To assess the proportion of clinically significant (cs) prostate cancer (PCa) found during follow‐up in patients with negative systematic biopsy (SB) followed by non‐suspicious multiparametric magnetic resonance imaging (mpMRI) and persistent clinical suspicion of PCa compared to the general population.

BACKGROUND:The role of targeted prostate biopsies (TBs) in patients with cancer suspicious lesions on multiparametric magnetic resonance imaging (mpMRI) following negative systematic biopsies (SBs) is undebated. However, whether they should be combined with repeated SBs remains unclear. OBJECTIVE:To evaluate the value of repeated SBs in addition to TBs in patients with a prior negative SB and a persistent suspicion of prostate cancer (PCa). DESIGN, SETTING, AND PARTICIPANTS:A prospective study as part of a multicenter randomized controlled trial conducted between 2014 and 2017, including 665 men with a prior negative SB and a persistent suspicion of PCa (suspicious digital rectal examination and/or prostate-specific antigen >4.0ng/ml). INTERVENTION:All patients underwent 3T mpMRI according to Prostate Imaging Reporting and Data System (PI-RADS) v2. Patients with PI-RADS ≥3 were randomized 1:1:1 for three TB techniques: MRI-TRUS fusion TB (FUS-TB), cognitive registration fusion TB (COG-TB), or in-bore MRI TB. FUS-TB and COG-TB were combined with repeated SBs. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS:Clinically significant prostate cancer (csPCa) was defined as Gleason ≥3+4. Differences in detection rates of csPCa, clinically insignificant PCa (cisPCa), and overall PCa between TBs (FUS-TB and COG-TB) and repeated SBs were compared using McNemar's test. RESULTS AND LIMITATIONS:In the 152 patients who underwent both TB and SB, PCa was detected by TB in 47% and by SB in 32% (p<0.001, 95% confidence interval [CI]: 6.0-22%). TB detected significantly more csPCa than SB (32% vs 16%; p<0.001, 95% CI: 11-25%). Clinically significant PCa was missed by TB in 1.3% (2/152). Combining SB and TB resulted in detection rate differences of 6.0% for PCa, 5.0% for cisPCa, and 1.0% for csPCa compared with TB alone. CONCLUSIONS:In case of a persistent suspicion of PCa following a negative SB, TB detected significantly more csPCa cases than SB. The additional value of SB was limited, and only 1.3% of csPCa would have been missed when SB had been omitted. PATIENT SUMMARY:We evaluated the role of systematic biopsies and magnetic resonance imaging (MRI)-targeted biopsies for the diagnosis of prostate cancer in patients with prior negative systematic biopsies. MRI-targeted biopsies perform better in detecting prostate cancer in these patients. The value of repeated systematic biopsies is limited.

De FUTURE-trial is een multicenter gerandomiseerde studie naar drie technieken van MRI-geleide target biopten (TB’s) bij patiënten na negatieve systematische biopten (SB’s) en blijvende verdenking op prostaatkanker. Tussen 2014 en 2017 ondergingen 665 patiënten een multiparametrische MRI. 234 (35 %) patiënten met een PIRADS-laesie ≥3 werden 1:1:1 gerandomiseerd voor TB: in-bore MRI (MRI-TB), MRI/TRUS-fusie (FUS-TB), en cognitieve fusie (COG-TB). Na FUS-TB en COG-TB werden tevens SB’s afgenomen. Tussen de drie technieken waren geen significante verschillen in detectie van prostaatkanker en significante prostaatkanker (MRI-TB 54,5 %/32,5 %, FUS-TB 49,4 %/34,2 %, en COG-TB 43,6 %/33,3 %, p = 0,39/p = 0,98). De combinatie van SB’s en TB’s versus enkel TB’s zorgde voor 1 % meer detectie van significante prostaatkanker (35 % versus 34 %). Er was geen significant voordeel voor één TB-techniek voor het detecteren van (klinisch significante) prostaatkanker. De toegevoegde waarde van een SB was beperkt.

BackgroundMycobacterium tilburgii is an opportunistic pathogen that has only been described 11 times in the literature. Hyperammonemia as a resulting symptom of a mycobacterial infection has only been reported once. We describe a patient with a disseminated M. tilburgii infection, leading to hyperammonemia. Case Presentation A 57-year-old man was referred with stupor, rapidly declining to coma. Hyperammonemia was found as the underlying cause. Ammonia-lowering interventions had an overall disappointing effect. Frequent causes of hyperammonemia were excluded. Finally, a disseminated opportunistic M. tilburgii infection was diagnosed by using 16sRNA sequencing. A combination of antimicrobial drugs was started, after which ammonia level declined and consciousness improved. Unfortunately, it failed to eradicate the infection. The patient died od pneumonia and multiorgan failure. Conclusions Hyperammonemia requires an urgent response to prevent cerebral damage. M. tilburgii is not cultivable and diagnosis is performed using 16S RNA sequencing. Long-term antimicrobial drugs are required for eradication.

