The skin has several functions as the largest and one of the most complex organs of the body. One of the skin’s primary functions is to prevent water loss by retaining water to allow the skin to function in dry environments. The outermost layer, stratum corneum (SC), retains water loss as rehydration by natural moisturizing factors (NMFs). In this project, HaCaT cells were incubated with commonly used skincare ingredients such as urea, glycerol, transcutol, salicylic acid, and polyethylene glycol 4000 Da (PEG-4000) to evaluate their impact on cell viability, MTT proliferation assay and gene expression measurements by qPCR. The relationship between cell viability, gene expression, and water activity was also studied. The excipients showed a dose-dependent decrease in cell viability because osmotic pressure increased. One finding was that transcutol exhibited a protective effect against concentrations where osmotic pressure harmed the cells. PEG-4000, with a concentration of 10 % (w/v), showed an upregulation of elongation of very long chain fatty acid 4 (ELOVL-4). Gene expression of serine palmitoyltransferase (SPT) was low, with 10 mM transcutol, even though the cells had a viability of >100 %. It should have conducted an upregulation of SPT from the high metabolic activity in the cells. In conclusion, the viability and gene expression were most likely related to osmotic stress but should be further analyzed with digital holographic microscopy (DHM) and Western blot. 

This study investigated the effect of mechanical infection control for periodontitis and periodontal surgery on the prevalence of well-established risk factors for atherosclerosis, and plasma levels of cytokines, antibodies against heat shock proteins and markers of systemic inflammation. Sixty-eight patients between 39 and 73 years of age with severe periodontitis who had been referred to four specialist periodontology clinics in Sweden were investigated. A fasting venous blood sample was taken at baseline and additional samples were collected after 3 and 12 months. A total of 54 patients underwent periodontal treatment. The periodontal treatment was successful, as pathogenic gingival pockets decreased significantly. Plasma glucose, lipids and markers of systemic inflammation were not significantly altered after 3 months. One year after the initial treatment, HDL-C concentrations were significantly increased (Delta0.08mmol/L) whereas LDL-C concentrations decreased (Delta0.23mmol/L). Haptoglobin concentrations were also lower. Interleukin-18 and interferon-gamma levels were also lower after 12 months (60ng/L (-23%) and 11ng/L (-97%) respectively). Treatment had no effect on plasma levels of IgA, IgG1, IgG2 antibodies against heat shock proteins. In conclusion, this study indicates that standard treatment for periodontal disease induces systemic changes in several biochemical markers that reflect the risk for atherosclerosis.

AIM: Studies have reported on an association between cardiovascular disease (CVD) and periodontitis. The purpose of this case-control study was to provide an insight into this association by determining the plasma levels of some risk markers for CVD in cases with periodontitis. MATERIALS AND METHODS: Sixty-eight cases with periodontitis, mean age 53.9 (SD 7.9) years, and 48 randomly selected healthy controls, mean age 53.1 (SD 7.9) years, were investigated. Fasting blood plasma was analysed for glucose, lipids, markers systemic inflammation, cytokines and antibodies against heat shock proteins (Hsp). The associations between periodontitis and the various substances analysed in plasma were calculated using a multivariate logistic regression model, which compensated for age, gender, smoking and body mass index. RESULTS: The regression analyses revealed a significant association between periodontitis and high levels of C-reactive protein (CRP) [odds ratio (OR) 4.0, confidence interval (CI) 1.4-11.4] and fibrinogen (OR 8.7, CI 2.6-28.4), IL-18 (OR 6.5, CI 2.2-19.5), and decreased levels of IL-4 (OR 0.12, CI 0.0-0.5). The study showed increased levels of antibodies against Hsp65 (OR 2.8, CI 1-7.6) and 70 (OR 2.9, CI 1.1-7.8) and decreased levels of antibodies against Hsp60 (OR 0.3, CI 0.1-0.8). CONCLUSIONS: Periodontitis was associated with increased levels of CRP, glucose, fibrinogen and IL-18, and with decreased levels of IL-4.

Objectives: To investigate the molecular pathogenesis of implant‐associated peripheral giant cell granuloma (IA‐PGCG). Methods: A convenience sample of 15 IA‐PGCG cases was selected. Hotspot mutations of KRAS, FGFR1, and TRPV4 genes, previously reported in conventional giant cell lesions of the jaws, were investigated by Sanger sequencing. As these mutations could activate MAPK/ERK pathway, the expression of phospho‐ERK1/2 was also evaluated by immunohistochemistry. Results: KRAS mutations were detected in 8/15 (53.4%) samples. Similar to conventional peripheral giant cell granuloma, the KRAS mutations most frequently occurred in codon 146 (p.A146V, n = 3), followed by codon 12 (p.G12A and p.G12D, n = 1 each) and codon 14 (p.V14L, n = 1). Variants of unknown significance (VUS) were also detected in two cases, affecting codons 37 (p.E37K) and 127 (p.T127I). All samples showed wild‐type (WT) sequences for FGFR1 and TRPV4 genes. Consistent with MAPK/ERK pathway activation, all mononuclear cells of the lesion showed strong staining for phospho‐ERK1/2 protein in the immunohistochemical analysis. Conclusions: KRAS mutations and activation of the MAPK‐ERK signaling pathway occur in IA‐PGCG. This is the first study to demonstrate cancer‐associated gene mutations in a non‐neoplastic reactive condition associated with dental implants.

High-density lipoproteins (HDLs) are responsible for removing cholesterol from arterial walls, through a process known as reverse cholesterol transport. The main protein in HDL, apolipoprotein A-I (ApoA-I), is essential to this process, and changes in its sequence significantly alter HDL structure and functions. ApoA-I amyloidogenic variants, associated with a particular hereditary degenerative disease, are particularly effective at facilitating cholesterol removal, thus protecting carriers from cardiovascular disease. Thus, it is conceivable that reconstituted HDL (rHDL) formulations containing ApoA-I proteins with functional/structural features similar to those of amyloidogenic variants hold potential as a promising therapeutic approach. Here we explored the effect of protein cargo and lipid composition on the function of rHDL containing one of the ApoA-I amyloidogenic variants G26R or L174S by Fourier transformed infrared spectroscopy and neutron reflectometry. Moreover, small-angle x-ray scattering uncovered the structural and functional differences between rHDL particles, which could help to comprehend higher cholesterol efflux activity and apparent lower phospholipid (PL) affinity. Our findings indicate distinct trends in lipid exchange (removal vs. deposition) capacities of various rHDL particles, with the rHDL containing the ApoA-I amyloidogenic variants showing a markedly lower ability to remove lipids from artificial membranes compared to the rHDL containing the native protein. This effect strongly depends on the level of PL unsaturation and on the particles' ultrastructure. The study highlights the importance of the protein cargo, along with lipid composition, in shaping rHDL structure, contributing to our understanding of lipid-protein interactions and their behavior.

