1,25-dihydroxyvitamin-D3 distinctly impacts the paracrine and cell-to-cell contact interactions between hPDL-MSCs and CD4+ T lymphocytes.

Introduction: Human periodontal ligament-derived mesenchymal stromal cells (hPDL-MSCs) possess a strong ability to modulate the immune response, executed via cytokine-boosted paracrine and direct cell-to-cell contact mechanisms. This reciprocal interaction between immune cells and hPDL-MSCs is influenced by 1,25-dihydroxyvitamin-D3 (1,25(OH)2D3). In this study, the participation of different immunomodulatory mechanisms on the hPDL-MSCs-based effects of 1,25(OH)2D3 on CD4+ T lymphocytes will be elucidated using different co-culture models with various cytokine milieus. Material and methods: hPDL-MSCs and CD4+ T lymphocytes were co-cultured indirectly and directly with inserts (paracrine interaction only) or directly without inserts (paracrine and direct cell-to-cell contact interaction). They were stimulated with TNF-α or IL-1ß in the absence/presence of 1,25(OH)2D3. After five days of co-cultivation, the CD4+ T lymphocyte proliferation, viability, and cytokine secretion were analyzed. Additionally, the gene expression of soluble and membrane-bound immunomediators was determined in hPDL-MSCs. Results: In the indirect and direct co-culture model with inserts, 1,25(OH)2D3 decreased CD4+ T lymphocyte proliferation and viability. The direct co-culture model without inserts caused the opposite effect. 1,25(OH)2D3 mainly decreased the CD4+ T lymphocyte-associated secretion of cytokines via hPDL-MSCs. The degree of these inhibitions varied between the different co-culture setups. 1,25(OH)2D3 predominantly decreased the expression of the soluble and membrane-bound immunomediators in hPDL-MSCs to a different extent, depending on the co-culture models. The degree of all these effects depended on the absence and presence of exogenous TNF-α and IL-1ß. Conclusion: These data assume that 1,25(OH)2D3 differently affects CD4+ T lymphocytes via the paracrine and direct cell-to-cell contact mechanisms of hPDL-MSCs, showing anti- or pro-inflammatory effects depending on the co-culture model type. The local cytokine microenvironment seems to be involved in fine-tuning these effects. Future studies should consider this double-edged observation by executing different co-culture models in parallel.

Comparative study of the McGrath™ videolaryngoscope blades and conventional laryngoscopy efficacy during mechanical chest compressions: Insights from a randomized trial with 90 anesthesiologists on objective and subjective parameters.

AIMS: This study aimed to compare the efficacy and utility of the McGrath™ videolaryngoscope, using the Macintosh-like McGrath™ MAC blade and the hyperangulated McGrath™ MAC Xblade with a conventional Macintosh blade under simulated resuscitation conditions. METHODS: A prospective, randomized study under conditions mimicking ongoing chest compressions was conducted with 90 anesthesiologists. Intubation success rates, time-to-vocal cords, time-to-intubate, and time-to-ventilate were measured. Additionally, the study assessed the subjective ratings and the perceived workload using the 'NASA-task-load-index' during the procedure. RESULTS: The overall intubation success rate was device dependent 99-100%. The McGrath™ MAC and McGrath™ MAC Xblade showed faster visualization times compared to conventional blades. The MAC blade demonstrated superior performance in time-to-intubate and time-to-ventilate compared to both conventional and MAC Xblades. Despite excellent visualization, the MAC Xblade posed challenges in tube placement, reflected in a prolonged intubation time of >120 seconds in one case. Both MAC and MAC Xblade reduced potential dental injuries and interruptions to chest compressions compared to conventional laryngoscopes. User experience significantly impacted intubation times with conventional laryngoscopes, but this effect was mitigated with videolaryngoscopy. Participants reported lower stress and effort when using videolaryngoscopes, with the MAC blade rated superior in perceived time pressure. CONCLUSION: The study supports the superiority of videolaryngoscopy with a Macintosh-like blade over conventional laryngoscopy during mechanical chest compressions, particularly for less experienced users. The McGrath™ MAC blade, in particular, offers advantages in intubation time, user-friendliness, and reduced stress. However, the MAC Xblade's challenges during tube placement highlight the need for further clinical validation. Continued research is essential to refine guidelines and improve resuscitation outcomes.

Quantitative bile acid profiling in healthy adult dogs and pups from serum, plasma, urine, and feces using LC-MS/MS.

