EPMA World Congress: Traditional Forum in Predictive, Preventive and Personalised Medicine for Multi-Professional Consideration and Consolidation

The breast cancer (BC) epidemic in the twenty-first century is evidenced by the approximately two million new cases and half-million related deaths annually worldwide [1]. The overall situation is particularly dramatic in specific subgroups such as those with triple-negative BC: more than 50% of these patients die of metastatic BC within the first 6 months of diagnosis. Consequently, breast cancer management in general should be thoroughly reconsidered, promoting a new paradigm based on predictive diagnostics, new screening programs, targeted prevention and individually tailored treatments. Due to the extremely high level of complexity of the field, the new paradigm represents a spectrum of complementary components that constitute the innovative PPPM concept. At the EPMA World Congress 2015, the dedicated working group elaborated the working hypothesis and concepts for a multi-centre pilot project; this action has consequently been taken [2], and the new knowledge collected will be summarised. The overall results substantially extend the Bseed and soil^ theory of metastasis with the finding that among individuals at risk, a strong predisposition toward the formation of the systemic hypoxic pre-metastatic niches can be established long before the breast malignancy is clinically manifested [3]. The BFlammer syndrome^ phenotype may strongly contribute to BC onset [4] and aggressive metastatic disease with particularly poor outcomes [4]. Both preand postmenopausal women may be affected [3-5]. Individualised patient profiles and the involvement of primary caregivers (family doctors) are essential for implementation of PPPM in BC management [5].

PPPM in Cardiovascular Disease (CVD) is a specialised session featuring a collection of innovative data regarding CVD prevention in stratified cohorts, as well as molecular pathways targeted for CVD prediction and prognosis. PPPM in Dentistry is a specialised session with multi-professional expertise which will present a whole spectrum of the field related innovation including the role of stem cells in tooth regeneration (keynote speech by Prof. Dr. Mahmood Mozaffari, University of Augusta, USA, September 16 th ), dental health in childhood, caries prevention in stratified patient cohorts, personalised orthopaedic treatments, and prevention of severe complications in CVD patients with dental pathologies, amongst others. Innovative Technologies in PPPM is a specialised session with multiprofessional expertise which will introduce unique technologies such as liquid biopsy (keynote speech by Prof. Dr. Evi S. Lianidou, University of Athens, Greece, September 15 th ), health condition-specific biomarker panels discovered in human tear fluid and saliva, and disease-specific multi-omics approaches, amongst others. Workshop of young professionals in PPPM (EPMA-YPS, September 14 th ) is the traditional session presenting new ideas developed by young researchers, including BMini-encyclopaedia of Wound Healing: Lessons for PPPM^, BLongevitygood luck or reasonable lifelong healthcare strategy?^The best presentations chosen by the international jury-board of prominent scientists will be awarded by the EPMA. The award ceremony will take place on September 16 th . The University of Malta, as the host of the EPMA World Congress 2017, has created an excellent platform for an effective congress and a pleasant atmosphere for the thematic sessions scheduled in the congress programme, as well as spontaneous professional meetings to exchange professional information and to plan new, successful projects in PPPM.
Welcome to the EPMA World Congress 2017 in Maltaenjoy the meeting! Objective: Triple-negative breast cancer (TNBC) patients derive little benefit from target-specific therapies due to a lack of favourable prognostic targets, ER, PR and HER2. The aim of this project was to define biomarkers to measure PP2A complex activity in breast cancer predicting a novel therapeutic class. Methods: A multiplex assay using branched DNA technology was used to measure the expression of 40 genes in RNA derived from cell lines (n=12) and formalin-fixed, paraffin-embedded (FFPE) archival material from breast cancer patients (2009)(2010)(2011). First, the PP2A activity biomarkers (PABs) were validated in breast cancer cellular models and compared to real-time PCR results. The QuantiGene 2.0 multiplex assay was used to measure expression of these markers in a cohort of laser-microdissected breast cancer (n=97) and normal breast tissue cases (n=30). Sensitivity of cell lines to FTY720 was measured using MTT assays. Results: Overexpression of these biomarkers in cell lines predicted sensitivity to FTY720 and were down-regulated in a dose-dependent manner.
Of interest, 37% of TNBC cases expressed high levels of PABs, and were associated with negative prognostic indicators (P = 0.023). Conclusion: TNBC cell lines sensitive to the PP2A activator FTY720 showed a high expression of PABs. Using patient material, PAB overexpression was common in the TNBC subtype. The novel biomarkers (PABs) provide a multiplex gene expression signature for a potential therapeutic group sensitive to PP2A activators.
Accelerated ageing in breast cancer aetiology and progression: Multi-factorial patient profiles for effective prediction and prevention of the disease Aviv Peer 1 and Olga Golubnitschaja 2,3,4 1 Abstract Diagnosis of breast cancer subtypes determines therapeutic options and prognosis. These subtypes are hallmarked by the expression of the human epidermal growth factor receptor 2, and oestrogen and progesterone receptors (HER2, ER and PgR). An audit of breast cancer diagnosed in Malta between 2009 and 2016 indicates that 85% were ER-positive, 11% were HER2-positive, and 10% were negative for all receptors. Current guidelines for diagnosis of HER2 are based on immunohistochemistry and confirmatory testing by fluorescent in situ hybridisation (FISH). HER2 copy number assessment on 960 breast cancer samples (cBIOPortal [1,2]) is estimated to detect 1.3% amplification in the absence of expression (false positives) and 4.2% diploid accompanied by overexpression of the gene (false negatives). A multiplex branched DNA assay has been validated for quantifying gene expression on degraded RNA derived from formalinfixed paraffin-embedded (FFPE) breast cancer tissue [3]. Using the diagnostic results as benchmark, this assay has average sensitivity and specificity of 100% and 97.8% for the determination of HER2 status and ER status, respectively. The gene expression assay is a novel, quick multiplex method that can accurately diagnose breast cancer subtypes, eliminating subjectivity of interpretation and minimising technical variation. This method has a wide range of possible applications in the diagnosis of tumours and is adapted to the current diagnostic workflow. Genentech), in conjunction with chemotherapy and hormonal therapy if required, as an adjuvant or neoadjuvant to surgery in operable cases. The rates of clinical response to treatment have not yet been recorded locally. A retrospective clinical audit is thus currently in progress in collaboration with the Sir Anthony Mamo Oncology Centre (SAMOC). All HER2-positive breast cancer patients tested between 2010 and 2016, extracted from Mater Dei Hospital laboratory records, were included for the review of patient clinical records from their treatment history files. The following information is being collected where available: All clinical and radiological responses are given a final assessment using the Response Evaluation Criteria In Solid Tumors (RECIST) 1.1. The findings to date will be presented and compared to published findings in other countries. This audit will allow treatment response rates to be correlated to laboratory findings on archival tissue specimens in the ongoing Breast Cancer Project at the University of Malta.

Abstract
Background. CA 19-9 is the only approved biomarker for monitoring tumor response in patients with pancreatic adenocarcinoma (PDAC), but it has several limitations, highlighting the need for new predictive biomarkers [1]. Mutant KRAS is a driving oncogene, occurring in 75-95% of PDAC [2]. Aim. To develop an analysis tool based on plasma to monitor the variation in mutant KRAS alleles in PDAC being treated with first-line chemotherapy.
Methods. Twenty-seven PDAC patients were enrolled in this study. Six milliliters of plasma were prospectively collected at baseline, after 15 days of chemotherapy and at each clinical follow-up. CftDNA was extracted from plasma with a specific kit and analyzed by the Droplet Digital™ PCR (ddPCR) for KRAS mutations in codon 12 (G12X) and G13D.
Results. At baseline, 19 out of 27 patients (70.3%) were carriers of one of the KRAS mutations. Eight patients had a negative cftDNA mutKRAS at baseline, and one out of eight became positive at day 15, and another one at the first radiological evaluation. There was a statistically significant difference in progressionfree survival (PFS) between patients with increased vs. stable/ reduced cftDNA at the 15-day sample (median PFS 2.5 vs. 7.5 months, p=0.03; see Fig. 1). A statistically significant difference was also observed in median overall survival (OS; 6.5 vs. 11.5 months in cftDNA increase vs. reduction, respectively, p=0.009).
Conclusions. cftDNA analysis of mutKRAS could be a reliable non-invasive approach for early prediction of tumor progression and responsiveness to chemotherapy. However, in order to assess these preliminary data, further analyses are needed.  Aim. To monitor the early variation in EGFR mutations in NSCLC patients using circulating cell-free tumor DNA (cftDNA) extracted from plasma. Methods. Seventy-nine NSCLC patients were enrolled in this ongoing explorative study. Six milliliters of plasma was prospectively collected at baseline and at each clinical follow-up. CftDNA was extracted from plasma with the QIAamp Circulating Nucleic Acid Kit (Qiagen®) and analyzed using the Droplet Digital PCR (ddPCR; (BioRad®) for EGFR p.L858R, exon 19 deletions and p.T790M mutations.
Results. Forty-four patients testing positive for EGFR activating mutation were treated with the first-generation EGFR-TKIs, while 35 patients testing positive for p.T790M at baseline were treated with AZD9291, the potent, irreversible EGFR-TKI selective for the p.T790M mutation. During the treatment with first-generation TKIs, nine patients experienced clinical progression after a median of 9 months and, testing positive for p.T790M, switched therapy to AZD9291. Interestingly, the appearance of low levels of activating EGFR mutations anticipated the appearance of both high levels of the resistance mutation and clinical progression occurring several months later.
Conclusions. CftDNA analysis is useful not only for monitoring clonal evolution of NSCL cancer cells under the stress of EGFR-TKIs, but also in evaluating treatment responses and choosing the more effective therapy in individual patients.
Detection and analysis of circulating tumour cells in small cell lung cancer utilizing a flexible rare cell scanning platform 1 Kilpatrick MW, 1 Kershnar ER, 1 Borgerding RH, 2,3 Robson P, 2 Sivakamasundari V. 1 Ikonisys Inc., New Haven, CT USA 2 The Jackson Laboratory for Genomic Medicine, Farmington, CT USA 3 Dept of Genetics & Genome Sciences, UConn School of Medicine, Farmington, CT USA flexible rare-cell imaging approach that could be applied to CTCs enriched by any desired method. Enrichment can be marker-based, size-based, or microfluidic, or large (two million per slide) numbers of cells can be scanned quickly and efficiently, requiring no enrichment. This approach was able to detect a single tumor cell in one million peripheral blood mononuclear cells (PBMC), with either filtration-based enrichment or no enrichment. CTCs were identified in patients with a variety of cancers [3], utilizing both tumorspecific antibodies and fluorescent in situ hybridization (FISH) markers for the detection of aneuploidy. Analysis of a 2-ml blood sample from an SCLC patient identified eight CTCs (Fig. 1). This uniquely flexible approach permits the interrogation of a CTC for up to five nucleic acid or protein markers, allowing, in addition to enumeration, the molecular characteristics of the detected CTCs to be explored.