Apolipoprotein A-I (ApoA-I) amyloidosis is a rare protein misfolding disease where fibrils of the N-terminal domain of the protein accumulate in several organs, leading to their failure. Although ApoA-I amyloidosis is systemic, the different amyloidogenic variants show a preferential tissue accumulation that appears to correlate with the location of the mutation in the protein sequence and with the local extracellular microenvironment. However, the factors leading to cell/tissues damage, as well as the mechanisms behind the observed organ specificity are mostly unknown. Therefore, we investigated the impact of ApoA-I variants on cell physiology and the mechanisms driving the observed tissue specificity. We focused on four ApoA-I amyloidogenic variants and analyzed their cytotoxicity as well as their ability to alter redox homeostasis in cell lines from different tissues (liver, kidney, heart, skin). Moreover, variant-specific interactions with extracellular matrix (ECM) components were measured by synchrotron radiation circular dichroism and enzyme-linked immunosorbent assay. Data indicated that ApoA-I variants exerted a cytotoxic effect in a time and cell-type-specific manner that seems to be due to protein accumulation in lysosomes. Interestingly, the ApoA-I variants exhibited specific preferential binding to the ECM components, reflecting their tissue accumulation pattern in vivo. While the binding did not to appear to affect protein conformations in solution, extended incubation of the amyloidogenic variants in the presence of different ECM components resulted in different aggregation propensity and aggregation patterns.

Digital holographic (DH) microscopy is a unique noninvasive method to analyze living cells. With DH microscopy, in vitro cell cultures can be imaged in 2D and pseudo-3D and measurements of size and morphology of the cells are provided. Here, a description of a novel methodology utilizing DH microscopy for the analysis of spheroids is presented. A cell culture protocol is introduced and morphological parameters of cell spheroids as measured by DH microscopy are presented. The study confirms the use of DH microscopy for the analysis of cell spheroids. In the future, organoids can be analyzed with DH microscopy, and it can also be used for drug response and cell death analyses. Method summary This method aims to optimize DH microscopy for the analysis of morphological parameters of spheroids in an easy and convenient way. Cell spheroids were cultured in culture dishes for 5 days. Then, the lid was replaced with HoloLid. After that, the spheroids were imaged and quantitative morphological information was collected and analyzed.

Patients diagnosed with diffuse large B-cell lymphoma (DLBCL) show varying responses to conventional therapy, and this might be contributed to the differentiation stage of the tumor B-cells. The aim of the current study was to evaluate a panel of kinases (ZAP70, PKC-beta I and II and phosphorylated PKB/Akt) and phosphatases (PTEN, SHP1 and SHP2) known to be frequently deregulated in lymphoid malignancies. De novo DLBCL cases were divided into two subgroups, the germinal center (GC) group (14/28) and the non-germinal center (non-GC) or activated B-cell (ABC) group (14/28). ZAP70 and PKC-beta II were expressed in a significantly higher percentage of tumor cells in the clinically more aggressive non-GC group compared with the prognostically favourable GC group. Also, the subcellular localization of PKC-beta I and II differed in DLBCL cells, with the PKC-beta I isoform being expressed in both the cytoplasm and nucleus, while PKC-beta II was found exclusively in the cytoplasm. Loss of nuclear PTEN correlated with poor survival in cases from both subgroups. In addition, five cell lines of DLBCL origin were analyzed for protein expression and for mRNA levels of PTEN and SHP1. For the first time, we show that ZAP70 is expressed in a higher percentage of tumor cells in the aggressive non-GC subgroup of DLBCL and that PKC-beta I and II are differently distributed in the two prognostic subgroups of de novo DLBCL.

PTEN and SHP1 are tumor suppressor genes involved in the regulation of cell cycle control and apoptosis. We have investigated the protein expression of PTEN and SHP 1, by immunohistochemistry in micro tissue arrays from bone marrow samples in children, diagnosed with acute lymphoblastic leukaemia and non-malignant controls. PTEN was over expressed in ALL samples, while SHP1 showed a low expression. PTEN showed a significant difference in expression compared to non-malignant controls. The role of PTEN and SHP1 are not well investigated in pediatric leukemias and could in future play a role as prognostic factors.

BACKGROUND AND OBJECTIVES: L was investigated both in normal and malignant skin cells.

METHODS: Antioxidant activity of the extracts was analyzed by using the Trolox Equivalent Antioxidant Capacity (TEAC) assay. High-Performance Thin-Layer Chromatography (HPTLC) was performed to demonstrate the phytochemical profile, and the total flavonoid content was analyzed with an aluminum chloride colorimetric method. The anti-inflammatory effect was investigated by cell treatments using the plant extracts. Thereafter, the possible suppression of induced IL-6 response was measured from the cultured skin cancer cell lines A2058 and A431, and normal primary keratinocytes with Enzyme-Linked Immunosorbent Assay (ELISA).

RESULTS: also had the highest flavonoid content and showed the highest antioxidant activity of the three extracts tested.

CONCLUSION: possess properties such as antioxidant and anti-inflammatory activities in both normal and malignant keratinocytes, and thus could be a promising agent controlling the pro-inflammatory IL-6 production.

Transcription factor activator protein-1 regulates genes involved in inflammation and repair. The aim of this study was to determine whether transcription factor activator protein-1 activity in carotid plaques is related to symptoms, lipid accumulation, or extracellular matrix composition. Methods: Twenty-eight atherosclerotic carotid plaques were removed by endarterectomy and divided into two groups based on the presence or absence of ipsilateral symptoms (b1 month ago). Activator protein-1 DNA binding activity was assessed, and subunit (c-Jun, JunD, JunB, c-Fos, FosB, Fra-1, Fra-2) protein levels analyzed by immunoblotting. Distribution of c-Jun in plaques was analyzed by immunohistochemistry. Results: Plaques associated with symptoms had increased activator protein-1 activity and increased expression of c-Jun and JunD, as compared to asymptomatic plaques. Fra-1 and Fra-2 were present in equal amounts in both groups, whereas JunB, FosB, and c-Fos were undetectable. Activator protein-1 activity correlated with cholesteryl ester and elastin in plaques and decreased with age. Activator protein-1 activity did not correlate with collagen, calcified tissue, or proteoglycan content. Conclusions: Activator protein-1 is increased in plaques associated with symptoms. The correlation between activator protein-1 and cholesteryl esters suggests that high activator protein-1 is a marker of plaque vulnerability. Activator protein-1 expression can also reflect the activation of repair processes.

Here we report about preliminary investigations on developing substrates for culturing osteoblasts, the cells responsible for production of mineralised bone, by lithographically modifying the surfaces of several materials. The proton beam writing system at the National University of Singapore was used to fabricate high aspect ratio structures in PMMA, while two-dimensional low aspect ratio structures were fabricated using conventional electron beam lithography (EBL) and UV lithography (UVL) in SU-8. It was found that oxygen plasma treatment of structured SU-8 surfaces changed the surface layer and significantly improved cell attachment and proliferation. Cells grown on patterned thick PMMA exhibit a remarkable geometry-dependent behaviour.