Synthesis and secretion of bile acids (BA) is a key physiological function of the liver. In pathological conditions like portosystemic shunt, hepatic insufficiency, hepatitis, or cirrhosis BA metabolism and secretion are disturbed. Quantification of total serum BA is an established diagnostic method to assess the general liver function and allows early detection of abnormalities, liver disease progression and guidance of treatment decisions. To date, data on comparative BA profiles in dogs are limited. However, BA profiles might be even better diagnostic parameters than total BA concentrations. On this background, the present study analyzed and compared individual BA profiles in serum, plasma, urine, and feces of 10 healthy pups and 40 adult healthy dogs using ultra-high performance liquid chromatography coupled to electrospray ionization mass spectrometry. Sample preparation was performed by solid-phase extraction for serum, plasma, and urine samples or by protein precipitation with methanol for the feces samples. For each dog, 22 different BA, including unconjugated BA and their glycine and taurine conjugates, were analyzed. In general, there was a great interindividual variation for the concentrations of single BA, mostly exemplified by the fact that cholic acid (CA) was by far the most prominent BA in blood and urine samples of some of the dogs (adults and pups), while in others, CA was under the detection limit. There were no significant age-related differences in the BA profiles, but pups showed generally lower absolute BA concentrations in serum, plasma, and urine. Taurine-conjugated BA were predominant in the serum and plasma of both pups (68%) and adults (74-75%), while unconjugated BA were predominant in the urine and feces of pups (64 and 95%, respectively) and adults (68 and 99%, respectively). The primary BA chenodeoxycholic acid and taurocholic acid and the secondary BA deoxycholic acid and lithocholic acid were the most robust analytes for potential diagnostic purpose. In conclusion, this study reports simultaneous BA profiling in dog serum, plasma, urine, and feces and provides valuable diagnostic data for subsequent clinical studies in dogs with different kinds of liver diseases.

Paracrine- and cell-contact-mediated immunomodulatory effects of human periodontal ligament-derived mesenchymal stromal cells on CD4+ T lymphocytes.

BACKGROUND: Mesenchymal stromal cells (MSCs) isolated from the periodontal ligament (hPDL-MSCs) have a high therapeutic potential, presumably due to their immunomodulatory properties. The interaction between hPDL-MSCs and immune cells is reciprocal and executed by diverse cytokine-triggered paracrine and direct cell-to-cell contact mechanisms. For the first time, this study aimed to directly compare the contribution of various mechanisms on this reciprocal interaction using different in vitro co-culture models at different inflammatory milieus. METHODS: Three co-culture models were used: indirect with 0.4 µm-pored insert, and direct with or without insert. After five days of co-culturing mitogen-activated CD4+ T lymphocytes with untreated, interleukin (IL)-1ß, or tumor necrosis factor (TNF)-α- treated hPDL-MSCs, the CD4+ T lymphocyte proliferation, viability, and cytokine secretion were investigated. The gene expression of soluble and membrane-bound immunomediators was investigated in the co-cultured hPDL-MSCs. RESULTS: Untreated hPDL-MSCs decreased the CD4+ T lymphocyte proliferation and viability more effectively in the direct co-culture models. The direct co-culture model without inserts showed a strikingly higher CD4+ T lymphocyte cell death rate. Adding IL-1ß to the co-culture models resulted in substantial CD4+ T lymphocyte response alterations, whereas adding TNF resulted in only moderate effects. The most changes in CD4+ T lymphocyte parameters upon the addition of IL-1ß or TNF-α in a direct co-culture model without insert were qualitatively different from those observed in two other models. Additionally, the co-culture models caused variability in the immunomediator gene expression in untreated and cytokine-triggered hPDL-MSCs. CONCLUSION: These results suggest that both paracrine and cell-to-cell contact mechanisms contribute to the reciprocal interaction between hPDL-MSCs and CD4+ T lymphocytes. The inflammatory environment affects each of these mechanisms, which depends on the type of cytokines used for the activation of MSCs' immunomodulatory activities. This fact should be considered by comparing the outcomes of the different models.

Do strength of zirconia-abutment-interfaces depend on cement, zirconia type and titanium abutment dimensions?

