Histopathological concordance between prostate biopsies and radical prostatectomy specimens-implications of transrectal and transperineal biopsy approaches.

BACKGROUND: This study aimed to evaluate the histopathological concordance rates between prostate biopsies and radical prostatectomy specimens according to the applied biopsy approach (transrectal or transperineal). METHODS: We studied patients who had been newly diagnosed with clinically significant prostate cancer and who underwent a radical prostatectomy between 2018 and 2022. Patients were included if they underwent a prebiopsy magnetic resonance imaging and if they had not been previously treated for prostate cancer. Histopathological grading on prostate biopsies was compared with that on radical prostatectomy specimens. Univariable and multivariable logistic regression analyses were performed to assess the effect of the applied biopsy approach on histopathological concordance. Additional analyses were performed to assess the effect of the applied biopsy approach on American Urological Association risk group migration, defined as any change in risk group after radical prostatectomy. RESULTS: In total, 1058 men were studied, of whom 49.3% (522/1058) and 50.7% (536/1058) underwent transrectal and transperineal prostate biopsies, respectively. Histopathological disconcordance was observed in 37.8% (400/1058) of men while American Urological Association risk group migration was observed in 30.2% (320/1058) of men. A transperineal biopsy approach was found to be independently associated with higher histopathological concordance rates (OR 1.33 [95% CI 1.01-1.75], p = 0.04) and less American Urological Association risk group migration (OR 0.70 [95% CI 0.52-0.93], p = 0.01). CONCLUSIONS: The use of a transperineal biopsy approach improved histopathological concordance rates compared to the use of a transrectal biopsy approach. A transperineal biopsy approach may provide more accurate risk stratification for clinical decision-making. Despite recent improvements, histopathologic concordance remains suboptimal and should be considered before initiating management.

Ultrastable, high-repetition-rate attosecond beamline for time-resolved XUV-IR coincidence spectroscopy.

The implementation of attosecond photoelectron-photoion coincidence spectroscopy for the investigation of atomic and molecular dynamics calls for a high-repetition-rate driving source combined with experimental setups characterized by excellent stability for data acquisition over time intervals ranging from a few hours up to a few days. This requirement is crucial for the investigation of processes characterized by low cross sections and for the characterization of fully differential photoelectron(s) and photoion(s) angular and energy distributions. We demonstrate that the implementation of industrial-grade lasers, combined with a careful design of the delay line implemented in the pump-probe setup, allows one to reach ultrastable experimental conditions leading to an error in the estimation of the time delays of only 12 as over an acquisition time of 6.5 h. This result opens up new possibilities for the investigation of attosecond dynamics in simple quantum systems.

Changes in the distribution of occlusal forces in the course of the orthodontic retention phase : A prospective cohort study.

PURPOSE: Aim of the present study was to assess the relative distribution of occlusal forces after orthodontic treatment and during the first 3 months of the retention phase using a computerized occlusal analysis system (T-Scan, Tekscan Inc., Norwood, MA, USA). MATERIALS AND METHODS: A total of 52 patients were included in this prospective cohort study and underwent analysis of occlusal forces on the level of tooth, jaw-half, and -quadrant during a 3-month period. Furthermore, differences between three retention protocols (group I: removable appliances in both jaws; group II: fixed 3-3 lingual retainers in both jaws; group III: removable appliance in the maxilla and fixed 3-3 lingual retainer in mandible) were assessed with Wilcoxon signed-rank tests at 5%. RESULTS: Directly after debonding, measured forces distribution were similar to published references for untreated samples. In the following, no significant difference was found between retention protocols II and III with regard to the asymmetry of the anterior occlusal forces. Both groups maintained an asymmetric force distribution in the anterior segment during the study period. There was also no difference between groups II and III in the distribution of occlusal forces for the posterior segments. Both retention concepts kept the symmetrical distribution of occlusal forces stable over the observation period. The retention concept of group I demonstrated a symmetrical distribution of occlusal forces in the anterior segment after debonding and this remained stable during the 3­month period. In the posterior segment, no improvement of the initially asymmetric masticatory force distribution could be observed. CONCLUSIONS: All three studied retention protocols showed stability in retaining their original symmetrical or asymmetrical occlusal force distribution posteriorly/anteriorly during the 3­month observation period. Therefore, an even distribution of occlusal forces should be the aim of the finishing phase, as no relative benefit of any single retention scheme in terms of post-debond improvement during the retention phase was seen.

