Identification of Lactoferricin B Intracellular Targets Using an Escherichia coli Proteome Chip

Lactoferricin B (LfcinB) is a well-known antimicrobial peptide. Several studies have indicated that it can inhibit bacteria by affecting intracellular activities, but the intracellular targets of this antimicrobial peptide have not been identified. Therefore, we used E. coli proteome chips to identify the intracellular target proteins of LfcinB in a high-throughput manner. We probed LfcinB with E. coli proteome chips and further conducted normalization and Gene Ontology (GO) analyses. The results of the GO analyses showed that the identified proteins were associated with metabolic processes. Moreover, we validated the interactions between LfcinB and chip assay-identified proteins with fluorescence polarization (FP) assays. Sixteen proteins were identified, and an E. coli interaction database (EcID) analysis revealed that the majority of the proteins that interact with these 16 proteins affected the tricarboxylic acid (TCA) cycle. Knockout assays were conducted to further validate the FP assay results. These results showed that phosphoenolpyruvate carboxylase was a target of LfcinB, indicating that one of its mechanisms of action may be associated with pyruvate metabolism. Thus, we used pyruvate assays to conduct an in vivo validation of the relationship between LfcinB and pyruvate level in E. coli. These results showed that E. coli exposed to LfcinB had abnormal pyruvate amounts, indicating that LfcinB caused an accumulation of pyruvate. In conclusion, this study successfully revealed the intracellular targets of LfcinB using an E. coli proteome chip approach

The role of released ATP in killing Candida albicans and other extracellular microbial pathogens by cationic peptides

A unifying theme common to the action of many cationic peptides that display lethal activities against microbial pathogens is their specific action at microbial membranes that results in selective loss of ions and small nucleotides chiefly ATP. One model cationic peptide that induces non-lytic release of ATP from the fungal pathogen Candida albicans is salivary histatin 5 (Hst 5). The major characteristic of Hst 5-induced ATP release is that it occurs rapidly while cells are still metabolically active and have polarized membranes, thus precluding cell lysis as the means of release of ATP. Other cationic peptides that induce selective release of ATP from target microbes are lactoferricin, human neutrophil defensins, bactenecin, and cathelicidin peptides. The role of released extracellular ATP induced by cationic peptides is not known, but localized increases in extracellular ATP concentration may serve to potentiate cell killing, facilitate further peptide uptake, or function as an additional signal to activate the host innate immune system at the site of infection

Mytilus galloprovincialis Myticin C: A Chemotactic Molecule with Antiviral Activity and Immunoregulatory Properties

Previous research has shown that an antimicrobial peptide (AMP) of the myticin class C (Myt C) is the most abundantly expressed gene in cDNA and suppressive subtractive hybridization (SSH) libraries after immune stimulation of mussel Mytilus galloprovincialis. However, to date, the expression pattern, the antimicrobial activities and the immunomodulatory properties of the Myt C peptide have not been determined. In contrast, it is known that Myt C mRNA presents an unusual and high level of polymorphism of unidentified biological significance. Therefore, to provide a better understanding of the features of this interesting molecule, we have investigated its function using four different cloned and expressed variants of Myt C cDNA and polyclonal anti-Myt C sera. The in vivo results suggest that this AMP, mainly present in hemocytes, could be acting as an immune system modulator molecule because its overexpression was able to alter the expression of mussel immune-related genes (as the antimicrobial peptides Myticin B and Mytilin B, the C1q domain-containing protein MgC1q, and lysozyme). Moreover, the in vitro results indicate that Myt C peptides have antimicrobial and chemotactic properties. Their recombinant expression in a fish cell line conferred protection against two different fish viruses (enveloped and non-enveloped). Cell extracts from Myt C expressing fish cells were also able to attract hemocytes. All together, these results suggest that Myt C should be considered not only as an AMP but also as the first chemokine/cytokine-like molecule identified in bivalves and one of the few examples in all of the invertebrates

Bioactive Molecules Released in Food by Lactic Acid Bacteria: Encrypted Peptides and Biogenic Amines