OBJECTIVES: Part 1 of this study investigates the influence of zirconia types, chimney heights, and gingival heights on the strength of the zirconia-abutment-interface. Part 2 extends the analysis to include adhesive brands and macro-retentions. METHODS: In Part 1, the study utilized three zirconia types (700 MPa, 1000 MPa, 1200 MPa) to fabricate 234 screw-retained zirconia crowns with varying chimney heights (3.5 mm, 4.1 mm, 5 mm) and gingival heights (0.65 mm, 1.2 mm, 3 mm) of the titanium abutments. All adherend surfaces underwent sandblasting with aluminum oxide before cementation with a specific resin cement. In Part 2, the investigation of 240 screw-retained zirconia crowns focused on a single zirconia type (1000 MPa) with chimney heights of 3.5 mm and 5 mm and a gingival height of 0.65 mm of the titanium abutments, cemented with three different resin cements. All adherent surfaces underwent sandblasting with aluminum oxide before cementation, whereas 120 out of 240 abutments received additional macro retentions. Storage in water at 37 °C for 24 h preceded the tensile test. RESULTS: The study revealed a substantial impact of chimney height and zirconia type on the bond strength of the zirconia-abutment-interface. Neither adhesive brands nor macro retentions significantly impacted the bond strength. Fracture incidence was significantly influenced by gingival height and zirconia type in part 1, whereas in part 2 smaller chimney heights correlated with a higher fracture incidence. SIGNIFICANCE: This study contributes insights into the complex interplay of factors influencing the zirconia-abutment-interface. The results provide a foundation for refining clinical approaches, emphasizing the importance of chimney height and zirconia type in achieving successful anterior gap implant restorations.

Video double-lumen tube for one lung ventilation: implementation and experience in 343 cases of routine clinical use during the first 20 months of the SARS-CoV-2 pandemic.

BACKGROUND: Double-lumen tubes (DLTs) are the preferred device for lung isolation. Conventional DLTs (cDLT) need a bronchoscopic position control. Visualisation of correct DLT positioning could be facilitated by the use of a video double-lumen tube (vDLT). During the SARS-CoV-2-pandemic, avoiding aerosol-generation was suggesting using this device. In a large retrospective series, we report both general and pandemic related experiences with the device. METHODS: All anesthesia records from patients aged 18 years or older undergoing surgery from April 1st, 2020 to December 31st, 2021 in the department of thoracic surgery requiring intraoperative lung isolation were analyzed retrospectively. RESULTS: During the investigation period 343 left-sided vDLTs (77.4%) and 100 left-sided cDLTs (22.6%) were used for one lung ventilation. In the vDLT group bronchoscopy could be reduced by 85.4% related to the cDLT group. Additional bronchoscopy to reach or maintain correct position was needed in 11% of the cases. Other bronchoscopy indications occured in 3.6% of the cases. With cDLT, in 1% bronchoscopy for other indications than conforming position was observed. CONCLUSIONS: The Ambu® VivaSight™ vDLT is an efficient, easy-to-use and safe airway device for the generation of one lung ventilation in patients undergoing thoracic surgery. The vDLT implementation was achieved easily with full interchangeability to the left-sided cDLT. Using the vDLT can reduce the need for aerosol-generating bronchoscopic interventions by 85.4%. Continuous video view to the carina enabling position monitoring of the DLT without need for bronchoscopy might be beneficial for both employee's and patient's safety.

Quercetin in the Prevention of Induced Periodontal Disease in Animal Models: A Systematic Review and Meta-Analysis.

BACKGROUND: Periodontitis is an inflammatory condition initiated by oral bacteria and is associated with several systemic diseases. Quercetin is an anti-inflammatory and anti-bacterial poly-phenol present in various foods. The aim of this meta-analysis was the evaluation of the effects of quercetin administration in animal models of experimental periodontitis. METHODS: A systematic search was performed in electronic databases using the following search terms: "periodontitis" or "periodontal disease" or "gingivitis" and "quercetin" or "cyanidanol" or "sophoretin" or "pentahydroxyflavone". In vivo preclinical animal models of experimental periodontal disease with a measurement of alveolar bone loss were included in the analysis. The risk of bias of the included studies was assessed using the SYRCLE tool. RESULTS: The systematic search yielded 335 results. Five studies were included, four of them qualified for a meta-analysis. The meta-analysis showed that quercetin administration decreased alveolar bone loss (τ2 = 0.31, 1.88 mm 95%CI: 1.09, 2.67) in experimental periodontal disease animal models. However, the risk of bias assessment indicated that four SYRCLE domains had a high risk of bias. CONCLUSIONS: Quercetin diminishes periodontal bone loss and prevents disease progression in animal models of experimental periodontal disease. Quercetin might facilitate periodontal tissue hemostasis by reducing senescent cells, decreasing oxidative stress via SIRT1-induced autophagy, limiting inflammation, and fostering an oral bacterial microenvironment of symbiotic microbiota associated with oral health. Future research will show whether and how the promising preclinical results can be translated into the clinical treatment of periodontal disease.

Clinical and Radiographic Outcomes of Single Implant-Supported Zirconia Crowns Following a Digital and Conventional Workflow: Four-Year Follow-Up of a Randomized Controlled Clinical Trial.

