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  • 1.
    Awad, Kassem
    Malmö University, Faculty of Health and Society (HS).
    Effects of skin care ingredients on keratinocytes: - Interplay between osmotic stress, cell viability, and gene expression towards increased understanding of keratinocyte differentiation2021Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    The epidermis is composed of multiple cell strata where viable keratinocytes, in the basal layer (stratum basale (SB)), go through a range of steps with the final stage of being dead corneocytes in the outer most layer (stratum corneum (SC)). The differentiation, which can be thought of as programmed cell death, include several key processes that are essential for an intact skin barrier. The route from SB to SC is accompanied by changes, such as osmotic pressure and pH, that are believed to trigger some of these processes. In this project, HaCaT cells were incubated with, commonly used, skin care substances (urea, glycerol, transcutol and salicylic acid) to assess their impact on cell viability, by MTT-assay, and gene expression, by qPCR. Further, the relationship between osmotic pressure, viability and gene expression was studied. The excipients showed a dose-dependent decrease of keratinocyte viability which also was explained by elevated osmotic pressure when concentration was increased. Exceptions were however observed for transcutol, which showed protective features against osmotic stress. Upregulation of the genes were mainly observed when cells were treated with high concentrations. Involucrin was affected by the substances to a greater extent when compared to other markers. The upregulation of involucrin was however seen to be driven by the osmotic pressure rather than biological effects of the molecules. The project conclude that the viability and gene expression of the keratinocytes are highly related to the osmotic pressure and probably influences the differentiation to a greater extent than the molecules themselves.

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  • 2.
    Barrantes, Alejandro
    et al.
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Camero, Sergio
    Garcia-Lucas, Angel
    Navarro, Pedro J.
    Benitez, Maria J.
    Jimenez, Juan S.
    Alzheimer’s Disease Amyloid Peptides Interact with DNA, As Proved by Surface Plasmon Resonance2012In: Current Alzheimer Research, ISSN 1567-2050, E-ISSN 1875-5828, Vol. 9, no 8, p. 924-934Article in journal (Refereed)
    Abstract [en]

    According to the amyloid hypothesis, abnormal processing of the β-amyloid precursor protein in Alzheimer's disease patients increases the production of β-amyloid toxic peptides, which, after forming highly aggregated fibrillar structures, lead to extracellular plaques formation, neuronal loss and dementia. However, a great deal of evidence has point to intracellular small oligomers of amyloid peptides, probably transient intermediates in the process of fibrillar structures formation, as the most toxic species. In order to study the amyloid-DNA interaction, we have selected here three different forms of the amyloid peptide: Aβ1-40, Aβ25-35 and a scrambled form of Aβ25-35. Surface Plasmon Resonance was used together with UV-visible spectroscopy, Electrophoresis and Electronic Microscopy to carry out this study. Our results prove that, similarly to the full length Aβ1-42, all conformations of toxic amyloid peptides, Aβ1-40 and Aβ25-35, may bind DNA. In contrast, the scrambled form of Aβ25-35, a non-aggregating and nontoxic form of this peptide, could not bind DNA. We conclude that although the amyloid-DNA interaction is closely related to the amyloid aggregation proneness, this cannot be the only factor which determines the interaction, since small oligomers of amyloid peptides may also bind DNA if their predominant negatively charged amino acid residues are previously neutralized.

  • 3.
    Bertelsen, Sissel Asser
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Hormonal and stress-related effects in men aged 25-35, from daily intake of Lactobacillus probiotic dietary supplements2021Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    Background: There is a bidirectional communication system between the gut-microbiota and the brain. Probiotics are live microorganisms that should have health benefits on the host. Modulations of the gut-microbiota using probiotics to target inflammation, stress and depression have shown promising results in rodent studies and are now to be proven in human subjects. 

    Method: 20 male participants aged 25-35, were aliquoted into three groups. All groups consumed placebo tablets for two weeks. For additionally six weeks, two of the groups were administered different commercially available probiotic tablets containing Lactobacillus reuteri and the last group continued the placebo tablets. Venous blood samples and a questionnaire about general wellbeing were collected at baseline, after two weeks, and at the end of the eight weeks study. Cortisol was analysed by electrochemiluminescence immunoassay (ECLIA). An enzymatic colorimetric test was used to determine glucose concentrations (GOD-PAP). Soluble urokinase plasminogen activator receptor (suPAR) concentrations determined with ELISA. Oxidative stress was carried out using Free Oxygen Radicals Testing (FORT).