You have accessJournal of UrologyProstate Cancer: Detection & Screening II1 Apr 2018PD23-02 THE FUTURE TRIAL; A MULTICENTER RCT ON THREE TECHNIQUES OF MRI TARGETED PROSTATE BIOPSY. Olivier Wegelin, Leonie Exterkate, Rik Somford, Jelle Barentsz, Marloes van der Leest, Alain Kummer, Willem Vreuls, Peter de Bruin, Ruud Bosch, and Harm van Melick Olivier WegelinOlivier Wegelin More articles by this author , Leonie ExterkateLeonie Exterkate More articles by this author , Rik SomfordRik Somford More articles by this author , Jelle BarentszJelle Barentsz More articles by this author , Marloes van der LeestMarloes van der Leest More articles by this author , Alain KummerAlain Kummer More articles by this author , Willem VreulsWillem Vreuls More articles by this author , Peter de BruinPeter de Bruin More articles by this author , Ruud BoschRuud Bosch More articles by this author , and Harm van MelickHarm van Melick More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1183AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Guidelines advise multiparametric MRI (mpMRI) in patients with prior negative prostate biopsies and a persisting suspicion of prostate cancer (PCa), enabling target biopsy of MRI suspicious lesions. Three techniques of MRI targeted biopsy (MRI-TB) are available and no consensus exists on which technique should be preferred. The FUTURE trial is a multicenter RCT comparing detection rates of (significant) PCa for three techniques of MRI-TB (cognitive TRUS, MR-TRUS fusion, in-bore MRI). METHODS Ethical board approval was granted for this study and all patients provided written informed consent. A total of 642 men were recruited between 2014 and 2017 with prior negative prostate biopsies and a suspicion on prostate cancer (based on PSA≥4 and/or abnormal DRE), and underwent mpMRI. Imaging was centrally evaluated by an expert radiologist using PIRADS v2. If mpMRI demonstrated PIRADS≥3 lesions patients were randomized 1:1:1 for MRI-TB. Overall PCa and significant PCa (Gleason≥7) detection rates were compared using Pearson Chi square test. RESULTS 231 patients (36.0%) had a PIRADS≥3 lesion on mpMRI, and 223 underwent MRI-TB. There were no significant differences in baseline characteristics (see table 1), nor were PIRADS scores significantly different between the three groups (p=0.87). Using cognitive TRUS biopsy PCa was detected in 44.0%, and significant PCa in 33.3%. Using MR-TRUS fusion biopsy PCa was detected in 49.3%, and significant PCa in 33.3%. Using in-bore MRI biopsy PCa was detected in 56.2%, and significant PCa in 34.2%. The detection rates of the three MRI-TB techniques were neither significantly different for overall PCa detection (p=0.33) nor for significant PCa (p=0.99). CONCLUSIONS Based on the results of this multicenter RCT there does not seem to be a significant advantage of one specific MRI-TB technique for the detection of (clinically significant) PCa following prior negative prostate biopsies. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e480-e481 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Olivier Wegelin More articles by this author Leonie Exterkate More articles by this author Rik Somford More articles by this author Jelle Barentsz More articles by this author Marloes van der Leest More articles by this author Alain Kummer More articles by this author Willem Vreuls More articles by this author Peter de Bruin More articles by this author Ruud Bosch More articles by this author Harm van Melick More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Differentiation between multiple primary lung cancers and pulmonary metastases (PM) has important implications in staging, prognosis, and treatment strategies. Clinical and immunohistopathologic criteria have been standardized; however, a substantial number of cases remain difficult to classify. Using next-generation sequencing, it is now possible to improve the classification of multiple lung cancer lesions. This study systematically investigated the value of routine morphologic and IHC characteristics, p53 protein expression, TP53 mutation analysis, and 50-gene panel sequencing (GPS) in 111 lesions from 50 patients with multiple lung lesions. Based on immunohistopathologic criteria, 32 paired lesions were classified as multiple primary lung cancer (MPLC) and 21 as PM. TP53 mutation analysis indicated MPLC in 23 and PM in 6 pairs, but in the majority of cases (n = 28, 49%) no mutation was observed and no conclusion could be drawn. In contrast, only 2 pairs were not conclusive using GPS. In a significant number of matching tumor samples (n = 19, 39%), sequencing results were contradictory to the initial immunohistopathology diagnosis. No separation in overall survival for classifications based on immunohistopathology was observed, while a clear but nonsignificant trend was observed concerning survival in MPLC patients (hazard ratio = 3.98) using 50-gene GPS. In about one-third of the patients, GPS provided additional information to improve the differentiation between MPLC and PM.