Plasma ESR1 mutations and outcome to first-line paclitaxel and bevacizumab in patients with advanced ER-positive/HER2-negative breast cancer.

BACKGROUND: ESR1 mutations have been identified as mechanism for endocrine resistance and are also associated with a decreased overall survival. We assessed ESR1 mutations in circulating tumor DNA (ctDNA) for impact on outcome to taxane-based chemotherapy in advanced breast cancer patients. METHODS: ESR1 mutations were determined in archived plasma samples from patients treated with paclitaxel and bevacizumab (AT arm, N = 91) in the randomized phase II ATX study. Samples collected at baseline (n = 51) and at cycle 2 (n = 13, C2) were analyzed using a breast cancer next-generation sequencing panel. This study was powered to detect a benefit in progression-free survival (PFS) at six months for patients treated with paclitaxel/bevacizumab compared to historical trials with fulvestrant. PFS, overall survival (OS), and ctDNA dynamics were exploratory analyses. RESULTS: PFS at six months was 86% (18/21) in patients with an ESR1 mutation detected and 85% (23/27) in wildtype ESR1 patients. In our exploratory analysis, median progression-free survival (PFS) was 8.2 months [95% CI, 7.6-8.8] for ESR1 mutant patients versus 8.7 months [95% confidence interval (CI), 8.3-9.2] for ESR1 wildtype patients [p = 0.47]. The median overall survival (OS) was 20.7 months [95% CI, 6.6-33.7] for ESR1 mutant patients versus 28.1 months [95% confidence interval (CI), 19.3-36.9] for ESR1 wildtype patients [p = 0.27]. Patients with ≥ two ESR1 mutations had a significantly worse OS, but not PFS, compared to those who did not [p = 0.003]. Change in ctDNA level at C2 was not different between ESR1 and other mutations. CONCLUSIONS: Presence of ESR1 mutations in baseline ctDNA might not be associated with inferior PFS and OS in advanced breast cancer patients treated with paclitaxel/bevacizumab.

The effect of attendance in the Dutch breast cancer screening program on breast tumor characteristics among migrant women.

BACKGROUND: In general, migrant women have a lower breast cancer (BC) incidence rate and higher BC mortality than autochthonous women. Further, migrant women show lower participation in the national BC screening program. To further investigate those aspects, we aimed to determine differences in incidence and tumor characteristics between autochthonous and migrant BC patients in Rotterdam, the Netherlands. METHODS: We selected women diagnosed with BC in Rotterdam during 2012-2015 from the Netherlands Cancer Registry. Incidence rates were calculated by migrant status (i.e., women with or without migration background). Multivariable analyses revealed adjusted odds ratios (OR) and 95% confidence intervals (CI) on the association between migration status and patient and tumor characteristics, additionally stratified by screening attendance (yes/no). RESULTS: In total 1372 autochthonous and 450 migrant BC patients were included for analysis. BC incidence was lower among migrants than among autochthonous women. Overall, migrant women were younger at BC diagnosis (53 vs. 64 years, p < 0.001), and had higher risks of positive lymph nodes (OR 1.76, 95% CI 1.33-2.33) and high grade tumors (OR 1.35, 95% CI 1.04-1.75). Especially non-screened migrant women had higher risk of positive nodes (OR 2.73, 95% CI 1.43-5.21). Among the subgroup of screened women, we observed no significant differences between migrant and autochthonous patients. CONCLUSION: Migrant women have lower BC incidence than autochthonous women, but diagnosis was more often at younger age and with unfavorable tumor characteristics. Attending the screening program strongly reduces the latter. Therefore, promotion of participation in the screening program is recommended.

The impact of endoxifen-guided tamoxifen dose reductions on endocrine side-effects in patients with primary breast cancer.

BACKGROUND: Tamoxifen is important in the adjuvant treatment of hormone-sensitive breast cancer and substantially reduces recurrence; however, almost 50% of patients are non-compliant mainly due to side-effects. The aim of this study was to investigate whether endoxifen-guided tamoxifen dose reduction could lead to fewer side-effects. MATERIALS AND METHODS: Effects of tamoxifen dose reduction were investigated in patients with bothersome side-effects and endoxifen levels ≥32 nM and compared to patients with side-effects who remained on tamoxifen 20 mg. Endocrine symptoms and health-related quality of life (HR-QOL) were assessed after 3 months with the Functional Assessment of Cancer Therapy-Endocrine Symptoms (FACT-ES) questionnaire. RESULTS: Tamoxifen dose was reduced in 20 patients, 17 of whom were assessable for side-effect analyses. A clinically relevant improvement of >6 points was observed in endocrine symptoms and HR-QOL in 41% and 65% of the patients, respectively. In total, there was a significant and clinically relevant improvement in endocrine symptoms [5.7, 95% confidence interval (CI) -0.5-11.5] and HR-QOL (8.2, 95% CI 0.9-15.4) after dose reduction. This was not seen in patients whose doses were not reduced (n = 60). In 21% of patients, endoxifen dropped slightly below the 16-nM threshold (12.8, 15.5, 15.8, 15.9 nM). CONCLUSIONS: Endoxifen-guided dose reduction of tamoxifen significantly improved tamoxifen-related side-effects and HR-QOL. Nearly 80% of patients remained above the most conservative endoxifen threshold.