Spherical, multicellular aggregates of tumor cells, or three-dimensional (3D) tumor models, can be grown from established cell lines or dissociated cells from tissues in a serum-free medium containing appropriate growth factors. Air–liquid interfaces (ALIs) represent a 3D approach that mimics and supports the differentiation of respiratory tract and skin 3D models in vitro. Many 3D tumor cell models are cultured in conjunction with supporting cell types, such as fibroblasts, endothelial cells, or immune cells. To further mimic the in vivo situation, several extracellular matrix models are utilized to support tumor cell growth. Scaffolds used for 3D tumor cell culture growth include both natural and synthetic hydrogels. Three-dimensional cell culture experiments in vitro provide more accurate data on cell-to-cell interactions, tumor characteristics, drug discovery, metabolic profiling, stem cell research, and diseases. Moreover, 3D models are important for obtaining reliable precision data on therapeutic candidates in human clinical trials before predicting drug cytotoxicity. This review focuses on the recent literature on three different tissue types of 3D tumor models, i.e., tumors from a colorectal site, prostate, and skin. We will discuss the establishment of 3D tumor cell cultures in vitro and the requirement for additional growth support.

BACKGROUND: The aim of this study was to evaluate the clinical outcome in patients with recurrent periodontal disease following treatment with 25% metronidazole gel. METHODS: Twenty subjects in a maintenance care program but with recurrent periodontal disease participated. Three months after scaling and root planing, a total of 40 sites, 2 in each patient, with probing depth > or = 5 mm were selected. One site randomly selected was treated with metronidazole gel (test) and the other site with a placebo gel (control). Baseline and follow-up measurements included plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), and clinical attachment level (CAL). RESULTS: There were no statistically significant differences in PI, GI, BOP, PD, or CAL between test and control sites. CONCLUSION: This study showed that local treatment with 25% metronidazole gel did not seem to influence the clinical healing in this group of subjects with recurrent periodontal disease.

The aim of this study was to evaluate the microbial outcome in patients with recurrent periodontal disease following treatment with 25% metronidazole gel using the polymerase chain reaction (PCR). Twenty subjects in a maintenance care program but with recurrent periodontal disease participated. Three months after scaling and root planing a total of 40 sites, 2 in each patient, with pocket probing depth of > or = 5 mm were selected. One site randomly selected was treated with 25% metronidazole gel (test) and the other site with a placebo gel (control). A bacterial sample was collected on paperpoint from each test and control site at baseline and 12 weeks after treatment. The following pathogens were analysed and detected with PCR:Actinobacillus actinomycetemcomitans (A.a.), Porphyromonas gingivalis (P.g.) and Prevotella nigrescens (P.n.). At baseline, A.a., P.g. and P.n. were detected in 30, 60 and 70% of all test sites and in 32, 58 and 21% of all control sites. There was a statistically significant difference between the test and control sites for P.n. at baseline. The major difference after treatment with 25% metronidazole gel was the increase of positive control sites for P.g. and P.n. However, there were no statistically significant differences in the occurrence rate of A.a., P.g. and P.n. at test and control sites after treatment. This study has shown that 25% metronidazole gel treatment did not seem to influence the microbial outcome, when PCR was used to analyse the presence/absence of A.a., P.g. and P.n. in this group of subjects with recurrent periodontal disease.

The aim of this study was to analyse whether the interleukin-1 (IL-1) and IL-6 gene polymorphisms were associated with the susceptibility of chronic periodontitis. Genomic DNA was obtained from 20 patients with chronic periodontitis and 31 periodontally healthy subjects. All subjects were of North European heritage. The test subjects were kept in a maintenance program after periodontal treatment but yet showing signs of recurrent disease. Genotyping of the IL-1alpha [+4845C>T], IL-1beta [-3954C>T] and IL-6 [-174G>C] polymorphisms was carried out using an allelic discrimination Assay-by-Design method on ABI PRISM 7900 Sequence Detection System. All genotypes were analyzed using the GeneMapper 2.0 software. A similar distribution of Single Nucleotide Polymorphism (SNP) was seen in both groups. Analysis by logistic regression including gender, IL-1alpha [+4845C>T], IL-1beta [-3954C>T], IL-6 [-174G>C] genotypes, the composite IL-1 genotype, the combination of the composite IL-1 genotype and the IL-6 -174G>C genotype and adjusting for smoking did not result in any statistically significant difference. SNPs in IL-1alpha [+4845C>T], IL-1beta [-3954C>T] and IL-6 [-174G>C] do not seem to increase the susceptibility to chronic periodontitis in this group of subjects.

The aim of this study was to evaluate a well-established periodontal risk assessment tool in patients with severe periodontitis included in a supportive periodontal treatment (SPT) program. In total 20 individuals were included in the analysis. All subjects were randomly selected after successful periodontal treatment and at least 5 years SPT. Clinical and radiographic measurements were collected from patient records and analyzed according to the periodontal risk assessment model. Using the periodontal risk assessment model all subjects were classified as low, moderate, or high-risk patients. According to the model 7 patients were classified as moderate risk patients and 13 as high-risk patients. When comparing all the patients using only bleeding on probing (BoP) mean prevalence of 20% as a cut-off point, 15 patients were categorised as having low-moderate risk for periodontitis progression and 5 subjects as having high-risk for disease progression. The periodontal risk assessment model seems to overestimate the risk for disease progression. However the model is a suitable tool to visualize for both the clinician and the patient different variables of importance for periodontal health. The model is also beneficial to show how periodontal treatment can reduce further risk for periodontal disease.

Department of Periodontology, Faculty of Odontology, Malmö University, SE-205 06 Malmö, Sweden. daniel.jonsson@od.mah.se OBJECTIVE: Periodontal ligament (PDL) cells express oestrogen receptors but the functional importance of oestrogen in PDL cells exposed to bacterial endotoxins is not known. Here we investigate if the inflammation promoter lipopolysaccharide (LPS) affects PDL cell production of interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), C-reactive protein (CRP) and/or normal functional PDL cell characteristics such as collagen synthesis and cell proliferation and if oestrogen modulates the effects of LPS. METHODS: PDL cells were obtained from periodontal ligament of premolars. PDL cells were treated with Escherichia coli LPS in the absence or presence of oestrogen (17beta-oestradiol, E2). Cellular concentration of IL-6, MCP-1 and CRP was determined by enzyme-linked immunosorbent assay (ELISA). Collagen synthesis was determined by l-[3H]proline incorporation. Cell proliferation was assessed by DNA synthesis measurement using [3H]thymidine incorporation. RESULTS: Stimulation with LPS (500 ng/ml to 10 microg/ml) increased IL-6 production in a concentration-dependent manner. Lower concentration (100 ng/ml) of LPS had no effect. LPS-induced stimulation of IL-6 was not reversed by a physiologically high concentration (100 nM) of E2. LPS increased also MCP-1 production which was unaffected by E2. Treatment with E2 alone had no effect on either IL-6 or MCP-1. Stimulation with LPS had no effect on CRP. LPS did not affect collagen synthesis and cell proliferation, reflecting normal physiological properties of PDL cells. CONCLUSIONS: LPS stimulates PDL cell IL-6 and MCP-1 production but has no effect on the normal physiological properties of PDL cells. LPS-induced IL-6 and MCP-1 is not reversed by oestrogen suggesting that oestrogen exerts no anti-inflammatory effect via this mechanism.