Abstract
Glioblastoma multiforme (GBM) represents one of the most malignant tumors of the primary central nervous system in adults, with considerably high mortality [1,2]. Standard therapies including neurosurgery, radiotherapy and chemotherapy with temozolomide have only a limited effect on the long-term survival of GBM patients [3,4]. A number of targeted anti-cancer therapeutics, including low-molecular-weight tyrosine kinase inhibitors or monoclonal antibodies, have been evaluated in numerous clinical trials with newly diagnosed as well as recurrent GBM. Unfortunately, until today, only one targeted anti-tumor drugthe antiangiogenic inhibitor bevacizumabhas been approved for the treatment of recurrent GBM, albeit not in the EU [5]. Moreover, its effect on the overall survival of patients with recurrent GBM remains controversial.
Major advances in cancer immunotherapy over the past few years now offer hope for the possible future use of such treatment strategies for patients with GBM as well [6,7]. This presentation will discuss the recent preclinical and early clinical results from studies evaluating anti-tumor immunotherapy using so-called immune checkpoint inhibitors that might improve the very unfavorable prognosis of patients with GBM in the near future.

Abstract
Most colorectal cancers are considered sporadic, and often the cause is not known. Biomarkers of individual repair capacity (IRC) play a role not only in the risk of developing cancer, but also in virtually every aspect of the course of the disease. In our case-control study of 273 colorectal cancer patients and 187 cancer-free controls, the 22 single-nucleotide polymorphisms (SNPs) in 12 genes (GSTP1, CYP1A2, CYP1B1, CYP2C8, CYP2C9, CYP2E1, CYP2W1*2, CYP19A, MTHFR, XRCC1, hOOG1, DPYD) were genotyped and analyzed for their correlation with the risk of sporadic colorectal cancer in multivariate logistic regression models. There were significant genotype associations of the following SNPs: rs1695 GSTP1, rs1801133 and rs1801131 MTHFR, rs25487 XRCC1 rs1052133 hOOG1 and rs1801160 and 1801159 DPYD. Haplotype analysis of four DPYD polymorphisms showed significant differences in the distribution of the IISt haplotype between cases and controls. In comparison to the most common haplotype (VISt), the IISt haplotype was associated with increased risk of colorectal cancer (p = 0.038, OR = 2.733, 95% CI = 1.019-7.326).

Abstract
Despite recent advances in clinical and experimental colorectal cancer (CRC) research, the optimal approach for stratifying CRC patients remains uncertain. We previously described amplification of TRIB1 and MYC in 14.5% and 7.4% of a CRC cohort (n=118), respectively, and found these amplifications significantly correlated (p = 0.0001) in a number of cases 1 . Our current aims are (i) to understand the prognostic role and function of TRIB1 in CRC, and ii) to identify a clinical and molecular signature for this subgroup of patients that can be used for prognostication and therapeutic targeting. To this end, we are validating TRIB1 expression/amplification in 500 Maltese patients diagnosed with CRC between 2008 and 2011. A tissue microarray platform was used to quantify gene amplification by fluorescence in situ hybridisation (FISH) and protein expression by immunofluorescence imaging. TRIB1 expression will be associated with several clinicopathological parameters. CRISPR/ Cas9 technology was used to create three stable TRIB1 knock-down CRC cell lines, which were used for functional analysis using the Celigo® image cytometer to measure cell cycle progression, proliferation, tumour growth, and invasion. Western blotting will be used to interrogate several pathways including MAPK, PI3K/AKT, NF-κB, cell cycle, and apoptosis. Our data show that Trib1 protein expression in the patient cohort is significantly positively correlated with Akt, BRCA1, Met, Erk, Stat3, MEK, Myc, caspase 3, PTEN, Stat3, and CDK2 (p =<0.05). These findings will fully characterise the TRIB1-positive subgroup, laying the foundation for therapeutic targeting and companion diagnostics. Correct patient stratification increases patient survival, reduces recurrence, and minimises overtreatment.

Abstract
Nanoparticles are gaining increasing popularity as medical tools against cancer. Their applications range from diagnostic purposes to possible therapeutic interventions. Although the ongoing research has revealed some promising results, a serious manifestation of nanoparticle-induced adverse effects has been reported by many researchers. Thus, the evaluation of the cytotoxicity that nanoparticles exert on cells must be thoroughly examined. This study focuses on the response of two cancer cell lines, HeLa cervical cancer and H460 lung cancer cells, to zinc oxide, gold and silicon dioxide nanoparticles. Cells were exposed to various doses over a 24-h period. Cytotoxicity levels were then evaluated by MTT and LDH cytotoxic assays. Results showed reduced cell viability over both cell lines and for all nanoparticle treatments. However, cytotoxicity levels varied, with correlations to dose-, nanoparticle-or celldependent cytotoxicity only in specific aspects of the experiment. Although further research is needed to reveal the full potential, along with the complete range of side effects, of nanoparticles, the versatile performance of these nanoscale materials can be a powerful tool for researchers. Thus far, gold nanoparticles have been the most popular in the modern face of personalised medicine, with applications in imaging, drug delivery and enhancement of conventional therapeutic strategies. In addition, zinc oxide and silica nanoparticles have been used in cancer diagnostics and therapeutics as well as imaging and drug monitoring, applications that make them promising candidates for individually targeted medicine [1][2][3][4][5].

Abstract
Psychiatry as a clinical discipline is largely driven by a symptom-based approach that underpins current diagnostic (e.g. DSM-V) and therapeutic decision-making processes. Reliable biological markers are largely absent from current diagnosis and treatment. In addition, evaluation of treatment efficacy rests solely on patient self-report and clinical judgement. Does psychiatry as a field have the opportunity to develop beyond a clinically driven Bwait-and-see^approach [1]? In this presentation, the concept of personalized psychiatry will be introduced, which integrates both biological systems and clinical markers, incorporating prevention, prediction and participation as key features of personalised medicine. The identification of distinct clinical and biological disease trajectories underscores a prospective and proactive preventive approach to mental illness for a large proportion of people at risk for a first or for subsequent episodes of mental illness [2,3]. Pre-treatment stratification into meaningful subgroups of patients is a necessary part of personalized psychiatry, in order to inform more effective treatment decisions [4]. A specific biologically determined clinical subgroup of patients with depressive symptomatology consists of patients presenting with increased peripheral inflammatory markers that may make these suitable diagnostic markers for clinical stratification into specific treatment groups. In addition, the genomics of treatment response to pharmacological and psychological interventions is crucial for a personalized approach in psychiatry. The prospective and predictive approach taken in personalized psychiatry requires bioinformatics modelling that considers both clinical and biological markers for more accurate prediction of disease onset and disease course, as well as prediction of treatment needs. Using our multimodal probabilistic prediction model, prediction of treatment needs in psychosis have been illustrated [5]. The outlined concept of personalized psychiatry as illustrated by several examples comprehensively describes key ingredients needed for personalizing the diagnosis and treatment of patients with psychiatric disorders. A clinical experimental validation process is essential and among the key challenges in personalized psychiatry for the translation of findings into clinical practice and guidelines and for patient participation.

Abstract
Proactive healthcare relies on the capacity to detect health risk and early disease states. The recent Institute of Medicine Report on diagnosis has identified diagnosis as a major source of error in the health system, even for established disease [1]. Diagnosis and the decisions that follow rely on good judgementoften involving the assessment of probability and comparisons of predictions over time for alternate scenarios. Work in psychology around cognitive bias, heuristics and decisionmaking show that humans are poor at these. Even the well-trained routinely fail at assessing probability and at predicting the outcome of even simple scenarios over time. Recognition of these weaknesses and training against them does not fix themwe seem unable to correct for our own biases. Indeed, it has been shown in well-constructed experiments that our confidence rises as we are more likely to predict the wrong outcome [2,3]. Informatics and its machines are important in all aspects of proactive health care, but helping to accommodate for the way that people think offers an important opportunity for the advancement of predictive, preventive, personalised medicine (PPPM) [4]. Machine assistance can help people avoid likely errors and correct for biases that will lead to a less rational outcome, but using the way that people think in a constructive way, they can also be used to reinforce the most appropriate interpretation and action. Scientific medicine could be made much easier, and informatics and its machines can help in new ways.