    Results: For all biomarkers no significant differences were found (p>0.05). For the questionnaires, a significant difference was found in the placebo group. Here, a general wellbeing improved throughout the study. 

    Conclusion: The present study was affected by the COVID-19 pandemic, which led to a decrease in number of participants, change of environment and sampling at different times of the day which could have had an impact on diurnal variation. Although none of the biomarkers showed statistically significant results, further investigations of Lactobacillus reuteri in human subjects are necessary. In addition, this study highlights the importance of developing a robust model for translating findings in rodent studies to healthy human subjects.

  • 4.
    Cabaleiro-Lago, Celia
    et al.
    Department of Bioanalysis, Faculty of Natural Sciences, Kristianstad University.
    Hasterok, Sylwia
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Gjörloff Wingren, Anette
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces. Department of Bioanalysis, Faculty of Natural Sciences, Kristianstad University.
    Tassidis, Helena
    Department of Bioanalysis, Faculty of Natural Sciences, Kristianstad University.
    Recent Advances in Molecularly Imprinted Polymers and Their Disease-Related Applications2023In: Polymers, E-ISSN 2073-4360, Vol. 15, no 21, p. 4199-4199Article, review/survey (Refereed)
    Abstract [en]

    Molecularly imprinted polymers (MIPs) and the imprinting technique provide polymeric material with recognition elements similar to natural antibodies. The template of choice (i.e., the antigen) can be almost any type of smaller or larger molecule, protein, or even tissue. There are various formats of MIPs developed for different medical purposes, such as targeting, imaging, assay diagnostics, and biomarker detection. Biologically applied MIPs are widely used and currently developed for medical applications, and targeting the antigen with MIPs can also help in personalized medicine. The synthetic recognition sites of the MIPs can be tailor-made to function as analytics, diagnostics, and drug delivery systems. This review will cover the promising clinical applications of different MIP systems recently developed for disease diagnosis and treatment.

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  • 5.
    Dabatos, Kyle Bhram
    Malmö University, Faculty of Health and Society (HS).
    UTVÄRDERING AV MALDI-TOF MS FÖR ARTBESTÄMNING AV NYA AEROCOCCUS-ARTER: KARTLÄGGNING AV DESS FÖREKOMST I URIN OCH BLOD I SKÅNE2024Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Aerococcus spp. are Gram-positive bacteria considered as human pathogens, historically challenging to identify due to their similarity with other bacteria. Previous research relied on 16S rRNA sequencing for species identification, until the revolutionary introduction of MALDI-TOF MS. Choi et al. utilized this method in 2023, combined with comprehensive whole genome analyses, and demonstrated that Aerococcus urinae can be classified into four taxonomic groups: Aerococcus tenax, Aerococcus mictus, Aerococcus loyalae, and Aerococcus urinae. The aim of this study was to evaluate the new MALDI-library for these species and map their occurrence in urine and blood in Skåne. This was made possible by 223 Aerococcus-positive strains from urine cultures and 130 from blood cultures, with a total of 276 previously called A. urinae strains analysed. The methodology comprised three main phases: establishing PCR as the reference method, conducting a pilot study running MALDI-TOF MS and PCR simultaneously, and performing a PCR screening to enhance the detection of new Aerococcus species. The results revealed that out of 276 strains, only 37 was successfully identified with PCR, with results matching that of MALDI-TOF MS (A. mictus n=15, A. loyalae n=20, A. urinae n=2). The study also indicated a potential difference in the occurrence of Aerococcus species between blood and urine, where A. mictus dominated both, followed by A. sangunicola and A. loyalae. In conclusion, the new MALDI-library cannot be reliably evaluated due to a lack of well-characterized strains and limitations of the reference method. Nevertheless, the study provides a basis for better evaluation of MALDI-TOF MS for Aerococcus identification.

