Early life adversity: Epigenetic regulation underlying drug addiction susceptibility.

Drug addiction is a leading cause of disability worldwide, with more than 70,000 Americans dying from drug overdose in 2019 alone. While only a small percentage of chronic drug users escalate to drug addiction, little is understood on the precise mechanisms of this susceptibility. Early life adversity is causally relevant to adult psychiatric disease and may contribute to the risk of addiction. Here we review recent pre-clinical evidence showing that early life exposure to stress and/or drugs regulates changes in behavior, gene expression, and the epigenome that persist into adulthood. We summarize the major findings and gaps in the preclinical literature, highlighting studies that demonstrate the often profound differences between female and male subjects.

Structure-function relationships of the disease-linked A218T oxytocin receptor variant.

Various single nucleotide polymorphisms (SNPs) in the oxytocin receptor (OXTR) gene have been associated with behavioral traits, autism spectrum disorder (ASD) and other diseases. The non-synonymous SNP rs4686302 results in the OXTR variant A218T and has been linked to core characteristics of ASD, trait empathy and preterm birth. However, the molecular and intracellular mechanisms underlying those associations are still elusive. Here, we uncovered the molecular and intracellular consequences of this mutation that may affect the psychological or behavioral outcome of oxytocin (OXT)-treatment regimens in clinical studies, and provide a mechanistic explanation for an altered receptor function. We created two monoclonal HEK293 cell lines, stably expressing either the wild-type or A218T OXTR. We detected an increased OXTR protein stability, accompanied by a shift in Ca2+ dynamics and reduced MAPK pathway activation in the A218T cells. Combined whole-genome and RNA sequencing analyses in OXT-treated cells revealed 7823 differentially regulated genes in A218T compared to wild-type cells, including 429 genes being associated with ASD. Furthermore, computational modeling provided a molecular basis for the observed change in OXTR stability suggesting that the OXTR mutation affects downstream events by altering receptor activation and signaling, in agreement with our in vitro results. In summary, our study provides the cellular mechanism that links the OXTR rs4686302 SNP with genetic dysregulations associated with aspects of ASD.

Chronic oxytocin-driven alternative splicing of Crfr2α induces anxiety.

The neuropeptide oxytocin (OXT) has generated considerable interest as potential treatment for psychiatric disorders, including anxiety and autism spectrum disorders. However, the behavioral and molecular consequences associated with chronic OXT treatment and chronic receptor (OXTR) activation have scarcely been studied, despite the potential therapeutic long-term use of intranasal OXT. Here, we reveal that chronic OXT treatment over two weeks increased anxiety-like behavior in rats, with higher sensitivity in females, contrasting the well-known anxiolytic effect of acute OXT. The increase in anxiety was transient and waned 5 days after the infusion has ended. The behavioral effects of chronic OXT were paralleled by activation of an intracellular signaling pathway, which ultimately led to alternative splicing of hypothalamic corticotropin-releasing factor receptor 2α (Crfr2α), an important modulator of anxiety. In detail, chronic OXT shifted the splicing ratio from the anxiolytic membrane-bound (mCRFR2α) form of CRFR2α towards the soluble CRFR2α (sCRFR2α) form. Experimental induction of alternative splicing mimicked the anxiogenic effects of chronic OXT, while sCRFR2α-knock down reduced anxiety-related behavior of male rats. Furthermore, chronic OXT treatment triggered the release of sCRFR2α into the cerebrospinal fluid with sCRFR2α levels positively correlating with anxiety-like behavior. In summary, we revealed that the shifted splicing ratio towards expression of the anxiogenic sCRFR2α underlies the adverse effects of chronic OXT treatment on anxiety.

Long-term microtensile bond strength of self-etch adhesives and influence of 7-s phosphoric acid etching on adhesion of a 3-step etch-and-rinse adhesive to the dentine of primary teeth.

