Association between Carbonic Anhydrase VI (CA VI) gene copy number and dental caries experience

The current study examined the association between the carbonic anhydrase VI (CA VI) copy number variations (CNVs) and dental caries experience in adults. In total, 202 of 35-72 years old subjects participating in the Lithuanian National Oral Health Survey (LNOHS) agreed to provide saliva samples, thus their data were included in the current study. Information about sociodemographic, environmental, and behavioural determinants was acquired via the self-administered World Health Organisation (WHO) questionnaire. Fluoride levels in the drinking water were recorded based on information provided by water suppliers. Dental caries experience was recorded by one calibrated examiner using the WHO criteria for recording caries on smooth (including proximal, buccal, and oral) or occlusal surfaces. Caries experience was measured as the total number of decayed (D3), missing (M), filled (F) surfaces (D3MFS). DNA was extracted from saliva samples to examine CA VI CNVs using the QX200 droplet digital PCR system. Negative binomial regression and Poisson regression analyses were employed for data analyses. Based on multivariable regression analyses, higher copy number of CA VI were associated with higher caries experience on smooth surfaces (IRR 1.04, 95% CI 1.005 - 1.08) and occlusal surfaces (IRR 1.02, 95% CI 1.003 - 1.04). Positive associations between higher copy number of CA VI and higher caries experience on smooth and occlusal surfaces were found, suggesting that the CA VI coding gene may be associated with caries development. Future studies are needed to validate our results and to examine the underlying mechanisms of such associations

Restorative treatment decisions for carious lesions: Do Russian dentists and dental students apply minimal intervention dentistry?

Background - The concept of minimal intervention dentistry (MID) includes both delayed restorative treatment and conservative caries removal, and is now recognised as an evidence-based approach for dental caries management. In order to determine if dental professionals in Russia are incorporating this concept into their clinical practice, we investigated the restorative treatment decisions of Russian dentists and dental students, and the factors associated with these decisions. Methods - We included 171 general dental practitioners and dental therapists (collectively referred to here as “dentists”) from North-West Russia, and 76 dental undergraduate students from the Northern State Medical University in Arkhangelsk (response rate of 11.5% and 67.9%, respectively). Participants completed a questionnaire, which collected background information (sex, region of work, place of dental school graduation, practice type, years of working experience, working in an urban or rural area, and specialisation in restorative dentistry) and information on restorative treatment decisions for proximal and occlusal carious lesions of permanent teeth. Treatment options in accordance with MID were defined as intervention at dentin level and minimally invasive cavity preparation. Multinomial logistic regression was used for statistical analysis. Results - For the proximal carious lesion, 9.4% of participants said they would employ both MID treatment options; 60.7% said they would choose only one; and 29.9% said they would use neither option. For the occlusal carious lesion, the corresponding figures were 37.2%, 52.1%, and 10.7%. No differences in restorative treatment options were observed among general dental practitioners, dental therapists, and dental students. For the proximal carious lesion, dentists from regions outside Arkhangelsk had 4.15 (95% confidence interval [CI] 1.13–15.27) times higher odds of following one versus both MID treatment options. For the occlusal carious lesion, working experience above 15 years was associated with higher odds of using only one versus both MID treatment options (adjusted odds ratio = 3.04, 95% CI 1.33–6.91). Almost all respondents preferred tooth-coloured materials for restorations; more than 75% chose resin-based composite. Conclusions - The majority of Russian dentists and dental students do not apply the MID concept when treating dental caries in permanent teeth. Clinical protocols on dental caries treatment and dental school curriculums should be updated to place an enhanced focus on evidence-based practice and preventive strategies. Further studies with larger samples of Russian dentists and dental students and alternative methods of recruitment are needed to validate our results

Identification of novel roles and new modes of regulation for the atypical MAP kinases ERK3 and ERK4