Abstract
We present the case of a 53-year-old male patient who had been suffering from symptoms of decompensated and chronified tinnitus for 4 years [1], most likely caused by work stress. In addition, the patient developed comorbid decompensated insomnia. Because of potential bidirectional connections between tinnitus and sleep disorders, an interdisciplinary approach to treatment was chosen. The treatment plan we developed consisted of ten sessions of repetitive transcranial magnetic stimulation (rTMS) for tinnitus [2], followed by ten sessions of cognitive behavioral therapy (CBT) for tinnitus and insomnia. We used the Tinnitus Questionnaire (TF) to assess tinnitus severity, the Beck Depression Inventory (BDI-II) for depressive symptoms, and the WHO Well-Being Index (WHO-5) for subjective well-being. Improvements were achieved with regard to everyday functioning, as the patient went from decompensated and severe to clinically negligible TF scores, from minimal to no depressive symptoms, and from just above critical to above average well-being. Combining equipment-based and psychological approaches to treatment proved successful in this case. We conclude that a combination of rTMS and CBT may be considered as an effective treatment for chronic tinnitus and comorbid sleep disorders.

Abstract
The aim of this paper is twofold. The first to assess the role of comorbidity as a risk factor in predicting suicidality among patients with depressive disorders and those with comorbidity or dual diagnosis. The second is to discuss implications for prevention. Within the framework of the UGD-supported project, the data were collected from Clinical Hospital, Stip, Macedonia, during the period January 2015 to March 2017. The sample consists of 140 patients (64% female and 36% male) aged 19-72 years. The respondents were divided on the basis of mono and comorbid diagnosis into four subgroups: 1) those with depressive disorder, 2) those with depressive disorder and physical illness, 3) those with depressive disorder and another mental disorder, and 4) those with depressive disorder, physical illness and mental disorder. Data were collected using questionnaires about sociodemographic data, structured interviews and medical documentation, while suicidal behavior was studied using the C-SSRS scale [1,2]. Written informed consent was obtained from all study participants.
The specific focus was on examining the relationship of suicidal thoughts with gender and comorbidity, frequency and intensity, as well as differences in preparation, trials, methods and number of suicides. Additionally, we examined the characteristics of suicide in 46 respondents and whether there was a difference among groups in terms of suicidal behavior. The results are in line with our expectations, and they show that comorbidity is a significant factor in predicting suicidal behavior [3][4][5]. A detailed statistical analysis is reported in the paper.
Although we are talking about a small sample, at the level of absolute numbers and percentages, where comorbidity is a clinical reality and suicides are related to the number of diagnoses, a larger number of diagnoses leads to a greater risk of suicidal behavior.

Abstract
Lamotrigine is a novel antiepileptic drug metabolised by UGT enzymes, mainly UGT1A4 [1,2] and UGT2B7 [2]. Genetic polymorphisms of UGTs result in different phenotypes by affecting the expression levels or activity of individual UGT enzymes, hence variability in drug metabolism and elimination [3,4,5]. Thus, pharmacogenetics is extremely useful in the individualisation of lamotrigine drug therapy. Moreover, paediatric populations with epilepsy offer a specific challenge in dose individualisation, since they undergo significant developmental changes from childhood to adulthood [6]. The aim of this study was to investigate the influence of genetic polymorphisms of UGT1A4 and UGT2B7 on drug concentration in Maltese paediatric patients with epilepsy. Blood samples from 18 Maltese paediatric patients were collected. Genomic DNA was extracted using well-established standard protocols. Primers were designed to span the entire coding sequence of the UGT1A4 and UGT2B7 genes. DNA amplification of the specific genes was performed by PCR on the respective patient samples. Gel electrophoresis was used to confirm that the desired DNA fragment was obtained. The PCR products were then purified and sequenced using the Sanger sequencing method.
Various mutations were identified in both genes of interest, leading to amino acid changes in the majority of patients. This may result in inter-individual variability in the pharmacokinetics of lamotrigine. This research will lead to improved therapeutic outcomes for paediatric populations with epilepsy, by providing a greater understanding of the contribution of pharmacogenetics to the individualisation of drug therapy and development of personalised medicine.

Abstract
Purpose: The aim of this study is to determine the effects of a physical activity program for first-stage Alzheimer's patients with regard to cognitive function, depression and quality of life. Materials and methods: This is single-blinded randomized controlled trial. The experimental group included 75 Alzheimer's patients (40 in the experimental group, 35 in the control group), as diagnosed by physicians, who were in the moderate stage of the disease (SMMSE score: 23-24) and who were allowed to undertake physical activity. An information form including the participants' sociodemographic characteristics and medical history, the Standardized Mini Mental State Examination (SMMSE), the Quality of Life-Alzheimer's Disease scale (QoL-AD) and Beck Depression scale were used for data collection. Statistical analysis was performed using SPSS 18.0 (IBM, Armonk, NY, USA). Continuous data were presented as medians (interquartile range), categorical data as counts and percentages. The researchers used descriptive statistics, including numbers, percentages, means and standard deviations.
Results: This study showed that the physical activity program improved cognitive function, depression and quality of life among first-stage Alzheimer's patients (p<0.05). These results suggest that regular physical activity programs should be used in routine practice to prevent cognitive decline and depression, and to the improve quality of life for first-stage Alzheimer's patients.
Conclusion: According to this study's findings, cognitive function, depression and quality of life among first-stage Alzheimer's patients improved as a result of a physical activity program.

Abstract
The overall objective of the Systems Medicine of Mitochondrial Parkinson's Disease (SysMedPD, 2017-2021) project is to identify novel drug candidates capable of slowing the progression of neurodegeneration in the subset of Parkinson's disease (PD) patients with overt mitochondrial dysfunction. Multi-modal phenotypic characterisation of cohorts of monogenic PD patients with overt mitochondrial dysfunction will be used as an anchor for the discovery of two extreme cohorts of idiopathic PD patients: with and without detectable mitochondrial dysfunction. A suite of personalised in vitro, in vivo, and in silico models will be generated using induced pluripotent stem cells (iPSCs) from selected subjects and controls. An industrial-quality 3D microfluidic cell culture product, specifically designed for the culture of iPSC-derived dopaminergic neurons, will be developed for use in a morphological and bioanalytical screen for lead compounds in order to reduce mitochondrial dysfunction. By monitoring motor behaviour and in situ striatal neurochemistry at high temporal resolution, the in vivo response to lead compounds will be characterised in humanised mouse models with striatal transplants of iPSC-derived dopaminergic neurons derived from PD patients. Personalised computational models of dopaminergic neuronal metabolism and mitochondrial morphology will be developed. These in silico models will be used to accelerate drug development by prioritising pathways for metabolomic assay optimisation, stratifying idiopathic PD patients by degree of mitochondrial dysfunction, predicting new targets for reducing mitochondrial dysfunction and mechanistic interpretation of in vitro and in vivo experimental results. SysMedPD unites a highly experienced multidisciplinary consortium in an ambitious project to develop and apply a systems biomedicine approach for preclinical identification of candidate neuroprotectants, for the estimated 1-2 million people worldwide who suffer from PD with mitochondrial dysfunction. An overview of the scientific rationale and main objectives of the project shall be presented, followed by preliminary results to date, particularly with respect to multi-scale mechanistic computational modelling of PD and the implications for systems medicine in other complex diseases.
Flammer syndrome, a potential risk factor for central serous chorioretinopathy? Josifova Tatjana  Abstract Interleukin-7 receptor-alpha (IL7-Ra) is involved in homeostasis of autoreactive T cells in multiple sclerosis (MS). Several studies have confirmed an association between the polymorphism rs6897932 C/T in the IL-7Ra gene and MS susceptibility. In our study, we tried to identify a possible association between this polymorphism and the risk of MS along with the rate of disease progression in a Slovak population. The rs6897932 polymorphism was detected in 219 clinically diagnosed MS patients and 218 healthy control subjects. We ascertained the Multiple Sclerosis Severity Score (MSSS) for each patient, and based on these scores, we chose 53 patients with rapidly progressing and 57 patients with slowly progressing disease. DNA samples were extracted from peripheral white blood cells and genotyped by PCR and restriction analysis. We found a significantly higher frequency of ancestral allele C in MS patients than in controls (p = 0.033).
We found no significant differences in genotype or allele frequencies of SNP rs6897932 between the slow-and rapid-progression subgroups. Allele C of rs6897932 seems to increase the risk of MS development in a Slovak population. We did not confirm an association between this gene polymorphism and the rate of MS progression.