Overall survival in the OlympiA phase III trial of adjuvant olaparib in patients with germline pathogenic variants in BRCA1/2 and high-risk, early breast cancer.

BACKGROUND: The randomized, double-blind OlympiA trial compared 1 year of the oral poly(adenosine diphosphate-ribose) polymerase inhibitor, olaparib, to matching placebo as adjuvant therapy for patients with pathogenic or likely pathogenic variants in germline BRCA1 or BRCA2 (gBRCA1/2pv) and high-risk, human epidermal growth factor receptor 2-negative, early breast cancer (EBC). The first pre-specified interim analysis (IA) previously demonstrated statistically significant improvement in invasive disease-free survival (IDFS) and distant disease-free survival (DDFS). The olaparib group had fewer deaths than the placebo group, but the difference did not reach statistical significance for overall survival (OS). We now report the pre-specified second IA of OS with updates of IDFS, DDFS, and safety. PATIENTS AND METHODS: One thousand eight hundred and thirty-six patients were randomly assigned to olaparib or placebo following (neo)adjuvant chemotherapy, surgery, and radiation therapy if indicated. Endocrine therapy was given concurrently with study medication for hormone receptor-positive cancers. Statistical significance for OS at this IA required P < 0.015. RESULTS: With a median follow-up of 3.5 years, the second IA of OS demonstrated significant improvement in the olaparib group relative to the placebo group [hazard ratio 0.68; 98.5% confidence interval (CI) 0.47-0.97; P = 0.009]. Four-year OS was 89.8% in the olaparib group and 86.4% in the placebo group (Δ 3.4%, 95% CI -0.1% to 6.8%). Four-year IDFS for the olaparib group versus placebo group was 82.7% versus 75.4% (Δ 7.3%, 95% CI 3.0% to 11.5%) and 4-year DDFS was 86.5% versus 79.1% (Δ 7.4%, 95% CI 3.6% to 11.3%), respectively. Subset analyses for OS, IDFS, and DDFS demonstrated benefit across major subgroups. No new safety signals were identified including no new cases of acute myeloid leukemia or myelodysplastic syndrome. CONCLUSION: With 3.5 years of median follow-up, OlympiA demonstrates statistically significant improvement in OS with adjuvant olaparib compared with placebo for gBRCA1/2pv-associated EBC and maintained improvements in the previously reported, statistically significant endpoints of IDFS and DDFS with no new safety signals.

Prospective Evaluation of a Circulating Tumor Cell Sensitivity Profile to Predict Response to Cisplatin Chemotherapy in Metastatic Breast Cancer Patients.

BACKGROUND: Cisplatin (cDDP) has regained interest for metastatic breast cancer (MBC) patients, given the platinum sensitivity in subtypes and better manageable toxicity. Here, the primary aim was to determine whether molecular characteristics of circulating tumor cells (CTCs) could identify patients responding to cDDP and to describe the outcomes to cDDP monotherapy in a large group of MBC patients pretreated with anthracycline- and taxane-based treatments. METHODS: Based on cell line data, a CTC-cDDP-sensitivity profile was generated. Applying an A'Herns single-stage phase II design, further investigation was considered worthwhile if 5/10 patients with a favorable profile responded to cDDP. Patients received 70mg/m2 cDDP every three weeks, CTCs were enumerated and the CTC-cDDP-sensitivity profile was determined. In total, 65 heavily pretreated MBC patients (77% received ≥2 lines of previous chemotherapy for MBC) were eligible for the per-protocol analysis. Primary endpoint was response rate, secondary endpoints included best observed response, progression-free survival (PFS) and overall survival (OS). RESULTS: The best observed response during cDDP therapy was a partial response in 7% and stable disease in 56% of the patients. None of the patients with a favorable CTC-cDDP-sensitivity profile had a response. The median baseline CTC count was 8 (range 0-3254). Patients with <5 CTCs had a better PFS and OS than patients with ≥5 CTCs (median PFS 4.5 months (95%CI 2.38-6.62) vs. 2.1 months [(95%CI 1.34-2.80)(p=0.009)] and median OS 13.1 months (95%CI 9.89-16.33) vs. 5.6 months [(95%CI 3.60-7.64)(p=0.003)]. No other factors than CTC count were associated with outcome to cDDP therapy, including triple-negative breast cancer versus ER-positive tumors. CONCLUSIONS: The CTC-cDDP-sensitivity profile was unable to select patients responding to cDDP monotherapy. In an unselected group of heavily pretreated MBC patients, cDDP yields outcomes comparable to other chemotherapeutic regimens for heavily pretreated MBC patients. CTC count was the only factor associated with outcome in these patients. CLINICAL TRIAL REGISTRATION: (https://www.trialregister.nl/trial/3885, identifier NTR4046).