BACKGROUND: The immediate microtensile bond strength (µTBS) of self-etch (SE) adhesives to primary dentine is promising, but evidence about the durability is scarce. AIM: To assess the long-term µTBS of SE adhesives to primary dentine and to examine the effect of 7-s etching on µTBS of a 3-step etch-and-rinse (ER) adhesive. DESIGN: The enamel of 115 caries-free human primary teeth was ground flat, and bonding was performed according to group assignment: G-aenial™ Bond/GC [GB], Clearfil™ SE Bond/Kuraray [CS], OptiBond™ XTR/Kerr [OX], Scotchbond™ Universal/3M™ [SU], Prime&Bond® NT™/Dentsply [PB], and OptiBond™ FL/Kerr [OF]. After storage (24 h, 6 mos., 12 mos.), 1,762 beams were cut for µTBS testing. Fracture analysis was performed using light/fluorescence microscopy. Resin-dentine interfaces were exemplarily analyzed using SEM. RESULTS: After 24 h, OX and SU showed significantly higher estimated mean µTBS than GB, which exhibited the lowest µTBS at all stages (p < .05). µTBS within each adhesive group showed slight variations over time (p > .05). OF_SE produced acceptable µTBS. Etching increased the mean immediate µTBS for OF (p > .05) and the percentage of cohesive fractures. CONCLUSIONS: SE adhesives achieved durable µTBS to primary dentine. 7-s etching may improve the immediate µTBS of a 3-step ER adhesive.

Longitudinal Study of the Signal Strength in two Serologic Assays for Detection of SARS-CoV-2 Specific Antibodies.

BACKGROUND: In the course of the current SARS-CoV-2 pandemic, antibody assays provide an important means for guidance of public health efforts. Thus, characterization of the course of antibody signals on different widely used assays is needed. METHODS: We selected 25 PCR-confirmed SARS-CoV-2 cases among 3,273 healthcare workers and measured the course of the antibody signal using the Abbott Architect SARS-CoV-2 IgG assay and the Roche Elecsys® Anti-SARS-CoV-2 immunoassay. The signal strength was then modelled using linear mixed models adjusted for age. RESULTS: Since first sampling, the assay signal decreased per day in the Abbott assay (standardized slope (ß) = -0.46, 95% CI = -0.54 to -0.39). In contrast, an increase in the signal was ascertained by the Roche immunoassay per day (ß = 0.25, 95% CI = 0.09 to 0.41). CONCLUSIONS: Roche Elecsys® Anti-SARS-CoV-2 immunoassay may exhibit greater sensitivity in detecting SARS-CoV-2-specific antibodies in individuals in late stages of postinfection.

Class II resin composite restorations-tunnel vs. box-only in vitro and in vivo.

PURPOSE: In a combined in vitro/in vivo approach, tunnel vs. box-only resin composite restorations should be evaluated using thermomechanical loading (TML) in vitro and a restrospective clinical trial in vivo. MATERIALS AND METHODS: For the in vitro part, box-only and tunnel cavities were prepared in 32 extracted human third molars under simulated intraoral conditions in a phantom head. Specimens were randomly assigned to four groups (n = 8; 16 box-only/16 tunnel) and received bonded resin composite restorations with Amelogen Plus (box A/tunnel A) or lining with Ultraseal and Amelogen plus (box B/tunnel B) both bonded using PQ1 (all Ultradent). Specimens were subjected to a standardized aging protocol, 1-year water storage (WS) followed by TML (100,000 × 50 N; 2500 × + 5/+ 55 °C). Initially and after aging, marginal qualities were evaluated using replicas at × 200 magnification (SEM). For the corresponding in vivo observational study, 229 patients received 673 proximal resin composite restorations. From 371 tunnel restorations, 205 cavities were filled without flowable lining (tunnel A), and 166 tunnels were restored using UltraSeal as lining (tunnel B). A total of 302 teeth received conventional box-only fillings. Restorations were examined according to modified USPHS criteria during routine recalls up to 5 years of clinical service. RESULTS: In vitro, all initial results showed 100% gap-free margins when a flowable lining was used. Tunnels without lining exhibited some proximal shortcomings already before TML and even more pronounced after TML (p < 0.05). After TML, percentages of gap-free margins dropped to 87-90% in enamel with lining and 70-79% without lining (p < 0.05). In vivo, annual failure rates for box-only were 2.2%, for tunnel A 6.1%, and for tunnel B 1.8%, respectively (p < 0.05). Tunnels had significantly more sufficient proximal contact points than box-only restorations (p < 0.05). Flowable lining was highly beneficial for clinical outcome of tunnel-restorations (p < 0.05). CONCLUSIONS: With a flowable lining, tunnel restorations proved to be a good alternative to box-only resin composite restorations. CLINICAL RELEVANCE: Class II tunnel restorations showed to be a viable alternative for box-only restorations, however, only when flowable resin composite was used as adaptation promotor for areas being difficult to access.