Abstract
Ataxia is a disorder which presents with varying degrees of loss of control of normal bodily movements. Although acquired ataxia may be encountered, it is more commonly found as a hereditary condition. A wide array of genetic variants have been identified as the cause of each of the multiple subtypes of inherited ataxia. Although several members of a Maltese family have been diagnosed with hereditary ataxia, little is known about the nature of the causative variant/s. The family history suggests an Xlinked recessive pattern of inheritance. This study undertook the identification and profiling of genetic variant/s causing this disorder in the affected family members by means of next-generation sequencing using a clinical exome panel coupled with bioinformatics analysis. The data are presently being followed up by PCR amplification of the identified variant/s or gene/s and subsequent Sanger sequencing of the PCR product. Once established, efforts can be made to understand the mechanisms underlying the causative variant/s such that a cure, or more likely a symptomatic treatment, may be provided. Additionally, genetic counselling can be provided to guide future generations of this family line with regard to aspects of family planning. Daily entities such as feeling of thirst, pain sensitivity and smell perception are generally conditioned by individual genetic set-up and post-genomic/epigenetic regulationstrongly influenced by social and environmental factors. On one hand, an abnormal (substantially deviating from the average) sense regulation is basically neglected at the standard level of medical care. On the other hand, an accumulated knowledge in the area of sense regulation demonstrates its relevance for development and progression of severe pathologies. For example, a significantly reduced feeling of thirst leading to low water intake results in body dehydration that is a well-acknowledged risk factor for breast cancer, amongst others [1]. Altered sense regulation attracts attention in bothdiabetes mellitus, leading to cascaded comorbidities [2], and Flammer syndrome, suboptimal health conditions relevant for several pathologies investigated [3][4][5] (see Fig. 1). Thirsty feeling Reduced in Flammer syndrome with possible adverse health effects: short-term: dehydration, headache, migraine, mood disturbances, amongst others long-term: association with breast cancer and aggressive metastatic disease [1,5]; Increased in type 2 diabetes mellitus, possibly linked to fatigue, nausea, cancer, preeclampsia, hip fractures, nephropathy, coronary heart diseases, hypertension, venous thromboembolism, cerebral infarct Pain sensitivity Increased in Flammer syndrome, possibly linked to multiple sclerosis, fibromyalgia, impaired wound healing [4]; Decreased in type 2 diabetes mellitus and linked to hypertension, neuropathy, impaired wound healing, amongst others Smell perception Increased in Flammer syndrome [4]; consequences are currently unclear Decreased in type 2 diabetes mellitus, possibly linked to cognitive impairments and neurodegenerative processes, also reduced in Alzheimer´s disease, Parkinson's disease and multiple sclerosis [ The cost of healthcare in the United States (US) is 50-100% more than other developed countries. Indeed, one of its closest neighbors -Cubaachieves an average lifespan equal to the US and lower infant mortality at one-tenth the cost. Californianicknamed the BGolden State^for both its nineteenth-century gold mining and the color of its landscapewould be ranked roughly sixth in the world by GDP. California is frequently on the Bcutting edge^of technology, climate change, and social policy. However, it is somewhat schizophrenic regarding healthcare.
In medical education, residents are increasingly burdened with ensuring maximize reimbursement for hospitals rather than lea rni ng the ir specifi c medical /surgical special ty a nd interacting with patients. Instruction on basic science and clinical medicine is being supplanted with instruction on enhancing the electronic medical record (e.g. maximizing the disease process Bcodes^for each patient) to ensure maximum reimbursement.
Another theme in both California and the US in general is the need for physicians to obtain advanced degrees such as the master of business administration (MBA) in order to participate in a healthcare organization. This highlights the difference between administering (i.e. enforcing a system's rules) and managing (i.e. optimizing a system's outcome), which is a fundamental difference between healthcare in the US and in the United Kingdom and elsewhere. However, the California state legislature in 2017 has introduced the BHealthy California Act^-a proposal for universal singlepayer healthcare coverage. In California it appears that legislators, no physicians, are playing Bcatch-up^with the rest of the developed world regarding healthcare! consequence, resistance to existing medicines continued to increase, while the number of new antibiotics receiving marketing authorization declined [1]. This study aims to identify major patient access barriers in PPPM, and to understand how innovative companies overcome these market access restrictions. The presenter suggests that there exist a number of ways for producers to overcome market access barriers, for example, through early-phase convergence with key stakeholders, introduction of miscellaneous risk-sharing agreements, policy shaping, differentiating value-added services, and Btailoring^evidence generation to stakeholder requirements. [1,3,4]. In personalized medicine, therapeutic intervention should be applied before the disease symptoms appear. This requires highly specific and highly sensitive biomarker arrays (BM) able to predict disease onset and therapeutic response [1]. Reliable, multiparametric semi-diagnostic and/or diseasepredictive biomarker arrays are urgently needed for different diseases to improve early diagnosis or for monitoring and improving new and existing therapeutic strategies. The employment of biochip technologies for the simultaneous measurement of several biomarkers (multiplex assay) is necessary in biomedicine, since common assays (ELISA, Western blot, etc.) allow only the quantification of a single analyte at nano-to picogram levels, and are inadequate for quantifying low-abundance biomarkers in a complex matrix such as serum, urine or saliva [2].

References
Aim of the institute: The proposed project, Christian Doppler -Laboratory for Personalized Medicine (CD-PM), is a multidisciplinary and collaborative effort aimed at performing innovative and novel research in the field of modern molecular diagnostics. The development and application of a novel, highly sensitive biochip will significantly enhance knowledge about biomarker detection in the early stage of diseases and thus their role in predictive medicine. The thinking behind this project includes both basic and oriented research, with a concrete commercial application.

Application areas, examples:
The proposed unique and novel, highly sensitive technique is currently being evaluated for the early detection of beta-cell dysfunction (diabetes mellitus) [3]. The CD-PM will validate other biomarker candidates already identified by key researchers from our cooperative partners.

PPPM outlook:
This novel technology has the capacity to significantly improve the sensitivity and specificity of existing analytical systems from femto to attogram levels. Once the assay platform is established, it can be retooled for nearly any diagnostic markers by replacing and optimizing the specific capture/detection conjugate, because the other components will be universal. Peripheral biomarkers for other metabolic disorders are currently under evaluation [4].
Aim. To analyze and discuss the retrospective medical data of long-term monitoring and observations at the Ukrainian Akademik Vernadsky Antarctica station and initial data from prospective studies.
Methods. We analyzed the multi-scale information of retrospective and prospective medical and biological studies, performed with the participation of all 22 Ukrainian wintering expeditions, based on surveys of over 220 men aged 20-60 years (mean age 37 years). Extensive medical examinations were carried out before the expedition, during the selection of candidates, and after returning, and particular functions were monitored during the entire stay in Antarctica. Medical records were analyzed to investigate microbiome, vascular dysfunction, hypoxia, mitochondrial dysfunction, and genetic polymorphism (hypoxia-inducible factor 1 (HIF-1) parameters.
Results. The examinations showed multi-level symptoms of the processes of dysregulation and dysadaptation, especially during adaptation to the Antarctic (1-month stay at the station) and, to a lesser extent, after returning from an expedition to Ukraine. A long stay in Antarctica evoked adaptive reactions associated with hypoxia and mitochondrial dysfunction, determined by a set of molecular-genetic mechanisms that trigger the expression of the corresponding genes and alter the mitochondria ultrastructure, leading to the death of organelles, and subsequently the cells, and are associated with pronounced oxidative stress [data in press]. Preliminary microbiome studies in the Antarctic setting have shown an increase in fungi on the skin of expedition participants, believed to be due to interference with local immunity and dysbiosis of the normal skin microbiome due to stress, recycled air, and antiseptic agents [1,2]. This region permits the study of conditions such as Flammer syndrome, amongst many others. Preliminary analysis demonstrated changes in responses on the Flammer syndrome questionnaire [3] carried out before and after the expedition; medical examinations showed stress dysadaptation via alteration the thyroid, gut function, and pain sensation [4].
Conclusions. The series of the retrospective and prospective studies demonstrated the strong influence of extreme environmental factors and stress in the absence of man-made factors on health parameters, with clearly defined individually patterned reactions. The translation of obtained results enables the determination of the a Bpre-pathology^condition for patient stratification that have a crucial impact on the management of chronic diseases in routine life by modifying stress-related conditions such as lifestyle, diet, and microbiome. In addition, since the region is essentially sterile, it permits unbiased research on the microbiome that may not be possible elsewhere.
Extension of laboratory medicine to provide complex health assessment in the context of PPPM Abstract Laboratory medicine is among the essential sources of reliable clinical information, providing the basis for objective disease/health assessment in individuals [1]. Predictive, preventive and personalized medicine (PPPM) emphasizes complex characteristics of health as a phenomenon [2]. In this context of complexity, laboratory medicine should not only significantly extend the range of tests and markers, but it should also significantly expand the range of distinct analyses, reflecting four principal categories of factors/materials related to health as outlined previously [3]. These principal categories comprise physical, chemical, biological and psychosocial factors, each of which can be of an internal and external nature. Complex clinical laboratory diagnostics/medicine should analyze factors/materials that have a potential impact on health and/or consistently reflect changes in health status. All data should then be interlinked with the patient's standard medical record in order to prepare a complex report on his/her health/disease state in the context of PPPM (Fig. 1). Such an extension of laboratory medicine, effectively supported by sophisticated information technology, will clearly represent a qualitative leap in healthcare and, in the long term, have the potential to significantly reduce healthcare costs through early prediction that is built on comprehensive, computer-assisted processing of reliable data acquired by complex analysis of various factors/materials, thus significantly reducing health deterioration in each individual. The vision of EPMA is to build such a pilot complex clinical laboratory dedicated to the personal health assessment of apparently healthy individuals [4]. Abstract Emergency and limited-resource subsets need sustainable, comprehensive and reliable diagnostic approaches [1][2][3]. Emergency physicians with imaging expertise can focus on further procedures, skipping timeconsuming and expensive steps [4][5]. Emergency clinical ultrasound, as an adjunct to other reliable biomarkers, enables early predictive information, preventive management of impending or worsening conditions, and personalized therapy. Acute abdominal emergency (appendicitis, small bowel obstruction, gallstones, cholecystitis, aortic aneurysm), and particularly acute chest disease (pleural-pericardial effusion, pneumonitis, pleura-pulmonary consolidation), may present discordant information between ultrasound (US) and chest-X-ray (CXR). An ongoing study is addressing some new concepts in clinical and ultrasound stratification of respiratory disease in 73 consecutive patients referred to the same emergency facility. Key symptoms (dyspnea, chest pain, cough), laboratory assays (CRP, total leukocyte and neutrophilic counts) were considered, and the relative odds of finding concordant or discordant US and CXR imaging in pleural effusion and lung consolidation are displayed. In general, the predictive accuracy of combined conventional non-imaging clues (symptoms and laboratory assays) is insufficient for lung density and pleural effusion. Accuracy in the detection of pleural effusion is exceedingly higher for US (98.5 vs. 77.5), while the combined use of US and CXR, with one and/or both procedures positive, provides greater accuracy for lung density detection. The implementation of a comprehensive culture of ultrasound imaging in medical school curricula and in continuous medical education is a sustainable advancement for the quality of our emergency departments and of the mobile medical units used in rescue activity during disaster or war crisis.

