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1.	Andersson, Johanna, 1983, et al. (author) Lifetime Heterogeneity of DNA-Bound dppz Complexes Originates from Distinct Intercalation Geometries Determined by Complex-Complex Interactions 2013 In: Inorganic Chemistry. - : American Chemical Society (ACS). - 0020-1669 .- 1520-510X. ; 52:2, s. 1151-1159 Journal article (peer-reviewed)abstract Despite the extensive interest in structurally explaining the photophysics of DNA-bound [Ru(phen)(2)dppz](2+) and [Ru(bpy)(2)dppz](2+), the origin of the two distinct emission lifetimes of the pure enantiomers when intercalated into DNA has remained elusive. In this report, we have combined a photophysical characterization with a detailed isothermal titration calorimetry study to investigate the binding of the pure Delta and Lambda enantiomers of both complexes with [poly(dAdT)](2). We find that a binding model with two different binding geometries, proposed to be symmetric and canted intercalation from the minor groove, as recently reported in high-resolution X-ray structures, is required to appropriately explain the data. By assigning the long emission lifetime to the canted binding geometry, we can simultaneously fit both calorimetric data and the binding-density-dependent changes in the relative abundance of the two emission lifetimes using the same binding model. We find that all complex complex interactions are slightly unfavorable for Delta-[Ru(bpy)(2)dppz](2+), whereas interactions involving a complex canted away from a neighbor are favorable for the other three complexes. We also conclude that Delta-[Ru(bpy)(2)dppz](2+) preferably binds isolated, Delta-[Ru(phen)(2)dppz](2+) preferably binds as duplets of canted complexes, and that all complexes are reluctant to form longer consecutive sequences than triplets. We propose that this is due to an interplay of repulsive complex complex and attractive complex-DNA interactions modulated by allosteric DNA conformation changes that are largely affected by the nature of the ancillary ligands.
2.	Fernandez, Yuri A. Diaz, 1978, et al. (author) The conquest of middle-earth: combining top-down and bottom-up nanofabrication for constructing nanoparticle based devices 2014 In: Nanoscale. - : Royal Society of Chemistry (RSC). - 2040-3372 .- 2040-3364. ; 6:24, s. 14605-14616 Journal article (peer-reviewed)abstract The development of top-down nanofabrication techniques has opened many possibilities for the design and realization of complex devices based on single molecule phenomena such as e. g. single molecule electronic devices. These impressive achievements have been complemented by the fundamental understanding of self-assembly phenomena, leading to bottom-up strategies to obtain hybrid nanomaterials that can be used as building blocks for more complex structures. In this feature article we highlight some relevant published work as well as present new experimental results, illustrating the versatility of self-assembly methods combined with top-down fabrication techniques for solving relevant challenges in modern nanotechnology. We present recent developments on the use of hierarchical self-assembly methods to bridge the gap between sub-nanometer and micrometer length scales. By the use of non-covalent self-assembly methods, we show that we are able to control the positioning of nanoparticles on surfaces, and to address the deterministic assembly of nano-devices with potential applications in plasmonic sensing and single-molecule electronics experiments.
3.	Fornander, Louise, et al. (author) Airway irritation among indoor swimming pool personnel : trichloramine exposure, exhaled NO and protein profiling of nasal lavage fluids 2013 In: International Archives of Occupational and Environmental Health. - : Springer. - 0340-0131 .- 1432-1246. ; 86:5, s. 571-580 Journal article (peer-reviewed)abstract PurposeOccurrence of airway irritation among indoor swimming pool personnel was investigated. The aims of this study were to assess trichloramine exposure levels and exhaled nitric oxide in relation to the prevalence of airway symptoms in swimming pool facilities and to determine protein effects in the upper respiratory tract.MethodsThe presence of airway symptoms related to work was examined in 146 individuals working at 46 indoor swimming pool facilities. Levels of trichloramine, as well as exhaled nitric oxide, were measured in five facilities with high prevalence of airway irritation and four facilities with no airway irritation among the personnel. Nasal lavage fluid was collected, and protein profiles were determined by a proteomic approach.Results17 % of the swimming pool personnel reported airway symptoms related to work. The levels of trichloramine in the swimming pool facilities ranged from 0.04 to 0.36 mg/m3. There was no covariance between trichloramine levels, exhaled nitric oxide and prevalence of airway symptoms. Protein profiling of the nasal lavage fluid showed that the levels alpha-1-antitrypsin and lactoferrin were significantly higher, and S100-A8 was significantly lower in swimming pool personnel.ConclusionsThis study confirms the occurrence of airway irritation among indoor swimming pool personnel. Our results indicate altered levels of innate immunity proteins in the upper airways that may pose as potential biomarkers. However, swimming pool facilities with high prevalence of airway irritation could not be explained by higher trichloramine exposure levels. Further studies are needed to clarify the environmental factors in indoor swimming pools that cause airway problems and affect the immune system.
4.	Fornander, Louise, et al. (author) Airway symptoms and biological markers in nasal lavage fluid in subjects exposed to metalworking fluids 2013 In: PLOS ONE. - : Public Library of Science. - 1932-6203. ; 8:12, s. e83089- Journal article (peer-reviewed)abstract BACKGROUNDS: Occurrence of airway irritation among industrial metal workers was investigated. The aims were to study the association between exposures from water-based metal working fluids (MWF) and the health outcome among the personnel, to assess potential effects on the proteome in nasal mucous membranes, and evaluate preventive actions.METHODS: The prevalence of airway symptoms related to work were examined among 271 metalworkers exposed to MWF and 24 metal workers not exposed to MWF at the same factory. At the same time, air levels of potentially harmful substances (oil mist, morpholine, monoethanolamine, formaldehyde) generated from MWF was measured. Nasal lavage fluid was collected from 13 workers and 15 controls and protein profiles were determined by a proteomic approach.RESULTS: Airway symptoms were reported in 39% of the workers exposed to MWF although the measured levels of MWF substances in the work place air were low. Highest prevalence was found among workers handling the MWF machines but also those working in the same hall were affected. Improvement of the ventilation to reduce MWF exposure lowered the prevalence of airway problems. Protein profiling showed significantly higher levels of S100-A9 and lower levels of SPLUNC1, cystatin SN, Ig J and β2-microglobulin among workers with airway symptoms.CONCLUSIONS: This study confirms that upper airway symptoms among metal workers are a common problem and despite low levels of MWF-generated substances, effects on airway immune proteins are found. Further studies to clarify the role of specific MWF components in connection to airway inflammation and the identified biological markers are warranted.