[Amalgam and alternatives-discussions on mercury reduction in the environment]. / Amalgam und Alternativen ­ Diskussionen zur Quecksilberreduktion in der Umwelt.

Dental amalgam has been successfully used for the restoration of carious lesions for more than 180 years. It is clinically characterized by high longevity and low technique sensitivity. For decades, dental amalgam has been discussed in the public, especially due to its roughly 50% mercury content. Since the Minamata Convention was published in 2013 with the primary goal of reducing the anthropogenic mercury release into the environment, the previously muted amalgam discussion has received fresh impetus. Another considerable disadvantage of amalgam is its silver/greyish color, which simply no longer matches patients' esthetic demands.The present paper describes the basic problems with amalgam against the background of multiple biological, clinical, and health policy factors. Possible consequences of the Minamata Convention concerning legal regulations as well as the use of dental biomaterials and therefore also relating to the future national healthcare system are discussed. Finally, possible amalgam alternatives and the urgent need for biomedical research towards restorative dentistry are presented, embedded into the crucial question of whether we are actually conducting the correct debate.

Prevalence of Severe Acute Respiratory Syndrome Coronavirus-2-specific Antibodies in German Blood Donors during the COVID-19 Pandemic.

BACKGROUND: Coronavirus disease-2019 (COVID-19) is a respiratory infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). While RT-PCR assays are used routinely to diagnose active COVID-19, serological testing offers a means of identifying individuals who previously experienced asymptomatic infections, as well as those who experienced symptomatic infections but no longer carry the virus. METHODS: The presence of SARS-CoV-2 IgG-positive antibodies in the sera of 673 blood donors residing in south-western Germany before and 3,880 donors after the advent of the COVID-19 pandemic was determined and confirmed using two highly sensitive serological tests. RESULTS: Approximately 0.40% of the donors assessed during the COVID-19 pandemic possessed SARS-CoV-2 IgG-positive antibodies, decidedly fewer than the percentage of SARS-CoV-2-infected individuals determined by real-time RT-PCR nationwide. CONCLUSIONS: These findings confirm the efficacy serological testing in identifying asymptomatic COVID-19 patients.

Myocyte Enhancer Factor 2A (MEF2A) Defines Oxytocin-Induced Morphological Effects and Regulates Mitochondrial Function in Neurons.

The neuropeptide oxytocin (OT) is a well-described modulator of socio-emotional traits, such as anxiety, stress, social behavior, and pair bonding. However, when dysregulated, it is associated with adverse psychiatric traits, such as various aspects of autism spectrum disorder (ASD). In this study, we identify the transcription factor myocyte enhancer factor 2A (MEF2A) as the common link between OT and cellular changes symptomatic for ASD, encompassing neuronal morphology, connectivity, and mitochondrial function. We provide evidence for MEF2A as the decisive factor defining the cellular response to OT: while OT induces neurite retraction in MEF2A expressing neurons, OT causes neurite outgrowth in absence of MEF2A. A CRISPR-Cas-mediated knockout of MEF2A and retransfection of an active version or permanently inactive mutant, respectively, validated our findings. We also identified the phosphatase calcineurin as the main upstream regulator of OT-induced MEF2A signaling. Further, MEF2A signaling dampens mitochondrial functioning in neurons, as MEF2A knockout cells show increased maximal cellular respiration, spare respiratory capacity, and total cellular ATP. In summary, we reveal a central role for OT-induced MEF2A activity as major regulator of cellular morphology as well as neuronal connectivity and mitochondrial functioning, with broad implications for a potential treatment of disorders based on morphological alterations or mitochondrial dysfunction.

[Kagami-Ogata Syndrome: An Anomaly of the Ribs as a Pathognomonic Feature for the Clinical Diagnosis of an (epi)Genetic Syndrome]. / Das Kagami-Ogata-Syndrom: Eine Rippenanomalie als pathognomonisches Korrelat für die klinische Diagnose eines (epi)genetischen Syndroms.