The atypical MAP kinases ERK3 and ERK4 are the main characters in this study. ERK3 and ERK4 were for long considered as orphan proteins without known upstream activators. However, in 2011 two studies had detected that group I p-21 protein activated kinase (PAKs) can constitutively phosphorylate ERK3 and ERK4 in their activation loop. The only well-characterized substrate for ERK3 and ERK4 at that time was MAP kinase-activated protein kinase 5 (MK5), also known by the name of PRAK. Recently, the steroid receptor co-activator 3 (SRC-3) has been identified as a substrate for ERK3. What was known about the biological function of ERK3 and ERK4 was limited and working with them was a challenge. In the last four years, more studies were focusing on ERK3 than ERK4. The first aim of this project was to identify novel regulators for both ERK3 and ERK4 in order to gain more knowledge to understand their physiological function. The second aim was to investigate the physiological function of ERK3 regarding cell morphology and migration. Here, in this study we focused mainly about the interaction of ERK3 and ERK4 with two large protein families that are involved in cancer and cancer treatment. The first protein family we investigated was the dual-specificity MAPK phosphatases (DUSPs), where we have used a yeast 2-hybrid assay to analyze all ten members of the mammalian MAP kinase phosphatase (MKP) family for their ability to interact with ERK3 and ERK4. We found that these kinases specifically interact with DUSP2. In paper I, we investigated the interaction between ERK3/ERK4 and DUSP2 in depth. The second protein family that we were interested in was the heat shock protein family. Preliminary data from co-precipitation studies done in our lab to identify interaction partners for ERK3 and ERK4, followed by mass-spectrometry analysis suggested that Hsp90 could be an interaction partner for ERK3 and ERK4. This observation was later supported by results from several high throughput analyses. The interaction between ERK3 and ERK4 with Hsp90 was investigated in paper III. Beside the detection of new targets for ERK3 and ERK4, we chose to investigate the role of ERK3 in cell morphology and migration as described in paper II. Our choice to study only the role of ERK3 in cell migration was based on previous studies, however, that does not exclude ERK4 from also might have a role in cell migration, adhesion and invasion

Design of tumour-specific immunotherapies using dendritic cells – analyses of IL15-DC

Immunotherapy of malignancies aims at activating the patient’s own immune system to fight the tumour affecting the patient. Even though the use of dendritic cells (DC) has shown promising results, the DC vaccination strategy needs improvement, as only few relevant clinical responses could be documented so far. Aim: In this study, the standard protocol to generate monocyte derived DC using GM-CSF and IL-4 was compared to the use of GM-CSF and IL-15. Methods: Monocytes were isolated by plastic adherence from peripheral blood mononuclear cells and cultured for 6 days with GM-CSF and either IL-4 (IL4-DC) or IL-15 (IL15-DC). A fraction of the IL4-DC was stimulated with a cytokine cocktail, while a fraction of the IL15-DC was stimulated by adding TNF-α 24 hours before harvesting. The phenotypes of the four DC populations were determined using flow cytometry. Intracellular signalling pathways were investigated using phospho-specific antibodies in a Western blot. IL-12 production was analysed in an ELISA. Migratory capacity was determined in a chemotaxis assay. Results: Monocytes cultured with GM-CSF and IL-15 developed a DC-like morphology. Phenotypic analyses revealed that both IL4-DC and IL15-DC had down-regulated CD14 expression and up-regulated CD1a expression, although IL15-DC to a lesser extent. IL15-DC showed an enhanced surface expression of co-stimulatory molecules CD80 and CD86 whereas no difference was observed in surface expression of MHC class II and CD40. Upon stimulation, an up-regulation of the maturation marker CD83 and CD86 were observed on IL4-DC only. The IL15-DC had more phosphorylated JNK and ERK than IL4-DC whereas the phosphorylation level of p38 in both IL4-DC and IL15-DC were approximately the same. None of the cell populations produced IL-12 or showed chemotaxis towards CCL19. Conclusions: The generation of IL15-DC turned out to be more problematic compared to the generation of IL4-DC. The stimulatory activity of IL-15 on T-cell proliferation resulted in a high degree of contamination with T -cells in the IL15-DC cultures as I did not have a pure monocyte population to start with. This problem might be overcome by using either alternative monocyte isolation protocols or by reducing the culture period from 5-6 to 3-4 days. Because of the variation in the results of the experiments the data needs to be confirmed with this approach

Associations of the activity and concentration of carbonic anhydrase VI with susceptibility to dental caries: A systematic review and meta‐analysis