Cardiotoxicity during long-term trastuzumab use in patients with HER2-positive metastatic breast cancer: who needs cardiac monitoring?

PURPOSE: Patients with HER2-positive metastatic breast cancer (MBC) usually receive many years of trastuzumab treatment. It is unknown whether these patients require continuous left ventricular ejection fraction (LVEF) monitoring. We studied a real-world cohort to identify risk factors for cardiotoxicity to select patients in whom LVEF monitoring could be omitted. METHODS: We included patients with HER2-positive MBC who received > 1 cycle of trastuzumab-based therapy in eight Dutch hospitals between 2000 and 2014. Cardiotoxicity was defined as LVEF < 50% that declined > 10%-points and was categorized into non-severe cardiotoxicity (LVEF 40-50%) and severe cardiotoxicity (LVEF < 40%). Multivariable Cox and mixed model analyses were performed to identify risk factors associated with cardiotoxicity. Additionally, we explored the reversibility of cardiotoxicity in patients who continued trastuzumab. RESULTS: In total, 429 patients were included. Median follow-up for cardiotoxicity was 15 months (interquartile range 8-31 months). The yearly incidence of non-severe + severe cardiotoxicity in the first and second year was 11.7% and 9.1%, respectively, which decreased thereafter. The yearly incidence of severe cardiotoxicity was low (2.8%) and stable over time. In non-smoking patients with baseline LVEF > 60% and no cardiotoxicity during prior neoadjuvant/adjuvant treatment, the cumulative incidence of severe cardiotoxicity was 3.1% after 4 years of trastuzumab. Despite continuing trastuzumab, LVEF decline was reversible in 56% of patients with non-severe cardiotoxicity and in 33% with severe cardiotoxicity. CONCLUSIONS: Serial cardiac monitoring can be safely omitted in non-smoking patients with baseline LVEF > 60% and without cardiotoxicity during prior neoadjuvant/adjuvant treatment.

Increased tooth mobility after fixed orthodontic appliance treatment can be selectively utilized for case refinement via positioner therapy - a pilot study.

BACKGROUND: Increased tooth mobility persists after fixed orthodontic appliance removal, which is therapeutically utilized for post-treatment finishing with positioners. As such a fine adjustment is only required for selected teeth, the aim of this pilot study was to investigate tooth mobility in vivo on corrected and uncorrected subgroups under positioner therapy. METHODS: Mobility was measured on upper teeth of 10 patients (mean age 16.8) by applying loadings for 0.1, 1.0 and 10.0 s with a novel device directly after multibracket appliance debonding as much as 2d, 1, 2 and 6 weeks later. Positioners were inserted at day 2. Specimens were divided into Group C (teeth corrected via positioner), Group N (uncorrected teeth adjacent to teeth from group C), and Group U (uncorrected teeth in an anchorage block). Untreated individuals served as controls (n = 10, mean age 22.4). Statistics were performed via Kolmogorov-Smirnov test and Welch's unequal variances t-test for comparisons between groups. P < 0.05 was considered statistically significant. RESULTS: After 1 week, tooth mobility in Group U almost resembled controls (13.0-15.7 N), and reached physiological values after 6 weeks (17.4 N vs. 17.3 N in controls). Group C (9.0-13.4 N) and Group N (9.2-14.7 N) maintained increased mobility after 6 weeks. Tooth mobility was generally higher by reason of long loading durations (10.0 s). CONCLUSIONS: Positioner therapy can selectively utilized increased tooth mobility upon orthodontic fixed appliance treatment for case refinements. Here, uncorrected teeth in anchorage blocks are not entailed by unwanted side effects and recover after 6 weeks post treatment. Corrected teeth and their neighbors exhibit enhanced mobility even after 6 weeks, which represents a necessity for the proper correction of tooth position, and concurrently arouses the requirement for an adequate retention protocol.