Abstract
Kashin-Beck disease (KBD) as an endemic disorder is clinically characterized by thickened and deformed joints. The etiology of KBD needs to be determined. The present report shows epidemiological characteristics of KBD via cross-sectional survey, national surveillance, casecontrol study, and intervention trial. X-ray images of the right hands of school children ages 7-12 were utilized to diagnose new KBD patients and disease severity.  [1], which in turn is recognised as a risk factor for developing cardiovascular (CV) pathology [2,3].

Objective. Evaluation of sleep and anxiety/depressive disorders (ADD) of medical students based on their additional work in hospitals.
Material and methods. Forty-nine students of five or six courses of medical faculty of StGMU were surveyed with a questionnaire regarding subjective sleep characteristics (Russian Somnological Centre) using the Epworth daytime sleepiness scale, Stanford and Berlin questionnaires, and the Hospital Anxiety and Depression Scale (HADS), in order to determine the presence of ADD. Students were divided into two groups: working students (23) and non-working students (26). Statistical analysis was conducted using Microsoft Excel.
Results. Working students slept on average 6.0 ± 1,1 hours at night, and nonworking students 7.06 ± 1.2 hours. Severe/moderate daytime drowsiness was found in 34% and 65% of students from the working group, but only 7% and 23% of the non-working group, respectively. Representation of apnea was similar between the two groups, but the structural characteristics of sleep quality differed significantly. The occurrence of pronounced/borderline values of ADD for the working students were 31% and 42%, and among the nonworking group were 15% and 23%, respectively. Conclusion. Additional work among medical students is associated with a decline in quality of sleep and development of ADD, which are in turn associated with reduced stress resistance. Working students must be included in the cardiovascular risk group [4] for timely implementation of preventive measures [5] as part of medical school student health centres.
The number of such centres in Russia has been steadily increasing.
Over the last decade, through the implementation of measures of surveillance, investigation and evaluation, adjusting iodine content in salt and supplying iodised oil pills or free iodized salt, and the establishment of an IDD information system, China has maintained sustainable IDD elimination on a national basis [3]. The results of a national IDD survey in 2014 show a goiter rate of 2.6% among children's, no deficient or excess urinary iodine was found among children in all provinces, and the coverage rate of qualified iodized salt had increased to 91.3%. No new cretinism cases have emerged since 2010. China has built a solid base of working experiences over the 20 years of its USI programme, including a mechanism of Bgovernment leadership, sectoral collaboration and participation^, and a strategy of Blocally appropriate and evidence-based iodine supplementation for various population groups^. Evaluations have shown enormous success achieved by China's USI programme in meeting global and national IDD targets [4].
However, there are serious challenges to upscaling of production, including quality control concerns and inconsistent clinical evidence, posing difficulties in regulatory registration and obtaining market authorization. Lessons learned will be presented from an actual rich R&D program that we performed in a biopharmaceutical company to screen for and further clinically develop a therapy against leishmaniasis.

Abstract
The Mediterranean diet has been associated with a reduced risk of developing major chronic diseases and with mortality [1]. Late evidence suggests, however, that people from Mediterranean areas are drifting away from this traditional diet, and has revealed emerging socioeconomic disparities in dietary habits even in epidemiological settings characterized by a relatively high socioeconomic homogeneity [2]. In this context, the economic crisis that started about 10 years ago poses a serious threat that the disparities in access to healthy foods will continue to grow. The decline in the Mediterranean diet began long ago, but a dramatic dropoff has been recorded in the most recent years. Data from the Moli-sani study showed that, beginning in 2007, material resources such as household income have become a strong determinant of the adherence to the Mediterranean diet, whereas they were not associated with diet in earlier years [3]. Although the shift was documented among the whole population, the decline is likely to be more evident across vulnerable groups and the elderly. Indeed, evidence from a recent Italian national survey (INHES study) revealed that undesirable dietary modifications possibly linked to the current economic crisis were mainly reported by lower socioeconomic groups, and that subjects perceiving a negative impact of the recession on their diet also showed lower adherence to the Mediterranean diet and reduced quality of grocery products. In light of this, a fruitful reasoning on the ways to revitalize the Mediterranean diet should necessarily deal with two key issues: First, the Mediterranean diet has become socioeconomically patterned, as had already been established for other quality eating models in non-Mediterranean settings, and the prominent role of financial over cultural resources in determining the adherence to this pattern should be taken into account. Second, the current economic crisis represents a major health threat for the general population, but in particular for the most vulnerable socioeconomic groups, possibly leading to wider gaps in terms of risk/protective factors across groups.

Abstract
The global epidemic of obesity and type 2 diabetes mellitus remains a pressing challenge for the scientific community and health care professionals (1). Chronic low-grade inflammation is believed to contribute significantly to the pathogenesis of target organ dysfunction/injury (e.g., nephropathy) in these conditions (1). Lifestyle interventions (e.g., diet and physical activity) are known to exert beneficial effects on glycemic control and general health in obese/type 2 diabetic individuals (2). Nonetheless, standard exercise programs are strenuous and time-consuming, and thus have low long-term compliance.
In recent years, there has been a surge of interest in whole-body vibration (WBV) as a preventive and/or interventional modality that can be applied to populations at risk of obesity/type 2 diabetes and associated sequelae. Indeed, utilizing the db/db mouse model of obesity/type 2 diabetes, we recently showed beneficial effects of WBV on glycemic status in association with changes in several indices of inflammatory response, suggestive of a reduction in inflammation (3). Importantly, skewing toward myeloid cell production is often observed in chronic inflammation and autoimmune diseases (4). In this context, myeloid-to-lymphoid skewing is defined by an increased number of granulocyte/macrophage lineage, associated with a decrease in the frequency of cells with lymphoid lineage. Thus, our recent studies examined the impact of WBV in db/db mice and their db/m controls in the context of assessing changes in myeloid and lymphoid cells in the blood, bone marrow, and kidney. The results indicate increased myeloid potential associated with obesity/type 2 diabetes and a developmental skewing in favor of myeloid precursors in diabetic bone marrow. Importantly, WBV altered myeloid skewing in blood, kidney, and bone marrow of db/db mice suggestive of anti-inflammatory effects. Collectively, our observations are consistent with the immunomodulatory capability of WBV. Thus, WBVoffers the promise of curtailing inflammation associated with obesity/type 2 diabetes, thereby positively influencing the outcome of these disorders.

Abstract
The rising prevalence of metabolic syndrome (MetS), a cluster of cardiometabolic risk factors predictive of type 2 diabetes, relates largely to increasing obesity and sedentary lifestyle, but also to early metabolic life events. The objective of the study was to build predictive models of evolution toward MetS 8 years later, and to bring new knowledge about this pathological state using a multilevel approach in an at-risk population (subjects with low birth weight). A case-control study (subjects free from MetS at baseline (n=92 born small vs. n=76 born adequate for gestational age) was designed within the French community-based Haguenau cohort. Serum signatures were determined and compared at baseline (age 20 years) to determine predictive biomarkers using both un-targeted metabolomics and targeted proteomics. Individual predictive models were first built using linear logistic regressions from the omics datasets and epidemiological data. All data were then integrated to determine whether multidimensional models improved prediction. Univariate statistical analyses enabled the identification of 93 discriminant metabolites and 37 proteins between cases and controls at baseline, with gender differences. The resulting models based on either four metabolites or four proteins showed good performance: 18% misclassification (19% crossvalidation error rate) vs. 11% misclassification (19% cross-validation), respectively. Multi-omic data integration improved the performance and robustness of the prediction (9% misclassification, 16% cross-validation). Correlation analyses also contributed to a better understanding of the role of these biomarkers in the pathological processes. These results should provide new tools for better stratification of at-risk populations. This project was supported by the Fondation Francophone de Recherche sur le Diabète. . Data analysis will be performed using reproducible online Galaxy workflows [2]. The metabolomic/lipidomic data will be processed to identify specific signatures of MetS and its components, and to study their stability over time. These data will then be analysed for evaluation of a molecular reclassification of the MetS phenotype. Finally, they will be integrated with available phenotypic and detailed nutritional data in order to better characterize sub-phenotypes (Fig. 1). The approach developed here will open a door for a more comprehensive understanding of the metabolic phenotype resulting from the complex interplay between intrinsic and extrinsic factors. Thus, this project will allow an improved description of MetS-associated characteristics and will offer new tools for better patient stratification in elderly populations.

