Analysis of the Drosophila melanogaster anti-ovarian response to honey bee queen mandibular pheromone.

Queen mandibular pheromone (QMP) is a potent reproductive signal to which honey bee workers respond by suppressing their ovaries and adopting alloparental roles within the colony. This anti-ovarian effect of QMP on workers can, surprisingly, be induced in other insects, including fruit flies, in which females exposed to synthetic QMP develop smaller ovaries with fewer eggs. In this study, we use the Drosophila melanogaster model to identify the components of synthetic QMP required for the anti-ovarian effect. We found that virgin females respond strongly to 9-oxo-2-decenoic acid and 10-hydroxy-2-decenoic acid (10HDA), suggesting that the decenoic acid components of QMP are essential for the anti-ovarian response. Further, a nuclear factor of activated T-cells reporter system revealed neurones expressing the olfactory receptors Or-56a, Or-49b and Or-98a are activated by QMP in the antenna. In addition, we used olfactory receptor GAL4 drivers and a neuronal activator (a neuronal activating bacterial sodium channel) to test whether the candidate neurones are potential labelled lines for a decenoic acid response. We identified Or-49b as a potential candidate receiver of the 10HDA signal. Finally, the anti-ovarian response to synthetic QMP is not mediated by decreasing the titre of the reproductive hormones ecdysone and juvenile hormone.

Dissecting dentine-pulp injury and wound healing responses: consequences for regenerative endodontics.

A thorough understanding of the biology of the dentine-pulp complex is essential to underpin new treatment approaches and maximize clinical impact for regenerative endodontics and minimally invasive vital pulp treatment (VPT) strategies. Following traumatic and carious injury to dentine-pulp, a complex interplay between infection, inflammation and the host defence responses will occur, which is critical to tissue outcomes. Diagnostic procedures aim to inform treatment planning; however, these remain clinically subjective and have considerable limitations. As a consequence, significant effort has focussed on identification of diagnostic biomarkers, although these are also problematic due to difficulties in identifying appropriate diagnostic fluid sources and selecting reproducible biomarkers. This is further compounded by the link between inflammation and repair as many of the molecules involved exhibit significant multifunctionality. The tertiary dentine formed in response to dental injury has been purposefully termed reactionary and reparative dentine to enable focus on associated biological processes. Whilst reactionary dentine produced in response to milder injury is generated from surviving primary odontoblasts, reparative dentine, in response to more intense injury, requires the differentiation of new odontoblast-like cells derived from progenitor/stem cells recruited to the injury site. These two diverse processes result in very different outcomes in terms of the tertiary dentine produced and reflect the intensity rather than specific nature (nonexposure versus exposure) of the injury. The subsequent identification of the odontoblast-like cell phenotype remains challenging due to lack of unique molecular or morphological markers. Furthermore, the cells ultimately lining the newly deposited dentine provide only a snapshot of events. The specific source and plasticity of the progenitor cells giving rise to the odontoblast-like cell phenotype are also of significant debate. It is likely that improved characterization of tertiary dentine may better clarify the influence of cell derivation for odontoblast-like cells and their diversity. The field of regenerative endodontics offers exciting new treatment opportunities, and to maximize outcomes, we propose that the term regenerative endodontics should embrace the repair, replacement and regeneration of dentine-pulp.

[Series: Communication in the dental practice. Low levels of literacy and limited health literacy]. / Serie: Communicatie in de tandartspraktijk. Laaggeletterdheid en beperkte gezondheidsvaardigheden.

Approximately 36% of the Dutch population has insufficient or limited health literacy. This disproportionately concerns patients with low socioeconomic status (SES), the elderly and migrants, but also highly-educated patients. Dentists are often unaware of this. A patient with limited health literacy might have trouble with a dentist's explanation or with reading written information. A dentist may not immediately notice this, but can be aware of signals indicating it. Such signals can vary from not completely filling out a questionnaire to wrongly interpreting instructions or systematically not showing up to appointments. The dentist needs to take this into account in his use of language and adapt any informational matter to make it understandable for everyone. Co-workers such as receptionists also need to be informed about signs indicating limited health literacy and how best to communicate with these patients.

[Series: Communication in the dental practice. Communicating in a culturally competent manner]. / Serie: Communicatie in de tandartspraktijk. Cultureel competent communiceren.

