Proton translocating nicotinamide nucleotide transhydrogenase from E. coli. Mechanism of action deduced from its structural and catalytic properties11This review is dedicated to the memory of Professor Lars Ernster.

AbstractTranshydrogenase couples the stereospecific and reversible transfer of hydride equivalents from NADH to NADP+ to the translocation of proton across the inner membrane in mitochondria and the cytoplasmic membrane in bacteria. Like all transhydrogenases, the Escherichia coli enzyme is composed of three domains. Domains I and III protrude from the membrane and contain the binding site for NAD(H) and NADP(H), respectively. Domain II spans the membrane and constitutes at least partly the proton translocating pathway. Three-dimensional models of the hydrophilic domains I and III deduced from crystallographic and NMR data and a new topology of domain II are presented. The new information obtained from the structures and the numerous mutation studies strengthen the proposition of a binding change mechanism, as a way to couple the reduction of NADP+ by NADH to proton translocation and occurring mainly at the level of the NADP(H) binding site

Change management within the public sector : A qualitative study of employees’ experiences

Syftet med denna studie är att undersöka hur medarbetare upplevde processen under implementeringen av ett nyligen infört löne- och personalsystem. Författarna till studien har valt en kvalitativ metod för att utforska empirin och ställa den i relation till tidigare forskning. Datainsamlingen skedde i form av intervjuer med sex lönekonsulter. Studien utgår ifrån fyra utvalda teman, hämtade från tidgare forksning vilka är kommunikation, motstånd, stress och delaktighet. I resultatet framhålls det som framträtt under intervjuerna grundat i de utvalda temana. Det visade sig att förändringen innebar mycket stress och en hög arbetsbelastning. Tidsbrist, kommunikationsbrist och kunskapsbrist framträdde också som tre huvudsakliga faktorer. Dessutom visar resultatet att förändringsprocessen är påverkad av de lagar och regler som styr organisationen. Studien avslutas med en sammanfattning av det som framträtt i resultatet och diskussionen. Ledningen bör finna strategier för att kommunicera förändringen till medarbetarna. Ledningen bör också se förändringen som en möjlighet istället för ett krav för att främja lärandet och motivera medarbetarna.The aim of this thesis is to investigate employees experiences of the process during the implementation of a newly introduced personnel- and salary system. The authors of this study have chosen a qualitative method in order to explore the empirics and connect this to previous research. The data collection method used was interviews with six salary consultants. This study is based on four themes that were chosen based on prevoius research which are communication, resistance, stress and participation. In the results the information from the respondents are presented, based on the four themes. It was shown that the change caused a lot of stress and a high workload. Lack of time, lack of communication and lack of knowledge were three other main factors that were shown. The results also demonstrate that the change process is affected by the law that controls the organization. Lastly a conclusion is made based on the results and the discussion. The managers should find strategies to communicate change to the employees. The managers shouls also see the change as a possibility instead of a demand in order to promote employees learning and motivation

Change management within the public sector : A qualitative study of employees’ experiences

Syftet med denna studie är att undersöka hur medarbetare upplevde processen under implementeringen av ett nyligen infört löne- och personalsystem. Författarna till studien har valt en kvalitativ metod för att utforska empirin och ställa den i relation till tidigare forskning. Datainsamlingen skedde i form av intervjuer med sex lönekonsulter. Studien utgår ifrån fyra utvalda teman, hämtade från tidgare forksning vilka är kommunikation, motstånd, stress och delaktighet. I resultatet framhålls det som framträtt under intervjuerna grundat i de utvalda temana. Det visade sig att förändringen innebar mycket stress och en hög arbetsbelastning. Tidsbrist, kommunikationsbrist och kunskapsbrist framträdde också som tre huvudsakliga faktorer. Dessutom visar resultatet att förändringsprocessen är påverkad av de lagar och regler som styr organisationen. Studien avslutas med en sammanfattning av det som framträtt i resultatet och diskussionen. Ledningen bör finna strategier för att kommunicera förändringen till medarbetarna. Ledningen bör också se förändringen som en möjlighet istället för ett krav för att främja lärandet och motivera medarbetarna.The aim of this thesis is to investigate employees experiences of the process during the implementation of a newly introduced personnel- and salary system. The authors of this study have chosen a qualitative method in order to explore the empirics and connect this to previous research. The data collection method used was interviews with six salary consultants. This study is based on four themes that were chosen based on prevoius research which are communication, resistance, stress and participation. In the results the information from the respondents are presented, based on the four themes. It was shown that the change caused a lot of stress and a high workload. Lack of time, lack of communication and lack of knowledge were three other main factors that were shown. The results also demonstrate that the change process is affected by the law that controls the organization. Lastly a conclusion is made based on the results and the discussion. The managers should find strategies to communicate change to the employees. The managers shouls also see the change as a possibility instead of a demand in order to promote employees learning and motivation

Is GPR146 really the receptor for proinsulin C-peptide?

Proinsulin C-peptide has previously been proposed to interact with a G-protein coupled receptor (GPCR), specifically the orphan receptor GPR146. To investigate the potential of C-peptide in treating complications of diabetes, such as kidney damage, it is necessary to understand its mode of action. We used CHO-K1 cells expressing human GPR146 to study human and murine C-peptide in dynamic mass redistribution and GPCR beta-arrestin assays, as well as with fluorescence confocal microscopy. Neither assay revealed any significant intracellular response to C-peptide at concentrations of up to 33 mu M. We observed no internalisation of C-peptide by fluorescence microscopy. Our results do not support GPR146 as the receptor for C-peptide, but suggest that further investigations of the mode of action of C-peptide should be undertaken

Potency Prediction of β‑Secretase (BACE-1) Inhibitors Using Density Functional Methods

Scoring potency is a main challenge for structure based drug design. Inductive effects of subtle variations in the ligand are not possible to accurately predict by classical computational chemistry methods. In this study, the problem of predicting potency of ligands with electronic variations participating in key interactions with the protein was addressed. The potency was predicted for a large set of cyclic amidine and guanidine cores extracted from β-secretase (BACE-1) inhibitors. All cores were of similar size and had equal interaction motifs but were diverse with respect to electronic substitutions. A density functional theory approach, in combination with a representation of the active site of a protein using only key residues, was shown to be predictive. This computational approach was used to guide and support drug design, within the time frame of a normal drug discovery design cycle

Optimised generation of iPSC-derived macrophages and dendritic cells that are functionally and transcriptionally similar to their primary counterparts.

Induced pluripotent stem cells (iPSC) offer the possibility to generate diverse disease-relevant cell types, from any genetic background with the use of cellular reprogramming and directed differentiation. This provides a powerful platform for disease modeling, drug screening and cell therapeutics. The critical question is how the differentiated iPSC-derived cells translate to their primary counterparts. Our refinement of a published differentiation protocol produces a CD14+ monocytic lineage at a higher yield, in a smaller format and at a lower cost. These iPSC-derived monocytes can be further differentiated into macrophages or dendritic cells (DC), both with similar morphological and functional profiles as compared to their primary counterparts. Transcriptomic analysis of iPSC-derived cells at different stages of differentiation as well as comparison to their blood-derived counterparts demonstrates a complete switch of iPSCs to cells expressing a monocyte, macrophage or DC specific gene profile. iPSC-derived macrophages respond to LPS treatment by inducing expression of classic macrophage pro-inflammatory response markers. Interestingly, though iPSC-derived DC show similarities to monocyte derived DC, they are more similar transcriptionally to a newly described subpopulation of AXL+ DC. Thus, our study provides a detailed and accurate profile of iPSC-derived monocytic lineage cells