Abstract
Cardiovascular diseases are the main reason for morbidity and mortality in diabetic patients, and cardiovascular risk is increased at least twofold in men and at least fourfold in women with diabetes compared to non-diabetic populations. Predictive medicine is of the utmost importance in the clinical care of diabetic patients, since predicting cardiovascular risk is essential for modification of risk factors aimed at prevention or delay of future cardiovascular events. The prediction of cardiovascular risk is a valuable tool within the context of patient-centered care, as it includes active participation of diabetic patients in the decision-making process, resulting in higher compliance with the treatments agreed. However, there are differences among the current guidelines of various international authorities, such as the International Diabetes Federation (IDF), European Society of Cardiology (ESC) / European Association for Study of Diabetes (EASD), American College of Cardiology (ACC) / American Heart Association (AHA), American Diabetes Association (ADA), and National Institute for Health and Care Excellence (NICE), for the prediction of cardiovascular risk in diabetic patients. Furthermore, the clinical use of models with classic risk factors and novel biomarkers that would predict cardiovascular risk in diabetic patients from various populations with acceptable precision poses a challenge. Taking into consideration the global diabetes pandemic and its close association with cardiovascular diseases, there is an urgent need for streamlining of current guidelines on the prediction of cardiovascular risk and its use in clinical practice.

Abstract
Vitamin D plays a pivotal role in functional processes that control immune function and mineral metabolism, as well as several chronic conditions [1,2].
Emerging data indicate a possible influence of vitamin D on glucose homeostasis [3,4,5]. This study sought to provide initial information on vitamin D status among a Ghanaian population with type 2 diabetes and its association with glucose homeostasis. This case-control study was conducted among 118 patients with clinically diagnosed type 2 diabetes mellitus (T2DM) attending diabetic clinic at the Nkawie Government Hospital between October and December 2015. One hundred healthy non-diabetics living in Nkawie district were selected as controls. Questionnaires were administered to obtain sociodemographic data. Venous blood samples were taken from both cases and controls to estimate their fasting blood glucose (FBG) and lipid profile by spectrophotometry and iPTH and 25(OH)D by enzyme-linked immunosorbent assay (ELISA). Statistical analyses were performed using SPSS 20.0 software. The average age of the study participants was 58.81 years for cases and 57.79 year for controls. Vitamin D deficiency was found in 92.  Abstract In resent years, plasma homocysteine (Hcy) level has been reported to be associated with vascular complications for patients with type 2 diabetes mellitus (DM2). In such patients, elevated Hcy levels were associated with insulin resistance and nephropathy.

Objective
To investigate the association of hyperhomocysteinemia with macro-and microvascular complications in patients with DM2 in a Macedonian population.

Materials and methods
In this study, 80 DM2 patients were enrolled for study and were classified into two groups: 30 patients with no associated complications (control), and 50 patients with complications, with diagnosed CAD. and with microalbuminuria. Homocysteine levels and HbA1c were determined in both investigated groups.

Results
Hcy levels were significantly higher (p<0.000) in DM2 with complications, CAD (p<0.000), and patients with microalbuminuria (p<0.000) compared with controls. There was a positive correlation between elevated HbA1c (r = 0.475) levels and Hcy concentration in DM2 patients.

Conclusion
The results show that homocysteine levels were higher in patients with microor macrovascular complications, and they were strongly positive correlated with HbA1c. The results also reveal that hyperhomocysteinemia is a risk factor in the etiology of vascular complications in DM2 patients.
Integrative selection of oral hygiene complex for pregnant women with diabetes mellitus and periodontal disease Orekhova Liudmila 1,2 , Musaeva Ramilya 1 , Aleksandrova Anna 1 , Posokhova Eleonora 2 . Abstract Introduction. It is scientifically proven that pregnant women with diabetes mellitus have a high risk of developing periodontal diseases, but prevention and treatment is difficult in this group of patients.
Objective: To investigate and analyze dental status of pregnant women with diabetes mellitus, to estimate the effect of individual oral hygiene complex on periodontal tissues in this group of patients. Materials and methods: A clinical and laboratory examination of the oral cavity was carried out among110 patients with different types of diabetes mellitus and those without it. Women were randomly separated into three groups depending on the proposed complex of individual oral hygiene. Patients of groups 1 and 2 were trained in cleaning teeth and using extra oral hygiene products. The first group used toothpaste with herbal extracts; the second group used toothpaste with herbal extracts, sodium bicarbonate and sodium fluoride. The third group continued oral care as always. After 2 weeks the results were evaluated.
Results. The study revealed a correlation between values of dental indices in these groups of patients. The highest intensity of caries, values of hygiene and periodontal indices were found in women with diabetes, especially with type 2 diabetes (TDI: 17.5%, OHI-S: 1.67%, PMA: 37.4%). The proposed complexes of individual oral hygiene demonstrated their effectiveness in groups 1 and 2.

Conclusion:
The results of the study dictate a need for developing schemes of individual oral hygiene complex based on the results of a full dental examination, including clinical and laboratory research.

Abstract
Transcription factor NF-κB plays a key role in inflammation and is also an important regulator of genes involved in coagulation and thrombosis. NF-κB is activated by a number of signaling pathways that converge, in the majority of cases, at the level of IκB kinase 2 (IKK2). Our study uses constitutive active IKK2 (caIKK2) in a conditional transgene mouse model to mimic chronic inflammation specifically in endothelial cells and to test a potential aggravating impact on the onset of atherosclerosis. Mice bearing inducible, aortic-EC-specific Cre recombinase on an ApoE-deficient background were crossed with a strain expressing caIKK2 downstream of a loxP-flanked stop cassette and fed a cholesterol-rich high-fat diet for 12 weeks. RNA sequencing analysis of the aortic transcriptome was performed 10 days after induction of Cre recombinase by tamoxifen, before starting the western-type diet, and revealed activation of inflammatory networks (involving TNF, IFNγ and IL1β), which were accompanied by increased immune cell infiltration. Flow cytometric analysis of lymph nodes and splenic tissue was used to detect infiltration and activation of B and T cells.
In summary, we conclude that endothelial NF-κB signaling orchestrates immune cell responses within the arterial wall and therefore plays an important role in the pathogenesis of atherosclerosis. Furthermore, our data indicate that inflammatory endothelial cell activation causes early B-cell and T-cell responses in the pathogenesis of atherosclerosis before the occurrence of cholesterol-rich lesions.
system. The effects of relevant miRNA on gene regulatory effects were also analysed. .001), respectively. miR-3691-3p reduced the luciferase activity of the A allele, but had no effect on the activity of the G allele. This result indicates that the mutation from G to A enhances the negative regulation of miR-3691-3p to NOTCH1, contributing to the development of CHD.
Conclusions: The polymorphism of rs6563G>A is related to hereditary susceptibility to CHD. The mutation of G>A may affect the regulation of miR-3691-3p to the NOTCH1 gene and alter CHD susceptibility.

Abstract
Introduction: Cold extremities (cold hypersensitivity in the hands and feet, CE) is a common symptom in East Asia, including Korea [1,2]. In Korean medicine, CE is used for pattern identification and is believed to be associated with various symptoms and diseases [3,4]. This study aimed to investigate whether quality of life (QOL) differs depending on the presence or absence of CE. Methods: In 2013, a national survey was conducted among the general population through a specialized survey institute. Subjects were selected randomly using a multistage stratified sampling method based on distribution by region, gender, and age (95% confidence level, sample error ± 3.0%). A total of 1101 subjects aged 20-84 years were asked to complete a questionnaire evaluating QOL (SF-12) and the extent of cold extremities [5]. Multiple regression analysis was used to evaluate the correlation between CE and QOL score, with adjustment for age, gender, and body mass index (BMI). Results: Among participants, 18.6% (n = 208) demonstrated CE and 72.7% (n = 800) no CE. CE was more prevalent in women than men, and those with warm hands and feet had higher BMI than those with cold hands and feet, regardless of gender or age. In multiple regression analysis, CE showed a significant independent effect on the SF-12 physical component summary (β = −0.11, p < 0.001) and mental component summary (β = −0.10, p < 0.01) when we adjusted for gender, age, and BMI.
Conclusion: This study showed that CE demonstrated significant independent effects on health-related QOL indicators.

Abstract
Background. Pregnancy can function as a load test for identifying women at high cardiovascular (CV) risk [1]. Improvement is needed in the system of preclinical diagnosis of CV disorders in pregnants [2].
Objective. To assess the significance of the parameters of central aortic pressure (CAP) and aortic pulse wave in the overall cardiac examination of pregnants with respect to preeclampsia (PE). Material and methods. A total of 253 pregnant women were surveyed, in whom risk factors and CV activity was evaluated using the BPLab diagnostic complex (Petr Telegin, Russia) with Vasotens Office software [3]. A comparison of central and peripheral pressure made it possible to identify various options for hypertension (H)hidden, false and systemic. The results were processed using Statistica 8.0 software. Conclusion. The data indicate the feasibility of including an assessment of CAP in CV examination protocols for pregnants [4,5]: first, the high prognostic value of this index was confirmed, and second, hidden AG was discovered more frequently in pregnant women than systemic and false forms of pressure buildup. The significance of this problem is evidenced by the increased aortic rigidity and more frequent development of PE.