Background: The objective of this study was to systematically review the evidence on the effectiveness of laser therapy as an adjunct to non-surgical periodontal treatment in adults with chronic periodontitis. Methods: A search was conducted for randomized controlled trials comparing the outcome of periodontitis with laser as an adjunct to scaling and root planing in the treatment of chronic periodontal disease. The electronic databases, PubMed and Cochrane Central Register of Controlled Trials, were used as data sources. Screening, data abstraction, and quality assessment were conducted independently by three reviewers (MK, HJ, and CDL). The primary outcome measures evaluated were changes in clinical attachment level, probing depth, and bleeding on probing. Results: The search resulted in 25 abstracts; four randomized controlled clinical trials were included. Four different laser methods were used; consequently, it was impossible to conduct a quantitative data synthesis leading to a meta-analysis. All studies included a limited number of subjects. Conclusions: No consistent evidence supports the efficacy of laser treatment as an adjunct to non-surgical periodontal treatment in adults with chronic periodontitis. More randomized controlled clinical trials are needed. PMID: 18980508 [PubMed - in process]

We have studied the expression of plasminogen activator inhibitor-1 (PAI-1) and urokinase receptor (uPAR) together with the gamma2-chain of laminin-5 (lam-gamma2) by immunohistochemistry in 20 cases with incipient oral squamous cell carcinoma (SCC). PAI-1-positive neoplastic cells located at the tip of the putative invasive front of grade 1 (incipient) carcinoma were seen in 16 of the 20 cases (75%), whereas adjacent normal and dysplastic epithelium was PAI-1-negative. Clusters of putative invasive neoplastic cells located in the lamina propria were PAI-1-positive in areas with grade 2 incipient carcinoma as were invasive cancer cells in areas of grade 3-4 invasive carcinoma. uPAR immunoreactivity was strongly expressed in numerous stromal cells in the carcinoma area in all 20 lesions, while a few uPAR-positive stromal cells were found in areas with normal and dysplastic epithelium. uPAR-positive neoplastic cell islands located at the front of the lesions were seen in 15 of the 20 cases. The expression pattern of lam-gamma2 was very similar to that of PAI-1; however, lam-gamma2-positive neoplastic cells were only detected in 11 of the 20 cases (55%) in areas of grade 1 incipient carcinoma. Direct comparison of the 3 components revealed colocalization in neoplastic cell islands in both incipient and invasive SCC. Our results suggest that PAI-1 is a novel potential marker of initial invasion in oral SCC, and that the coordinated expression of PAI-1 with uPAR and lam-gamma2 sustain the features of the early invasive cancer cells.

Spheroids are three-dimensional (3D) cell structures of aggregated cells that more closely mimic the cell-cell interactions and microenvironment of tumors. In this study 0,1% v/v of nano fibrillar cellulose hydrogel solubilized in spheroid medium was tested against static suspension culture. The aim was to determine whether either of the mediums was suitable to make the cell lines MCF-7, MDAMB-231, HMEC-1 as well as co-cultures of MCF-7/HMEC-1 and MDA-MB-231to form growth as spheroids. This study used holographic digital microscopy as well as light microscopy with image processing to analyze the cells. This study also examined the possible effect that the receptor-binding domain (RBD) of SARSCov2 might have on cell motility by interacting with angiotensin-converting enzyme 2 (ACE2). This study also included cross-linking of the RBD-ACE2complex to strengthen the RBD-ACE2 interaction. To this end, a motility assay was conducted on monolayers of the cell lines. This study found that among the cell lines and co-cultures only MCF-7 and MCF-7/HMEC-1 formed spheroids. The other cell lines and co-culture formed loose or tight aggregates. This study also showed that there was a significant difference in motility between the cell lines and indicated upon the difference between cells treated these results implicate decreased invasiveness by tumors in cancer patients due to toRBD-ACE2 interactions. This merits further experiments to determine whether-ACE2 treatment in vitro can serve to decrease the motility of tumor cells in cultures. 

Aims/hypothesis: Roux-en-Y gastric bypass surgery (RYGB) frequently results in remission of type 2 diabetes as well as exaggerated secretion of glucagon-like peptide-1 (GLP-1). Here, we assessed RYGB-induced transcriptomic alterations in the small intestine and investigated how they were related to the regulation of GLP-1 production and secretion in vitro and in vivo.

Methods: Human jejunal samples taken perisurgically and 1 year post RYGB (n=13) were analysed by RNA-seq. Guided by bioinformatics analysis we targeted four genes involved in cholesterol biosynthesis, which we confirmed to be expressed in human L cells, for potential involvement in GLP-1 regulation using siRNAs in GLUTag and STC-1 cells. Gene expression analyses, GLP-1 secretion measurements, intracellular calcium imaging and RNA-seq were performed in vitro. OGTTs were performed in C57BL/6j and iScd1-/- mice and immunohistochemistry and gene expression analyses were performed ex vivo.

Results: Gene Ontology (GO) analysis identified cholesterol biosynthesis as being most affected by RYGB. Silencing or chemical inhibition of stearoyl-CoA desaturase 1 (SCD1), a key enzyme in the synthesis of monounsaturated fatty acids, was found to reduce Gcg expression and secretion of GLP-1 by GLUTag and STC-1 cells. Scd1 knockdown also reduced intracellular Ca2+ signalling and membrane depolarisation. Furthermore, Scd1 mRNA expression was found to be regulated by NEFAs but not glucose. RNA-seq of SCD1 inhibitor-treated GLUTag cells identified altered expression of genes implicated in ATP generation and glycolysis. Finally, gene expression and immunohistochemical analysis of the jejunum of the intestine-specific Scd1 knockout mouse model, iScd1-/-, revealed a twofold higher L cell density and a twofold increase in Gcg mRNA expression.

Conclusions/interpretation: RYGB caused robust alterations in the jejunal transcriptome, with genes involved in cholesterol biosynthesis being most affected. Our data highlight SCD as an RYGB-regulated L cell constituent that regulates the production and secretion of GLP-1.

The aim of this study was to investigate changes in knowledge before and after a mass media campaign, in relation to social attributes, care system attributes and oral health aspects. The study was based on a questionnaire in a cohort design, sent out to 900 randomly sampled people aged 50–75 in Sweden. The response rate to the questionnaire before and after the campaign was 70% and 65% respectively. Sixty-four percent answered both questionnaires. Two questions addressed knowledge, while 10 questions aimed to measure social attributes, care system attributes and oral health aspects. Data were analysed for bivariate relations as to change in knowledge and social attributes, care system attributes and oral health aspects. Data were also analysed in multiple regression analysis with knowledge before, knowledge after and knowledge differences as dependent variables. The results showed that there were a number of independent variables with influence on the dependent variables. Of the social attributes, secondary education gave almost 10% (P< 0.001) better knowledge both before and after the campaign. Among care system attributes, high care utilization was related to knowledge both before and after the campaign. The most important factors for knowledge about periodontitis were education, care utilization and perceived importance of oral health. In conclusion, this study demonstrates that mass media might increase knowledge about periodontitis as a health promotion strategy.

