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Yongjie Liu, Fujun Hou, Zhaoying Qian, Xiaolin Liu
The clade B serpins contain predominantly intracellular proteins and were reported to be involved in multiple biological functions, especially in inflammation and immune system function. However, studies about the role of the invertebrate intracellular serpins in immune responses were still deficient. Therefore, this paper focused on the functional characterization of LvserpinB3 in white shrimp Litopenaeus vannamei. The pAc5.1-LvserpinB3-EGFP vector was transfected into Drosophila Schneider 2 (S2) cells to analyze the subcellular localization of LvserpinB3, and fluorescent imaging showed that LvserpinB3 were mainly localized to the mitochondria...
March 14, 2018: Developmental and Comparative Immunology
Theresa Stolle, Freya Grondinger, Andreas Dunkel, Thomas Hofmann
Recently, studies on human salt taste sensitivity demonstrated that sodium chloride (NaCl) sensitive and non-sensitive subjects differed in their salivary proteome and, in particular, in endopeptidase activity. In order to investigate individual's NaCl sensitivity and the role of endoprotease activity in salt taste perception, 20 panellists were classified according to NaCl sensitivity and saliva samples collected. A targeted protein quantitation by means of selected-reaction-monitoring (SRM) mass spectrometry and stable-isotope incorporation revealed the joint abundance of lysozyme C and lipocalin-1 to be indicative for non-sensitive subjects...
July 15, 2018: Food Chemistry
Valeria Messina, Mauro Valtieri, Mercedes Rubio, Mario Falchi, Francesca Mancini, Alfredo Mayor, Pietro Alano, Francesco Silvestrini
The gametocytes of Plasmodium falciparum , responsible for the transmission of this malaria parasite from humans to mosquitoes, accumulate and mature preferentially in the human bone marrow. In the 10 day long sexual development of P. falciparum , the immature gametocytes reach and localize in the extravascular compartment of this organ, in contact with several bone marrow stroma cell types, prior to traversing the endothelial lining and re-entering in circulation at maturity. To investigate the host parasite interplay underlying this still obscure process, we developed an in vitro tridimensional co-culture system in a Matrigel scaffold with P...
2018: Frontiers in Cellular and Infection Microbiology
Zhu Yi-Shen, Sun Shuai, Richard FitzGerald
To date, no extensive literature review exists regarding potential uses of mung bean proteins and peptides. As mung bean has long been widely used as a food source, early studies evaluated mung bean nutritional value against the Food and Agriculture Organization of the United Nations (FAO)/the World Health Organization (WHO) amino acids dietary recommendations. The comparison demonstrated mung bean to be a good protein source, except for deficiencies in sulphur-containing amino acids, methionine and cysteine...
2018: Food & Nutrition Research
S M Hayen, A M Ehlers, C F den Hartog Jager, J Garssen, E F Knol, A C Knulst, W Suer, L E M Willemsen, H G Otten
BACKGROUND: Screening for specific IgE against 2S albumin proteins Ara h 2 and 6 has good positive predictive value in diagnosing peanut allergy. From the third 2S member Ara h 7, three isoforms have been identified. Their allergenicity has not been elucidated. OBJECTIVE: This study investigated the allergenicity of Ara h 7 isoforms compared to Ara h 2 and 6. METHODS: Sensitization of 15 DBPCFC confirmed peanut allergic patients to recombinant Ara h 2...
March 14, 2018: Clinical and Experimental Allergy: Journal of the British Society for Allergy and Clinical Immunology
Antonio Junior Lepedda, Gabriele Nieddu, Silvia Rocchiccioli, Nadia Ucciferri, Michela Idini, Pierina De Muro, Marilena Formato
Background: Diabetes mellitus is a global health problem representing the fifth leading cause of mortality and a major risk factor for cardiovascular diseases. In the last years, we reported an association among urinary trypsin inhibitor (UTI), a small proteoglycan that plays pleiotropic roles in many inflammatory processes, and both type 1 and 2 diabetes and developed a method for its direct quantitation and structural characterization. Methods: Urine from 39 patients affected by type 1 diabetes, 32 patients with type 2 diabetes, and 52 controls were analysed...
2018: Journal of Diabetes Research
Sakihito Kitajima, Wataru Aoki, Daisuke Shibata, Daisuke Nakajima, Nozomu Sakurai, Kazufumi Yazaki, Ryosuke Munakata, Toki Taira, Masaru Kobayashi, Shunsuke Aburaya, Eric Hyrmeya Savadogo, Susumu Hibino, Haruna Yano
Latexes in immature fruit, young petioles and lignified trunks of fig trees protect the plant using toxic proteins and metabolites in various organ-dependent ways. Latexes from plants contain high amounts of toxic proteins and metabolites, which attack microbes and herbivores after exudation at pest-induced wound sites. The protein and metabolite constituents of latexes are highly variable, depending on the plant species and organ. To determine the diversity of latex-based defense strategies in fig tree (Ficus carica) organs, we conducted comparative proteomic, transcriptomic and metabolomic analyses on latexes isolated from immature fruit, young petioles and lignified trunks of F...