Dentists will increasingly have to treat patients with a variety of cultural backgrounds. They will consequently have to deal with various preconceptions about health and how people ought to behave with each other. In order to develop a good dentist-patient relationship and provide appropriate care, the dentist will have to learn to communicate in a culturally competent manner. That requires an open, empathetic attitude and an awareness of one's own norms, values and views. The dentist will have to consider how they differ from those of people from another (sub) culture. With the help of a number of examples, this article will provide an impulse to do that.

[Series: Communication in the dental practice. Motivating change in behaviour]. / Serie: Communicatie in de tandartspraktijk. Motiveren tot gedragsverandering.

Why people display certain behaviour is determined by a number of factors. If a dentist wants to change behaviour harmful to the patient's health, he will have to identify the factors determining this behaviour, taking into account factors like social acceptability. The dentist can then counsel the patient to change his or her behaviour using motivational interviewing techniques fitting the patient's motivational phase. A lot of frustration over non-compliance with (failing to follow-up on) recommended advice couldbe avoided this way.

[Series: Communication in the dental practice. Delivering bad news]. / Serie: Communicatie in de tandartspraktijk. Het slechtnieuwsgesprek.

In general practice a dentist will from time to time have to convey bad news to his patients. Often they feel burdened by this because of the emotional impact of such a conversation. This article discusses the way in which patients prefer to have bad news relayed to them. Using cases suggestions are made how to structure the bad news conversation.

[Series: Communication in the dental practice. Dealing with conflict and intimidation in the dental practice]. / Serie: Communicatie in de tandarts­praktijk. Omgaan met conflicten en intimidatie in de tandartspraktijk.

In the dental practice, several situations that might lead to conflict are imaginable. How a dentist deals with a conflict depends in part on the importance he places on the content of the conflict in relation to the importance of the relationship with the patient. This article briefly describes 5 conflict management styles. On the basis of case histories, suggestions are made for handling situations when a conflict leads to aggression, you are facing sexual intimidation or a conflict develops with a demanding patient. Finally, some advice is given on how to deal with errors/complications in treatment and dealing with criticism.

[Series: Communication in the dental practice. Building a good dentist-patient relationship with communication]. / Serie: Communicatie in de tandartspraktijk. Met communicatie bouwen aan een goede tandarts-patiëntrelatie.

A good dentist-patient relationship promotes better oral health of the patient. In order to build and maintain such a relationship a dentist should possess good communication skills. First of all, an open, empathic attitude is important, in which the dentist is aware that he can make mistakes interpreting patient behaviour. Good observational skills are necessary to detect (non-)verbal signals indicating how to interpret the patient's story. By demonstrating non-verbal, paralingual and verbal listening behaviour,he can show patients that he is paying attention to them. When discussing findings, it is important for patients to receive a clear explanation and be given the opportunity to ask questions. It is also wise to pay attention to the patient's expectations of the treatment outcome, elaborate on the advantages and disadvantages of the treatment alternatives and to be transparent in case of complications.

[Series: Communication in the dental practice. Dealing with anxiety in the dental office]. / Serie: Communicatie in de tandartspraktijk. Omgaan met angst in de tandartspraktijk.

Dentists frequently have to deal with anxious children and adults. Whether these patients can be treated successfully in a regular dental office depends on the type of fear and its severity. Proper diagnostic procedures are therefore paramount. This article gives an overview of how dental fear develops, is sustained, and can be treated. It elaborates on the communication skills important in reducing anxiety and preventing children from developing dental fear.

[Communication in the dental practice. Communicating with older patients]. / Serie: Communicatie in de tandartspraktijk. Communiceren met de oudere patiënt.

The ageing of the population in the Netherlands and in Belgium and the growing number of older people who still have their own teeth mean that dentists are seeing older patients in their dental practices increasingly often. Loss of cognitive abilities can hinder communication with the patients. Dementia and care-resistant behaviour demand a special patient-centred approach. In addition, frail older people are often dependent on carers or volunteers who actively participate in the conversation when the patient visits the dentist. This article addresses issues dentists should take into account in such cases. It also makes suggestions about how most effectively to direct communications with older patients and/or their companions.

Hydro-alcoholic media effects on theophylline release from sesamum polysaccharide gum matrices.