Abstract
Introduction: Hypertensive nephropathy (HN) is a medical condition characterized by damage to the kidney due to chronic high blood pressure, and it is second common cause of end-stage renal disease. Podocytopathies have a role in pathogenesis of HN, thus the podocyte proteins may have significance in early detection of disease. The purpose of this paper is to test the significance of nephrin as a marker for early detection of HN.
Material and methods: This study included 64 people with chronic hypertension (CH; 33 male/31 female) with an average age of 54.1 ± 5.1 years. Thirty healthy subjects (10 male/20 female) with an average age of 48.7 ± 9.4 years were included as a control group. All patients were divided into three groups: patients with normoalbuminuria, microalbuminuria and macroalbuminuria, according to urinary microalbumin/creatinine ratios. As material, we used fresh urine and venous blood. In urine we measured nephrin by the ELISA method, creatinine photometrically and microalbumin turbidimetrically. In blood we measured urea, creatinine, albumin and total protein using a photometric method.
Results: In 69.7% of normoalbuminuric subjects with CH, we found elevated urinary nephrin. Nephrin was significantly elevated in all groups of participants with HN compared with the control group (p<0.05). Nephrin was significantly elevated in patients with normoalbuminuria compared with the control group (p<0.05). We found a positive correlation between urinary concentration of nephrin and serum creatinine and a negative correlation between urinary nephrin and eGFR. Conclusion: Nephrin can be a useful marker for early and non-invasive detection of HN.
Abstract Impression cytology (IC) is a method which has been used to assess ocular surface cells from patients, e.g. conjunctival and corneal epithelial cells. IC is a minimally invasive technique which utilises biopore, cellulose acetate or polyethersulfone (PES) filters to sample cells, and has traditionally been used to ascertain morphological changes to aid in the diagnosis of squamous metaplasia and dry eye disease (DED). 1,2 More recently, IC has been utilised for the investigation of gene expression of ocular surface cells. 3 We demonstrate the use of IC to identify gene and protein expression changes between DED subjects and healthy controls (HC). Conjunctival epithelial cells from DED patients (n=8) and HC (n=7) were harvested by IC using the Eyeprim™ device. Quantitative PCR (qPCR) was performed to quantify differences in target gene expression: p38-α, IL-1β, IL-8, MCP-1 and MMP-9. In addition, the spatial localisation of proteins p38-α and phospho p38 were probed in immunofluorescence assays. Using the Eyeprim device, the average yield of conjunctival epithelial cells (CEC) retrieved was 1.1 x 10 5 cells/mm 2 . qPCR analysis demonstrated significantly higher p38-α, IL-1β, IL-8, MCP-1 (all p<0.0001) and MMP-9 (p<0.002) expression in CE from DED patients versus HC. Immunofluorescence studies significantly increased p38-α and phospho p38-α protein levels in DED subjects versus HC (p<0.0001 and p<0.01, respectively). As IC retrieval procedures have improved and the sensitivity of detection has increased, 4 gene and protein analysis of IC-retrieved CEC may be used to identify novel biomarkers of ocular surface diseases.

Abstract
Omics and systems biology have driven the rapid development of predictive, preventive, and personalized medicine (PPPM). Variations exist in the entire process of healthcare. Extensive genomic, transcriptomic, peptidomic, proteomic, and metabolomic data have been achieved in the field of human omics. Of these, proteomic variations are the final presentation of the genomic and transcriptomic variations. Metabolomic variations are the comprehensive results originating from proteins, nucleic acids, sugars, and lipids. Proteins and metabolites are two essential elements to differentiate the phenotypes. Moreover, many aspects in the proteome including splicing, post-translational modifications, translocation, conformation, redistributions, and variants, and in metabolome including entire component of metabolites, pattern variations, and disease-specific metabolomic variations, remain unknown. Clarification of variations in the phenome is a key step toward realizing PPPM practice. Proteome and metabolome are important components that contribute to the phenome. Therefore, studies of variations in proteomes and metabolomes have important scientific value in PPPM, including resolution of disease molecular mechanisms, determination of effective and reliable biomarkers for personalized prediction/prevention, diagnosis/ therapy, and prognostic assessment, and discovery of effective therapeutic targets to achieve precision medicine.

Abstract
IgG N-glycan profiles can serve as dynamic indicators of the ageing process and are able to discriminate between normal and accelerated ageing by highlighting a discrepancy between a body's age in years of life (chronological age) and its age in terms of health status (biological age). Therefore, IgG Nglycome represents a real-time indicator of the interaction between genetic/epigenetic predisposition and the environment. IgG N-glycans serve as predictive biomarkers when health is evaluated in the context of age, based on the proposition that specific IgG N-glycans i) identify variations in responses to maturation, ageing, and environment across the life course at a population level; ii) facilitate early risk prediction of chronic disease development; and iii) serve as prognostic indicators for the benefit of targeted preventive and disease treatment interventions. A new challenge, the ability to develop an Bageing biomarker kit^to be used in a (pre)clinical setting as a tool of predictive, preventive and personalised medicine, is presented here.
Novel approach for preventing implant failure: programming of anti-inflammatory macrophages using self-standing release systems with a phenotype-fixing cytokine cocktail formulation Julia Kzhyshkowska 1

Abstract
Nondegradable metallic and polymeric implants are used as mainstream solutions in regenerative medicine. Chronic inflammation is the most common complication leading to implant intolerance and failure. Key cells that control local inflammation and foreign body response are tissue macrophages (1). After implantation, macrophages induce acute inflammatory reactions to trauma and foreign material, which must be followed by the resolution of inflammation, wound healing phase and restoration of homeostasis. Personalized therapeutic solutions are required to improve implant integration and acceptance. Macrophages are highly plastic cells that define the reactions of the immediate tissue microenvironment to the implanted material. However, frustrated phagocytosis and inability of macrophages to resolve inflammation and restore tissue-specific homeostatic balance can lead to the chronic inflammation around implants. The prevention of implant-induced chronic inflammation by programming of the macrophage phenotype is a promising strategy for improving implant acceptance. The aims of such programming include controlling the phenotype of local macrophages by long-term fixation of their healing activities and suppression of inflammatory reactions. We have developed a cytokine cocktail formula (M2Ct) that induces a stable M2-type phenotype in human primary macrophages characterized by significant suppression of induced pro-inflammatory reactions and increased secretion of anti-inflammatory cytokines (M2Ct) (2). The positive effect of the M2Ct was demonstrated in an in vitro wound healing model. Using this model for induction of inflammation by LPS, we demonstrated that the M2Ct phenotype was stable for at least 12 days. However, in the absence of M2Ct components in the medium, macrophages underwent rapid proinflammatory reprogramming upon IFNg stimulation. In order to overcome the plasticity of macrophages and stabilize their phenotype, for direct application of this cocktail on implants and in tissue engineering in vivo, the loading and release of the cytokine cocktail from a selfstanding, transferable gelatin/tyraminated hyaluronic acid based release system was developed. The cytokine cocktail demonstrated its antiinflammatory activity in controlled-release conditions. Our data suggest that the direct application of a potent M2-inducing cytokine cocktail in a transferable release system can be used as a personalized solution to improve the long-term acceptance of implanted materials. Funding: EU FP7 for research and technological development and demonstration (Grant number 606294, IMMODGEL).

Abstract
Main phases of the physiologic wound healing (WH) are the Bearly wound^(hemostasis characterised by fibrin clot formation, bleeding stoppage and creation of the hypoxic microenvironment) followed by the inflammatory phase well controlled by the wound healing mechanisms. That proceeds into the Blate wound^featuring the proliferative phase comprising stem cell recruitment, angiogenesis/neovascularisation and wound contraction, followed by tissue remodelling and scar formation, finalising the physiologic WH [1]. Major players in the physiologic WH are the cellular stakeholders operating the repair processes, namely platelets, DECTs, neutrophils, macrophages, T lymphocytes, fibroblasts, myofibroblasts, keratinocytes, epithelial cells, endothelial cells, endothelial progenitor cells, stem cells, peripheral neurons, and extracellular matrix [2]. Potential biomarker panels includes growth factors (PDFGF, VEGF, TGF-ß, FGF protein core, EGF, IGF), cytokines/chemokines and transcription factors (IL-1, IL-6, TNF-α, IL-10, IL-4, IL-13, SDF-1α, HIF-1), neuropeptides (substance P, NGF, NPY, CGRP), matrix-proteases/inhibitors (MMPs/TIMPs), hormones and nutrients, amongst others [3,4]. Impaired WH leads to post-surgical complications frequently observed in the elderly, chronic ulcers in diabetic patients, hindered and ineffective pain management, etc. However, these well-acknowledged examples are just the Btip of the iceberg^. The entire spectrum of potential consequences of the impaired WH is much mor broader, further extended by the chronic inflammation and cell/tissue transformation processes causing tissue dysfunction and calcification as well as (neuro)degeneration, progressive formation of pre/cancerous lesions and pre-metastatic niches. That is a subject for predictive diagnostics, targeted prevention and personalised medical approaches [5].