Objective. Non-human primates, dogs, rats, hamsters and ferrets, have frequently been used as laboratory animals in periodontal biology and pathology. In the past, mice have been used less for this purpose, but nowadays attract a lot of interest because gene knockout and transgenic technologies utilize mice primarily. In this study, we investigate the effects of ovariectomy and aging on tooth attachment in female mice. Material and methods. Eight-week-old mice (n=15) were divided into three experimental groups (control, n=5; sham-operated, n=5; ovariectomy, n=5) and ovaries removed bilaterally. Attachment level, assessed by measuring alveolar bone height and apical termination of the junctional epithelium, was determined 6 weeks post-ovariectomy by digital morphometric analysis in sagittal sections of the mandible. The plasma level of the inflammation marker serum amyloid A (SAA) was determined by ELISA. In another series of experiments, tooth attachment was determined in female mice (n=7) at 8–26 weeks of age. Results. Withdrawal of female sex hormone production by ovariectomy had no effect on alveolar bone height and apical termination of the junctional epithelium. The SAA level in plasma was unaffected by removal of the ovaries, suggesting that systemic inflammation is not induced by ovariectomy. Bone height was similar in mice sacrificed at 8–26 weeks of age and apical termination of the junctional epithelium was at the cemento-enamel junction at all ages. Conclusions. Removal of ovarian production of female sex hormones by ovariectomy has no influence on tooth attachment, and further tooth attachment is preserved with age in female mice.

Background: The periodontal ligament cells (PDL cells) play a key role in the formation of the periodontal ligament but these cells have other functions as well. The PDL cells express estrogen receptors but the functional importance is not known. Estrogen modulates inflammation and our hypothesis is that estrogen protects the periodontium via an anti-inflammatory effect on PDL cells. Aim: To identify genes regulated by estrogen in LPS-treated human PDL cells by whole genome arrays. Materials and methods: PDL cells were obtained from human premolars extracted for orthodontic reasons. The cells were divided into two groups. One group was pre-treated with 17β-estradiol (E2, 100 nM) for 2 h and then with Escherichia coli LPS (500ng/ml) for 24 h. The other group was treated with LPS only. Total RNA was extracted and purified by RNeasy Mini Kit (Qiagen®, USA). A whole genome microarray was performed (Affymetrix®, USA) comparing gene expression in the two groups. The cut-off limit was set to a twofold change. Results and Conclusion: Estrogen caused an up-regulation of 38 genes, while 28 genes were down-regulated. Estrogen regulated genes associated with cell-metabolism and cell-signalling but also genes associated with early the inflammatory response. The functional significance of these findings is now determined by e.g. measuring protein levels with ELISA. We conclude that estrogen regulates gene expression in human PDL cells exposed to LPS.

Background: The periodontal ligament cells (PDL cells) play a key role in the formation of the periodontal ligament but these cells have other functions as well. The PDL cells express estrogen receptors but the functional importance is not known. Estrogen modulates inflammation and our hypothesis is that estrogen protects the periodontium via an anti-inflammatory effect on PDL cells. Aim: To identify genes regulated by estrogen in LPS-treated human PDL cells by whole genome arrays. Materials and methods: PDL cells were obtained from human premolars extracted for orthodontic reasons. The cells were divided into two groups. One group was pre-treated with 17ÿ-estradiol (E2, 100 nM) for 2 h and then with Escherichia coli LPS (500ng/ml) for 24 h. The other group was treated with LPS only. Total RNA was extracted and purified by RNeasy Mini Kit (Qiagen®, USA). A whole genome microarray was performed (Affymetrix®, USA) comparing gene expression in the two groups. The cut-off limit was set to a twofold change. Results and Conclusion: Estrogen caused an up-regulation of 38 genes, while 28 genes were down-regulated. Estrogen regulated genes associated with cell-metabolism and cell-signalling but also genes associated with early the inflammatory response. The functional significance of these findings is now determined by e.g. measuring protein levels with ELISA. We conclude that estrogen regulates gene expression in human PDL cells exposed to LPS.

The oral microbiota, which is known to include at least 600 different bacterial species, is found on the teethand mucosal surfaces as multi-species communities or biofilms. The oral surfaces are covered with a pellicleof proteins absorbed from saliva, and biofilm formation is initiated when primary colonizers, which expresssurface adhesins that bind to specific salivary components, attach to the oral tissues. Further developmentthen proceeds through co-aggregation of additional species. Over time, the composition of oral biofilms,which varies between different sites throughout the oral cavity, is determined by a combination ofenvironmental factors such as the properties of the underlying surface, nutrient availability and oxygenlevels, and bacterial interactions within the community. A complex equilibrium between biofilm communities and the host is responsible for the maintenance of a healthy biofilm phenotype (eubiosis). In the faceof sustained environmental perturbation, however, biofilm homeostasis can break down giving rise todysbiosis, which is associated with the development of oral diseases such as caries and periodontitis.In vitro models have an important part to play in increasing our understanding of the complex processesinvolved in biofilm development in oral health and disease, and the requirements for experimental system,microbial complexity, and analysis techniques will necessarily vary depending on the question posed. In thischapter we describe some current and well-established methods used in our laboratory for studying oralbacteria in biofilm models which can be adapted to suit the needs of individual users. 

Apolipoprotein A-I (ApoA-I) of high density lipoproteins (HDLs) is essential for the transportation of cholesterol between peripheral tissues and the liver. However, specific mutations in ApoA-I of HDLs are responsible for a late-onset systemic amyloidosis, the pathological accumulation of protein fibrils in tissues and organs. Carriers of these mutations do not exhibit increased cardiovascular disease risk despite displaying reduced levels of ApoA-I/HDL cholesterol. To explain this paradox, we show that the HDL particle profiles of patients carrying either L75P or L174S ApoA-I amyloidogenic variants show a higher relative abundance of the 8.4-nm versus 9.6-nm particles and that serum from patients, as well as reconstituted 8.4- and 9.6-nm HDL particles (rHDL), possess increased capacity to catalyze cholesterol efflux from macrophages. Synchrotron radiation circular dichroism and hydrogen-deuterium exchange revealed that the variants in 8.4-nm rHDL have altered secondary structure composition and display a more flexible binding to lipids than their native counterpart. The reduced HDL cholesterol levels of patients carrying ApoA-I amyloidogenic variants are thus balanced by higher proportion of small, dense HDL particles, and better cholesterol efflux due to altered, region-specific protein structure dynamics.

Gastric MUC5AC and MUC6 mucins were studied using polyclonal antibodies. Immunohistochemistry showed MUC5AC to originate from the surface epithelium, whereas MUC6 was produced by the glands. Mucins from the surface epithelium or glands of corpus and antrum were purified using CsCl/4M guanidinium chloride density-gradient centrifugation. MUC5AC appeared as two distinct populations at 1.4 and 1.3 g/ml, whereas MUC6, which was enriched in the gland tissue, appeared at 1.45 g/ml. Reactivity with antibodies against the Le(b) structure (where Le represents the Lewis antigen) followed the MUC5AC distribution, whereas antibodies against the Le(y) structure and reactivity with the GlcNAc-selective Solanum tuberosum lectin coincided with MUC6, suggesting that the two mucins are glycosylated differently. Rate-zonal centrifugation of whole mucins and reduced subunits showed that both gastric MUC5AC and MUC6 are oligomeric glycoproteins composed of disulphide-bond linked subunits and that oligomeric MUC5AC was apparently smaller than MUC6. A heterogeneous population of 'low-density' MUC5AC mucins, which were smaller than the 'high-density' ones both before and after reduction, reacted with an antibody against a variable number tandem repeat sequence within MUC5AC, suggesting that they represent precursor forms of this mucin. Following ion-exchange HPLC, both MUC5AC and MUC6 appeared as several distinct populations, probably corresponding to 'glycoforms' of the mucins, the most highly charged of which were found in the gland tissue.