PURPOSE: This study aimed to compare the clinical and radiographic outcomes of single posterior screw-retained monolithic implant crowns following a digital and conventional workflow and to report on the survival/complication rate after a mean 4-year follow-up. MATERIALS AND METHODS: Thirty patients with a single posterior tooth missing were rehabilitated with a bone-level implant. After a healing period of ≥3 months, they were subjected to both a digital and conventional workflow to fabricate two screw-retained monolithic implant crowns. The quantitative clinical adjustments to both crowns (intrasubject comparison) and a questionnaire were recorded at try-in. Thereafter, a crown of the digital and conventional workflows was randomly inserted. At the last follow-up, the marginal bone level (MBL), peri-implant health-related parameters (bleeding on probing (BoP), plaque, pocket probing depth (PPD)), and functional implant prosthodontic score (FIPS) were assessed. Furthermore, the implant survival and success rates and technical complications were evaluated. RESULTS: A total of 27 patients were followed for a mean period of 4.23 ± 1.10 years. There was no significant difference between the digital and conventional workflows regarding clinical adjustments and questionnaire outcomes. More than twice as many participants recommended digital (n = 16) compared to conventional impressions (n = 7) to friends. The implant survival and success rate were 100% and 96.3%, respectively. Furthermore, two de-cementations and one fracture of the ti-base abutment occurred. There were no significant differences in BoP, plaque, and PPD metrics between the two groups. The changes in the MBL between implant crown insertion (baseline) and the last follow-up were 0.07 ± 0.19 mm and 0.34 ± 0.62 mm in the digital and conventional groups, respectively (p = 0.195). The mean overall FIPS score was 8.11 ± 1.37 (range: 5-10). CONCLUSIONS: The clinical and radiographic outcomes of single screw-retained monolithic implant crowns were similar between both workflows after a mean of 4 years of service. The patients did not clearly prefer an impression technique for their restoration, although they would recommend the digital impression more often to friends. Thus, decision regarding clinical workflows may be based on the patient's and/or clinician's preference.

Pharmacodynamic effects of molidustat on erythropoiesis in healthy cats.

BACKGROUND: Inhibition of hypoxia-inducible factor prolyl hydroxylase (HIF-PH) stimulates erythropoiesis in rats, dogs, monkeys, and humans. HYPOTHESIS/OBJECTIVE: Determine if molidustat, a novel HIF-PH inhibitor, stimulates erythropoiesis in healthy cats. ANIMALS: Seventeen healthy adult laboratory cats. METHODS: Randomized, placebo-controlled study. Cats were treated PO once daily with suspensions of 0 (Group 1; n = 6), 5 (Group 2; n = 6), or 10 (Group 3; n = 5) mg/kg of molidustat. Effects on red blood cell parameters, reticulocyte indices and plasma erythropoietin (EPO) concentrations were evaluated. Molidustat treatment was stopped when hematocrit (HCT) exceeded 60%. RESULTS: Compared to placebo, a significant increase in mean HCT was evident starting on Day 14 (Group 2:54.4% vs 40.3%, P < .001, 95% confidence interval [CI] for the difference [8.95-19.28]; Group 3:61.2% vs 40.3%, P < .001, 95% CI [15.48-26.43]) and remained significantly higher for the entire treatment period. In molidustat-treated groups, HCT exceeded 60% on Day 21 (Group 2) and Day 14 (Group 3). Mean HCT in molidustat-treated cats returned to within the reference range (29%-45%) after Day 56 and was numerically comparable to placebo from Day 70 onwards. Red blood cell count and hemoglobin concentrations followed a similar pattern as HCT. Mean EPO concentrations significantly increased after molidustat administration on all assessment days. Molidustat treatments were well tolerated. CONCLUSIONS AND CLINICAL IMPORTANCE: Marked erythropoietic effects were identified after daily administration of molidustat to healthy cats and additional studies are warranted to evaluate the effects in anemic cats.

Effects of enamel matrix derivative in nonsurgical periodontal therapy on pro-inflammatory profiles, microbial environment and clinical outcome: a randomized clinical trial.