Concomitant ingestion of alcohol and medications can greatly affect drug plasma concentrations as dose dumping or failure may occur as a result of the fact that formulation excipients may not always be resistant to alcohol. In this study, a natural polysaccharide (Sesamum radiatum gum) (SG) was extracted, characterized and used to formulate sustained release theophylline compacts to study the effect of varying alcohol concentrations (v/v) in dissolution media on drug release from these compacts. X-ray powder diffraction showed that the extracted gum was amorphous in nature with the powder having excellent compaction properties as observed with its compact being significantly harder than those prepared with pure hydroxypropyl methyl cellulose (HPMC) K4M. X-ray microtomography showed that the compacts produced were homogenous in nature, however, swelling studies showed failure of the compacts at the highest concentration of absolute ethanol used (40% v/v). Dissolution studies showed similarity at all levels of alcohol tested (f2 = 57-91) in simulated gastric (0.1 N HCl, pH 1.2) and intestinal fluids (phosphate buffer, pH 6.8) for the HPMC compacts whereas dissimilarity only occurred for the SG compacts at the highest alcohol concentration in both media (f2 = 35). The suitability of SG as a matrix former that can resist alcoholic effects therefore makes it suitable as an alternative polymer with wider applications for drug delivery.

Growth factor release from dentine matrix by pulp-capping agents promotes pulp tissue repair-associated events.

AIM: To characterize growth factor release from dentine by pulp-capping agents and to determine the effects of liberated dentine extracellular matrix (dECM) components on pulp cells in the key wound healing processes of migration and cell growth. METHODOLOGY: Powdered human dentine was exposed to solutions of calcium hydroxide, white and grey mineral trioxide aggregate (MTA) (ProRoot, (Dentsply Tulsa, Tulsa, OK, USA) over 14 days. The solubilized dECM components were dialysed and lyophilized and characterized using multiplex quantitative ELISA. Following dECM component extraction dentine was analysed using Fourier-transformed infrared spectroscopy (FTIR). Primary rat dental pulp cells (RDPCs) were exposed to dECM components (0.1-100 µg mL-1 ) released by calcium hydroxide, white and grey MTA, and cell growth and chemotactic responses were assessed. Statistical differences between the experimental and control groups were determined using one-way anova. RESULTS: A broad range of growth factors, many not previously reported in dentine, were liberated by these pulp-capping agents, including SCF, M-CSF, GM-CSF, IGFBP-1, NGF and GDNF. White and grey MTA liberated more growth factors than calcium hydroxide. FTIR analysis of dentine exposed to pulp-capping agents showed partial depletion of amide bands I, II and III, with little alteration in phosphate peaks compared to untreated dentine. dECM components released by white and grey MTA induced significantly more cell growth at low-to-moderate concentrations (P â‰¦ 0.05) examined in this study and significantly enhanced cell chemotaxis at all concentrations compared with controls (P â‰¦ 0.05). CONCLUSIONS: White and grey MTA solubilize a broad range of bioactive molecules from dentine, which can induce proliferation and chemotaxis in pulp cells.

Release of bio-active dentine extracellular matrix components by histone deacetylase inhibitors (HDACi).

AIM: To characterize dentine matrix component (DMC) release and smear layer removal by histone deacetylase inhibitors (HDACis). METHODOLOGY: DMCs were extracted from powdered human dentine over 14 days using three HDACis, valproic acid (VPA), trichostatin A (TSA) and suberoylanilide hydroxamic acid (SAHA) and compared with a control extractant, 10% (w/v) EDTA. Protein compositions of the resultant extracts were analysed by 1D-polyacrylamide gel electrophoresis (1D-PAGE), TGF-ß-1 and MMP-9 ELISAs and a high-throughput growth factor antibody array. Dentine discs with a standardized smear layer were prepared from human molars and treated with EDTA (17% w/v), polyacrylic acid (PA) (20% v/v) and the experimental HDACis prior to analysis by scanning electron microscopy. Parametric ELISA data were analysed using one-way anova and Tukey's post hoc test, whilst nonparametric smear layer data were analysed by Kruskal-Wallis test and Mann-Whitney U-test (P < 0.05). RESULTS: HDACis did not remove smear layer in the presence or absence of PA pre-treatment (P ≥ 0.478). 1D-PAGE analysis demonstrated different protein profiles for EDTA and HDACi extracts. All HDACi solutions released TGF-ß-1 although less effectively than EDTA (P < 0.001), whilst MMP-9 was extracted in significantly higher concentration by EDTA and VPA compared with TSA (P < 0.012). Antibody array analysis demonstrated the ability of HDACis to extract a complex cocktail of established/novel growth factors from dentine, albeit significantly less efficiently than EDTA for certain cytokines (TGF-ß-1, PDGF-AA, VEGF-A) and significantly more effectively for others (GDF-15, IGF-1, EGRF-1, NGFR, BDNF, SCF-R). CONCLUSIONS: HDACi release a range of bioactive DMCs that could promote dentine repair processes in vivo; however, they are ineffective at removing smear layer.