  • 6.
    Dalla-Riva, Jonathan
    et al.
    Department of Experimental Medical Science, Lund University, Lund, Sweden .
    Lagerstedt, Jens O.
    Department of Experimental Medical Science, Lund University, Lund, Sweden .
    Petrlova, Jitka
    Department of Experimental Medical Science, Lund University, Lund, Sweden .
    Structural and Functional Analysis of the ApolipoproteinA-I A164S Variant2015In: PLOS ONE, E-ISSN 1932-6203, Vol. 10, no 11, p. e0143915-e0143915Article in journal (Refereed)
    Abstract [en]

    There is a clinical need for conceptually new treatments that target the excessive activation of inflammatory pathways during systemic infection. Thrombin-derived C-terminal peptides (TCPs) are endogenous anti-infective immunomodulators interfering with CD14-mediated TLR-dependent immune responses. Here we describe the development of a peptide-based compound for systemic use, sHVF18, expressing the evolutionarily conserved innate structural fold of natural TCPs. Using a combination of structure- and in silico-based design, nuclear magnetic resonance spectroscopy, biophysics, mass spectrometry, cellular, and in vivo studies, we here elucidate the structure, CD14 interactions, protease stability, transcriptome profiling, and therapeutic efficacy of sHVF18. The designed peptide displays a conformationally stabilized, protease resistant active innate fold and targets the LPS-binding groove of CD14. In vivo, it shows therapeutic efficacy in experimental models of endotoxin shock in mice and pigs and increases survival in mouse models of systemic polymicrobial infection. The results provide a drug class based on Nature ' s own anti-infective principles.

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  • 7.
    Danielsson, Ravi
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Eriksson, Håkan
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Aluminium adjuvants in vaccines: A way to modulate the immune response2021In: Seminars in Cell and Developmental Biology, ISSN 1084-9521, E-ISSN 1096-3634, Vol. 115, p. 3-9, article id S1084-9521(20)30202-0Article in journal (Refereed)
    Abstract [en]

    Aluminium salts have been used as adjuvants in vaccines for almost a century, but still no clear understanding of the mechanisms behind the immune stimulating properties of aluminium based adjuvants is recognized. Aluminium adjuvants consist of aggregates and upon administration of a vaccine, the aggregates will be recognized and phagocytosed by sentinel cells such as macrophages or dendritic cells. The adjuvant aggregates will persist intracellularly, maintaining a saturated intracellular concentration of aluminium ions over an extended time. Macrophages and dendritic cells are pivotal cells of the innate immune system, linking the innate and adaptive immune systems, and become inflammatory and antigen-presenting upon activation, thus mediating the initiation of the adaptive immune system. Both types of cell are highly adaptable, and this review will discuss and highlight how the occurrence of intracellular aluminium ions over an extended time may induce the polarization of macrophages into inflammatory and antigen presenting M1 macrophages by affecting the: endosomal pH; formation of reactive oxygen species (ROS); stability of the phagosomal membrane; release of damage associated molecular patterns (DAMPs); and metabolism (metabolic re-programming). This review emphasizes that a persistent intracellular presence of aluminium ions over an extended time has the potential to affect the functionality of sentinel cells of the innate immune system, inducing polarization and activation. The immune stimulating properties of aluminium adjuvants is presumably mediated by several discrete events, however, a persistent intracellular presence of aluminium ions appears to be a key factor regarding the immune stimulating properties of aluminium based adjuvants.

  • 8.
    Hallan, Supandeep Singh
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces. Department of Chemical and Pharmaceutical Sciences, University of Ferrara, I-44121 Ferrara, Italy.
    Sguizzato, Maddalena
    Department of Chemical and Pharmaceutical Sciences, University of Ferrara, I-44121 Ferrara, Italy.
    Mariani, Paolo
    Department of Life and Environmental Sciences, Polytechnic University of Marche, I-60131 Ancona, Italy.
    Cortesi, Rita
    Department of Chemical and Pharmaceutical Sciences, University of Ferrara, I-44121 Ferrara, Italy.
    Huang, Nicolas
    CNRS, Institut Galien Paris-Saclay, Université Paris-Saclay, 92296 Châtenay-Malabry, France.
    Simelière, Fanny
    CNRS, Institut Galien Paris-Saclay, Université Paris-Saclay, 92296 Châtenay-Malabry, France.
    Marchetti, Nicola
    Department of Chemical and Pharmaceutical Sciences, University of Ferrara, I-44121 Ferrara, Italy.
    Drechsler, Markus
    Bavarian Polymer Institute (BPI) Keylab "Electron and Optical Microscopy" University of Bayreuth, D-95440 Bayreuth, Germany.
    Ruzgas, Tautgirdas
    Malmö University, Biofilms Research Center for Biointerfaces. Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Esposito, Elisabetta
    Department of Chemical and Pharmaceutical Sciences, University of Ferrara, I-44121 Ferrara, Italy.
    Design and Characterization of Ethosomes for Transdermal Delivery of Caffeic Acid.2020In: Pharmaceutics, E-ISSN 1999-4923, Vol. 12, no 8, article id E740Article in journal (Refereed)
    Abstract [en]

