forms of these proteins only marginally altered B. afzelii infection in I. ricinus-challenged mice for one of the proteins. Collectively, our data show that identification of tick salivary genes upregulated in the presence of pathogens could serve to identify potential pathogen-blocking vaccine candidates.
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Pérez-Sánchez, Ricardo, Ángel Carnero-Morán, Beatriz Soriano, Carlos Llorens, and Ana Oleaga. "RNA-seq analysis and gene expression dynamics in the salivary glands of the argasid tick Ornithodoros erraticus along the trophogonic cycle." Parasites & Vectors 14, no. 1 (March 20, 2021). http://dx.doi.org/10.1186/s13071-021-04671-z.
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Abstract Background The argasid tick Ornithodoros erraticus is the main vector of tick-borne human relapsing fever (TBRF) and African swine fever (ASF) in the Mediterranean Basin. Tick salivary proteins secreted to the host at the feeding interface play critical roles for tick feeding and may contribute to host infection by tick-borne pathogens; accordingly, these proteins represent interesting antigen targets for the development of vaccines aimed at the control and prevention of tick infestations and tick-borne diseases. Methods To identify these proteins, the transcriptome of the salivary glands of O. erraticus was de novo assembled and the salivary gene expression dynamics assessed throughout the trophogonic cycle using Illumina sequencing. The genes differentially upregulated after feeding were selected and discussed as potential antigen candidates for tick vaccines. Results Transcriptome assembly resulted in 22,007 transcripts and 18,961 annotated transcripts, which represent 86.15% of annotation success. Most salivary gene expression took place during the first 7 days after feeding (2088 upregulated transcripts), while only a few genes (122 upregulated transcripts) were differentially expressed from day 7 post-feeding onwards. The protein families more abundantly overrepresented after feeding were lipocalins, acid and basic tail proteins, proteases (particularly metalloproteases), protease inhibitors, secreted phospholipases A2, 5′-nucleotidases/apyrases and heme-binding vitellogenin-like proteins. All of them are functionally related to blood ingestion and regulation of host defensive responses, so they can be interesting candidate protective antigens for vaccines. Conclusions The O. erraticus sialotranscriptome contains thousands of protein coding sequences—many of them belonging to large conserved multigene protein families—and shows a complexity and functional redundancy similar to those observed in the sialomes of other argasid and ixodid tick species. This high functional redundancy emphasises the need for developing multiantigenic tick vaccines to reach full protection. This research provides a set of promising candidate antigens for the development of vaccines for the control of O. erraticus infestations and prevention of tick-borne diseases of public and veterinary health relevance, such as TBRF and ASF. Additionally, this transcriptome constitutes a valuable reference database for proteomics studies of the saliva and salivary glands of O. erraticus.