Xenogeneic collagen matrix for periodontal plastic surgery procedures: a systematic review and meta-analysis.

Several clinical trials describe the effectiveness of xenogeneic collagen matrix (XCM) as an alternative option to surgical mucogingival procedures for the treatment of marginal tissue recession and augmentation of insufficient zones of keratinized tissue (KT). The aim of this systematic review and meta-analysis was to evaluate the clinical and patient-centred outcomes of XCM compared to other mucogingival procedures. Applying guidelines of the Preferred Reporting Items for Systematic Reviews and Meta analyses statement, randomized controlled trials were searched for in electronic databases and complemented by hand searching. The risk of bias was assessed using the Cochrane Collaboration's Risk of Bias tool and data were analysed using statistical software. A total of 645 studies were identified, of which, six trials were included with 487 mucogingival defects in 170 participants. Overall meta-analysis showed that connective tissue graft (CTG) in conjunction with the coronally advanced flap (CAF) had a significantly higher percentage of complete/mean root coverage and mean recession reduction than XCM. Insufficient evidence was found to determine any significant differences in width of KT between XCM and CTG. The XCM had a significantly higher mean root coverage, recession reduction and gain in KT compared to CAF alone. No significant differences in patient's aesthetic satisfaction were found between XCM and CTG, except for postoperative morbidity in favour of XCM. Operating time was significantly reduced with the use of XCM compared with CTG but not with CAF alone. There is no evidence to demonstrate the effectiveness of XCM in achieving greater root coverage, recession reduction and gain in KT compared to CTG plus CAF. Superior short-term results in treating root coverage compared with CAF alone are possible. There is limited evidence that XCM may improve aesthetic satisfaction, reduce postoperative morbidity and shorten the operating time. Further long-term randomized controlled trials are required to endorse the supposed advantages of XCM.

Epigenetic modulation of dental pulp stem cells: implications for regenerative endodontics.

Dental pulp stem cells (DPSCs) offer significant potential for use in regenerative endodontics, and therefore, identifying cellular regulators that control stem cell fate is critical to devising novel treatment strategies. Stem cell lineage commitment and differentiation are regulated by an intricate range of host and environmental factors of which epigenetic influence is considered vital. Epigenetic modification of DNA and DNA-associated histone proteins has been demonstrated to control cell phenotype and regulate the renewal and pluripotency of stem cell populations. The activities of the nuclear enzymes, histone deacetylases, are increasingly being recognized as potential targets for pharmacologically inducing stem cell differentiation and dedifferentiation. Depending on cell maturity and niche in vitro, low concentration histone deacetylase inhibitor (HDACi) application can promote dedifferentiation of several post-natal and mouse embryonic stem cell populations and conversely increase differentiation and accelerate mineralization in DPSC populations, whilst animal studies have shown an HDACi-induced increase in stem cell marker expression during organ regeneration. Notably, both HDAC and DNA methyltransferase inhibitors have also been demonstrated to dramatically increase the reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) for use in regenerative therapeutic procedures. As the regulation of cell fate will likely remain the subject of intense future research activity, this review aims to describe the current knowledge relating to stem cell epigenetic modification, focusing on the role of HDACi on alteration of DPSC phenotype, whilst presenting the potential for therapeutic application as part of regenerative endodontic regimens.

The effect of piezoelectric ultrasonic instrumentation on titanium discs: a microscopy and trace elemental analysis in vitro study.