    in ethosome after six months, while in water, an almost complete degradation occurred within one month. The addition of poloxamer slightly modified vesicle structure and size, while it decreased the vesicle deformability. Caffeic acid diffusion coefficients from ethosome and ethosome gel were, respectively, 137- and 33-fold lower with respect to the aqueous solution. At last, the caffeic acid permeation and antioxidant power of ethosome were more intense with respect to the simple solution.

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  • 9. Miletic Dahlström, Ruzica
    TOPICAL SAMPLING OF POTENTIAL SKIN CANCER BIOMARKERS, KYNURENINE AND TRYPTOPHAN: STUDY ON A 3D MELANOMA MODEL2022Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    Background: Malignant melanoma continues to be one of the deadliest forms of skin cancer. Despite immense international efforts, it remains a major clinical challenge. To date, the golden standard for skin cancer diagnosis is through visual inspection and biopsies. UVB (280-320 nm) and IFN-γ have been proven to induce IDO-1 expression in tumor cells, increasing the ratio between tryptophan (Trp) and kynurenine (Kyn) in the tumor microenvironment. Non-invasive sampling of kyn and trp could thereby serve as an alternative for skin cancer diagnostics. 

    Objective: This study aimed to investigate the clinical relevance of monitoring trp and kyn by stimulation of skin cancer development in a 3D melanoma model.

    Methods: Polystyrene scaffolds were used to create 3D melanoma and melanocyte models respectively. Monolayers were used to examine keratinocytes, fibroblasts, melanoma cells, and melanocytes' response to stimuli. After treatment with UVB and IFN-g, the release of cytokines was measured with ELISA, and gene expression was analyzed with quantitative reverse transcription PCR. Secreted trp and kyn were quantified by high-performance liquid chromatography. The 3D models were then sectioned into 3 µm thick pieces for histological analysis and immunohistochemistry staining.

    Results: Significantly increased gene expression of IDO-1 was seen in all monolayers, including the 3D models stimulated with IFN-g. Released IL-6 was induced after UVB exposure in the 3D models, and IL-1a was only detected in the melanocyte model. Both models showed an increase in kyn levels after stimulation with IFN-g. No induced IDO-1 gene expression could be detected in the models after UVB exposure, and no significant change in Trp/Kyn ratio was detected in those samples. 

    Conclusion: Our results suggest that IFN-g induces IDO-1 gene expression in 3D models, leading to an increased trp/kyn ratio. Topical sampling of kyn and trp may be possible as an alternative non-invasive method to present diagnostics. UVB seemed to induce inflammation in the 3D models, but no significant signs of malignant transformation.

  • 10.
    Startaite, Lauryna
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Effects of Buffer Composition on DNase I Formulation in Disordered Mesoporous Silica Particles2024Independent thesis Advanced level (degree of Master (Two Years)), 80 credits / 120 HE creditsStudent thesis
    Abstract [en]