5.	Fornander, Louise, 1984 (author) Biophysical studies of DNA binding – by the large filament-forming protein Rad51 and the small minor-groove binder Hoechst 33258 2015 Doctoral thesis (other academic/artistic)abstract Mechanistic insight into the nature of DNA-binding ligands is crucial for both drug development as well as understanding more complex biological reactions that take place in the cell. In this Thesis, two rather different DNA-binding molecules are considered: 1. the large, filament-forming eukaryotic recombination protein Rad51, which is essential in the strand exchange reaction during homologous recombination, the most accurate repair system of DNA double‑strand breaks, and 2. the small, synthetic DNA ligand Hoechst 33258, which is a model drug for DNA minor groove interactions. The Rad51 filament formation, reflected in the length of short individual Rad51 filament patches on long DNA strands, has been examined by nanofluidics in combination with fluorescence microscopy. Analyses of the dynamics of the Rad51-DNA complex in the nanochannel reveal structural variations that depend on the filament formation conditions; the choice of divalent cations (Mg2+ or Ca2+), the DNA substrate (single- or double‑stranded), and the Rad51 nucleation concentration affected the macroscopic structure of the filament. The structural effects that the divalent cations Mg2+ and Ca2+, and the accessory protein Swi5-Sfr1 exert on the Rad51-single-stranded (ss) DNA filament at a microscopic level have also been examined by linear dichroism (LD). The naturally unordered bases in ssDNA become preferentially perpendicularly oriented relative to the DNA backbone in presence of Rad51 with Ca2+ alone, or Mg2+ in combination with the accessory protein Swi5-Sfr1. A preferentially perpendicular base organization is proposed to mechanistically relate to an efficient strand exchange reaction, supposedly due to more critical base matching with the invading double‑stranded DNA. To aid future spectroscopic structural analyses of proteins that contain tyrosine residues, such as Rad51, a combined spectroscopic and in silico study of the chromophore in tyrosine has been conducted. It is demonstrated how the spectroscopic properties of tyrosine are sensitively dependent on the polarity of the environment, mainly through the ability to form hydrogen bonds and the rotation of the hydroxyl group. The last part of the Thesis deals with spectroscopic and thermodynamic studies of the binding of Hoechst 33258 to three different DNA oligonucleotides with AT‑tracts of various lengths. The binding at high drug‑to‑DNA ratio is especially considered, and an important conclusion is that two Hoechst 33258 molecules bind in parallel, a slight distance apart, in the minor groove of an oligonucleotide with 8 consecutive adenines/thymines.
6.	Fornander, Louise, 1984, et al. (author) Ca2+ improves organization of single-stranded DNA bases in human Rad51 filament, explaining stimulatory effect on gene recombination 2012 In: Nucleic Acids Research. - : Oxford University Press (OUP). - 1362-4962 .- 0305-1048. ; 40:11, s. 4904-4913 Journal article (peer-reviewed)abstract Human RAD51 protein (HsRad51) catalyses the DNA strand exchange reaction for homologous recombination. To clarify the molecular mechanism of the reaction in vitro being more effective in the presence of Ca2+ than of Mg2+, we have investigated the effect of these ions on the structure of HsRad51 filament complexes with single- and double-stranded DNA, the reaction intermediates. Flow linear dichroism spectroscopy shows that the two ionic conditions induce significantly different structures in the HsRad51/single-stranded DNA complex, while the HsRad51/double-stranded DNA complex does not demonstrate this ionic dependence. In the HsRad51/single-stranded DNA filament, the primary intermediate of the strand exchange reaction, ATP/Ca2+ induces an ordered conformation of DNA, with preferentially perpendicular orientation of nucleobases relative to the filament axis, while the presence of ATP/Mg2+, ADP/Mg2+ or ADP/Ca2+ does not. A high strand exchange activity is observed for the filament formed with ATP/Ca2+, whereas the other filaments exhibit lower activity. Molecular modelling suggests that the structural variation is caused by the divalent cation interfering with the L2 loop close to the DNA-binding site. It is proposed that the larger Ca2+ stabilizes the loop conformation and thereby the protein–DNA interaction. A tight binding of DNA, with bases perpendicularly oriented, could facilitate strand exchange.
7.	Fornander, Louise, 1984 (author) DNA-Complexes with Drugs and Proteins 2012 Licentiate thesis (other academic/artistic)abstract DNA is fundamental for all living cells; the DNA holds the genetic code, which is more or less the instruction book for how all cells are built and function. Several diseases are also linked to DNA, emerging either from a mutation in our genome, which could result in a malfunctioning protein, or that the transcription of genes is somehow affected by structural changes in the DNA, induced by mutations or DNA binding molecules. Research concerning how both small molecules and larger protein assemblies bind to the DNA are therefore of great interest since these could be used as future drugs in for example gene therapy.In the first part of this Thesis the non-covalent binding to DNA of a small minor groove binder, Hoechst 33258, is examined. The molecule is rather well-studied, but there are still questions concerning its multiple binding modes to DNA sequences rich in adenines (A) and thymines (T) that remain unanswered. An increased understanding of the nature of the multiple binding modes could benefit the future design and development of sequence specific drugs. Using the thermodynamic characterization of the binding through Isothermal Titration Calorimetry (ITC) in combination with the spectroscopic properties of the formed complexes through Circular Dichroism (CD) we have analyzed the experimental results in a global dataset. We conclude that two molecules of Hoechst 33258 can bind next to each other in AT-rich sequences that consist of eight AT base pairs, but not in sequences consisting of six or less AT base pairs. They do not bind on top of one another, in the form of a sandwich, as previously proposed, nor contiguously, but with distinct separation between monomeric units.The second part of this Thesis reports how the structure and activity of the human recombination protein RAD51 (HsRad51) depends on presence of cofactors: ATP and divalent cations. The eukaryotic HsRad51 is one of the evolutionarily best-conserved proteins and homologues to it can be found in both Bacteria and Archaea. HsRad51 is involved in the strand exchange reaction of homologous recombination, which takes place during meiosis and repair of double-strand breaks in eukaryotes. With further understanding of the strand exchange reaction we might find ways to utilize it in the medicinal field, such as for correction and repair of defective genes in gene therapy, or as a potential target in cancer treatment. We confirm that the first intermediate of this reaction, in which HsRad51 forms a helical filament around a single strand of DNA, demonstrates a perpendicular organization of the DNA bases relative the filament axis when ATP and Ca2+ are present. This organization is most probably related to the observed high strand exchange activity of the HsRad51/ssDNA complex in with ATP and Ca2+. By contrast, in presence of Mg2+ we observe both poor base organization and strand exchange activity.