OBJECTIVES: This study aimed to evaluate the impact of enamel matrix derivative (EMD) application following subgingival instrumentation of residual pockets in periodontitis patients on inflammatory host response, microbiological composition, and clinical outcome. METHODS: In this double-blinded randomized controlled trial, a total of 22 patients with generalized periodontitis stage III or IV presenting with ≥ 6 mm probing pocket depth (PPD) at re-evaluation after initial periodontal therapy were included. Participants were randomly allocated at a 1:1 ratio to subgingival instrumentation with (EMD +) or without (EMD-) non-surgical EMD application into the pocket. PPD, clinical attachment level (CAL), bleeding on probing (BoP), plaque index (PI), as well as a panel of pro-inflammatory cytokines and periodontal pathogen count in the gingival crevicular fluid (GCF) of the respective sites were evaluated at baseline (T0) and six months afterwards (T1). RESULTS: Both treatment groups showed a significant PPD reduction (EMD + 1.33 ± 1.15 mm, p < 0.001; EMD- 1.32 ± 1.01 mm, p < 0.001) as well as CAL gain (EMD + 1.13 ± 1.58 mm, p < 0.001; EMD- 0.47 ± 1.06 mm, p = 0.005) from T0 to T1. While no intergroup differences for PPD reduction were observed, CAL gain was higher in EMD + sites compared to EMD- (p = 0.009). No essential effects on cytokine expression as well as bacterial count were detected. CONCLUSIONS: Application of EMD as an adjunct to subgingival instrumentation of residual pockets yielded benefits regarding CAL gain; however, effects on PPD reduction, inflammatory cytokines, and bacterial count were negligible. TRIAL REGISTRATION: ClinicalTrials.gov (NCT04449393), registration date 26/06/2020. CLINICAL RELEVANCE: Based on the obtained results, additional non-surgical EMD application compared to subgingival instrumentation alone showed no clinically relevant effects on treatment outcome and underlying biological mechanisms.

Optimized Erbium-Doped Yttrium Aluminum Garnet (Er:YAG) Laser Parameters for the Removal of Dental Ceramic Restorations.

OBJECTIVES: The use of lasers for debonding adhesively luted ceramic restorations is a rather recent oral laser application in dentistry. The removal of all-ceramic restorations in the mouth can often be a troublesome task. A novel method for the debonding of ceramic restorations without damaging the restorations is Er:YAG laser irradiation. The aim of this study was to evaluate the Er:YAG laser for debonding procedures of different dental ceramics and to identify appropriate laser settings. MATERIAL AND METHODS: Lithium disilicate, zirconium-reinforced lithium silicate, feldspatic ceramic, and zirconium dioxide were investigated. Ten ceramic rectangular-shaped specimens with 1 and 2 mm thickness were produced from each material. All specimens were irradiated with four different power settings 1.5; 2.5; 3.5; 4.5 W, pulse duration 50 µs, laser repetition rate 10 Hz, time of irradiation 10 s. The transmitted energy was measured with a powermeter. Additionally the suitability of the Er:YAG laser to remove the adhesively bonded ceramic and the time until loss of retention was evaluated. RESULTS: The transmission rate for 1 and 2 mm platelets was determined for zirconium-reinforced lithium silicate at 54.6%/35.6%, lithium disilicate at 53.2%/35.7%, zirconium dioxide at 40.6%/32.4%, and for the feldspathic ceramic at 19.4%/10.1%. For zirconium-reinforced lithium silicate and zirconium dioxide 2.5 W (250 mJ/10 Hz) was an appropriate energy level for effective debonding. Whereas for lithium disilicate and for feldspathic ceramic, 4.5 W (450 mJ/10 Hz) is required for efficient debonding. CONCLUSIONS: There are differences regarding transmission rates between ceramic types for the Er:YAG laser light and additionally depending on the type of ceramic different energy settings should be used for adequate debonding. Based on our in-vitro experiments we recommend 2.5 W for zirconium-reinforced lithium silicate and zirconium dioxide and 4.5 W for lithium disilicate and feldspatic ceramic. Transmission rates of different ceramic types and varying influences of thicknesses and bonding materials should be considered to adjust the laser parameters during laser debonding of adhesively luted all-ceramic restorations.

An approach to difficult airway in infants: Comparison of GlideScope® Spectrum LoPro, GlideScope® Spectrum Miller and conventional Macintosh and Miller blades in a simulated Pierre Robin sequence performed by 90 anesthesiologists.