A novel serum-free medium for the isolation, expansion and maintenance of stemness and tissue-specific markers of primary human periodontal ligament cells.

PURPOSE: Periodontal ligament (PDL) cell cultures are classically maintained in serum-containing media. However, unwanted side-effects of these conditions on cellular and molecular characteristics demand a serum-free alternative. Even though these limitations are well known and efforts for the development of adequate serum-free alternatives have been made, these approaches for replacement remained unsuccessful so far. This study aimed at developing a well-defined, serum-free formulation supporting both isolation from tissue samples and efficient expansion of PDL cells. Here, of particular focus was the perpetuation of tissue-characteristic markers detectable in primary tissues and of stemness features. BASIC PROCEDURES: Primary PDL cell cultures from generally healthy human donors (n = 3) were maintained in basal media N2B27 and E6 together with different concentrations of growth and attachment factors. Cell proliferation was recorded via microscopy and WST assay. Gene expression of RUNX2, Periostin, ALP, CD73, CD90, CD105, CD45, SOX10 and SOX2 was compared to primary PDL explants via qRT-PCR. Immunocytochemistry was performed for anti-CD105, SSEA-3, CD271, HNK1. Serum-containing sDMEM medium served as control. MAIN FINDINGS: N2B27 medium substituted with 25 ng/mL EGF, 25 ng/mL IGF1, 0.5 mg/mL Fetuin plus gelatine coating (designated N2B27-PDLsf) emerged as potent serum-free formulation ensuring adequate culture isolation and expansion. Here, PDL primary tissue signature markers RUNX2 and Periostin remained stable in N2B27-PDLsf compared to controls (229.0-fold ±101.0 and 83.2-fold ±9.6 increase). Additionally, stemness markers ALP and CD105 were significantly upregulated on transcriptional, and CD105 and SOX2 on protein level. PRINCIPAL CONCLUSIONS: This investigation identified a novel serum-free medium for the isolation, and expansion of primary human PDL cells with constantly high proliferation rates. Here, purity and stemness properties are maintained. Thus, N2B27-PDLsf represents a valid replacement for serum-containing media in PDL cultures.

Regulation of the autophagy-marker Sequestosome 1 in periodontal cells and tissues by biomechanical loading.

PURPOSE: Orthodontic treatment is based on the principle of force application to teeth and subsequently to the surrounding tissues and periodontal cells. Sequestosome 1 (SQSTM1) is a well-known marker for autophagy, which is an important cellular mechanism of adaptation to stress. The aim of this study was to analyze whether biomechanical loading conditions regulate SQSTM1 in periodontal cells and tissues, thereby providing further information on the role of autophagy in orthodontic tooth movement. METHODS: Periodontal ligament (PDL) fibroblasts were exposed to cyclic tensile strain of low magnitude (3%, CTSL), and the regulation of autophagy-associated targets was determined with an array-based approach. SQSTM1 was selected for further biomechanical loading experiments with dynamic and static tensile strain and assessed via real-time polymerase chain reaction (RT-PCR) and immunoblotting. Signaling pathways involved in SQSTM1 activation were analyzed by using specific inhibitors, including an autophagy inhibitor. Finally, SQSTM1 expression was analyzed in gingival biopsies and histological sections of rats in presence and absence of orthodontic forces. RESULTS: Multiple autophagy-associated targets were regulated by CTSL in PDL fibroblasts. All biomechanical loading conditions tested increased the SQSTM1 expression significantly. Stimulatory effects of CTSL on SQSTM1 expression were diminished by inhibition of the c­Jun N­terminal kinase (JNK) pathway and of autophagy. Increased SQSTM1 levels after CTSL were confirmed by immunoblotting. Orthodontic force application also led to significantly elevated SQTSM1 levels in the gingiva and PDL of treated animals as compared to control. CONCLUSIONS: Our in vitro and in vivo findings provide evidence of a role of SQSTM1 and thereby autophagy in orthodontic tooth movement.

Radiological complete remission in HER2-positive metastatic breast cancer patients: what to do with trastuzumab?