8.	Fornander, Louise Helena, et al. (author) Using Nanofluidic Channels to Probe the Dynamics of Rad51-DNA Filaments 2014 In: Biophysical Journal. - : Elsevier BV. - 0006-3495 .- 1542-0086. ; 106:2, s. 692A-693A Journal article (other academic/artistic)abstract Rad51 is a key protein involved in the strand exchange reaction, a reaction where genetic material is transferred between two homologous DNA strands. Strand exchange is initiated by Rad51 forming a helical filament around single-stranded DNA (ssDNA), and the strand exchange is thereafter executed with a homologous double-stranded DNA (dsDNA). The structure of Rad51-DNA filaments, and also the activity of the strand exchange reaction, is dependent on the presence of ATP and dications, where Ca2+ has been shown to promote a higher degree of strand exchange than Mg2+.
9.	Fornander, Louise, et al. (author) Innate immunity proteins and a new truncated form of SPLUNC1 in nasopharyngeal aspirates from infants with respiratory syncytial virus infection 2011 In: PROTEOMICS CLINICAL APPLICATIONS. - : Wiley-VCH Verlag Berlin. - 1862-8346. ; 5:9-10, s. 513-522 Journal article (peer-reviewed)abstract Purpose: Respiratory syncytial virus (RSV) is the most common cause of severe respiratory tract infection in infants. The aim was to identify host defence components in nasopharyngeal aspirate (NPA) from infants with RSV infection and to study the expression of the novel 25 kDa innate immunity protein SPLUNC1. less thanbrgreater than less thanbrgreater thanExperimental design: NPAs from infants were analyzed with 2-DE and MS in a pilot study. The levels of SPLUNC1 were analyzed with immunoblotting in 47 NPAs, admitted for RSV diagnosis. less thanbrgreater than less thanbrgreater thanResults: Totally, 35 proteins were identified in NPA, including several innate immunity proteins such as group X phospholipase A(2), different S100 proteins and SPLUNC1. In addition, a new truncated 15 kDa form of SPLUNC1 was identified that was detected in about 50% of the aspirates admitted for RSV diagnosis. RSV-positive boys had significantly less 25 kDa SPLUNC1 than RSV-negative boys while there were no significant differences among girls. less thanbrgreater than less thanbrgreater thanConclusions and clinical relevance: Several important innate immunity proteins were identified in NPA. Notably, a new truncated form of the newly suggested anti-bacterial protein SPLUNC1 was found. It is possible that a decrease in SPLUNC1 in the upper airways may increase the risk for severe pneumonia in boys.
10.	Fornander, Louise, 1984, et al. (author) Minor-Groove Binding Drugs: Where Is the Second Hoechst 33258 Molecule? 2013 In: Journal of Physical Chemistry B. - : American Chemical Society (ACS). - 1520-6106 .- 1520-5207. ; 117:19, s. 5820-5830 Journal article (peer-reviewed)abstract Hoechst 33258 binds with high affinity into the minor groove of AT-rich sequences of double-helical DNA. Despite extensive studies of this and analogous DNA binding molecules, there still remains uncertainty concerning the interactions when multiple ligand molecules are accommodated within close distance. Albeit not of direct concern for most biomedical applications, which are at low drug concentrations, interaction studies for higher drug binding are important as they can give fundamental insight into binding mechanisms and specificity, including drug self-stacking interactions that can provide base-sequence specificity. Using circular dichroism (CD), isothermal titration calorimetry (ITC), and proton nuclear magnetic resonance (1H NMR), we examine the binding of Hoechst 33258 to three oligonucleotide duplexes containing AT regions of different lengths: [d(CGCGAATTCGCG)]2 (A2T2), [d(CGCAAATTTGCG)]2 (A3T3), and [d(CGAAAATTTTCG)]2 (A4T4). We find similar binding geometries in the minor groove for all oligonucleotides when the ligand-to-duplex ratio is less than 1:1. At higher ratios, a second ligand can be accommodated in the minor groove of A4T4 but not A2T2 or A3T3. We conclude that the binding of the second Hoechst to A4T4 is not cooperative and that the molecules are sitting with a small separation apart, one after the other, and not in a sandwich structure as previously proposed.
11.	Fornander, Louise, 1984, et al. (author) Swi5-Sfr1 protein stimulates Rad51-mediated DNA strand exchange reaction through organization of DNA bases in the presynaptic filament 2014 In: Nucleic Acids Research. - : Oxford University Press (OUP). - 0305-1048 .- 1362-4962. ; 42:4, s. 2358-2365 Journal article (peer-reviewed)abstract The Swi5-Sfr1 heterodimer protein stimulates the Rad51-promoted DNA strand exchange reaction, a crucial step in homologous recombination. To clarify how this accessory protein acts on the strand exchange reaction, we have analyzed how the structure of the primary reaction intermediate, the Rad51/single-stranded DNA (ssDNA) complex filament formed in the presence of ATP, is affected by Swi5-Sfr1. Using flow linear dichroism spectroscopy, we observe that the nucleobases of the ssDNA are more perpendicularly aligned to the filament axis in the presence of Swi5-Sfr1, whereas the bases are more randomly oriented in the absence of Swi5-Sfr1. When using a modified version of the natural protein where the N-terminal part of Sfr1 is deleted, which has no affinity for DNA but maintained ability to stimulate the strand exchange reaction, we still observe the improved perpendicular DNA base orientation. This indicates that Swi5-Sfr1 exerts its activating effect through interaction with the Rad51 filament mainly and not with the DNA. We propose that the role of a coplanar alignment of nucleobases induced by Swi5-Sfr1 in the presynaptic Rad51/ssDNA complex is to facilitate the critical matching with an invading double-stranded DNA, hence stimulating the strand exchange reaction.