Abstract
Physiologic wound healing (WH) is a well-controlled, highly orchestrated process involved in numerous ordinary (sporadic finger cut during meal preparation, regular manicure, etc.) and extraordinary (e.g. acute or planned surgery) conditions. WH is life-important. Physiologic WH is initiated by tissue injury and resolved within a reasonable time frame by the restoration of tissue integrity. The entire process follows the essential sequence of attributed phases: hemostasis, inflammation, proliferation, and tissue remodelling ( Fig. 1) [1]. In contrast, impaired WH is characterised by low quality of phase-performance, stagnation (such as chronic inflammation), prolonged WH process or even non-healing wounds [2]. The quality of WH is highly individual, resulting from both non-modifiable (genetic predisposition, age) and modifiable (e.g. nutrition, lifestyle, preventable collateral pathologies such as type 2 diabetes) risk factors. Contextually, the quality of WH is considered n essential parameter in individualised patient profiling and the subject for advanced approaches by systems medicine, predictive diagnostics, targeted preventive measures, and treatments tailored to the personaltogether innovative approaches by predictive, preventive and personalised medicine as the Bmedicine of the future^. For that, multilevel diagnostics utilising comprehensive biomarker panels is proposed. The potential application is multifaceted: early diagnosis of related pathologies and disease predisposition, effective prevention and treatment of collateral pathologies, prediction and prognosis of post-surgical complications, improved individual outcomes in plastic surgery, amongst others [3].
pregnant women are required to travel to the corresponding location, and wait for the test and then the diagnosis. In the case of high-risk pregnancy, CTG is a very useful diagnostics tool. An abnormal CTG indicates serious abnormalities, and in late pregnancy may lead to emergency caesarian section. However, even in low-risk pregnancy, it gives the comfort of the proper fetal care what undoubtedly influences a well-being of a future mother. There are some attempts to allow the in situ CTG diagnosis without the necessity to visit health care unit. Here, we report on an innovative integrated device based on a microprocessor and a dedicated operating system for in-home use by pregnant women. GSM/ GPRS/3G-based communication enables a territorially almost unlimited CTG signal transfer from the woman to the telecare center. The pregnant woman is instructed by the doctor or nurse on how to use the device and how to transmit the recordings. She then receives the recorded signals in the form of diagrams, along with explanations relating to the fetal heart rate, fetal movements, mother's HR, and warning of (if present) decelerations, signal variability, etc. She is also instructed what to do, and in the case of emergency, she can be transferred to the nearest hospital as quickly as possible. Keywords: Predictive preventive personalized medicine, Personalized dentistry, Tooth regeneration, Stem cells

Abstract
Clinical dentistry continues to rely heavily on traditional restorative approaches and biomaterials to treat/replace damaged and diseased teeth and the periodontium. More recently, implant dentistry has also made major strides in techniques for replacement of missing teeth. Importantly, the recognition of the pivotal role of stem cells in tissue repair and regeneration has led to intense research to identify toothderived stem cells. Studies have subsequently shown not only the ability of tooth-derived stem cells to generate various components of the tooth and periodontal structures, but also their utility in regenerative medicine, as exemplified by the generation of cells of organs such as the heart, liver and brain. More recent studies have demonstrated (functional) tooth regeneration in animal models that display physical properties seemingly similar to those of natural teeth. Even more exciting is the advent of induced pluripotent stem cells (iPSCs) which, similar to embryonic stem cells, have the hallmark feature of differentiation to cells of the three germ layers; iPSCs make it feasible to generate epithelial and mesenchymal stem cells for eventual in vitro generation of a tooth bud, followed by implantation into the tooth socket. Since the creation of iPSCs requires the use of somatic cells, investigators have explored the possibility of using integration-free human urine-induced pluripotent stem cells for tooth regeneration. Clearly, this approach has the potential to ultimately lead to Bcustom-making^of tooth and supporting structures, thereby bringing true meaning to the term Bpersonalized dentistry^. Other highly relevant research areas involve the identification of growth factors and delivery options as well as scaffolding material that would be most conducive to tooth regeneration. These aspects and their relevance to PPPM are the focus of this presentation.
Comparison of dental fear and anxiety in relation to dental health in Estonian and Vietnamese children Jana Olak* 2 ; Minh Son Nguyen 1,2 ; Thuy Trang Nguyen 1 ; Bui Bao Tien Nguyen 1 ; Mare Saag 2 1 Danang University of Medical Technology and Pharmacy, Danang, Vietnam 2 Institute of Dentistry, University of Tartu, Tartu, Estonia *Correspondence: Assoc. Prof. Jana Olak, University of Tartu, Raekoja plats 6, Tartu, Estonia; e-mail: Jana.Olak@ut.ee Keywords: Schoolchildren, Dental anxiety, Dental fear, Dental caries Abstract Objectives. We hypothesized was that differences in culture and oral health care system in Estonia and Vietnam might have an impact on dental health (dmft+DMFT) and dental fear and anxiety (DFA) in children of the two countries. The aim of this study was to compare dental fear and anxiety (DFA) in relation to dental health between Estonian and Vietnamese schoolchildren using the modified Dental Subscale of the Children's Fear Survey Schedule (MCFSS-DS). Methods. Among a total of 900 schoolchildren 8 to 10 years of age, 344 from Estonia and 556 from Vietnam agreed to participate in this study.
Dental health was recorded based on dental caries experience index of mixed dentition (dmft+DMFT). DFA was measured using the MCFSS-DS with five Likert-scale response options (1 = not afraid at all, 5 = very afraid). Results. The mean dmft+DMFT score in Estonian and Vietnamese schoolchildren was 5.2 ± 3.1 and 4.1 ± 3.2, respectively. The mean score of 11 fear items among Vietnamese children (20.8 ± 9.1) was significantly higher than that of Estonian children (15.4 ± 4.4). The prevalence of high DFA in Estonian schoolchildren (30.7%) was statistical equivalent to that of Vietnamese schoolchildren (28.0%, p>0.05).
Conclusion. There were significant differences in DFA between Estonian and Vietnamese schoolchildren. Differences in oral health care systems between the two countries might be considered as a factor affecting dental health and DFA among schoolchildren. preferred the empathic, non-referential type of doctor. A doctor of this type listens to the patient, builds partnerships, and discusses treatment strategies with the patient. Most patients (65%) preferred a female dentist. Findings. The majority of respondents noted that dental health was important and they needed treatment, but they postponed it. Patients preferred a doctor who discussed the course of treatment. Conclusion. The study revealed the need to develop a plan for individual approaches for patients of different age groups.
Improving the effectiveness of dental caries prevention using therapeutic toothpastes Anatoly A. Kunin Abstract A number of authors consider rational oral hygiene an integral component of etiopathogenetic prevention of dental caries and periodontal disease. For in vitro studies and the evaluation of physical and chemical properties, the structure and substructure of 30 toothpaste samples were examined using a Libra 120 transmission electron microscope (ZEISS) with different techniques: fast electron diffraction, and bright-field and darkfield imaging. In vivo studies included clinical assessment of the state of hard dental tissues after the application of ROCS Sensitive Instant Relief toothpaste during the month before and after electromagnetic field exposure. The in vitro results revealed certain changes in the microstructure of the ROCS Sensitive Instant Relief toothpaste after electromagnetic field exposure consisting of compound particle enlargement and corresponding changes in density. According to the in vivo results, a reliable positive effect after the application of therapeutic toothpaste was achieved in 98% of cases. However, when it was exposed to the electromagnetic field, a greater anti-caries effect was observed, which was confirmed by numerous diagnostic techniques. Thus, as a result of the application of ROCS Sensitive Instant Relief toothpaste in the study group, the mean value of the dental hygiene index was reduced from 0.80 ± 0.06 to 0.50 ± 0.02 towards an improvement in oral hygiene status. The cariogenicity of dental plaque disappeared in most patients of the study group, which was confirmed by the statistical data, from 40.4 ± 2.53% before to 13.73 ± 1.82% after. There was no significant reduction in the control group indices. The studies thus revealed certain changes in the microstructure of ROCS Sensitive Instant Relief toothpaste caused by the electromagnetic field and in the ion-exchange processes in hard dental tissues during their use, which leads to a significant increase in their therapeutic and prophylactic efficacy.
The effectiveness of a personal approach to learning manual skills during dental hard tissue preparation Abstract The development of practical skills using simulation training eliminates the risk to patient's health and life, provides a personalized approach to learning and the possibility of developing practical skills and bringing them to automatism, and ensures the objective control of the quality of their execution. Objective: to determine the effectiveness of a simulation method of teaching manual skills when handling removable dentures in a clinical setting.
Methods: The first phase of the study involved 82 third-year students, divided into a control group where the training was conducted with the use of standard phantoms, and a study group that used a fifth-generation CDS 100 dental simulator. The second phase included 48 young specialists who had participated in the first phase, and were divided into groups similar to those of the first phase. The quality of the preparation for the both stages was objectively assessed with a Zirkozahn S600 ARTI scanner. Results: In the control group, the main reason for the need for additional correction of the abutment teeth at both stages was inadequacy of the dental ledge, with an ANOVA factor indicator of 1.1 for the first phase and of 1.15 for the second, which indicates statistical identity of predictors in this group. The study group found no statistically significant difference between the quality of the dental ledges executed at the first and second stages of the study, with an ANOVA factor indicator of 0.9, which indicates the statistical significance of such a predictor for preliminary acquisition of automatic movements acquired while working with the simulator. Conclusion: The use of the CDS 100 interactive dental simulator provides a higher level of practical skills development and, most significantly, their "survival" compared with the classical form of education, which is confirmed by subsequent clinical practice.
Prediction of the development of dental caries in adolescents based on the functional state of the masseter muscle

Abstract
The scientific data proving that enamel is a mineral and protein tissue with a system of microcirculation in the form of enamel tubules served as the basis for this study. The orifices of the enamel tubules open onto the enamel surface. The dental pulp fluid rushes out through enamel and dentinal canals because of the pressure gradient in the pulp. The chewing function is the only physiological mechanism to provide the pressure gradient. The aim of the study was to evaluate the functional state of superficially localized masseter and temporal muscles in adolescents aged 13-16 years with high and medium caries resistance, by means of electromyography. Electromyography was conducted to evaluate the masseter and temporal muscles in 62 adolescents aged 13-16. The assessment was carried out using the Synapsis apparatus. The degree of tooth caries development was determined by intraoral roentgen diagnostics using the de Götzen S.r.l. electrometric diagnostics dental apparatus and light-induced diagnostics. The electrobiological activity of the masseter muscles was found to be significantly lower in patients with medium caries resistance than in adolescents with high caries resistance. There was an imbalance between the electrobiological activity indices in the masseter and temporal muscles; an increase in masseter fatigue in response to a static load in adolescents with medium caries resistance was observed. That situation may lead to a reduced pressure gradient of the dental pulp fluid in the pulp cavity, its failure to enter enamel tubules, and consequently to lower enamel resistance, with the risk of a cariogenic situation in adolescents.