AIM: To evaluate in vitro topographical and composition changes by piezoelectric ultrasonic instrumentation with metallic and plastic tips on machined and moderately roughened titanium surfaces. METHODS: Twenty machined and moderately roughened laser-marked titanium discs were ultrasonically instrumented with metallic and plastic tips. Surface instrumentation was carried out with controlled pressure for 20 and 30 seconds at two power settings. For each time and power setting, instrumentation was repeated four times with one instrumentation per disc quadrant. Surface topography analysis was performed using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Surface roughness measurements were compared between instrumented and non-instrumented surfaces. Surface element composition and rinsing solutions were evaluated using energy-dispersive spectroscopy (EDS) and trace elemental analysis using inductively coupled plasma mass spectrometry (ICPMS), respectively. RESULTS: SEM photomicrographs and CLSM 3D surface plot images of instrumented machined and moderately roughened surfaces demonstrated severe surface topographical alterations with metallic tips and mild to moderate changes for plastic tip instrumented sites. ICPMS analysis of the rinsing solutions identified titanium and other metal traces with the use of metallic tips, and mainly titanium and carbon when plastic tips were used. Surface EDS analysis showed elemental traces of the ultrasonic tips. CONCLUSION: Ultrasonic instrumentation with metallic or plastic tips created surface topographical and compositional changes. Different changes in surface topography were noted between the surfaces, as the roughness of the machined surfaces increased while the extent of roughness of the moderately roughened surfaces decreased. The clinical relevance of these changes is yet to be determined.

A comparison of the in vitro mineralisation and dentinogenic potential of mesenchymal stem cells derived from adipose tissue, bone marrow and dental pulp.

Stem-cell-based therapies provide a biological basis for the regeneration of mineralised tissues. Stem cells isolated from adipose tissue (ADSCs), bone marrow (BMSCs) and dental pulp (DPSCs) have the capacity to form mineralised tissue. However, studies comparing the capacity of ADSCs with BMSCs and DPSCs for mineralised tissue engineering are lacking, and their ability to regenerate dental tissues has not been fully explored. Characterisation of the cells using fluorescence-activated cell sorting and semi-quantitative reverse transcription PCR for MSC markers indicated that they were immunophenotypically similar. Alizarin red (AR) staining and micro-computed tomography (µCT) analyses demonstrated that the osteogenic potential of DPSCs was significantly greater than that of BMSCs and ADSCs. Scanning electron microscopy and AR staining showed that the pattern of mineralisation in DPSC cultures differed from ADSCs and BMSCs, with DPSC cultures lacking defined mineralised nodules and instead forming a diffuse layer of low-density mineral. Dentine matrix components (DMCs) were used to promote dentinogenic differentiation. Their addition to cultures resulted in increased amounts of mineral deposited in all three cultures and significantly increased the density of mineral deposited in BMSC cultures, as determined by µCT analysis. Addition of DMCs also increased the relative gene expression levels of the dentinogenic markers dentine sialophosphoprotein and dentine matrix protein 1 in ADSC and BMSC cultures. In conclusion, DPSCs show the greatest potential to produce a comparatively high volume of mineralised matrix; however, both dentinogenesis and mineral volume was enhanced in ADSC and BMSC cultures by DMCs, suggesting that these cells show promise for regenerative dental therapies.

The effects of cryopreservation on cells isolated from adipose, bone marrow and dental pulp tissues.

The effects of cryopreservation on mesenchymal stem cell (MSC) phenotype are not well documented; however this process is of increasing importance for regenerative therapies. This study examined the effect of cryopreservation (10% dimethyl-sulfoxide) on the morphology, viability, gene-expression and relative proportion of MSC surface-markers on cells derived from rat adipose, bone marrow and dental pulp. Cryopreservation significantly reduced the number of viable cells in bone marrow and dental pulp cell populations but had no observable effect on adipose cells. Flow cytometry analysis demonstrated significant increases in the relative expression of MSC surface-markers, CD90 and CD29/CD90 following cryopreservation. sqRT-PCR analysis of MSC gene-expression demonstrated increases in pluripotent markers for adipose and dental pulp, together with significant tissue-specific increases in CD44, CD73-CD105 following cryopreservation. Cells isolated from different tissue sources did not respond equally to cryopreservation with adipose tissue representing a more robust source of MSCs.