12.	Fornander, Louise, 1983- (author) Upper Airway Mucosal Inflammation : Proteomic Studies after Exposure to Irritants and Microbial Agents 2015 Doctoral thesis (other academic/artistic)abstract People are, in their daily lives, exposed to a number of airborne foreign compounds that do not normally affect the body. However, depending on the nature of these compounds, dose and duration of exposure, various airway symptoms may arise. Early symptoms are often manifested as upper airway mucosal inflammation which generates changes in protein composition in the airway lining fluid.This thesis aims at identifying, understanding mechanisms and characterizing protein alterations in the upper airway mucosa that can be used as potential new biomarkers for inflammation in the mucosa. The protein composition in the mucosa was studied by sampling of nasal lavage fluid that was further analyzed with a proteomic approach using twodimensional gel electrophoresis and mass spectrometry. Additionally, by studying factors on site through environmental examination, health questionnaires and biological analyses, we have tried to understand the background to these protein alterations and their impact on health.Respiratory symptoms from the upper airways are common among people who are exposed to irritative and microbial agents. This thesis have focused on personnel in swimming pool facilities exposed to trichloramine, metal industry workers exposed to metalworking fluids, employees working in damp and moldy buildings and infants diagnosed with respiratory syncytial virus infection. The common denominator in these four studies is that the subjects experience upper airway mucosal inflammation, which is manifested as cough, rhinitis, phlegm etc. In the three occupational studies, the symptoms were work related. Notably, a high prevalence of perceived mucosal symptoms was shown despite the relatively low levels of airborne irritants revealed by the environmental examination. Protein profiling verified an ongoing inflammatory response by identification of several proteins that displayed altered levels. Interestingly, innate immune proteins dominated and four protein alterations occurred in most of the studies; SPLUNC1, protein S100A8 and S100A9 and alpha-1-antitrypsin. Similarly, these proteins were also found in nasal fluid from children with virus infection and in addition a truncated form of SPLUNC1 and two other S100 proteins (S100A7-like 2 and S100A16), not previously found in nasal secretion, were identified.Altogether, the results indicate the potential use of a proteomic approach for identifying new biomarkers for the upper respiratory tract at an early stage in the disease process after exposure to irritant and microbial agents. The results indicate an effect on the innate immunity system and the proteins; SPLUNC1, protein S100A8 and S100A9 and alpha-1-antitrypsin are especially promising new biomarkers. Moreover, further studies of these proteins may help us to understand the molecular mechanisms involved in irritant-induced airway inflammation.
13.	Fornander, Louise, 1984, et al. (author) Using nanofluidic channels to probe dynamics of RAD51-Filaments 2015 In: 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2014. - 9780979806476 ; , s. 1826-1828 Conference paper (peer-reviewed)abstract Using nanochannels, passivated with a lipid bilayer to avoid sticking of proteins, we study Rad51 filaments bound to single- and double stranded DNA. We demonstrate how we can discern different properties of the filaments by studying them at different degrees of confinement. Unlike the bacterial homologue RecA, that forms homogeneous filaments along DNA, Rad51 forms heterogeneous filaments containing both rigid kinks as well as flexible regions. Varying the counterion, the DNA substrate as well as the initial protein concentration, we try to understand the factors governing the structure of the filaments.
14.	Fornander, Louise, 1984, et al. (author) Using Nanofluidic Channels to Probe the Dynamics of Rad51-DNA Filaments 2014 In: Biophysical Journal. - 0006-3495 .- 1542-0086. ; 106:2, s. 692A-693A Journal article (other academic/artistic)
15.	Fornander, Louise, 1984, et al. (author) UV Transition Moments of Tyrosine 2014 In: Journal of Physical Chemistry B. - : American Chemical Society (ACS). - 1520-5207 .- 1520-6106. ; 118:31, s. 9247-9257 Journal article (peer-reviewed)abstract To assist polarized-light spectroscopy for protein-structure analysis, the UV spectrum of p-cresol, the chromophore of tyrosine, was studied with respect to transition moment directions and perturbation by solvent environment. From linear dichroism (LD) spectra of p-cresol aligned in stretched matrices of poly(vinyl alcohol) and polyethylene, the lowest pi-pi* transition (L-b) is found to have pure polarization over its entire absorption (250-300 nm) with a transition moment perpendicular to the symmetry axis (C-1-C-4), both in polar and nonpolar environments. For the second transition (L-a), polarized parallel with the symmetry axis, a certain admixture of intensity with orthogonal polarization is noticed, depending on the environment. While the L-b spectrum in cydohexane shows a pronounced vibrational structure, it is blurred in methanol, which can be modeled as due to many microscopic polar environments. With the use of quantum mechanical (QM) calculations, the transition moments and solvent effects were analyzed with the B3LYP and omega B97X-D functionals in cyclohexane, water, and methanol using a combination of implicit and explicit solvent models. The blurred L-b band is explained by solvent hydrogen bonds, where both accepting and donating a hydrogen causes energy shifts. The inhomogeneous solvent-shift sensitivity in combination with robust polarization can be exploited for analyzing tyrosine orientation distributions in protein complexes using LD spectroscopy.
16.	Fornander, Louise, 1984, et al. (author) Visualizing the Nonhomogeneous Structure of RAD51 Filaments Using Nanofluidic Channels 2016 In: Langmuir. - : American Chemical Society (ACS). - 0743-7463 .- 1520-5827. ; 32:33, s. 8403-8412 Journal article (peer-reviewed)abstract RAD51 is the key component of the homologous recombination pathway in eukaryotic cells and performs its task by forming filaments on DNA. In this study we investigate the physical properties of RAD51 filaments formed on DNA using nanofluidic channels and fluorescence microscopy. Contrary to the bacterial ortholog RecA, RAD51 forms inhomogeneous filaments on long DNA in vitro, consisting of several protein patches. We demonstrate that a permanent "kink" in the filament is formed where two patches meet if the stretch of naked DNA between the patches is short. The kinks are readily seen in the present microscopy approach but would be hard to identify using conventional single DNA molecule techniques where the DNA is more stretched. We also demonstrate that protein patches separated by longer stretches of bare DNA roll up on each other and this is visualized as transiently overlapping filaments. RAD51 filaments can be formed at several different conditions, varying the cation (Mg2+ or Ca2+), the DNA substrate (single-stranded or double-stranded), and the RAD51 concentration during filament nucleation, and we compare the properties of the different filaments formed. The results provide important information regarding the physical properties of RAD51 filaments but also demonstrate that nanofluidic channels are perfectly suited to study protein-DNA complexes.