You have accessJournal of UrologyProstate Cancer: Detection & Screening II1 Apr 2018PD23-03 THE FUTURE TRIAL; A RCT ON MRI TARGETED PROSTATE BIOPSY. COMPARISON OF TARGETED AND SYSTEMATIC BIOPSY OUTCOMES. Leonie Exterkate, Olivier Wegelin, Harm van Melick, Jelle Barentsz, Marloes van der Leest, Alain Kummer, Willem Vreuls, Peter de Bruin, Ruud Bosch, and Rik Somford Leonie ExterkateLeonie Exterkate More articles by this author , Olivier WegelinOlivier Wegelin More articles by this author , Harm van MelickHarm van Melick More articles by this author , Jelle BarentszJelle Barentsz More articles by this author , Marloes van der LeestMarloes van der Leest More articles by this author , Alain KummerAlain Kummer More articles by this author , Willem VreulsWillem Vreuls More articles by this author , Peter de BruinPeter de Bruin More articles by this author , Ruud BoschRuud Bosch More articles by this author , and Rik SomfordRik Somford More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1184AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Guidelines advise multiparametric MRI (mpMRI) in patients with prior negative systematic biopsies (SB) and a persisting suspicion of prostate cancer (PCa), enabling targeted biopsy (TB) if tumour suspicious lesions are present. In addition to TB repeated SB can be performed. In this analysis of the FUTURE trial the detection rates of (significant) PCa using TB are compared with the detection rates using SB. METHODS Ethical board approval was granted for this study and all patients provided written informed consent. A total of 642 men were recruited between 2014 and 2017 with prior negative prostate biopsies and a suspicion of prostate cancer (based on PSA ≥4 and/or abnormal DRE) and underwent mpMRI. Imaging was centrally evaluated by an expert radiologist using PIRADS v2. If mpMRI demonstrated PIRADS ≥3 lesions, patients were randomized 1:1:1 for TB (cognitive TRUS, MR-TRUS fusion, in-bore MRI). In both the cognitive TRUS and MR-TRUS fusion groups, additional SB was performed. TB was performed initially, followed by SB using a standardized template irrespective of the location of MRI lesions. Overall PCa and significant PCa (Gleason ≥7) detection rates were compared using McNemar test. RESULTS 231 patients (36.0%) had a PIRADS ≥3 lesion on mpMRI, and 223 underwent MRI-TB. Of these, 143 underwent both TB and SB. See table 1 for baseline characteristics. Using TB PCa was detected in 48% (n=68) and significant PCa in 34% (n=48). Using SB PCa was detected in 34% (n=48) and significant PCa in 16% (n=23). See table 2 for a cross tabulation of biopsy outcomes. TB detected significantly more PCa (p=0.002) as well as significantly more significant PCa (p=0.000) than SB. In patients in whom SB did not detect PCa, TB detected PCa in 21% (n=30) and significant PCa in 18.9% (n=27). Alternatively, in patients in whom TB did not detect PCa, SB detected PCa in 7% (n=10) and significant PCa in 1.4% (n=2). SB detected insignificant PCa in 5.6% (n=8) of patients in which TB detected no cancer. CONCLUSIONS Targeted biopsy has significantly increased detection rates for overall and significant PCa compared to systematic biopsy in subjects with prior negative prostate biopsies and a persisting suspicion of PCa. Few significant PCa were missed when systematic biopsies would have been omitted, while less insignificant PCa would have been detected. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e481 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Leonie Exterkate More articles by this author Olivier Wegelin More articles by this author Harm van Melick More articles by this author Jelle Barentsz More articles by this author Marloes van der Leest More articles by this author Alain Kummer More articles by this author Willem Vreuls More articles by this author Peter de Bruin More articles by this author Ruud Bosch More articles by this author Rik Somford More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Alexander JJ Smits1,2, Paul Roepman1, Niels JM Claessens 3, Franz M Schramel4 and J Alain Kummer 1*