PURPOSE: Patients with HER2-positive metastatic breast cancer (MBC) treated with trastuzumab may experience durable tumor response for many years. It is unknown if patients with durable radiological complete remission (rCR) can discontinue trastuzumab. We analyzed clinical characteristics associated with rCR and overall survival (OS) in a historic cohort of patients with HER2-positive MBC and studied the effect of stopping trastuzumab in case of rCR. METHODS: We included patients with HER2-positive MBC treated with first or second-line trastuzumab-based therapy in eight Dutch hospitals between 2000 and 2014. Data were collected from medical records. We used multivariable regression models to identify independent prognostic factors for rCR and OS. Time-to-progression after achieving rCR for patients who continued and stopped trastuzumab, and breast cancer-specific survival were also evaluated. RESULTS: We identified 717 patients with a median age of 53 years at MBC diagnosis. The median follow-up was 109 months (IQR 72-148). The strongest factor associated with OS was achievement of rCR, adjusted hazard ratio 0.27 (95% CI 0.18-0.40). RCR was observed in 72 patients (10%). The ten-year OS estimate for patients who achieved rCR was 52 versus 7% for patients who did not achieve rCR. Thirty patients with rCR discontinued trastuzumab, of whom 20 (67%) are alive in ongoing remission after 78 months of median follow-up since rCR. Of forty patients (58%) who continued trastuzumab since rCR, 13 (33%) are in ongoing remission after 68 months of median follow-up. Median time-to-progression in the latter group was 14 months. CONCLUSIONS: Achieving rCR is the strongest predictor for improved survival in patients with HER2-positive MBC. Trastuzumab may be discontinued in selected patients with ongoing rCR. Further research is required to identify patients who have achieved rCR and in whom trastuzumab may safely be discontinued.

The stability of different housekeeping genes in human periodontal ligament cells under inflammatory conditions.

PURPOSE: Stability of housekeeping genes as internal reference for RT-qPCR analyses is mandatory for a correct interpretation of results. As no normalization benchmark exists and reference gene validation is highly specific for individual experiments, it was the purpose of this study to identify stable candidates for investigations on periodontal inflammation. BASIC PROCEDURES: Human PDL cells from one cell line (Lonza) and three primary donors were challenged with IL-1ß (5 ng/ml) or centrifugation (170 × g) for 6 h under serum-free conditions. Unstimulated cells represented controls. qRT-PCR was performed with a TaqMan® array of 32 housekeeping genes (n = 3). Transcriptional stability was analyzed for (i) mean absolute CT values and (ii) relative fold changes. Finally, stability of mean CT values across specimens was evaluated for most stable candidates. Statistics were performed with one-way ANOVA and Bonferroni correction and one sample t-test, with 95% confidence level. Values represent mean ± SEM. MAIN FINDINGS: 18S was constant in experimental groups and specimens for mean absolute CT values and relative fold changes, and MT-APT6 for mean absolute CT values. Both genes exhibited low CT thresholds ranging from 20.2 ± 0.1 to 25.9 ± 0.2 for 18S, and from 18.9 ± 0.0-23.7 ± 0.1 for MT-APT6. Likewise stable YWHAZ ranged between 32.6 ± 0.2 and 37.2 ± 0.2 cycles. However, candidates were unstable across specimes. PRINCIPAL CONCLUSIONS: Reference validation is mandatory for RT-qPCR analyses in new experimental designs. Here, only three genes out of 32 turned out to be appropriate candidates. Due to low CT values and stability, 18S and MT-APT6 are most valid genes for data normalization in experiments with PDL cells under inflammatory conditions and are recommended as standards under these premises.

Inhibitory control in BALB/c mice sub-strains during extinction learning.

Dysregulation of executive function (EF) involves alterations in cognitive flexibility / control and is underscored by learning impairments in neurodevelopmental disorders. Here, we examine cognitive inflexibility in BALB/cJ mice (a mouse model showing diminished sociability, increased anxiety and inattentive behaviour) and closely related "reference" BALB/cByJ mice. We used an appetitive extinction paradigm to investigate if cognitive flexibility measures are different between learning acquisition and extinction. The two BALB/c sub-strains learned to respond to a stimulus in a touchscreen operant chamber, after which the reward was removed and responses should be inhibited. Both mice sub-strains showed a different rate of learning while acquiring the task, in which the BALB/cJ mice were faster learners compared to the BALB/cByJ mice. This was not observed during the extinction phase, in which the BALB/cJ mice were able to extinguish responding to unrewarded stimuli equally. Within the BALB/cJ sub-strain, variation in the ability to inhibit a learnt response was observed when comparing them to similar grouped BALB/cByJ mice: BALB/cJ animals that reached the criterion were more reward driven, while BALB/cJ mice failing to reach the set criterion during extinction processing make more mistakes. Additionally, the changes observed during acquisition, were driven by animals not reaching the extinction criterion. Our results suggest that the BALB/c mice sub-strains may use different strategies to learn during appetitive extinction. This may be useful in the phenotypic dissection of cognitive flexibility in BALB/c sub-strains and their mapping on genetic variance revealed by next-generation sequencing in future studies.