17.	Frykholm, Karolin, 1977, et al. (author) Probing physical properties of DNA-protein complexes using nanofluidic channels 2013 In: 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013; Freiburg; Germany; 27 October 2013 through 31 October 2013. - 9781632666246 ; 2, s. 1311-1313 Conference paper (peer-reviewed)abstract We present the use of nanofluidic channels as a tool for determining physical properties of single DNA-protein complexes. By coating the nanochannels with a lipid bilayer we avoid sticking of proteins to the channel walls. RecA is a prokaryotic protein involved in recombination and DNA repair. We study filaments of RecA, bound to both double stranded (ds) and single stranded (ss) DNA. We determine the persistence length of RecA filaments on both dsDNA and ssDNA and obtain values in agreement with the literature. Neither the DNA nor the protein has to be attached to handles or surfaces, and the technique is directly transferable to Lab-on-a-Chip technologies for high throughput measurements in solution.
18.	Gunnarsson, Peter, et al. (author) Sialic acid residues play a pivotal role in alpha(1)-acid glycoprotein (AGP)-induced generation of reactive oxygen species in chemotactic peptide pre-activated neutrophil granulocytes 2010 In: INFLAMMATION RESEARCH. - : Springer Science and Business Media LLC. - 1023-3830 .- 1420-908X. ; 59:2, s. 89-95 Journal article (peer-reviewed)abstract We have recently shown that terminal sialic acid residues are essential for alpha(1)-acid glycoprotein (AGP)-induced Ca2+ mobilization in neutrophils. The aim of the present study was to establish the importance of sialic acid residues on AGP in modulating human neutrophil functions, with emphasis on the generation of reactive oxygen species (ROS). ROS were measured by luminol-enhanced chemiluminescence in isolated human neutrophils. We found that AGP did not provoke ROS generation in resting or L-selectin presensitized neutrophils. Moreover, AGP did not affect the N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced ROS generation, but it slightly suppressed opsonized zymosan-induced responses. However, when the neutrophils were prestimulated with fMLP, the subsequent addition of AGP provoked a marked ROS response. Dose-response studies and time studies revealed that the ROS generating capacity of AGP was highest at a concentration of 0.05 mg/ml and when given 3-10 min after addition of fMLP. A desialylated form of AGP or pretreatment of neutrophils with 3- and 6-sialyllactose caused a substantially lower ROS response in neutrophils prestimulated with fMLP. Our data show that AGP can stimulate a second ROS response in fMLP preactivated neutrophils and that terminal sialic acid residues on AGP play a crucial role in this regard.
19.	Levels, Johannes HM, et al. (author) High-density Lipoprotein proteome dynamics in human endotoxemia 2011 In: Proteome Science. - : BiOMed Central. - 1477-5956. ; 9:34 Journal article (peer-reviewed)abstract Background: A large variety of proteins involved in inflammation, coagulation, lipid-oxidation and lipid metabolism have been associated with high-density lipoprotein (HDL) and it is anticipated that changes in the HDL proteome have implications for the multiple functions of HDL. Here, SELDI-TOF mass spectrometry (MS) was used to study the dynamic changes of HDL protein composition in a human experimental low-dose endotoxemia model. Ten healthy men with low HDL cholesterol (0.7+/-0.1 mmol/L) and 10 men with high HDL cholesterol levels (1.9+/-0.4mmol/L) were challenged with endotoxin (LPS) intravenously (1ng/kg bodyweight). We previously showed that subjects with low HDL cholesterol are more susceptible to an inflammatory challenge. The current study tested the hypothesis that this discrepancy may be related to differences in the HDL proteome.Results: Plasma drawn at 7 time-points over a 24 hour time period after LPS challenge was used for direct capture of HDL using antibodies against apolipoprotein A-I followed by subsequent SELDI-TOF MS profiling. Upon LPS administration, profound changes in 21 markers (adjusted p-value<0.05) were observed in the proteome in both study groups. These changes were observed 1 hour after LPS infusion and sustained up to 24 hours, but unexpectedly were not different between the 2 study groups. Hierarchical clustering of the protein spectra at all time points of all individuals revealed 3 distinct clusters, which were largely independent of baseline HDL cholesterol levels but correlated with paraoxonase 1 activity. The acute phase protein serum amyloid A-1/2 (SAA-1/2) was clearly upregulated after LPS infusion in both groups and comprised both native and N-terminal truncated variants that were identified by two-dimensional gel electrophoresis and mass spectrometry. Individuals of one of the clusters were distinguished by a lower SAA-1/2 response after LPS challenge and a delayed time-response of the truncated variants.Conclusions: This study shows that the semi-quantitative differences in the HDL proteome as assessed by SELDI-TOF MS cannot explain why subjects with low HDL cholesterol are more susceptible to a challenge with LPS than those with high HDL cholesterol. Instead the results indicate that hierarchical clustering could be useful to predict HDL functionality in acute phase responses towards LPS.
20.	Ljunggren, Stefan, et al. (author) Biomonitoring of Metal Exposure During Additive Manufacturing (3D Printing) 2019 In: SH@W Safety and Health at Work. - : Elsevier. - 2093-7911 .- 2093-7997. ; 10:4, s. 518-526 Journal article (peer-reviewed)abstract Background: Additive manufacturing (AM) is a rapidly expanding new technology involving challenges to occupational health. Here, metal exposure in an AM facility with large-scale metallic component production was investigated during two consecutive years with preventive actions in between.Methods: Gravimetric analyzes measured airborne particle concentrations, and filters were analyzed for metal content. In addition, concentrations of airborne particles <300 nm were investigated. Particles from recycled powder were characterized. Biomonitoring of urine and dermal contamination among AM operators, office personnel, and welders was performed.Results: Total and inhalable dust levels were almost all below occupational exposure limits, but inductively coupled plasma mass spectrometry showed that AM operators had a significant increase in cobalt exposure compared with welders. Airborne particle concentrations (<300 nm) showed transient peaks in the AM facility but were lower than those of the welding facility. Particle characterization of recycled powder showed fragmentation and condensates enriched in volatile metals. Biomonitoring showed a nonsignificant increase in the level of metals in urine in AM operators. Dermal cobalt and a trend for increasing urine metals during Workweek Year 1, but not in Year 2, indicated reduced exposure after preventive actions.Conclusion: Gravimetric analyses showed low total and inhalable dust exposure in AM operators. However, transient emission of smaller particles constitutes exposure risks. Preventive actions implemented by the company reduced the workers' metal exposure despite unchanged emissions of particles, indicating a need for careful design and regulation of the AM environments. It also emphasizes the need for relevant exposure markers and biomonitoring of health risks.