March 13, 2018: Planta
Itay Cohen, Si Naftaly, Efrat Ben-Zeev, Alexandra Hockla, Evette S Radisky, Niv Papo
High structural and sequence similarity within protein families can pose significant challenges to the development of selective inhibitors, especially towards proteolytic enzymes. Such enzymes usually belong to large families of closely similar proteases and may also hydrolyze, with different rates, protein or peptide-based inhibitors. To address this challenge, we employed a combinatorial yeast surface display library approach complemented with a novel pre-equilibrium, competitive screening strategy for facile assessment of the effects of multiple mutations on inhibitor association rates and binding specificity...
March 13, 2018: Biochemical Journal
Jonathan James Weldrick, Mohammad Abdul-Ghani, Lynn A Megeney, Patrick G Burgon
The capacity to isolate and study single cardiomyocytes has dramatically enhanced our understanding of the fundamental mechanisms of the heart. Currently, two primary methods for the isolation of cardiomyocytes are employed; i) The neonatal isolation protocol and, ii) the Langendorff isolation method. A major limiting feature of both procedures is the inability to isolate cardiomyocytes between 3 days and 3 weeks post-birth. Herein we report the establishment and validation of a new method for the rapid and efficient isolation of mouse cardiomyocytes, regardless of age...
March 13, 2018: Canadian Journal of Physiology and Pharmacology
Petra Bilić, Nicolas Guillemin, Alan Kovačević, Blanka Beer Ljubić, Ines Jović, Asier Galan, Peter David Eckersall, Richard Burchmore, Vladimir Mrljak
Idiopathic dilated cardiomyopathy (iDCM) is a primary myocardial disorder with an unknown aetiology, characterized by reduced contractility and ventricular dilation of the left or both ventricles. Naturally occurring canine iDCM was used herein to identify serum proteomic signature of the disease compared to the healthy state, providing an insight into underlying mechanisms and revealing proteins with biomarker potential. To achieve this, we used high-throughput label-based quantitative LC-MS/MS proteomics approach and bioinformatics analysis of the in silico inferred interactome protein network created from the initial list of differential proteins...
March 9, 2018: Journal of Proteomics
Ramanjeet Kaur, Santosh Kumar Tiwari
Bacteriocin LB44 was purified from cell-free supernatant (CFS) of Pediococcus pentosaceus LB44 using activity-guided chromatography techniques. It was stable up to 121 °C, pH 2.0-6.0, sensitive to proteinase K, papain and trypsin, and retained complete activity in the presence of organic solvents tested. The molecular weight of bacteriocin was ∼6 kDa and initial ten amino acid residues (GECGMCXECG) suggested a new compound. The loss in viable cell count and K+ ion efflux of target cells of Micrococcus luteus suggested bactericidal activity...
March 9, 2018: Biochemical and Biophysical Research Communications
Melanie Christine Föll, Matthias Fahrner, Victor Oginga Oria, Markus Kühs, Martin Lothar Biniossek, Martin Werner, Peter Bronsert, Oliver Schilling
Background: Proteomic analyses of clinical specimens often rely on human tissues preserved through formalin-fixation and paraffin embedding (FFPE). Minimal sample consumption is the key to preserve the integrity of pathological archives but also to deal with minimal invasive core biopsies. This has been achieved by using the acid-labile surfactant RapiGest in combination with a direct trypsinization (DTR) strategy. A critical comparison of the DTR protocol with the most commonly used filter aided sample preparation (FASP) protocol is lacking...
2018: Clinical Proteomics
Xin Wang, Chuanan Shen, Zhe Li, Shengbo Xu, Dawei Li
Cultured keratinocytes play important roles in burn wound healing and scientific research studies. We aimed to modify the isolation method to avoid over-digestion, maximize the number of isolated epidermal cells and establish a more efficient and innocuous way of cell isolation. Compared to the conventional method, the modified method combines the more dynamic process of enzymatic digestion with multiple harvestings of dissociated cells via digestion. The cells from each harvesting were immediately re-suspended in culture medium with serum to avoid extended trypsinization and then pooled for further analysis...