Within 4 years (2014-2017), we genetically diagnosed 2 newborns with Kagami-Ogata syndrome (OMIM #608149). As fetuses they exhibited prenatal polyhydramnios and in 1 case hepatomegaly. After birth, the newborns suffered from respiratory distress. Typical phenotypic features, such as muscular hypotonia, a protruding philtrum, full cheeks and a depressed nasal bridge, were present. Chest X-rays revealed coat-hanger ribs and a bell-shaped thorax, suggestive of the entity. Kagami-Ogata syndrome is caused by an aberrant gene expression of chromosome 14 and was first described in 1991. Possible causes are paternal uniparental disomy of chromosome 14, epimutations and microdeletions. Approximately 70 cases have been reported in the literature, with 34 comprising the original cohort of M. Kagami and T. Ogata. The incidence of the disease is unknown. Patients often manifest a developmental delay and an intellectual disability, although in the meantime cases with milder clinical courses have been described. In the cohort of Kagami and Ogata 3 patients developed hepatoblastoma, which is a common feature in another imprinting disorder, namely the Beckwith-Wiedemann syndrome. Therefore, hepatoblastoma should be considered in follow-up examinations.

The interplay between oxytocin and the CRF system: regulation of the stress response.

Oxytocin (OT) has drawn the attention of researchers since 1930. Since then, many aspects of oxytocin have been uncovered, such as reproductive functions, dampening anxiety, enhancing socioemotional behavior, or regulating genomic effects on a cellular level. Here, we want to focus on the interaction between the OT system and the stress/corticotropin-releasing factor (CRF)-system of the brain. Depending on the nature of the stressor, OT is released simultaneously or directly after the stress from the neurohypophysis into the periphery and/or via somato-dendritic release in stress-sensitive brain areas. This stress-induced OT release might serve to modulate or dampen the stress response; however, the functional relevance is not yet fully understood. In this review, we will describe the effects of OT and discuss the interplay between OT and CRF on a cellular, physiological, and behavioral level.

Association between a New Sugar Index and Caries Experience: Results of a Cross-Sectional Field Study.

OBJECTIVES: The aim of this study was to test the relationship between a newly developed sugar index and caries experience in 4th- and 6th-graders. MATERIALS AND METHODS: A total of 1,019 fourth-graders and 925 sixth-graders from 2 regions in Northern Germany participated in the study. Caries experience was recorded by applying the ICDAS criteria. Tooth-brushing habits and other independent variables were examined psychometrically. Structured questions were formulated that were related to specific everyday eating situations, especially between main meals, which proved to be relevant for a higher caries risk. To compare mean caries scores, nonparametric tests were applied. The Pearson coefficient was calculated in order to verify the correlation between the sugar index and the caries experience. The influence of various independent variables on caries experience was assessed by stepwise backward logistic regression analysis. RESULTS: With regard to the primary teeth of the 4th-graders, significant correlations with values > 0.1 were found for all ICDAS outcome variables. With regard to the permanent teeth of the 6th-graders, the only significant correlations with the sugar index were for the ICDAS outcome variables D3-6MFT and D3-6FS. CONCLUSION: The results of our study show that not only the correlation between the sugar index and caries experience was stronger for 4th-graders than for 6th-graders, but also more distinctly visible differences were observed when comparing extreme groups. This indicates that protective factors, such as fluoridation and the preventive sealing of fissures, help level out the differences in caries occurrence in older children when compared to younger children.

Implementation and evaluation of an interdisciplinary preventive program to prevent early childhood caries.

OBJECTIVES: Since early childhood caries (ECC) is a major disease in the public health sector, an interdisciplinary prevention concept to avert this disease was developed, launched in a project region, and evaluated. It was examined whether the emergence of ECC could be prevented or curbed with the newly developed program. MATERIALS AND METHODS: The program encompasses seven pulses of information from an interdisciplinary team (gynecologists, midwives, pediatricians, dentists, municipal social services, and the public health office). The primary dependent variables were caries experience and prevalence among the 3- to 4-year-olds who took part in the ECC prevention program, compared to children of the same age without such measures. The effect of the prevention program was assessed by means of a "historical" comparison. The dental diagnoses were performed by two calibrated dentists based on the "ICDAS collapsed." Data on diet and preventive behavior were gathered using a standardized parental questionnaire. The significance was tested using the nonparametric Mann-Whitney U test. RESULTS: There were 661 3- to 4-year-olds in the control group and 706 in the test group. The mean d3-6mf-t for the control group (0.92) was significantly higher than that for the test group (0.46). While 78.8% of the children examined in the control group exhibited predominantly sound primary dentition, this was the case for 86.3% of the test group. The proportion of children with S-ECC was 7.9% in the test group and 14.5% in the control group. The test group differed significantly from the control group both in terms of reported diet and dental hygiene habits. CONCLUSIONS AND CLINICAL RELEVANCE: The interdisciplinary prevention program presented here is an effective concept for preventing ECC and considerably helps reduce problems in the treatment of small children.