Lactic acid bacteria (LAB) can produce a huge amount of bioactive compounds. Since their elective habitat is food, especially dairy but also vegetal food, it is frequent to find bioactive molecules in fermented products. Sometimes these compounds can have adverse effects on human health such as biogenic amines (tyramine and histamine), causing allergies, hypertensive crises, and headache. However, some LAB products also display benefits for the consumers. In the present review article, the main nitrogen compounds produced by LAB are considered. Besides biogenic amines derived from the amino acids tyrosine, histidine, phenylalanine, lysine, ornithine, and glutamate by decarboxylation, interesting peptides can be decrypted by the proteolytic activity of LAB. LAB proteolytic system is very efficient in releasing encrypted molecules from several proteins present in different food matrices. Alpha and beta-caseins, albumin and globulin from milk and dairy products, rubisco from spinach, beta-conglycinin from soy and gluten from cereals constitute a good source of important bioactive compounds. These encrypted peptides are able to control nutrition (mineral absorption and oxidative stress protection), metabolism (blood glucose and cholesterol lowering) cardiovascular function (antithrombotic and hypotensive action), infection (microbial inhibition and immunomodulation) and gut-brain axis (opioids and anti-opioids controlling mood and food intake). Very recent results underline the role of food-encrypted peptides in protein folding (chaperone-like molecules) as well as in cell cycle and apoptosis control, suggesting new and positive aspects of fermented food, still unexplored. In this context, the detailed (transcriptomic, proteomic, and metabolomic) characterization of LAB of food interest (as starters, biocontrol agents, nutraceuticals, and probiotics) can supply a solid evidence-based science to support beneficial effects and it is a promising approach as well to obtain functional food. The detailed knowledge of the modulation of human physiology, exploiting the health-promoting properties of fermented food, is an open field of investigation that will constitute the next challenge

The Potential of Antimicrobial Peptides as Biocides

Antimicrobial peptides constitute a diverse class of naturally occurring antimicrobial molecules which have activity against a wide range of pathogenic microorganisms. Antimicrobial peptides are exciting leads in the development of novel biocidal agents at a time when classical antibiotics are under intense pressure from emerging resistance, and the global industry in antibiotic research and development stagnates. This review will examine the potential of antimicrobial peptides, both natural and synthetic, as novel biocidal agents in the battle against multi-drug resistant pathogen infections

Besvär från rörelseapparaten hos anställda på Elektroniken, Peltor

Syftet med undersökningen var att kartlägga omfattningen av besvär från rörelseapparaten hos de anställda på den undersökta produktionsenheten för hörselskydd. Ytterligare syften var att se om man tidigt kan identifiera personer eller arbetsgrupper med likartade besvär, där åtgärder behövs på individ- eller gruppnivå. Den undersökta gruppen bestod av 86 personer, varav 59 kvinnor och 27 män. Produktionsenheten var fördelad på tre avdelningar. En avdelning skilde sig något i arbetsförutsättningar med lägre tempokrav och större rörlighet. Den bestod enbart av män. Ergonomiska förutsättningar var i övrigt ensartade med dominerande sittande monteringsarbete (70-80 procent av arbetstiden) men med möjlighet att stå. Belastning i arbetet förkom främst på händer, handleder, armar och nacke. Metoden var en enkätundersökning där värkbesvär redovisades från nio olika anatomiska lokalisationer. Värken graderades i en fem-gradig skala från de kroppsområden som drabbats under den senaste 12 månadersperioden. Hänsyn togs även till om man varit sjukskriven för värken. Värkpoängen sammanräknades från de nio anatomiska områdena enligt den fem-gradiga skalan och kunde ge totalt mellan 0-45 poäng. Arbetstrivsel belystes med separata frågor och uppdelades i hög och låg arbetstrivsel där skillnader i värkrapportering studerades. Svarsfrekvensen på enkätundersökningen var 95 procent. Smärtprevalens den senaste veckan, samt de senaste 12 månaderna var högst för nacke, ländrygg och skuldror. Genomgående högre värkprevalens påvisades hos kvinnor jämfört män, vilket också rapporterats i litteraturen. Dock skilde sig en produktionsenhet från detta då männen vid enheten genomgående rapporterade mer värk än kvinnorna. Värkintensiteten var högst i ländryggen hos båda könen, samt i handleder/händer hos kvinnor. Värk i länd- och bröstrygg var de lokalisationer som oftast medförde sjukskrivning. Skillnader i värkrapportering fanns mellan de olika avdelningarna. Det gällde även mellan de två avdelningar som hade jämförbara ergonomiska förutsättningar. Vid analys av sambandet mellan arbetstrivsel och totala värkpoäng fann man klart högre värkpoäng i gruppen med låg arbetstrivsel hos kvinnor, men inte hos män. För de som varit sjukskrivna för sin värk fanns en klar övervikt för låg arbetstrivsel hos båda könen. Slutsatsen av enkätundersökningen bedöms vara att värkrapporteringen hos den undersökta gruppen är högre än den som rapporteras i tidigare studier av allmän befolkning. Det får betonas att man i denna undersökning inte har kunnat jämföra med studier av liknande produktionsenheter. Undersökningen kunde identifiera vissa individer med hög värkrapportering och avdelningar med ensartade besvär där behov av vidare uppföljning på såväl individ- som gruppnivå finns. Resultaten tyder på en koppling mellan arbetstrivsel och värkomfattning, samt mellan arbetstrivsel och sjukskrivning för värken. Dessa fynd pekade på att enkäten uppnådde syftet med att kartlägga omfattningen av värkproblematiken, samt fånga upp individer och grupper med behov av vidare utredning och åtgärder