21.	Löfgren, Anton, et al. (author) Nerve Function Impairment after Acute Vibration Exposure 2020 In: Journal of Occupational and Environmental Medicine. - : Lippincott Williams & Wilkins. - 1076-2752 .- 1536-5948. ; 62:2, s. 124-129 Journal article (peer-reviewed)abstract OBJECTIVE: This study was to investigate the acute effects of hand-arm vibrations on the nerve functions of the hands, and the impact of the grip force applied to the vibrating tool during exposure.METHODS: Grip strength and perception of vibration, touch, and temperature were evaluated using QST before and after vibration exposure in 21 occupationally unexposed individuals. The procedure was performed twice, with a higher grip force being applied during exposure on the second occasion.RESULTS: Vibration perception was significantly impaired after both exposures. Grip strength, perception of touch and temperature were only significantly affected after the high grip force exposure.CONCLUSIONS: Exposure to hand-arm vibrations has acute effects on hand nerve function that are sensitive to the grip force applied during exposure.
22.	Montén, Adam, et al. (author) Occupational Quartz Exposure in a Population of Male Individuals-Association With Risk of Developing Atrial Fibrillation 2020 In: Journal of Occupational and Environmental Medicine. - : Lippincott Williams & Wilkins. - 1076-2752 .- 1536-5948. ; 62:6, s. e267-e272 Journal article (peer-reviewed)abstract OBJECTIVE: Occupational quartz exposure is a health risk, with increased risk of developing lung, autoimmune diseases, and elevated mortality in cardiovascular diseases.METHODS: The population was obtained from the period 2005 to 2016 and consisted of 5237 cases of patients with atrial fibrillation (AF). Quartz exposure information was obtained through a Swedish job exposure matrix.RESULTS: The risk of developing AF was increased for the quartz-exposed male population who were within a year of having commenced employment OR 1.54; (95% CI 1.06-2.24); this increased in the age group 20 to 55 (OR 2.05; CI 95% 1.02-4.10).CONCLUSION: Our main conclusion is that quartz dust exposure may be related to increased risk of AF in high exposed (above 0.05 mg/m mean quartz dust) in men aged 20 to 55 years.
23.	Mowitz, Martin, et al. (author) Patch Testing with Isobornyl Acrylate in 16 Swedish Patients with Contact Dermatitis from Glucose Sensors and/or Insulin Pumps 2019 In: Acta Dermato-Venereologica. - : Society for Publication of Acta Dermato-Venereologica. - 0001-5555 .- 1651-2057. ; 99:13, s. 1286-1287 Journal article (other academic/artistic)
24.	Pettersson, Hans, 1980-, et al. (author) Introducing a new design of digital tool to increase vibration risk assessments : challenges with education-based interventions 2022 In: International Journal of Occupational Safety and Ergonomics. - : Taylor & Francis. - 1080-3548 .- 2376-9130. ; 28:3, s. 1705-1710 Journal article (peer-reviewed)abstract Objectives: This study aimed to investigate whether introducing a digital risk assessment tool, the Swedish National Vibration Database, would increase the number of risk assessments on hand–arm and whole-body vibration. Employer and safety representatives from companies where vibration exposure is common were invited.Methods: Of the 2953 invited companies, 1916 were selected for educational intervention and the remaining 1037 companies served as a control group with no intervention. For the educational intervention, participating companies were further divided into two groups (group A, n = 26; group B, n = 47) that both received information regarding risk assessment, but group B was also informed about the digital tool. Both groups answered a questionnaire on risk assessment before the intervention and at the follow-up, 6 months later; the control group received the same questionnaire but no education (group C, n = 22).Results: Of the invited companies, only 2% chose to participate and 7% at follow-up. Seventy-eight percent of the participants had made some kind of risk assessment of vibration at follow-up.Conclusion: Due to the low participation rate among invited companies, this study is not able to draw any conclusions on whether the digital tool can be used to increase the number of risk assessments.
25.	Ragnebro, Oskar, et al. (author) Chloroform exposure in air and water in Swedish indoor swimming pools : urine as a biomarker of occupational exposure 2023 In: Annals of Work Exposures and Health. - : Oxford University Press. - 2398-7308 .- 2398-7316. ; 67:7, s. 876-885 Journal article (peer-reviewed)abstract INTRODUCTION: Disinfection by-products are produced in water disinfected with chlorine-based products. One such group is trihalomethanes, and chloroform is the most abundant trihalomethane in swimming pool areas. Chloroform can be absorbed by inhalation, ingestion, and dermal absorption, and is classified as possibly carcinogenic.AIM: To investigate if chloroform concentrations in air and water affect the chloroform concentration in urine samples of exposed swimming pool workers. METHODS: Workers from 5 adventure indoor swimming pools carried personal chloroform air samplers and provided up to 4 urine samples during one workday. Chloroform concentrations were analyzed with a linear mixed model analysis to investigate a possible correlation between air and urine concentrations.RESULTS: The geometric mean chloroform concentration was 11 μg/m3 in air and 0.009 µg/g creatinine in urine among individuals with ≤2 h at work, 0.023 µg/g creatinine among those with >2-5 working hours, and 0.026 µg/g creatinine in the group with >5-10 working hours. A risk of higher levels of chloroform in urine was associated with longer hours at work (≤2 h versus >5-10 h, odds ratio [OR] 2.04, 95% confidence interval [CI] 1.25-3.34), personal chloroform concentrations in air (≤17.00 µg/m3 versus >28.00 µg/m3, OR 9.23, 95% CI 3.68-23.13) and working at least half the working day near the swimming pools (OR 3.16, 95% CI 1.33-7.55). Executing work tasks in the swimming pool water was not associated with higher chloroform concentrations in urine compared to only working on land (OR 0.82, 95% CI 0.27-2.45).CONCLUSION: There is an accumulation of chloroform concentrations in urine during a workday and a correlation between personal air and urine concentrations of chloroform among workers in Swedish indoor swimming pools.