Glucocorticoids promote a glioma stem cell-like phenotype and resistance to chemotherapy in human glioblastoma primary cells: Biological and prognostic significance.

Glioma stem cells (GSCs) are glioblastoma (GBM) cells that are resistant to therapy and can give rise to recurrent tumors. The identification of patient-related factors that support GSCs is thus necessary to design effective therapies for GBM patients. Glucocorticoids (GCs) are used to treat GBM-associated edema. However, glucocorticoids participate in the physiological response to psychosocial stress, which has been linked to poor cancer prognosis. This raises concern that glucocorticoids affect the tumor and GSCs. Here, we treated primary human GBM cells with dexamethasone and evaluated GC-driven changes in cell morphology, proliferation, migration, gene expression, secretory activity and growth as neurospheres. Dexamethasone treatment of GBM cells appeared to promote the development of a GSC-like phenotype and conferred resistance to physiological stress and chemotherapy. We also analyzed a potential correlation between GC treatment and tumor recurrence after surgical excision in a population-based consecutive cohort of 48 GBM patients, adjusted for differences in known prognostic factors concerning baseline and treatment characteristics. In this cohort, we found a negative correlation between GC intake and progression-free survival, regardless of the MGMT methylation status. In conclusion, our findings raise concern that treatment of GBM with GCs may compromise the efficacy of chemotherapy and may support a GSC population, which could contribute to tumor recurrence and the poor prognosis of the disease.

Long-term effect of intensive prevention on dental health of primary school children by socioeconomic status.

OBJECTIVES: Children in a German region took part in regular toothbrushing with fluoride gel during their time in primary school after having received a preventive program in kindergarten. The study aimed at determining the dental health of the students as a function of prevention in kindergarten and at school while taking into account their socioeconomic status and other confounders. MATERIALS AND METHODS: The subjects were in six groups: groups 1 and 2, intensive prevention in kindergarten with and without fluoride gel at school; groups 3 and 4, basic prevention in kindergarten with and without fluoride gel at school; groups 5 and 6, no organized prevention in kindergarten with and without fluoride gel at school. Two dental examinations were performed for assessing caries experience and calculating caries increment from second grade (7-year-olds) to fourth grade (9-year-olds). A standardized questionnaire was used to record independent variables. To compare caries scores and preventive measures of various subgroups, non-parametric tests and a binary logistic regression analysis were performed. RESULTS: A significant difference was found in the mean decayed, missing, and filled tooth/teeth (DMFT) depending on socioeconomic status (no prevention in kindergarten, fluoride gel at school in children with low SES: DMFT = 0.47 vs. DMFT = 0.18 in children with high SES; p = 0.023). Class-specific differences were no longer visible among children who had taken part in an intensive preventive program combining daily supervised toothbrushing in kindergarten and application of fluoride gel in school. CONCLUSIONS: Early prevention, focusing on professionally supported training of toothbrushing in kindergarten and at school, has a positive effect on dental health and is able to reduce class-specific differences in caries distribution. CLINICAL RELEVANCE: Early training of toothbrushing and fissure sealing of first permanent molars are the most important factors for the dental health of primary school children.

Weaning Premature Infants from Nasal Continuous Positive Airway Pressure - Current Practice and Scientific Evidence.

Objective In the past decade, a number of trials have been conducted to determine the optimal strategy of weaning premature infants from nasal continuous airway pressure (nCPAP). However, a paucity of information exists on how weaning is actually performed in clinical routine. Aim of this study was to investigate the current practice of weaning premature infants from nCPAP in Germany. Methods An online survey was performed in German tertiary care neonatal units. Results All 160 German tertiary care units were contacted. Replies were retrieved from 85/160 (53%) units, of which 83/160 (52%) completed the questionnaire. 66/83 (80%) respondents indicated to wean without the use of formal written policies. In 44/83 (53%) units weaning decisions are made jointly between physicians and nurses, whereas physicians are the sole decision makers in 33/83 (40%) as are nurses in 6/83 (7%) units. Many units use more than one weaning strategy. 81/83 units (98%) gradually reduce nCPAP pressure as the initial step in the weaning process. 9/83 (11%) units stop nCPAP at standard criteria [CICADA (CeasIng nCpap At standarD criteriA) method] and 58/83 (70%) units use a cycling nCPAP on/off strategy. 52/83 (63%) of the responding units use nasal high flow at least at some point during the weaning process, either as a gradual weaning method or during nCPAP breaks. Conclusion Weaning strategies from nCPAP vary widely in German tertiary care neonatal units. It appears that evidence is still insufficient to promote a distinct weaning strategy which in turn highlights the urgent need for further adequately powered clinical trials.