Breast cancer is the second most common cancer type worldwide and breast cancer metastasis accounts for the majority of breast cancer-related deaths. Tumour cells produce increased levels of sialic acid (SA) that terminates the monosaccharide on glycan chains of the glycosylated proteins. SA can contribute to cellular recognition, cancer invasiveness and increase the metastatic potential of cancer cells. SA-templated molecularly imprinted polymers (MIPs) have been proposed as promising reporters for specific targeting of cancer cells when deployed in nanoparticle format. The sialic acid-molecularly imprinted polymers (SA-MIPs), which use SA for the generation of binding sites through which the nanoparticles can target and stain breast cancer cells, opens new strategies for efficient diagnostic tools. This study aims at monitoring the effects of SA-MIPs on morphology and motility of the epithelial type MCF-7 and the highly metastatic MDAMB231 breast cancer cell lines, using digital holographic cytometry (DHC). DHC is a label-free technique that is used in cell morphology studies of e.g., cell volume, area and thickness as well as in motility studies. Here, we show that MCF-7 cells move slower than MDAMB231 cells. We also show that SA-MIPs have an effect on cell morphology, motility and viability of both cell lines. In conclusion, by using DH microscopy, we could detect SA-MIPs impact on different breast cancer cells regarding morphology and motility.

This thesis investigates responses to external cues in oral bacteria on a molecular level. Paper I maps Ser/Thr/Tyr phosphorylated proteins in relation to the general proteome in an oral commensal streptococcus (Streptococcus gordonii DL1). The identified phosphoproteins were involved in various bacterial processes, several associated to dysbiosis and development of biofilm-induced disease. Comparison against phosphoproteomes of other bacteria showed many similarities. This is of interest for the identification of shared phosphorylation profiles. 

Paper II studies differences between the S. gordonii DL1 general proteomes in planktonic and biofilm growth phases, and the regulatory effects of salivary mucin MUC5B on protein expression in the biofilm cells. Regulations in protein expression between the different growth conditions provides insights in bacterial mechanisms for adaptation to the biofilm lifestyle. 

Paper III examines the regulatory roles of salivary MUC5B on biofilm attachment and metabolic output in two clinical isolates of oral commensals, S. gordonii CW and Actinomyces naeslundii CW. S. gordonii facilitated adhesion of A. naeslundii to MUC5B during early attachment. Both bacteria were also able to utilize MUC5B as a sole nutrient source during early biofilm formation, individually and synergistically in a dual species biofilm. The specific responses elicited by MUC5B in paper II-III seem to promote commensal colonization while down-regulating dysbiosis-related biofilm activities. 

Microbiological studies are often focused on dysbiosis and development of disease. However, mechanisms that promote eubiosis are equally important to understand how health can be maintained. Findings associated with responses to external cues in oral bacteria may contribute to future development of novel preventative strategies and identification of predictive biomarkers for oral health. 

Dental implants have become an often used alternative to replace missing teeth, resulting in an increasing percentage of the adult population with implant supported prosthesis. Although favourable longtermresults of implant therapy have been reported, infections occur.Until recently few reports included data on peri-implant infections,possibly underestimating this complication of implant treatment. It is possible that some infections around implants develop slowly andthat with time peri-implantitis will be a common complication to implant therapy as an increasing number of patients have had theirimplants for a long time (>10 years). Data on treatment of periimplant lesions are scarce leaving the clinician with limited guidance regarding choice of treatment.The aim of this thesis was to study the frequency of implant loss and presence of peri-implant lesions in a group of patients supplied with Brånemark implants 9-14 years ago, and to relate these events to patient and site specific characteristics. Moreover three surgical treatment modalities for peri-implantitis were evaluated. The thesis is based on six studies; Studies I-III included 218 patients and 1057 implants followed for9-14 years evaluating prevalence of, and factors related to implantloss (Paper I) and prevalence of peri-implant infections and relatedfactors (Paper II-III).Study IV is a review describing different treatment modalities ofperi-implant infections.Study V is a prospective cohort study involving 36 patients and 65implants, evaluating the use of a bone substitute with or withoutthe use of a resorbable membrane. Study VI is a case series with 12patients and 16 implants, evaluating a bone substitute in combinationwith a resorbable membrane and submerged healing.This thesis demonstrated that:After 9-14 years the survival rates of dental implants are high(95.7%). Implant loss seems to cluster within patients and arerelated to periodontitis evidenced as bone loss on radiographsat remaining teeth before implant placement. (Paper I)Peri-implantitis is a common clinical entity after 9-14 years.(Paper II)Using the implant as the statistical unit the level of keratinizedmucosa and pus were explanatory for a bone level at ≥3 threads(1.8 mm). When the patient was used as a statistical unit ahistory of periodontitis and smoking were explanatory for periimplantitis.(Paper III)Animal research has demonstrated that re-osseointegration canoccur. The majority of human studies were found to be casereports. Using submerged healing and bone transplants, bonefill can occur in peri-implant defects. (Paper IV)Surgical treatment of peri-implantitis using a bone substitute withor without a resorbable membrane resulted in similar pocketdepth reduction, attachment gain and defect fill. (Paper V)Bone substitute in combination with a resorbable membraneand a submerged healing resulted in defect fill ≥2 threads (1.2mm) in 81% of the implants. (Paper VI)In conclusion: 9-14 years after implant installation peri-implantlesions are a common clinical entity. Smokers and patients with ahistory of periodontal disease are at higher risk to develop periimplantitis.Clinical improvements and defect fill can be obtainedwith various surgical techniques using a bone substitute.

Three-dimensional (3D) spheroid culture can promote the osteogenic differentiation and bone regeneration capacity of mesenchymal stromal cells (MSC). Gingiva-derived progenitor cells (GPC) represent a less invasive alternative to bone marrow MSC (BMSC) for clinical applications. The aim of this study was to test the in vivo bone forming potential of human GPC and BMSC cultured as 3D spheroids or dissociated cells (2D). 2D and 3D cells encapsulated in constructs of human platelet lysate hydrogels (HPLG) and 3D-printed poly (L-lactide-co-trimethylene carbonate) scaffolds (HPLG-PLATMC) were implanted subcutaneously in nude mice; cell-free HPLG-PLATMC constructs served as a control. Mineralization was assessed using micro-computed tomography (µCT), histology, scanning electron microscopy (SEM) and in situ hybridization (ISH). After 4–8 weeks, µCT revealed greater mineralization in 3D-BMSC vs. 2D-BMSC and 3D-GPC (p < 0.05), and a similar trend in 2D-GPC vs. 2D-BMSC (p > 0.05). After 8 weeks, greater mineralization was observed in cell-free constructs vs. all 2D- and 3D-cell groups (p < 0.05). Histology and SEM revealed an irregular but similar mineralization pattern in all groups. ISH revealed similar numbers of 2D and 3D BMSC/GPC within and/or surrounding the mineralized areas. In summary, spheroid culture promoted ectopic mineralization in constructs of BMSC, while constructs of dissociated GPC and BMSC performed similarly. The combination of HPLG and PLATMC represents a promising scaffold for bone tissue engineering applications. 