26.	Sun, Lu, 1982, et al. (author) Sensing Conformational Changes in DNA upon Ligand Binding Using QCM-D. Polyamine Condensation and Rad51 Extension of DNA Layers 2014 In: Journal of Physical Chemistry B. - : American Chemical Society (ACS). - 1520-5207 .- 1520-6106. ; 118:41, s. 11895-11904 Journal article (peer-reviewed)abstract Biosensors, in which binding of ligands is detected through changes in the optical or electrochemical properties of a DNA layer confined to the sensor surface, are important tools for investigating DNA interactions. Here, we investigate if conformational changes induced in surface-attached DNA molecules upon ligand binding can be monitored by the quartz crystal microbalance with dissipation (QCM-D) technique. DNA duplexes containing 59–184 base pairs were formed on QCM-D crystals by stepwise assembly of synthetic oligonucleotides of designed base sequences. The DNA films were exposed to the cationic polyamines spermidine and spermine, known to condense DNA molecules in bulk experiments, or to the recombination protein Rad51, known to extend the DNA helix. The binding and dissociation of the ligands to the DNA films were monitored in real time by measurements of the shifts in resonance frequency (Δf) and in dissipation (ΔD). The QCM-D data were analyzed using a Voigt-based model for the viscoelastic properties of polymer films in order to evaluate how the ligands affect thickness and shear viscosity of the DNA layer. Binding of spermine shrinks all DNA layers and increases their viscosity in a reversible fashion, and so does spermidine, but to a smaller extent, in agreement with its lower positive charge. SPR was used to measure the amount of bound polyamines, and when combined with QCM-D, the data indicate that the layer condensation leads to a small release of water from the highly hydrated DNA films. The binding of Rad51 increases the effective layer thickness of a 59bp film, more than expected from the know 50% DNA helix extension. The combined results provide guidelines for a QCM-D biosensor based on ligand-induced structural changes in DNA films. The QCM-D approach provides high discrimination between ligands affecting the thickness and the structural properties of the DNA layer differently. The reversibility of the film deformation allows comparative studies of two or more analytes using the same DNA layer as demonstrated here by spermine and spermidine.
27.	Thorneus, Erik, et al. (author) Occupational Exposure to Metalworking Fluid and the Effect on Health Symptoms-An Intervention Study 2021 In: Journal of Occupational and Environmental Medicine. - : LIPPINCOTT WILLIAMS & WILKINS. - 1076-2752 .- 1536-5948. ; 63:10, s. E667-E672 Journal article (peer-reviewed)abstract Exposure to metalworking fluid has been shown to cause health problems among workers. The aim of this study was to compare health outcomes and levels of exposure among workers exposed to metalworking fluid before and after the implementation of preventive measures. The frequencies of self-reported symptoms, as well as the concentrations of measured substances were lower after the implementation of preventive measures. Logistic regression showed statistically significant differences in the report of irritations, a stuffy or runny nose, eye irritation, a hoarse or dry throat, and a cough with odds ratios of 0.31 (95% confidence interval [CI] 0.21 to 0.47), 0.12 (95% CI 0.05 to 0.29), 0.13 (95% CI 0.06 to 0.28), and 0.24 (95% CI 0.12 to 0.46), respectively. This confirms the efficiency of the performed intervention and highlights the importance of eliminating the recirculation of contaminated air.
28.	Wallden, Albin, et al. (author) Risks of developing ulcerative colitis and Crohn's disease in relation to silica dust exposure in Sweden : a case-control study 2020 In: BMJ Open. - : BMJ Publishing Group Ltd. - 2044-6055. ; 10:2 Journal article (peer-reviewed)abstract Objective: To determine whether occupational exposure to silica dust causes an increased risk of developing Crohn's disease (CD) and ulcerative colitis (UC).Design: Case-control study of CD (K50) and UC (K51) from 2007 through 2016. Controls were matched to cases (2:1) based on age, sex and county at the time of diagnosis. A job exposure matrix was used to estimate the occupational silica exposure of all cases and controls.Setting: Medical and occupational data from the National Outpatient Register were used to implement a case-control analysis, while the two controls used for each case were selected from the National Register of the Total Population.Participants: All men and women aged 20-65 years old who were diagnosed with CD (K50) and UC (K51) during the years of study were included and assigned two controls, resulting in 58 136 cases and 116 272 controls.Main outcomes: Silica dust exposure correlates with an increased risk of developing UC in men and CD in women.Results: The prevalence of UC was significantly higher in the group exposed to silica dust (OR 1.13, 95% CI 1.06 to 1.21) than in controls, particularly in individuals with over 5 years exposure. When stratified by sex, a significantly increased OR was detected for men (OR 1.33, 95% CI 1.05 to 1.22). This trend was also consistent with longer exposure times. The prevalence of UC was not increased in exposed women. The prevalence of CD was significantly increased among exposed women (OR 1.29, 95% CI 1.01 to 1.65), but not for exposed men.Conclusions: Silica dust exposure correlates with an increased risk of developing UC, especially in men, and the risk seems to increase with the duration and degree of exposure. Conversely, silica dust exposure correlates positively with the risk of developing CD in women.
29.	Westerlund, Jessica, 1983- (author) Exposure to trichloramine, trihalomethanes and endotoxins : adverse respiratory and ocular effects among Swedish indoor swimming pool workers 2022 Doctoral thesis (other academic/artistic)abstract Indoor swimming pool facilities often use chlorine for pool water disinfection. Chlorine can also contribute to the formation of unwanted disinfection byproducts (DBPs). Trichloramine and trihalomethanes (THMs) are DBPs formed in swimming pool water and swimming pool air causing occupational exposure. Trichloramine can cause ocular and airway irritation, while some THMs are potentially carcinogenic. Bacterial endotoxins may also cause adverse respiratory effects among swimming pool workers. The overall aim of the study was to measure levels of trichloramine, THMs and endotoxins in the air at conventional, habilitation and rehabilitation, and adventure swimming pool facilities, and to investigate adverse ocular and respiratory effects among indoor swimming pool workers.Trichloramine and THM levels in air were comparable to previous studies. Endotoxin levels in air were low compared to the reference value. Trichloramine concentrations in personal samples were approximately half as high as those measured in stationary samples - an important consideration for a future Swedish occupational exposure limit. The adventure facilities had the highest trichloramine levels while the conventional facilities had the highest THM levels. The adventure workers also reported more ocular and respiratory symptoms compared to referents (office workers) and had a lower FEV1/FVC ratio indicating airway obstruction. Workers both in conventional, and in habilitation and rehabilitation facilities exhibited a difference in FeNO levels compared to referents, implying airway inflammation.The occupational exposure and adverse health effects found in indoor swimming pool environments emphasizes the importance of a personal based Swedish occupational exposure limit for trichloramine in air.