Objectives: A number of studies have claimed that carbonic anhydrase VI (CA VI) is associated with dental caries. The aim of this systematic review and meta‐analysis was to systematically review and analyze the literature on the association of CA VI (in terms of concentration and activity) with dental caries. Materials and Methods: A systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta‐Analyses (PRISMA) guidelines. Relevant search terms were employed to search the following databases: PubMed, Web of Science, Scopus, China National Knowledge Infrastructure (CNKI), and Cochrane Library databases. Eligible publications from inception to August 2022 were included. The relevant records were assessed independently by two reviewers, and a meta‐analysis was performed using RevMan 5.3. Results: Out of 237 relevant records from the initial search, 9 met the criteria for this review. The 9 papers, including 477 participants, were qualitatively analyzed. Seven studies with 411 participants (203 caries‐free) were included in the meta‐analysis on CA VI activity, and 2 studies with 141 participants (71 caries‐free) were included in the meta‐analysis on CA VI concentration. The results showed that CA VI activity was significantly higher among participants with caries than their caries‐free counterparts (standardized mean difference (SMD) = 0.894, 95% confidence interval (CI95%): 0.386 and 1.392; p < 0.001), whereas the CA VI concentration was significantly lower among participants with caries than their caries‐free counterparts (SMD = −0.672, CI95%: −1.011 and −0.332; p < 0.001). Conclusions: This meta‐analysis of a relatively small number of studies suggests that the CA VI concentration is lower and CA VI activity is higher in patients with dental caries than in caries‐free individuals; however, further studies are needed to determine the exact role of CA VI in dental caries

Comparison of antimicrobial prescribing for dental and oral infections in England and Scotland with Norway and Sweden and their relative contribution to national consumption 2010–2016

Background - Prescribing in dental practice has a relatively small but important contribution to the quantity of antibiotics prescribed in primary care. This study aimed to analyse antibiotic prescribing in dentistry over time (2010–2016) in 4 different Northern European countries and their relative contribution to national outpatients consumption. Methods - This retrospective study evaluated the frequency and number of national antibiotic prescriptions written by dentists in England, Scotland, Norway and Sweden. The consumption of such antibiotics was measured using WHO defined daily doses (DDDs), DDDs per 100,000 inhabitants per day (DIDs100,000). Results - A total of more than 27 million prescriptions (27,026,599) archived between 2010 and 2016 from the four countries were analysed. The national contribution of Norwegian dentists to the total primary care prescription during this period was 8%. The corresponding figures for Sweden, Scotland and England were 7, 6, and 8%. Dental contribution to National antibiotic use in all four countries has decreased over the study time period for commonly prescribed antibiotics in dentistry, i.e., the beta-lactams (Phenoxymethyl penicillin/Amoxicillin) and metronidazole. There were less numbers of prescriptions by dentists in Norway and Sweden compared to England and Scotland. Marked differences in some classes of antibiotics were noted with Phenoxymethyl penicillin dominating in Sweden/Norway compared to Amoxicillin and Metronidazole in England/Scotland. In England and Scotland, dentists were the largest prescribers of metronidazole in primary care. Clindamycin prescriptions was higher in Norway and Sweden. Conclusion - Noticeable differences exist in prescribing patterns for the management of oral infections. High levels of metronidazole use in England and Scotland also require further analysis. All countries over the study period showed a decrease in total numbers of antibiotics prescribed

A novel role for atypical MAPK kinase ERK3 in regulating breast cancer cell morphology and migration