Gingiva has been identified as a minimally invasive source of multipotent progenitor cells (GPCs) for use in bone tissue engineering (BTE). To facilitate clinical translation, it is important to characterize GPCs in xeno-free cultures. Recent evidence indicates several advantages of three-dimensional (3D) spheroid cultures of mesenchymal stromal cells (MSCs) over conventional 2D monolayers. The present study aimed to characterize human GPCs in xeno-free 2D cultures, and to test their osteogenic potential in 3D cultures, in comparison to bone marrow MSCs (BMSCs). Primary GPCs and BMSCs were expanded in human platelet lysate (HPL) or fetal bovine serum (FBS) and characterized based onin vitroproliferation, immunophenotype and multi-lineage differentiation. Next, 3D spheroids of GPCs and BMSCs were formed via self-assembly and cultured in HPL. Expression of stemness- (SOX2, OCT4, NANOG) and osteogenesis-related markers (BMP2, RUNX2, OPN, OCN) was assessed at gene and protein levels in 3D and 2D cultures. The cytokine profile of 3D and 2D GPCs and BMSCs was assessed via a multiplex immunoassay. Monolayer GPCs in both HPL and FBS demonstrated a characteristic MSC-like immunophenotype and multi-lineage differentiation; osteogenic differentiation of GPCs was enhanced in HPL vs. FBS. CD271(+)GPCs in HPL spontaneously acquired a neuronal phenotype and strongly expressed neuronal/glial markers. 3D spheroids of GPCs and BMSCs with high cell viability were formed in HPL media. Expression of stemness- and osteogenesis-related genes was significantly upregulated in 3D vs. 2D GPCs/BMSCs; the latter was independent of osteogenic induction. Synthesis of SOX2, BMP2 and OCN was confirmed via immunostaining, andin vitromineralization via Alizarin red staining. Finally, secretion of several growth factors and chemokines was enhanced in GPC/BMSC spheroids, while that of pro-inflammatory cytokines was reduced, compared to monolayers. In summary, monolayer GPCs expanded in HPL demonstrate enhanced osteogenic differentiation potential, comparable to that of BMSCs. Xeno-free spheroid culture further enhances stemness- and osteogenesis-related gene expression, and cytokine secretion in GPCs, comparable to that of BMSCs.

Aim: The aim of this stydy was to assess whether supplementation of Lactobacillus reuteri could have an impact on the oral microbiota. Material and Methods: Twent-three healthy people aged 29 to 63 years were included. A randomised double-blind placebo-controlled study was executed consisting of 12 persons in the test group which was given the study product twice a day for twelve weeks containing L. reuteri (an equal mix of ATCC 55730 and PTA 5289 at a total of 2x108 CFU/dose) and the control group of 11 persons having corresponding placebo. Pre and post of study period plaque index and oral health status (gingivitis, probing pocket depth, bleeding on probing) were measured together with sampling of supra-, subgingival plaque and saliva collection. Microbiological analysis was done by using checkerboard DNA-DNA hybridization technique and selective culturing for lactobacilli determination. Four weeks after the last intake of the product reassessments of plaque and saliva was performed. Results: No difference in general oral health could be observed between the groups after L. reuteri supplementation. Plaque index increased significantly in the control group after twelve weeks (p= 0.023). A significant increase in total Lactobacillus counts in saliva occurred in both groups (p<0.05). The probiotic intervention resulted in a significant increase of L. reuteri (p=0.008) corresponding to 13.8% of the total lactobacilli couont. A distribution ratio of 1:4 (ATCC 55730/ATCC PTA 5289) between the two installed L. reuteri strains in saliva was noticed. Termination of intervention resulted in a wash out of L. reuteri. A significant increase was found for most bacterial species in both groups and both in supra- and subgingival plaque during the test period. There was no significant difference detected for any of the bacterial species between the groups neither in plaque location. The ratio between "bad/good" supragingival bacteria decreased for the test group, however, not significant. The corresponding ratio for subgingival bacteria decreased significantly for both groups (p= 0.05)with no significant difference between the groups. Conclusions: The supplementation of L. reuteri did not affect general oral health. Presence of L. reuteri in saliva is only temporary and washed out after termination of intervention. Microbiologically no significant effect of the probiosis was observed. It can not be concluded whether L. reuteri was established in the plaque of the test group or not.

The aims of this study were to investigate alexithymia in relation to facial imitation, attachment, and empathy. Pictures of facial expressions were presented to 102 participants. Electromyographic activity was measured. Self-report questionnaires were used to measure alexithymia, attachment, and empathy. High-alexithymic participants showed less imitative responses and less empathy and less secure pattern of attachment.

Title Clinical evaluation of local antibiotic treatment in smokers with chronic periodontitis in a maintenance care program. Authors Kerstin Stagnér, Gunilla Bratthall Institution Department of Periodontology, Faculty of Odontology, Malmö University Aim To evaluate if local antibiotics, 8.8 % doxycycline gel, can improve periodontal healing in smoking adults in a maintenance care program. Material and Methods In a double-blind randomized controlled study 20 smoking patients were selected from a maintenance care program. Each subject demonstrated at least 5 probing pocket depths (PPD) ≥ 5mm with bleeding on probing (BOP) in a minimum of 5 teeth. The patients were divided into 2 groups, the test group (8.8 % doxycycline gel, Atridox®) and the control group (placebo gel). At baseline and 3 months after treatment a clinical registration was performed, including PPD, clinical attachment level (CAL), BOP, and plaque index (PlI) at 4 sites of test- and control teeth. Gingival index (GI) was performed using Periotron® at 2 sites of all index-teeth. At baseline all subjects received full-mouth scaling followed with application of test or placebo gel in all index pockets. After gel application 0.1 % chlorhexidine was used for mouth rinsing during 7-10 days, since no mechanical oral hygiene was allowed during that period. Results There were 16 patients left in the end of the study. Four subjects in the test group had to be excluded because of disease. Three months after treatment significant improvement for PPD was shown both in the test (p=0.0313) and the control group (p=0.0020). Significant improvement for the control group was also demonstrated for CAL and BoP. There were no significant differences in any parameter in the change after 3 months between test - and control groups. When the results from all teeth (including index teeth) were analyzed significant improvement was shown in PPD both in the test (p=0.0313) and the control group (p=0.0039). Significant improvement for the control group was also demonstrated for CAL, BOP and GI.. There were no significant differences between test and placebo treated dentitions. Conclusion This study could not show any statistically significant clinical differences between the test - and control groups based on results of index teeth as well as all teeth. This means that local antibiotic treatment with 8.8 % doxycycline gel (Atridox®) did not show any additional effects to scaling. The results, however, showed significant improvement of periodontal pockets after subgingival scaling in both test- and control groups.