Snow depth measurements and predictions : Reducing environmental impact for artificial grass pitches at snowfall

Rubber granulates, used at artificial grass pitches, pose a threat to the environment when leaking into the nature. As the granulates leak to the environment through rain water and snow clearances, they can be transported by rivers and later on end up in the marine life. Therefore, reducing the snow clearances to its minimum is of importance. If the snow clearance problem is minimized or even eliminated, this will have a positive impact on the surrounding nature. The object of this project is to propose a method for deciding when to remove snow and automate the information dispersing upon clearing or closing a pitch. This includes finding low powered sensors to measure snow depth, find a machine learning model to predict upcoming snow levels and create an application with a clear and easy-to-use interface to present weather information and disperse information to the responsible persons. Controlled experiments is used to find the models and sensors that are suitable to solve this problem. The sensors are tested on a single snow quality, where ultrasonic and infrared sensors are found suitable. However, fabricated tests for newly fallen snow questioned the possibility of measuring snow depth using the ultrasonic sensor in the general case. Random Forest is presented as the machine learning model that predicts future snow levels with the highest accuracy. From a survey, indications is found that the web application fulfills the intended functionalities, with some improvements suggested

Snow depth measurements and predictions : Reducing environmental impact for artificial grass pitches at snowfall

Rubber granulates, used at artificial grass pitches, pose a threat to the environment when leaking into the nature. As the granulates leak to the environment through rain water and snow clearances, they can be transported by rivers and later on end up in the marine life. Therefore, reducing the snow clearances to its minimum is of importance. If the snow clearance problem is minimized or even eliminated, this will have a positive impact on the surrounding nature. The object of this project is to propose a method for deciding when to remove snow and automate the information dispersing upon clearing or closing a pitch. This includes finding low powered sensors to measure snow depth, find a machine learning model to predict upcoming snow levels and create an application with a clear and easy-to-use interface to present weather information and disperse information to the responsible persons. Controlled experiments is used to find the models and sensors that are suitable to solve this problem. The sensors are tested on a single snow quality, where ultrasonic and infrared sensors are found suitable. However, fabricated tests for newly fallen snow questioned the possibility of measuring snow depth using the ultrasonic sensor in the general case. Random Forest is presented as the machine learning model that predicts future snow levels with the highest accuracy. From a survey, indications is found that the web application fulfills the intended functionalities, with some improvements suggested

Induced resistance to the antimicrobial peptide lactoferricin B in Staphylococcus aureus

AbstractThis study was designed to investigate inducible intrinsic resistance against lactoferricin B in Staphylococcus aureus. Serial passage of seven S. aureus strains in medium with increasing concentrations of peptide resulted in an induced resistance at various levels in all strains. The induced resistance was unstable and decreased relatively rapidly during passages in peptide free medium but the minimum inhibitory concentration remained elevated after thirty passages. Cross-resistance to penicillin G and low-level cross-resistance to the antimicrobial peptides indolicidin and Ala3,13,18-magainin was observed. No cross-resistance was observed to the human cathelicidin LL-37. In conclusion, this study shows that S. aureus has intrinsic resistance mechanisms against antimicrobial peptides that can be induced upon exposure, and that this may confer low-level cross-resistance to other antimicrobial peptides