March 8, 2018: Burns: Journal of the International Society for Burn Injuries
Yafei Duan, Yun Wang, Jiasong Zhang, Qingsong Liu, Xian Ding
The shrimp intestine barrier serves as the first line of the host defense against pathogen infection. Lipopolysaccharide (LPS) is the cell wall component of gram-negative bacteria, which known as endotoxin and induce the intestine inflammation. In this study, the acute toxicity effects of LPS injection on the morphology, digestive enzymes and immunological responses of intestine from Litopenaeus vannamei was investigated. HE stain showed that LPS injection damaged the intestine connective and epithelium tissue...
March 8, 2018: Journal of Invertebrate Pathology
Andrzej Rafal Hellmann, Salvatore Paiella, Justyna Kostro, Iwona Marek, Krystian Adrych, Zbigniew Śledziński, Stanisław Hać, Claudio Bassi
OBJECTIVES: The primary aim of this study was to determine the blood levels of SPINK1 in patients with chronic pancreatitis (CP) submitted to surgical or endoscopic decompression of pancreatic duct (PD). Additionally, we measured trypsin activity levels. METHODS: Two groups were identified, surgical (group A) and endoscopic (group B). Levels of SPINK1 and trypsin activity were measured at baseline and 6 months after pancreatic duct decompression and then compared within the groups...
March 3, 2018: Pancreatology: Official Journal of the International Association of Pancreatology (IAP) ... [et Al.]
Yu Bin Lee, Eun Mi Kim, Hayeon Byun, Hyung-Kwan Chang, Kwanghee Jeong, Zachary M Aman, Yu Suk Choi, Jungyul Park, Heungsoo Shin
Numerous methods have been reported for the fabrication of 3D multi-cellular spheroids and their use in stem cell culture. Current methods typically relying on the self-assembly of trypsinized, suspended stem cells, however, show limitations with respect to cell viability, throughput, and accurate recapitulation of the natural microenvironment. In this study, we developed a new system for engineering cell spheroids by self-assembly of micro-scale monolayer of stem cells. We prepared synthetic hydrogels with the surface of chemically formed micropatterns (squares/circles with width/diameter of 200 μm) on which mesenchymal stem cells isolated from human nasal turbinate tissue (hTMSCs) were selectively attached and formed a monolayer...
March 1, 2018: Biomaterials
Claudia Miersch, Katja Stange, Monika Röntgen
BACKGROUND: Satellite cells (SC) and their descendants, muscle precursor cells (MPC), play a key role in postnatal muscle development, regeneration, and plasticity. Several studies have provided evidence that SC and MPC represent a heterogeneous population differing in their biochemical and functional properties. The identification and characterization of functionally divergent SC subpopulations should help to reveal the precise involvement of SC/MPC in these myogenic processes. The aim of the present work was therefore to separate SC subpopulations by using Percoll gradients and to characterize their myogenic marker profiles and their functional properties (adhesion, proliferation, and differentiation)...
March 9, 2018: BMC Cell Biology
Rezeda A Akhmadishina, Ruslan Garifullin, Natalia V Petrova, Marat I Kamalov, Timur I Abdullin
Although delocalized lipophilic cations have been identified as effective cellular and mitochondrial carriers for a range of natural and synthetic drug molecules, little is known about their effects on pharmacological properties of peptides. The effect of triphenylphosphonium (TPP) cation on bioactivity of antioxidant tetrapeptides based on the model opioid YRFK motif was studied. Two tetrapeptide variants with L-arginine (YRFK) and D-arginine (YrFK) were synthesized and coupled with carboxyethyl-TPP (TPP-3) and carboxypentyl-TPP (TPP-6) units...
2018: Frontiers in Pharmacology
Haijin Liu, Renata Servan de Almeida, Patricia Gil, Emmanuel Albina
Newcastle disease, caused by infection with virulent strains of Newcastle disease virus (NDV), poses a risk for the poultry industry. The virulence of NDV is mainly determined by the cleavage site of F protein. Lentogenic NDV can become velogenic after passages in SPF chicken brain and air sac based on some strains isolated from water birds, because the proportion of virulent-related strains gradually increases. In contrast, this proportion remains unchanged if NDV is passaged via 10-day-old SPF chicken embryos...
March 2018: Veterinary Microbiology
Awen Menou, JanWillem Duitman, Bruno Crestani
Idiopathic Pulmonary Fibrosis (IPF) is a devastating chronic, progressive and irreversible disease that remains refractory to current therapies. Matrix metalloproteinases (MMPs) and their inhibitors, tissue inhibitors of MMPs (TIMPs), have been implicated in the development of pulmonary fibrosis since decades. Coagulation signalling deregulation, which influences several key inflammatory and fibro-proliferative responses, is also essential in IPF pathogenesis, and a growing body of evidence indicates that Protease-Activated Receptors (PARs) inhibition in IPF may be promising for future evaluation...
March 5, 2018: Matrix Biology: Journal of the International Society for Matrix Biology
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