Role of tryptophan residues of Erv1: Trp95 and Trp183 are important for its folding and oxidase function

Erv1 is an FAD-dependent sulphydryl oxidase of the ERV/ALR sub-family, and an essential component of the mitochondrial import and assembly pathway. Erv1 contains six tryptophan residues, which are all located in the highly conserved C-terminal FAD-binding domain. Though important structural roles were predicted for the invariable Trp95, no experimental study has been reported. In this study, we investigated the structural and functional roles of individual Trp residues of Erv1. Six single Trp-to-Phe yeast mutant strains were generated and their effects on cell viability were tested at various temperatures. Then, the mutants were purified from E. coli. Their effects on folding, FAD-binding, and Erv1 activity were characterised. Our results showed that Erv1W95F has the strongest effect on the stability and function of Erv1, and followed by Erv1W183F. Erv1W95F results in a decrease of the Tm of Erv1 by 23°C, a significant loss of the oxidase activity, and thus causing cell growth defects at both 30°C and 37°C. Erv1W183F induces changes in the oligomerisation state of Erv1, along with a pronounced effect on the stability of Erv1 and its function at 37°C, whilst the other mutants had no clear effect on the function of Erv1 including the highly conserved Trp157 mutant. Finally, computational analysis indicates that Trp95 plays a key role in stabilising the isoalloxazine ring to interact with Cys133. Taken together, this study provided important insights into the molecular mechanism of how sulfhydryl oxidases use FAD in catalyzing disulfide bond formation

Cytosolic Fe-S cluster protein maturation and iron regulation are independent of the mitochondrial Erv1/Mia40 import system

The sulfhydryl oxidase Erv1 partners with the oxidoreductase Mia40 to import cysteine-rich proteins in the mitochondrial intermembrane space. In Saccharomyces cerevisiae, Erv1 has also been implicated in cytosolic Fe-S protein maturation and iron regulation. To investigate the connection between Erv1/Mia40-dependent mitochondrial protein import and cytosolic Fe-S cluster assembly, we measured Mia40 oxidation and Fe-S enzyme activities in several erv1 and mia40 mutants. Although all the erv1 and mia40 mutants exhibited defects in Mia40 oxidation, only one erv1 mutant strain (erv1-1) had significantly decreased activities of cytosolic Fe-S enzymes. Further analysis of erv1-1 revealed that it had strongly decreased glutathione (GSH) levels, caused by an additional mutation in the gene encoding the glutathione biosynthesis enzyme glutamate cysteine ligase (GSH1). To address whether Erv1 or Mia40 plays a role in iron regulation, we measured iron-dependent expression of Aft1/2-regulated genes and mitochondrial iron accumulation in erv1 and mia40 strains. The only strain to exhibit iron misregulation is the GSH-deficient erv1-1 strain, which is rescued with addition of GSH. Together, these results confirm that GSH is critical for cytosolic Fe-S protein biogenesis and iron regulation, whereas ruling out significant roles for Erv1 or Mia40 in these pathways

Keratin-based particles for protection and restoration of hair properties

Objective Human hair is an element with unquestionable relevance in society both for women and men. Therefore, it is of great importance to develop new cosmetic products for hair care capable to restore and improve hair's characteristics. Here we explore the potential of keratinbased particles in the protection and recovery of hair mechanical properties and thermal stability. Methods Keratinbased particles were obtained by high pressure homogenization (HPH) using keratin and silk fibroin. The particles were characterized regarding size, superficial charge and polydispersity index. Their safety to cells was assessed using human skin keratinocytes. Virgin and overbleached Asian hair were treated with eight keratinbased formulations. The effect of particles on hair's mechanical properties was evaluated in terms of stiffness and tensile strength. The impact of treatments in hair thermal performance was studied using differential scanning calorimetry (DSC). Results Keratinbased particles were capable to recover and/or improve the mechanical properties of virgin and overbleached hair. Virgin hair treated with K80SF20P particles presented an improvement of the mechanical properties around 40%. An increase in keratin helix denaturation enthalpy and in surface smoothness for both types of hair was also verified after treatment. These particles demonstrated stability over time and proved to be safe when tested in human keratinocytes. Conclusion The keratinbased particles here presented have the potential to be incorporated in the development of new and effective hair care cosmetic formulations.This study was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE2020 (POCI01-0145-FEDER-006684) and BioTecNorte operation (NORTE-010145-FEDER-000004) and Nanotechnology Based Functional Solutions (NORTE-01-0145- FEDER-000019) funded by the European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. We also acknowledge the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER007569) funded by national funds through Fundação para a Ciência e a Tecnologia (FCT) and by the ERDF through the COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI). Artur Ribeiro and Ana Tinoco thank FCT for funding the scholarships with the references SFRH/BPD/98388/2013 and SFRH/BD/ 114035/2015, respectively.info:eu-repo/semantics/publishedVersio

Evaluation of the Efficacy of Two Amide Group Local Intrapocket Anesthetic Gels in Non Invasive Periodontal Therapy - A Clinical Study