30.	Westerlund, Jessica, 1983-, et al. (author) Occupational Exposure to Trichloramine and Endotoxins : Adverse Health Effects Among Personnel in Adventure and Rehabilitation Swimming Pool Facilities 2022 In: Journal of Occupational and Environmental Medicine. - : Lippincott Williams & Wilkins. - 1076-2752 .- 1536-5948. ; 64:5, s. 361-369 Journal article (peer-reviewed)abstract OBJECTIVE: To study occupational exposure to trichloramine and endotoxins in air at adventure and rehabilitation swimming pool facilities from an adverse health effects perspective.METHODS: Air concentrations of trichloramine and endotoxins were measured in five adventure and 10 rehabilitation facilities. Respiratory and ocular symptoms were self-reported, and spirometry and fraction of exhaled nitric oxide (FENO) were measured.RESULTS: Compared to rehabilitation facilities, the mean trichloramine concentrations in the adventure facilities were higher, both personal (80 μg/m3 (n = 41) vs 19 μg/m3 (n = 21)) and stationary (183 μg/m3 (n = 51) vs 23 μg/m3 (n = 32)), with higher frequency of ocular and respiratory symptoms. Low stationary endotoxin levels (<0.64 to 25 EU/m3) were found, compared to the reference value (90 EU/m3).CONCLUSIONS: Higher trichloramine concentrations in air and more ocular and respiratory symptoms in adventure facilities call for adequate occupational exposure limits.
31.	Westerlund, Jessica, 1983-, et al. (author) Occupational exposure to trichloramine and endotoxins : Adverse health effects among personnel in adventure and rehabilitation swimming pool facilities Other publication (other academic/artistic)
32.	Wrangel, Oscar, et al. (author) Silica Dust Exposure Increases Risk for Rheumatoid Arthritis : A Swedish National Registry Case-Control Study 2021 In: Journal of Occupational and Environmental Medicine. - : Lippincott Williams & Wilkins. - 1076-2752 .- 1536-5948. ; 63:11, s. 951-955 Journal article (peer-reviewed)abstract OBJECTIVE: Rheumatoid arthritis (RA) is an inflammatory disease with unknown etiology. This study examines if silica dust exposure increases the risk for seropositive and seronegative RA.METHODS: A nationwide registry case-control study was conducted that included all cases of RA in Sweden between 2005 and 2016. In total, 31,139 cases with two matched controls were included. A JEM was used to estimate exposure.RESULTS: Silica dust exposure was associated with a statistically significant increase in odds ratio (OR) for seropositive (OR 1.22, 95% CI 1.05 to 1.40) and seronegative (OR 1.23, 95% CI 1.04 to 1.46) RA among men.CONCLUSION: This study found an increased OR for RA in silica-exposed men. The OR was equal for seropositive and seronegative RA. These findings further support the hypothesis that silica dust may be a trigger for RA.
33.	Wåhlén, Karin, et al. (author) Protein profiles of nasal lavage fluid from individuals with work-related upper airway symptoms associated with moldy and damp buildings 2016 In: Indoor Air. - Hoboken, USA : Wiley-Blackwell Publishing Inc.. - 0905-6947 .- 1600-0668. ; 26:5, s. 743-754 Journal article (peer-reviewed)abstract Upper airway irritation is common among individuals working in moldy and damp buildings. The aim of this study was to investigate effects on the protein composition of the nasal lining fluid. The prevalence of symptoms in relation to work environment was examined in 37 individuals working in two damp buildings. Microbial growth was confirmed in one of the buildings. Nasal lavage fluid was collected from 29 of the exposed subjects and 13 controls, not working in a damp building. Protein profiles were investigated with a proteomic approach and evaluated by multivariate statistical models. Subjects from both workplaces reported upper airway and ocular symptoms. Based on protein profiles, symptomatic subjects in the two workplaces were discriminated from each other and separated from healthy controls. The groups differed in proteins involved in inflammation and host defense. Measurements of innate immunity proteins showed a significant increase in protein S100-A8 and decrease in SPLUNC1 in subjects from one workplace, while alpha-1-antitrypsin was elevated in subjects from the other workplace, compared with healthy controls. The results show that protein profiles in nasal lavage fluid can be used to monitor airway mucosal effects in personnel working in damp buildings and indicate that the profile may be separated when the dampness is associated with the presence of molds.
34.	Åmand, Helene, 1983, et al. (author) Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans 2012 In: Biochimica et Biophysica Acta - Biomembranes. - : Elsevier BV. - 1879-2642 .- 0005-2736. ; 1818:11, s. 2669-2678 Journal article (peer-reviewed)abstract Cell surface proteoglycans (PGs) appear to promote uptake of arginine-rich cell-penetrating peptides (CPPs), but their exact functions are unclear. To address if there is specificity in the interactions of arginines and PGs leading to improved internalization, we used flow cytometry to examine uptake in relation to cell surface binding for penetratin and two arginine/lysine substituted variants (PenArg and PenLys) in wildtype CHO-K1 and PG-deficient A745 cells. All peptides were more efficiently internalized into CHO-K1 than into A745. but their cell surface binding was independent of cell type. Thus, PGs promote internalization of cationic peptides, irrespective of the chemical nature of their positive charges. Uptake of each peptide was linearly dependent on its cell surface binding, and affinity is thus important for efficiency. However, the gradients of these linear dependencies varied significantly. Thus each peptide's ability to stimulate uptake once bound to the cell surface is reliant on formation of specific uptake-promoting interactions. Heparin affinity chromatography and clustering experiments showed that penetratin and PenArg binding to sulfated sugars is stabilized by hydrophobic interactions and result in clustering, whereas PenLys only interacts through electrostatic attraction. This may have implications for the molecular mechanisms behind arginine-specific uptake stimulation as penetratin and PenArg are more efficiently internalized than PenLys upon interaction with PGs. However, PenArg is also least affected by removal of PGs. This indicates that an increased arginine content not only improve PG-dependent uptake but also that PenArg is more adaptable as it can use several portals of entry into the cell.

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