    Cystic fibrosis, a genetic disorder affecting multiple organs in the body, including the lungs, remains a significant threat to patients due to inadequate treatment options. Treatment includes aerosolized deoxyribonuclease I which bolsters pulmonary function and improves affected patient condition. However, taking the liquid formulations requires prolonged inhalation times and nebulization equipment. Conversely, dry powder inhalers are handheld devices, delivering fine particles deep into the lung on a single inhalation. Dry formulations may be enhanced through the use of mesoporous silica particles which have an optimal size for inhalation, are light in weight and have a large surface area. Loading deoxyribonuclease I into mesoporous silica particles could potentially improve drug delivery to cystic fibrosis patients with reduced administration frequency when taken with dry powder inhalers. The incorporation of buffers into this system is crucial for ensuring efficient drug loading and stability at the biointerface during dry powder preparation. Thus, the objective of this project was to ascertain the most suitable buffer composition for loading deoxyribonuclease I into mesoporous silica particles. Protein size and activity were evaluated in different buffers prior to adsorption. Subsequently, dry formulations were prepared by freeze drying, and studied by thermogravimetric analysis and dynamic vapour sorption. Cumulative release analysis in simulated lung fluid was performed, followed by released protein enzymatic activity evaluation. Findings indicated the necessity of incorporating Ca2+ into buffers to increase protein loading efficiency and stability in dry formulations. Highest level of adsorption, and adequate remaining deoxyribonuclease I activity was observed in formulations prepared with calcium doped mesoporous silica particles in pH 5.0 50 mM sodium acetate buffer with added 5 mM CaCl2.

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  • 11.
    Vasiliu, Alina
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Interactions of constituents of topical formulations with skin microbiota: Effects of propylene glycol on relevant skin microbiota isolates2023Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    One of the lesser explored research areas is the influence factors such as personal care products, cosmetics, and everyday routines have on skin microbiota. This study investigated the effects of propylene glycol, a widely used ingredient in cosmetics and self-care products, on a staphylococcal system ubiquitously distributed on human skin. The system, comprised of Staphylococcus hominis, Staphylococcus epidermidis, and Staphylococcus aureus, comes from a healthy donor, devoid of any skin afflictions. Because Staphylococcus aureus is part of this microbial community, it was of great interest to contribute to the understanding of the manner in which its growth is kept in check.

    To fulfill this task, a new methodology was developed. Purposely intended to be facile and easily scalable in laboratories around the world, it can be used for the study of microbial systems of variable dimensions alone or with the complementary use of other methods.

    Results indicate that the effects of propylene glycol are complex, as it acts on the skin, the resident microbiota, and at the microbiota-skin interface. Commensals such as Staphylococcus hominis and Staphylococcus epidermidis seem to have synergy of action with propylene glycol, increasing each other’s power in reducing the number of viable colonies of Staphylococcus aureus. Lastly, results seem to also reveal the incompletely understood role of Staphylococcus hominis on human skin. While Staphylococcus epidermidis and Staphylococcus aureus ravenously compete with each other, it is the contribution of Staphylococcus hominis that seems to limit the latter’s overgrowing. This speaks volumes of the extent, complexities, and unknowns of microbial interactions.

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  • 12.
    Wakil, Bashir
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces. x.
    OPERATION OF AN ELECTROCHEMICAL BIOSENSOR DEVELOPED FOR COVID-19 DETECTIONIN SARS-COV-2 FREE AND INFECTED HUMAN SALIVA2022Independent thesis Advanced level (degree of Master (Two Years)), 80 credits / 120 HE creditsStudent thesis
    Abstract [en]

    The demand for the improvement of currently available tests for qualitative non-invasive diagnostic of COVID-19, i.e., the development of new methods for fast, low-cost and accurate tests for the conformation of SARS-CoV-2, is increasing rapidly. Among many different approaches, electrochemical biosensors, which have the capability of miniaturization and could be available globally in most remote areas, may also help in avoiding the transmission of COVID-19 disease. When properly designed, electrochemical tests might have higher sensitivity, specificity, and accuracy, which is very important for COVID-19 diagnostics. In this work a saliva based electrochemical biosensor developed for COVID-19 detection was tested using real human samples. First, 41 saliva samples from volunteers were collected during January-February 2022, when the rate of SARS-CoV-2 infection was the highest in Skåne region, Sweden. Second, cyclic voltammograms of SARS-CoV-2 biomodified electrodes were recorded in buffers with and without SARS-CoV-2 positive control, as well as in saliva samples. Third, the samples were analyzed using commercially available COVID-19 salivary tests, viz., rapid antigen test and RT-qPCR (quantitative reverse transcription polymerase chain reaction). It was shown that 8 samples were collected from COVID-19 positive volunteers. Based on the analysis of all experimental results it was concluded that compared to rapid antigen and RT-qPCR tests, the sensitivity and reproducibility of the biosensor is not enough for real practical applications. Thus, some suggestions for further improvement of basic parameters of the developed biodevice were made.

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