Potential impact of lingual retainers on oral health: comparison between conventional twistflex retainers and CAD/CAM fabricated nitinol retainers : A clinical in vitro and in vivo investigation.

PURPOSE: To evaluate the impact of a novel computer-fabricated lingual nitinol retainer compared to a conventional lingual flexible spiral wire twistflex retainer on oral health. METHODS: The study was based on a retrospective controlled clinical study with pilot character, an in vitro investigation of material-dependent biofilm formation and an analysis of biofilm formation after intraoral incubation. Sixty-one patients with completed fixed orthodontic treatment and retention phase for at least 6 months with twistflex retainers (group 1, n = 31) or computer-aided design/computer-aided manufacturing (CAD/CAM) nitinol retainers (group 2, n = 30) were included and examined regarding plaque index (PI), gingival index (GI), probing depths, bleeding on probing (BOP) and marginal recessions (MR). Material-dependent biofilm formation of twistflex, untreated nitinol and electropolished nitinol wire samples were assessed (1) in vitro: using optical density (OD) measurement of 10 samples of each and (2) in vivo: using histomorphometric analysis of 18 samples of each. RESULTS: Patients treated with nitinol retainers had significant better oral health indices (PI1 = 1.29 ± 0.06, PI2 = 0.94 ± 0.06; GI1 = 0.71 ± 0.05, GI2 = 0.56 ± 0.04; BOP1 = 0.11 ± 0.01, BOP2 = 0.08 ± 0.01; PD1 = 1.79 ± 0.03 mm, PD2 = 1.59 ± 0.04 mm) except for MR (0.08 ± 0.03 mm versus 0.08 ± 0.02 mm) compared to twistflex retainers. After 24 h intraoral incubation nitinol retainers demonstrated significant less biofilm formation compared to twistflex retainers. In the in vitro investigation the temporary significant differences between the groups were compensated in the end. CONCLUSIONS: Based on the results it can be assumed that nitinol-made CAD/CAM developed lingual retainers have a positive effect on oral health.

Aggression in BALB/cJ mice is differentially predicted by the volumes of anterior and midcingulate cortex.

Anterior cingulate cortex (ACC) and midcingulate cortex (MCC) have been implicated in the regulation of aggressive behaviour. For instance, patients with conduct disorder (CD) show increased levels of aggression accompanied by changes in ACC and MCC volume. However, accounts of ACC/MCC changes in CD patients have been conflicting, likely due to the heterogeneity of the studied populations. Here, we address these discrepancies by studying volumetric changes of ACC/MCC in the BALB/cJ mouse, a model of aggression, compared to an age- and gender-matched control group of BALB/cByJ mice. We quantified aggression in BALB/cJ and BALB/cByJ mice using the resident-intruder test, and related this to volumetric measures of ACC/MCC based on Nissl-stained coronal brain slices of the same animals. We demonstrate that BALB/cJ behave consistently more aggressively (shorter attack latencies, more frequent attacks, anti-social biting) than the control group, while at the same time showing an increased volume of ACC and a decreased volume of MCC. Differences in ACC and MCC volume jointly predicted a high amount of variance in aggressive behaviour, while regression with only one predictor had a poor fit. This suggests that, beyond their individual contributions, the relationship between ACC and MCC plays an important role in regulating aggressive behaviour. Finally, we show the importance of switching from the classical rodent anatomical definition of ACC as cingulate area 2 and 1 to a definition that includes the MCC and is directly homologous to higher mammalian species: clear behaviour-related differences in ACC/MCC anatomy were only observed using the homologous definition.

Selecting the optimal position of CDK4/6 inhibitors in hormone receptor-positive advanced breast cancer - the SONIA study: study protocol for a randomized controlled trial.