Background: The efficacy of two intrapocket local anesthetics of amide group, Lignocaine 5% and Bupivacaine 5% were analyzed and compared to each other in the procedure of Scaling and root planing.Methods: Forty patients were divided in two equal groups of Lignocaine 5% and Bupivacaine 5% .Gels of each drug were obtained and used for the procedure of scaling and root planing (SRP). The gels were applied in the pocket with a 2mm diameter plastic canula attached to a syringe. Every 1 minute probing was done for anesthesia assessment and on ‘no pain’ of probing SRP done. Every 5 minutes probing was continued to find out the time for which the anesthetic drug effect lasted, SRP was done by an independent blind observer and pain assessment and gel application by another to eliminate the bias. Pain assessment was done on 100mm Visual analog scale (VAS), and Verbal Response Scale (VRS) on a four point scale having points labeled from none pain to severe pain, which was used at the end of the procedure.Results: The data analyzed showed that Bupivacaine 5% gel takes a longer time to start its action and similarly the peak effect is reached late when compared with Lignocaine 5% gel (p< 0.01).But the duration for which the anesthesia lasts is significantly higher for Bupivacaine (average 30.25 mins). And the overall pain experienced by patients is also significantly less than that by patients in Lignocaine group.Conclusion: Bupivacaine 5% is more effective topical intrapocket anesthetic for Scaling and Root Planing procedure and other time taking Periodontal procedures as compared to Lignocaine 5% ,and it may be a better alternative to Infiltration local anesthesia

SAP Flexible Real Estate Management [electronic resource] /

Learn SAP's real estate management integrated solution to effectively manage the real estate portfolio at your organization. You will configure SAP REFX for business scenarios covering solutions from master data to financial posting and reporting. You will address all phases of the real estate life cycle, including real estate acquisition or disposal, portfolio management, and property and technical management. To succeed in today's global and highly competitive economy, asset optimization in real estate management has become a strategic task. Organizations need to ensure insight into their property portfolio to make informed decisions, improve portfolio performance, and reduce compliance costs. Sophisticated solutions are needed to manage changing consumer demands and the global workforce as well as information management, compliance adherence, and leasing and property management. SAP Flexible Real Estate Management by Daithankar is a full-featured book that integrates REFX with Controlling (CO), Plant and Maintenance (PM), CRM, SAP AA (ssset accounting), and SAP PS (project systems). You will refer to real-world, practical examples to illustrate configuration concepts and processes, and learn in an interactive, hands-on way through the use of screenshots, menu paths, and transaction codes throughout the book. What You Will Learn: Understand the SAP REFX Solutions landscape and industry best practices for SAP REFX implementation Configure SAP REFX Integrate REFX with other modules Understand how processes are supported by SAP REFX Who This Book Is For: CIOs/CEOs of organizations with real estate portfolios, SAP REFX purchasing decision makers, SAP REFX pre-sales teams, SAP REFX implementation/AMS consultants.Learn SAP's real estate management integrated solution to effectively manage the real estate portfolio at your organization. You will configure SAP REFX for business scenarios covering solutions from master data to financial posting and reporting. You will address all phases of the real estate life cycle, including real estate acquisition or disposal, portfolio management, and property and technical management. To succeed in today's global and highly competitive economy, asset optimization in real estate management has become a strategic task. Organizations need to ensure insight into their property portfolio to make informed decisions, improve portfolio performance, and reduce compliance costs. Sophisticated solutions are needed to manage changing consumer demands and the global workforce as well as information management, compliance adherence, and leasing and property management. SAP Flexible Real Estate Management by Daithankar is a full-featured book that integrates REFX with Controlling (CO), Plant and Maintenance (PM), CRM, SAP AA (ssset accounting), and SAP PS (project systems). You will refer to real-world, practical examples to illustrate configuration concepts and processes, and learn in an interactive, hands-on way through the use of screenshots, menu paths, and transaction codes throughout the book. What You Will Learn: Understand the SAP REFX Solutions landscape and industry best practices for SAP REFX implementation Configure SAP REFX Integrate REFX with other modules Understand how processes are supported by SAP REFX Who This Book Is For: CIOs/CEOs of organizations with real estate portfolios, SAP REFX purchasing decision makers, SAP REFX pre-sales teams, SAP REFX implementation/AMS consultants

Moisturizing efficiency of silk protein hydrolysate: Silk fibroin

115-121A biomimetic approach of composition and natural function of natural moisturizing factor (NMF) with the amino acid content of silk fibroin was advantageously used to reconstruct the skin moisturizing system. The isolation of silk hydrolysate with water and sodium chloride treatment was complete in one hour. Lithium ion from LiBr effectively penetrated crystal domains of fibroin and gave desired solubility. Silk fibroin from Bombyx mori cocoons was non-allergic and biocompatible in skin and rabbit eye tests. The concentration dependent moisturizing efficacy of fibroin (1-5% w/v) in solution and cream form has been demonstrated by TEWL in vitro and in volunteers. As compared to dry and normal skin the fibroin containing cream revealed increased substantivity. The increased hydroxproline content was responsible for retaining higher moisture in the skin. This in turn maintained the skin in soft and supple state. The significant drop in impedance was observed within 1 hr of the application of fibroin and the effect was sustained for more than 6 hrs. Thus, increased hydration level in stratum corneum was achieved by fibroin treatment. The SEM of fibroin treated skin replicas showed a desired attribute of soft, smooth skin texture and improved flexibility. The increased state of hydration caused interdigitating of cell edges as evident in microphotographs. The rapid and sustained moisturizing efficiency observed with silk fibroin was well substantiated by the results of skin substantivity and impedance tests

Synthesis, antimicrobial and antitubercular activity of some novel [3-isonicotinoyl-5-(4-substituted)-2,3-dihydro-1,3,4-oxadiazol-2-yl] and substituted 5-(pyridin-4-yl)-1,3,4-oxadiazole-2-thiol derivatives

293-299A series of aryloxadiazole and mercaptooxadiazoles have been synthesized and evaluated for antibacterial, antifungal and antitubercular activities. The newly synthesized compounds have been characterized by IR, 1H NMR and elemental analyses. All the compounds have shown promising antibacterial, antifungal and antitubercular activity when compared with standard drug Ciprofloxacin, Griseofulvin and Streptomycin respectively