ERK3 is an atypical Mitogen-activated protein kinase (MAPK6). Despite the fact that the Erk3 gene was originally identified in 1991, its function is still unknown. MK5 (MAP kinase- activated protein kinase 5) also called PRAK is the only known substrate for ERK3. Recently, it was found that group I p21 protein activated kinases (PAKs) are critical effectors of ERK3. PAKs link Rho family of GTPases to actin cytoskeletal dynamics and are known to be involved in the regulation of cell adhesion and migration. In this study we demonstrate that ERK3 protein levels are elevated as MDA-MB-231 breast cancer cells adhere to collagen I which is concomitant with changes in cellular morphology where cells become less well spread following nascent adhesion formation. During this early cellular adhesion event we observe that the cells retain protrusive activity while reducing overall cellular area. Interestingly exogenous expression of ERK3 delivers a comparable reduction in cell spread area, while depletion of ERK3 expression increases cell spread area. Importantly, we have detected a novel specific endogenous ERK3 localization at the cell periphery. Furthermore we find that ERK3 overexpressing cells exhibit a rounded morphology and increased cell migration speed. Surprisingly, exogenous expression of a kinase inactive mutant of ERK3 phenocopies ERK3 overexpression, suggesting a novel kinase independent function for ERK3. Taken together our data suggest that as cells initiate adhesion to matrix increasing levels of ERK3 at the cell periphery are required to orchestrate cell morphology changes which can then drive migratory behavior

A Quantitative Analysis of Colonic Mucosal Oxylipins and Endocannabinoids in Treatment-Naïve and Deep Remission Ulcerative Colitis Patients and the Potential Link With Cytokine Gene Expression

Background - The bioactive metabolites of omega 3 and omega 6 polyunsaturated fatty acids (ω-3 and ω-6) are known as oxylipins and endocannabinoids (eCBs). These lipid metabolites are involved in prompting and resolving the inflammatory response that leads to the onset of inflammatory bowel disease (IBD). This study aims to quantify these bioactive lipids in the colonic mucosa and to evaluate the potential link to cytokine gene expression during inflammatory events in ulcerative colitis (UC). Methods - Colon biopsies were taken from 15 treatment-naïve UC patients, 5 deep remission UC patients, and 10 healthy controls. Thirty-five oxylipins and 11 eCBs were quantified by means of ultra-high-performance liquid chromatography coupled with tandem mass spectrometry. Levels of mRNA for 10 cytokines were measured by reverse transcription polymerase chain reaction. Results - Levels of ω-6-related oxylipins were significantly elevated in treatment-naïve patients with respect to controls, whereas the levels of ω-3 eCBs were lower. 15S-Hydroxy-eicosatrienoic acid (15S-HETrE) was significantly upregulated in UC deep remission patients compared with controls. All investigated cytokines had significantly higher mRNA levels in the inflamed mucosa of treatment-naïve UC patients. Cytokine gene expression was positively correlated with several ω-6 arachidonic acid–related oxylipins, whereas negative correlation was found with lipoxin, prostacyclin, and the eCBs. Conclusions - Increased levels of ω-6-related oxylipins and decreased levels of ω-3-related eCBs are associated with the debut of UC. This highlights the altered balance between pro- and anti-inflammatory lipid mediators in IBD and suggests potential targets for intervention

Association of Polymorphism of the Methyl Tetrahydrofolate Reductase (MTHFR) Gene with Anti-Seizure Medication Response in Pediatric Patients in Jeddah, Saudi Arabia

Background and Objectives: Epilepsy is a chronic brain disease, with inherent and noninherent factors. Although over 20 anti-seizure medications (ASMs) are commercially available, nearly one-third of patients develop drug-resistant epilepsy. We evaluated the association between the clinical features and the methyl tetrahydrofolate (MTHFR) rs1801133 polymorphism and ASMs response among pediatric patients with epilepsy. Materials and Methods: This was a multicenter, retrospective, case&ndash;control study of 101 children with epilepsy and 59 healthy children in Jeddah. The MTHFR rs1801133 polymorphism was genotyped using the real-time polymerase chain reaction TaqMan Genotyping Assay. Results: Among the patients with epilepsy, 56 and 45 showed good and poor responses to ASMs, respectively. No significant genetic association was noted between the single-nucleotide polymorphism (SNP) rs1801133 within the MTHFR gene and the response to ASMs. However, a significant association was noted between reports of drug-induced toxicity and an increase in allele A frequencies. The MTHFR rs1801133 genotype was significantly associated with the development of electrolyte disturbance among good and poor responders to ASMs. Conclusions: This is the first pharmacogenetic study of MTHFR in patients with epilepsy in Saudi Arabia that found no significant association between the MTHFR SNP rs1801133 and gene susceptibility and drug responsiveness. A larger sample size is needed for testing gene polymorphisms in the future