Clinical evaluation of local antibiotic treatment in smokers with chronic periodontitis in a maintenance care program. Authors: Kerstin Stagnér, Gunilla Bratthall. Institution Department of Periodontology, Faculty of Odontology, Malmö University. Aim. To evaluate if local antibiotics, 8.8 % doxycycline gel, can improve periodontal healing in smoking adults in a maintenance care program. Material and Methods. In a double-blind randomized controlled study 20 smoking patients were selected from a maintenance care program. Each subject demonstrated at least 5 probing pocket depths (PPD) ≥ 5mm with bleeding on probing (BOP) in a minimum of 5 teeth. The patients were divided into 2 groups, the test group (8.8 % doxycycline gel, Atridox®) and the control group (placebo gel). At baseline and 3 months after treatment a clinical registration was performed, including PPD, clinical attachment level (CAL), BOP, and plaque index (PlI) at 4 sites of test- and control teeth. Gingival index (GI) was performed using Periotron® at 2 sites of all index-teeth. At baseline all subjects received full-mouth scaling followed with application of test or placebo gel in all index pockets. After gel application 0.1 % chlorhexidine was used for mouth rinsing during 7-10 days, since no mechanical oral hygiene was allowed during that period. Results. There were 16 patients left in the end of the study. Four subjects in the test group had to be excluded because of disease. Three months after treatment significant improvement for PPD was shown both in the test (p=0.0313) and the control group (p=0.0020). Significant improvement for the control group was also demonstrated for CAL and BoP. There were no significant differences in any parameter in the change after 3 months between test - and control groups. When the results from all teeth (including index teeth) were analyzed significant improvement was shown in PPD both in the test (p=0.0313) and the control group (p=0.0039). Significant improvement for the control group was also demonstrated for CAL, BOP and GI. There were no significant differences between test and placebo treated dentitions. Conclusion. This study could not show any statistically significant clinical differences between the test - and control groups based on results of index teeth as well as all teeth. This means that local antibiotic treatment with 8.8 % doxycycline gel (Atridox®) did not show any additional effects to scaling. The results, however, showed significant improvement of periodontal pockets after subgingival scaling in both test- and control groups.

Molecularly imprinted polymers (MIPs) against sialic acid (SA) have been developed as a detection tool to target cancer cells. Before proceeding to in vivo studies, a better knowledge of the overall effects of MIPs on the innate immune system is needed. The aim of this study thus was to exemplarily assess whether SA-MIPs lead to inflammatory and/or cytotoxic responses when administered to phagocytosing cells in the innate immune system. The response of monocytic/macrophage cell lines to two different reference particles, Alhydrogel and PLGA, was compared to their response to SA-MIPs. In vitro culture showed a cellular association of SA-MIPs and Alhydrogel, as analyzed by flow cytometry. The reference particle Alhydrogel induced secretion of IL-1β from the monocytic cell line THP-1, whereas almost no secretion was provoked for SA-MIPs. A reduced number of both THP-1 and RAW 264.7 cells were observed after incubation with SA-MIPs and this was not caused by cytotoxicity. Digital holographic cytometry showed that SA-MIP treatment affected cell division, with much fewer cells dividing. Thus, the reduced number of cells after SA-MIP treatment was not linked to SA-MIPs cytotoxicity. In conclusion, SA-MIPs have a low degree of inflammatory properties, are not cytotoxic, and can be applicable for future in vivo studies.

Rate-zonal centrifugation of a reduced and alkylated respiratory mucin preparation identified a protein-rich fraction. This was subjected to trypsin treatment and one of the many liberated peptides was purified and its N-terminal sequence determined. The peptide was identical to a 14 amino acid sequence from the scavenger receptor cysteine-rich domain containing glycoprotein gp-340. A polyclonal antiserum, raised against the peptide, stained the serous cells in the submucosal glands of human tracheal tissue. The glycoprotein was purified from respiratory mucus by density-gradient centrifugation, gel chromatography, and anion exchange chromatography. The molecule exhibited a heterogeneous distribution of buoyant density (1.28-1.46 g/ml) that overlapped with the gel-forming mucins, was included on Sepharose CL-2B and was quite highly anionic. SDS-PAGE indicated a mass greater than 208 kDa and measurements performed across the molecular size distribution indicated an average M(r) of 5 x 10(5) with a range of M(r) from 2 x 10(5) to 1 x 10(6). Gel chromatography of respiratory mucus extracts ("associative" and "dissociative") indicated that this glycoprotein forms complexes that may involve the large gel-forming mucins MUC5AC and MUC5B. Rate zonal centrifugation suggested such complexes are more likely to involve MUC5B rather than MUC5AC mucins.

Proteases are capable of tissue breakdown. Plasma and gingival crevicular fluid (GCF) contain antiproteases, such as alfa-1-antitirypsin (AAT). Lack of AAT may lead to periodontal destruction. The aim was to study if periodontal parameters and elastase in GCF and plasma are different in AAT deficient subjects compared to subjects without AAT deficiency. Material & methods: 30 subjects were included, 20 of whom with severe AAT deficiency. Ten of them suffered from chronic obstructive pulmonary disease (group 1) and 10 were asymptomatic (group 2). Ten control subjects (group 3) were recruited from a public dental clinic. The examination comprised GCF, Gingival index (GI), Plaque Index (PlI), probing pocket depth (PPD) and radiography. GCF was collected with paper strips (Periopaper®). Plasma AAT concentration was measured by nephelometry and AAT in GCF with ELISA. Elastase activity and protein in plasma and GCF were determined by spectrophotometry. Results: The mean values for GI, PlI, PPD and the radiological measurements did not show any statistically significant differences between the groups. AAT in GCF and plasma did not show any significant difference between group 1 and 2 but a statistical difference in comparison with group 3. Elastase in GCF and plasma did not show any difference between the three groups. In conclusion no differences were found between AAT deficient subjects and healthy controls in this limited material.

Research shows that physical activity promotes good health and reduces the risk of developing various types of diseases that are primarily correlated with a sedentary lifestyle. At present, it is difficult to describe in detail how the body is genetically affected by different types of physical exertion. In recent years, it has become clearer that epigenetic modification plays a significant role in how the body's tissues adapt to the prevailing external factors. Epigenetics, which is a relatively new field in molecular biology, describes how the genome can be regulated without changing the nucleotides. Which means a change in the gene expression that takes place on top of the genome. The purpose of this literature study is to investigate how physical activity affects individual’s epigenome specifically on the skeletal muscle cells in the age groups 18-65 years, in both men and women. The articles that were used in this literature study were searched for by using the PubMed database with specifically selected keywords that were in line with the purpose of this literature study. The keywords used were: Epigenetic AND exercise AND skeletal muscle AND Epigenetic skeletal muscle regulation. This literature study showed that a reduction of methylations on several promoter regions on DNA is formed after an intensive training session. Which in turn leads to an increased transcription of the affected genes. Methylation patterns that are formed are highly correlated with training form / intensity. An increase in acetylation on specific genes could also be observed. Only specific microRNAs were regularly upregulated after intensive training that has a link to muscle proliferation and differentiation. A down-regulation of the same genes could also be observed during chronic training. Most of the results obtained were in direct correlation with the training intensity. This is the most decisive factor for how the methylation pattern will look like and that these patterns are formed faster than previously speculated.