BACKGROUND: Airway management can be challenging in neonates and infants. The Pierre Robin sequence (PRS) is a condition characterized by micrognathia, glossoptosis and airway obstruction. The airway management of these patients poses great challenges for anesthesiologists and pediatricians alike. To date, there has been no direct comparison of the hyperangulated GlideScope® Spectrum LoPro (GLP), the straight GlideScope® Spectrum Miller (GSM), a conventional Macintosh (MC) and a conventional Miller blade (ML) in patients with PRS. METHODS: For this purpose, 90 anesthesiologists (43 with limited experience, 47 with extensive experience) performed orotracheal intubation on an Air-Sim® Pierre Robin X manikin using GLP, GSM, MC and ML in randomized order. 'Time-to-vocal-cords', 'time-to-intubate', 'time-to-ventilate', the severity of oral-soft-tissue-trauma and the subjective evaluation of each device were recorded. RESULTS: A significantly faster and better view of the vocal cords and lower oral-soft-tissue-trauma was achieved using the GLP (p<0.001). Though, there were no significant differences in the 'time-to-intubate' or 'time-to-ventilate'. The highest intubation success rate was found with GSM and the lowest with GLP (GSM 100%, ML 97.8%, MC 96.7%, GLP 93.3%). When using the videolaryngoscopes, there were no undetected esophageal intubations but in six cases prolonged attempts of intubation (>120s) with the GLP. In the sub-group with extensive experience, we found significantly shorter intubation times for the GSM and ML. The GLP was the tool of choice for most participants, while the conventional MC received the lowest rating. CONCLUSIONS: Videolaryngoscopy leads to increased safety for the prevention of undetected esophageal intubation in the airway management in a PRS manikin. Hyperangulated blades may ensure a good and fast view of the vocal cords and low oral-soft-tissue-trauma but pose a challenge during the placement of the tube. Specific skills and handling seem to be necessary to ensure a safe tube placement with this sort of blades.

Evaluation of canine and feline leukocyte differential counts obtained with the scil vCell 5 compared to the Advia 2120 hematology analyzer and a manual method.

The vCell 5 (scil Animal Care), a point-of-care hematology analyzer (POCA), was recently introduced to veterinary laboratories. This laser- and impedance-based analyzer is capable of providing a CBC with 5-part WBC differential count (Diff) along with WBC cytograms and flags serving as interpretation aids for numerical results. We compared the scil POCA-Diff to reference methods (i.e., manual differential count, Advia 2120 hematology analyzer [Siemens]) for canine and feline blood samples and considered WBC cytograms and flags. Total observed error (TEo), calculated from CV and bias%, was compared to total allowable error (TEa). Data were analyzed before and after a review process (exclusion of flagged and samples with invalid cytograms). For both species, correlation was good-to-excellent (rs = 0.81-0.97) between both analyzers for all variables, except for feline monocytes (rs = 0.21-0.63) and canine monocyte% (rs = 0.50). Smallest biases were seen for neutrophils (dog: -5.7 to 0.8%; cat: 1.5-9.4%) with both reference methods. Quality requirements (TEo < TEa) were fulfilled for canine and feline neutrophils (TEo = 5.3-10.6%, TEa = 15%) and eosinophils (TEo = 67.1-83%, TEa = (90)-50%) considering at least one reference method. Our review process led to mildly higher rs-values for most variables. Although not completely satisfactory, the scil POCA provides reliable results in compliance with ASVCP quality goals for canine and feline neutrophils and eosinophils. Analyzer flag and cytogram analysis served as useful tools for QA, indicating the necessity for manual review of blood smears, and contributed to improvement of scil POCA performance.

The effect of combination treatment of CO2-laser irradiation and tetracalcium phosphate/dicalcium phosphate anhydrate on dentinal tubules blockage: an in vitro study.

The aim of this study was the evaluation of the in vitro efficacy of a carbon dioxide (CO2) laser, a tetracalcium phosphate/dicalcium phosphate anhydrate (TP/DP) desensitizer and the combination of the desensitizer and additional CO2 laser irradiation as a treatment modality for cervical dentin hypersensitivity. A total of 48 dental specimens, prepared from extracted human premolars and molars, were divided into four groups: a control group, a TP/DP desensitizer paste group, a CO2 laser (10.600-nm wavelength) group, and a paste and laser group. The specimens were coated with nail varnish except in the marked area and were then immersed in 2% methylene blue dye for 1 h. The specimens were then washed, dried, and cut longitudinally. Thereafter, photos of 40 dentin specimens were taken and evaluated. The area of penetration was assessed and reported as percentage of the dentin surface area. Additionally eight dental specimens were examined with the aid of a scanning electron microscope and evaluated. Significant differences in the penetration depth were found for all experimental groups compared to the control group. The lowest penetration area was detected in the paste-laser group (16.5%), followed by the laser (23.7%), the paste (48.5%), and the control group (86.2%). The combined treatment of the CO2 laser and a TP/DP desensitizer was efficient in sealing the dentinal surface and could be a treatment option for cervical dentin hypersensitivity.

An ultra-high-throughput screen for the evaluation of peptide HLA-Binder interactions.