BACKGROUND: Combining cyclin-dependent kinases 4 and 6 (CDK4/6) inhibitors with endocrine therapy is an effective strategy to improve progression-free survival in hormone receptor-positive (HR+), human epidermal growth factor receptor 2 (HER2)-negative advanced breast cancer. There is a lack of comparative data to help clinicians decide if CDK4/6 inhibitors can best be added to first- or second-line endocrine therapy. Improvement in median progression-free survival in first-line studies is larger than in second-line studies, but CDK4/6 inhibitors have not consistently shown to improve overall survival or quality of life. They do come with added toxicity and costs, and many patients have lasting disease remission on endocrine therapy alone. No subgroup has been identified to select patients who are most likely to benefit from the addition of CDK4/6 inhibition in any line of treatment. Altogether, these factors make that the optimal strategy for using CDK4/6 inhibitors in clinical practice is unknown. METHODS: The SONIA study is an investigator-initiated, multicenter, randomized phase III study in patients with HR+/HER2-negative advanced breast cancer. Patients are randomly assigned to receive either strategy A (first-line treatment with a non-steroidal aromatase inhibitor combined with CDK4/6 inhibition, followed on progression by fulvestrant) or strategy B (first-line treatment with a non-steroidal aromatase inhibitor, followed on progression by fulvestrant combined with CDK4/6 inhibition). The primary objective is to test whether strategy A is more effective than strategy B. The primary endpoint is time from randomization to second objective progression (PFS2). Secondary endpoints include overall survival, safety, quality of life, and cost-effectiveness. Five-hundred seventy-four events yield 89% power to show that strategy A has statistically significant, clinically meaningful superior PFS2 (according to ESMO-MCBS) in a log-rank test at the two-sided 95% confidence level. Given an accrual period of 42 months and an additional 18 months follow-up, inclusion of 1050 evaluable patients is required. DISCUSSION: This study design represents daily clinical practice, and the results will aid clinicians in deciding when adding CDK4/6 inhibitors to endocrine therapy will benefit their patients most. Additional biomarker analyses may help to optimize patient selection. TRIAL REGISTRATION: http://clinicaltrials.gov: NCT03425838 (8 February 2018). EudraCT-number: 2017-002334-23 (29 September 2017).

Three-dimensional ultrasonography of the breast; An adequate replacement for MRI in neoadjuvant chemotherapy tumour response evaluation? - RESPONDER trial.

BACKGROUND: Accurate measurement of tumour response during and after neoadjuvant chemotherapy (NAC) is important and may influence treatment decisions in invasive breast cancer patients. Breast MRI forms the gold standard but is more burdensome, time consuming and costly. In this study response measurement was done with 3-D ultrasound by Automated Breast Volume Scanner (ABVS) and compared to breast MRI. Moreover, patient satisfaction with both techniques was compared. METHODS AND MATERIALS: A single-institution, prospective observational pilot study evaluating tumour response by ABVS in addition to breast MRI (standard care) was performed in 25 invasive breast cancer patients receiving NAC. Tumour response was evaluated comparing longest tumour diameters as well as tumour volumes at predefined time points using Bland-Altman analysis. Volume measurements for breast MRI were obtained using a fully immersive virtual reality system (a Barco I-Space) and V-Scope software. Same software was used to obtain ABVS volume measurements using an in-house developed desktop VR system. Inter- and intra-observer agreement was evaluated by Intraclass Correlation Coefficient (ICC). RESULTS: Twenty-five patients were eligible for baseline measurement, 20 for a mid-NAC response evaluation, and five for a post-NAC response evaluation. MRI and ABVS showed absolute concordance in 73% of patients for the mid-NAC evaluation, with a 'good' correlation for the difference in longest diameter measurement (ICC 0.73, p < 0.01) as compared to baseline assessment. Concerning difference in volume measurement in the mid-NAC response evaluation showed a 'fair' correlation (ICC 0.52, p < 0.01) and in the post-NAC response evaluation an 'excellent' correlation (ICC 0.98, p < 0.01). 'Excellent' inter- and intra-observer agreement was found (ICC 0.88, p < 0.01) with comparable limits of agreement (LOA) for observer 1 and 2 in both diameter and volume measurement. Patient satisfaction was higher for ABVS compared to breast MRI, 93% versus 12% respectively. CONCLUSION: ABVS showed 'good' correlation with MRI tumour response evaluation in breast cancer patients during NAC with 'excellent' inter- and intra-observer agreement. ABVS has patients' preference over breast MRI and could be considered as alternative to breast MRI, in case results on an on-going prospective trial confirm these results (NTR6799).