Viral Infections in Neonates with Suspected Late-Onset Bacterial Sepsis-A Prospective Cohort Study.

Objective The aim of our study was to evaluate the occurrence of viral infections in infants with suspected late-onset bacterial sepsis in a neonatal intensive care unit. Methods In a prospective study, infants with suspected late-onset bacterial sepsis underwent viral testing alongside routine blood culture sampling. Using a multiplex reverse transcription-polymerase chain reaction enzyme-linked immunosorbent assay, nasopharyngeal aspirates were analyzed for adenovirus, respiratory syncytial virus (RSV), influenza virus A and B, H1N1 virus, parainfluenza virus 1 to 4, metapneumovirus, coronavirus, and picornavirus. Stools were examined for adenovirus, rotavirus, norovirus, and enterovirus. Results Between August 2010 and March 2014, data of 88 infants with 137 episodes of suspected late-onset bacterial sepsis were analyzed. Six infants were diagnosed with a respiratory viral infection (2 × RSV, 4 × picornavirus). Blood culture-proven bacterial sepsis was detected in 15 infants. Neither viral-bacterial coinfections nor polymerase chain reaction positive stool samples were found. Conclusion Respiratory viruses can be detected in a considerable number of neonates with suspected late-onset bacterial sepsis. In contrast, gastrointestinal viral or enterovirus infections appear uncommon in such cases.

Effect of supervised brushing with fluoride gel during primary school, taking into account the group prevention schedule in kindergarten.

OBJECTIVES: In one region of Germany, a group of children took part in regular fluoride gel applications during primary school following intensified prevention in kindergarten. This observational study aimed to ascertain whether the dental health of primary school children can be improved by introducing a group prevention program based on applications of fluoride gel. MATERIALS AND METHODS: The subjects were distributed among six groups with varying preventive measures in kindergarten and at school. The basis for determining caries experience and calculating the caries increment consisted of dental findings gathered in the second and fourth grade. RESULTS: While second graders without professionally supported daily toothbrushing in kindergarten exhibited an average d3-6mft of 2.17, in those who had enjoyed intensive dental prevention, the corresponding value was 19% lower (d3-6mft = 1.74). The caries increment was significantly lower mainly among children who had received the maximum of group prevention (intensive prevention in kindergarten and gel program at school). CONCLUSIONS: The results show that intensified preventive programs in kindergartens and schools, based mainly on supervised toothbrushing, have a positive effect on the dental health of primary school children. CLINICAL RELEVANCE: Such programs are efficient in reducing caries experience especially in socially deprived areas.

Prevention in kindergartens with 500 ppm fluoride toothpaste-a randomized clinical trial.

OBJECTIVES: The aim of the study was to evaluate the effect of an intensified preventive programme in kindergartens. MATERIALS AND METHODS: Two thousand two hundred twenty-eight 2- to 4-year-old children attending kindergartens in two counties in northern Hesse were randomly allocated to two groups. Children in the test group participated in daily tooth brushing with 500 ppm fluoride toothpaste, which was supervised by specially trained dental nurses. Children in the control group received tooth brushing instructions three to four times a year. Caries experience was recorded using WHO criteria. The basic survey was performed from October 2006 to April 2007, with the final examinations between March and July 2009. The statistical evaluation was performed by means of the software package SPSS 17.0. To compare the mean Δdmf values of the test and control groups, Mann-Whitney U tests were performed. RESULTS: Two thousand one hundred twelve children (test group, 1096; control group, 1048) participated in the final examinations. The caries increment of the test group (Δdmf-s = 1.55) was 24 % lower than the increment of the control group (Δdmf-s = 2.02, p = 0.043). Based on Δdmf-t values, the corresponding figures were 0.79 for the test group and 1 for the control group (p = 0.44). High increment values (Δdmf-s >7) were observed in the control group twice as often as in the test group. CONCLUSION: The results show that an intensified preventive programme in kindergartens, based on supervised daily tooth brushing, has a positive effect on the dental health of preschool children. CLINICAL RELEVANCE: It is sufficient to offer such programmes in socially deprived areas.