Peptide human leukocyte antigen (pHLA) targeting therapeutics like T-cell receptor based adoptive cell therapy or bispecific T cell engaging receptor molecules hold great promise for the treatment of cancer. Comprehensive pre-clinical screening of therapeutic candidates is important to ensure patient safety but is challenging because of the size of the potential off-target space. By combining stabilized peptide-receptive HLA molecules with microarray printing and screening, we have developed an ultra-high-throughput screening platform named ValidaTe that enables large scale evaluation of pHLA-binder interactions. We demonstrate its potential by measuring and analyzing over 30.000 binding curves for a high-affinity T cell Engaging Receptor towards a large pHLA library. Compared to a dataset obtained by conventional bio-layer interferometry measurements, we illustrate that a massively increased throughput (over 650 fold) is obtained by our microarray screening, paving the way for use in pre-clinical safety screening of pHLA-targeting drugs.

25-hydroxyvitamin D3 generates immunomodulatory plasticity in human periodontal ligament-derived mesenchymal stromal cells that is inflammatory context-dependent.

Introduction: Human periodontal ligament-derived mesenchymal stromal cells (hPDL-MSCs) exhibit a tight bi-directional interaction with CD4+ T lymphocytes. The hPDL-MSCs' immunomodulatory abilities are drastically enhanced by pro-inflammatory cytokines via boosting the expression of various immunomediators. 25-hydroxyvitamin D3 (25(OH)D3), the major metabolite of vitamin D3 in the blood, affects both hPDL-MSCs and CD4+ T lymphocytes, but its influence on their interaction is unknown. Methods: Therefore, primary hPDL-MSCs were stimulated in vitro with tumor necrosis factor (TNF)-α a or interleukin (IL)-1ß in the absence and presence of 25(OH)D3 followed by an indirect co-culture with phytohemagglutinin-activated CD4+ T lymphocytes. The CD4+ T lymphocyte proliferation, viability, and cytokine secretion were analyzed. Additionally, the expression of various immunomediators in hPDL-MSCs was investigated, and their implication was verified by using pharmacological inhibitors. Results: 25(OH)D3 significantly counteracted the suppressive effects of IL-1ß-treated hPDL-MSCs on CD4+ T lymphocyte proliferation, whereas no effects were observed in the presence of TNF-α. Additionally, 25(OH)D3 significantly increased the percentage of viable CD4+ T lymphocytes via TNF-α- or IL-1ß-treated hPDL-MSCs. It also caused a significant decrease in interferon-γ, IL-17A, and transforming growth factor-ß productions, which were triggered by TNF-α-treated hPDL-MSCs. 25(OH)D3 significantly decreased the production of various immunomediators in hPDL-MSCs. Inhibition of two of them, prostaglandin E2 and indoleamine-2,3-dioxygenase-1, partially abolished some of the hPDL-MSCs-mediated effects of 25(OH)D3 on CD4+ T lymphocytes. Conclusion: These data indicate that 25(OH)D3 influences the immunomodulatory activities of hPDL-MSCs. This modulatory potential seems to have high plasticity depending on the local cytokine conditions and may be involved in regulating periodontal tissue inflammatory processes.

Effects of customized CAD/CAM abutments on cytokine levels in peri-implant crevicular fluid during early implant healing: a pilot study.

OBJECTIVES: This study aimed to assess levels of biomarkers associated with inflammation and tissue destruction in peri-implant crevicular fluid (PICF) of implants provided with customized or standard healing abutments during early implant healing. MATERIALS AND METHODS: Thirty implants were placed in 22 patients with partial posterior edentulism. Subsequently, test group implants (n=15) received one-piece titanium abutments that were fabricated using computer-aided design/computer-aided manufacturing (CAD/CAM). Control group implants (n=15) were provided with standard abutments. PICF collection and standardized periapical radiographs were carried out at suture removal one week later, following crown delivery after 3 months and at 6 months. Expression of C-reactive protein (CRP), interferon-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-1α, IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12A, IL-17A, macrophage inflammatory protein (MIP)-1α, matrix metalloproteinase (MMP)-13, osteopontin, osteoactivin, Receptor Activator of NF-κB (RANK), and TGF-ß were analyzed using a multiplex ELISA kit. RESULTS: Both groups showed a significant decrease in protein expression of CRP, IL-1ß, IL-6, IL-8, MIP-1α, osteopontin, osteoactivin, and TGF-ß, while MMP-13 levels increased during the observation period. A rise in OPG and RANK levels was detected among customized abutments. Expression of CRP was higher, whereas IL-1ß, IL-1α, and MIP-1α were decreased in control compared to test group implants after 6 months. Marginal bone loss did not depend on abutment modality. CONCLUSIONS: Both abutment types showed distinctive temporal expression of inflammatory biomarkers during 6 months following implant placement. TRIAL REGISTRATION: ISRCTN98477184, registration date 18/05/2022 CLINICAL RELEVANCE: Customized healing abutments exert similar effects on inflammation during early implant healing compared to standard healing abutments.