Systemic azithromycin versus amoxicillin/metronidazole as an adjunct in the treatment of periodontitis: a systematic review and meta-analysis.

BACKGROUND: The use of systemic azithromycin (AZT) and amoxicillin/metronidazole (AMX/MTZ) as adjuncts provided additional clinical and microbiological benefits over subgingival instrumentation alone. However, the superiority of one antibiotic regimen over another has not been proven. Therefore, the aim of this systematic review and meta-analyses was to evaluate the clinical efficacy and safety of subgingival instrumentation (SI) in conjunction with the systemic use of AZT or AMX/MTZ for the treatment of periodontitis from current published literature. METHODS: Electronic databases were searched to identify randomized controlled trials (RCTs), controlled clinical trials, prospective and retrospective human studies that compared the adjunctive use of systemic AZT to AMX/MTZ with SI in the treatment of periodontitis. The eligibility criteria were defined based on the participant (who had periodontitis), intervention (SI with adjunctive use of systemic AZT), comparison (SI with adjunctive use of systemic AMX/MTZ), outcomes (primary outcome: changes in probing pocket). The risk of bias was assessed using the Cochrane Collaboration's Risk of Bias tool. Data were analysed using a statistical software program. RESULTS: Five studies with 151 participants with periodontitis were included in the present review. Of these, 74 participants received adjunctive AZT, while the remaining participants received AMX/MTZ as an adjunct to SI. The adjunctive use of AZT and AMX/MTZ had comparable changes in probing pocket depths at 1-3 months with no statistically significant difference (mean difference (MD) 0.01; 95% CI -0.20 to 0.22; P = 0.94). The adjunctive use of AZT had significantly fewer number of residual sites with probing pocket depths of ≥5 mm at 1-3 months compared to the adjunctive use of AMX/MTZ (MD -3.41; 95% CI -4.73 to -2.10; P < 0.0001). The prevalence rates of adverse events among participants who received AZT and AMX/MTZ were 9.80% and 14.8%, respectively. The meta-analysis showed that the difference between the two groups was not statistically significant (risk ratio 0.69; 95% CI 0.28 to 1.72; P = 0.43). CONCLUSIONS: Within the limitation of this review, there was no superiority between AZT and AMX/MTZ in terms of mean changes in probing pocket depths, clinical attachment level, bleeding on probing at 1-3 months. AZT seem to be associated with less sites with residual probing pocket depths of ≥5 mm at 1-3 months and fewer adverse events compared with AMX/MTZ. © 2023 Australian Dental Association.

Association of a polymorphism in the indoleamine- 2,3-dioxygenase gene and interferon-α-induced depression in patients with chronic hepatitis C.

Interferon (IFN)-α treatment for infectious diseases and cancer is associated with significant depressive symptoms that can limit therapeutic efficacy. Multiple mechanisms have been implicated in IFN-α-induced depression including immune, neuroendocrine and neurotransmitter pathways. To further explore mechanisms of IFN-α-induced depression and establish associated genetic risk factors, single nucleotide polymorphisms in genes encoding proteins previously implicated in IFN-α-induced depression were explored in two self-reported ethnic groups, Caucasians (n=800) and African Americans (n=232), participating in a clinical trial on the impact of three pegylated IFN-α treatment regimens on sustained viral response in patients with chronic hepatitis C. Before treatment, all subjects were free of psychotropic medications and had a score ≤20 on the Center for Epidemiologic Studies Depression Scale (CES-D), which was used to assess depressive symptom severity throughout the study. In Caucasians, a polymorphism (rs9657182) in the promoter region of the gene encoding indoleamine-2,3-dioxygenase (IDO1) was found to be associated with moderate or severe IFN-α-induced depressive symptoms (CES-D>20) at 12 weeks of IFN-α treatment (P=0.0012, P<0.05 corrected). Similar results were obtained for treatment weeks 24, 36 and 48. In subjects homozygous for the risk allele (CC, n=150), the odds ratio for developing moderate or severe depressive symptoms at treatment week 12 was 2.91 (confidence interval: 1.48-5.73) compared with TT homozygotes (n=270). rs9657182 did not predict depression in African Americans, who exhibited a markedly lower frequency of the risk allele at this locus. The findings in Caucasians further support the notion that IDO has an important role in cytokine-induced behavioral changes.