Evaluation of a novel moving threshold gating strategy for assessment of reticulated platelets in dogs using the ADVIA 2120 analyzer.

BACKGROUND: A novel method using a moving threshold (r-PLTmt) to determine canine r-PLTs (reticulated platelets) has been introduced for ADVIA 2120 software v6.11.7. OBJECTIVES: We aimed to evaluate absolute (ar-PLTmt) and percent (%r-PLTmt) prior to and after visual inspection of scattergrams (ar-PLTmtv, %rPLTmtv) compared with flow cytometry (flow) and to determine reference intervals (RIs) in 120 dogs. METHODS: For method comparison, 42 blood samples of healthy and thrombocytopenic dogs were included. Calculation of Spearman's rho, Bland-Altman, and Passing-Bablok analysis was performed. Coefficients of variation (CVs) were determined for three concentration levels. RESULTS: Moderate correlations between %r-PLTmt and %r-PLTmtv (rs 0.75-0.76) were seen compared with flow cytometry. The CV for medium %r-PLTs counts assessed with flow cytometry was 12.9%. Comparable CVs were obtained for ar-PLTmt (14.4%) and %r-PLTmt (15.7%), and ar-PLTmtv and %r-PLTmtv (10.9% and 12.9%, respectively). At low and high concentration levels, CVs for % and absolute r-PLTmt/rPLTmtv ranged between 23%-30% and 15%-20%. In patients with microcytic hypochromic erythrocytes, CVs for ar-PLTmt and %r-PLTmt were 36%-66%. Visual inspection of scattergrams resulted in a marked decrease in CV ranging between 15% and 20%. A proportional bias of 10.8% between %r-PLTmt and flow cytometry became lower (9.7%) after visual validation of scattergrams. Passing-Bablok analysis showed proportional and constant error. RIs for r-PLTmt and r-PLTmtv were 0.2%-3.8% and 0.6-10.2 × 109 /L and 0.3%-4.5% and 1.1-10.3 × 109 /L, respectively. Median values for %r-PLTmtv were higher in young adults (≤2 years) than in older dogs (P = 0.03). CONCLUSIONS: r-PLTmt and r-PLTmtv were moderately correlated with flow cytometry. Visual inspection of scattergrams is recommended.

[99m-Technetium-pertechnetate- and 99m-Technetium-sestamibi-scintigraphy for visualization of hypofunctioning thyroid tissue and staging in a dog with thyroid carcinoma]. / 99m-Technetium-Pertechnetat- und 99m-Technetium-Sestamibi-Szintigrafie zur Darstellung von hypofunktionellem Schilddrüsengewebe und Staging bei einem Hund mit Schilddrüsenkarzinom.

A 10-year-old female mixed breed dog was presented for thyroid scintigraphy due to a cervical mass. Apart from 99m-Technetium-pertechnetate (Tc-pertechnetate) scintigraphy, a second scintigraphy using 99m-Technetium sestamibi (Tc-MIBI) was performed because of additional hypothyroidism suspective for a "cold" nodule and as screening for metastases.Twenty minutes following intravenous injection of 38 MBq Tc-pertechnetate, a "hot" cervical as well as a "hot" intrathoracal nodule were seen with an uptake of 8.40 and 0.25 %, respectively. The second scintigraphy was performed 20 minutes after intravenous injection of 364 MBq Tc-MIBI and 70 minutes following the first. After subtraction of pertechnetate activity and decay correction, both nodules showed an uptake of 0.99 and 0.03 %. Additionally, both thyroid lobes were visible in the thyroid loge with a weak MIBI-uptake. For both lesions, the ratio Tc-uptake/Tc-MIBI-uptake was 8.48 and 8.33, respectively.Following the extirpation of the cervical mass, histopathology revealed atrophied healthy thyroid tissue almost completely displaced by a well-differentiated follicular thyroid carcinoma.This case report describes performance, utility and calculative correction of consecutive pertechnetate- and MIBI-scan, that enable a visualization of hypofunctional thyroid tissue.Therefore and because of their similar MIBI metabolic activity, both nodules were considered to be dystopic tissue/metastases so that this dog had to be classified as prognostically less favorable WHO stabe IV. Different from human patients, both scintigraphies should be performed shortly after another in dogs in order to avoid the necessity of a second anesthetic procedure. A reliable qualitative/visual evaluation of the MIBI-scan is therefore not possible, so that a quantitative assessment using the uptake after calculative correction of the pertechnetate activity is recommended.