Aims Molecular PCR-based clonality analysis is helpful for identification of monoclonal B-cell or T-cell populations and to distinguish malignant lymphoma from reactive lymphoid hyperplasia. Typically, clonality assessment on fine-needle aspiration cytology (FNAC) requires freshly obtained aspirates, but the collection and processing of these samples are often challenging in daily practice. In this study, we assessed the routine diagnostic value of the EuroClonality/BIOMED-2 assay for B-cell clonality on air-dried archived Giemsa-stained smears. Methods This study comprised a retrospective analysis of a consecutive diagnostic cohort of 192 FNAC samples from 184 patients with at least 2-year follow-up. The results from the clonality assay were integrated with cytomorphological assessment and evaluated for their accuracy in detecting malignant disease. EuroClonality expert re-evaluation was performed for all cases with ambiguous results and for cases in which the diagnosis did not match the follow-up data. Results The clonality assay showed a high accuracy of 93% for detection of malignancy, with a sensitivity of 93% and a specificity of 92%. All 64 cases with monoclonal Ig heavy chain (IGH)/Ig kappa chain (IGK) rearrangements were confirmed malignant by histology or clinical follow-up. Expert re-evaluation changed the definite diagnosis for five cases (3%), mainly because of low signals or no proper duplicate results. We discuss and elucidate all cases for which the clonality results did not match the disease follow-up. Conclusions This study showed that EuroClonality/BIOMED-2 assay can successfully be performed on cytological Giemsa-stained smears and inclusion in daily practice can assist in better identification of malignant lymphoma.

With the use of more specific treatments and targeted therapies for non–small cell lung carcinoma, distinction between adenocarcinoma (ADC) or squamous cell carcinoma (SCC) becomes increasingly important. For this, the key technique is an immunohistochemical panel in which a thyroid transcription factor-1 (TTF-1) antibody is often used. Two different TTF-1 clones (8G7G3/1 and SPT24) are used in daily practice, which appear to have different sensitivities and specificities. The aim of this study was to assess the differences between these clones and to identify the optimal cutoff value for correctly diagnosing primary or metastatic lung ADC. 182 pulmonary (109 lung ADCs, 62 lung SCCs, 11 lung metastases) and 115 extrapulmonary (36 metastatic lung ADCs, 79 nonpulmonary tumors) samples were stained with both TTF-1 antibodies. The percentage of tumor cells with nuclear staining was scored in categories of <1%, 1% to 5%, 6% to 25%, 26% to 50%, 51% to 75%, and 76% to 100%. The staining was further assessed as weak or strong. The sensitivity and specificity were calculated at different cutoff values. Applying the same cutoff value for positivity to both clones resulted in a significant difference between the clones at all cutoff values, with a lower sensitivity of 8G7G3/1 at high cutoff values and a lower specificity of SPT24 at low cutoff values. However, when the optimal cutoff value was used for each clone (>5% staining for 8G7G3/1 and >50% strong staining for SPT24), no significant difference in sensitivity (0.79 vs. 0.82) or specificity (0.98 vs. 0.98) was detected, making the clones equally useful for reliably diagnosing lung ADC.

UNLABELLED:Histology is an important outcome variable in basic science and pre-clinical studies regarding intervertebral disc degeneration (IVD). Nevertheless, an adequately validated histological classification for IVD degeneration is still lacking and the existing classifications are difficult to use for inexperienced observers. OBJECTIVE:Therefore the aim of this study was to develop and to validate a new histological classification for IVD degeneration. Moreover, the new classification was compared to the frequently used non-validated classification. METHODS:The new classification was applied to human IVD sections. The sections were scored twice by two independent inexperienced observers, twice by two experienced IVD researchers and once by a pathologist. For comparison, the sections were also scored according to the classification described by Boos et al. by two experienced IVD researchers. Macroscopic grading according Thompson et al., glycosaminoglycan (GAG) content and age were used for validation. RESULTS:The new classification had an excellent intra- and a good inter-observer reliability. Intraclass Correlation Coefficients (ICC) were 0.83 and 0.74, respectively. Intra- and inter-observer reliability were comparable for experienced and inexperienced observers. Statistically significant correlations were found between the new classification, macroscopic score, GAG content in the nucleus pulposus (NP) and age; Correlation coefficient (CC) 0.79, -0.62 and 0.68, respectively. The CCs of the Boos classification were all lower compared to the new classification. CONCLUSION:the new histological classification for IVD degeneration is a valid instrument for evaluating IVD degeneration in human IVD sections and is suitable for inexperienced and experienced researchers.

Molecular pathology is becoming more and more important in present day pathology. A major challenge for any molecular test is its ability to reliably detect mutations in samples consisting of mixtures of tumor cells and normal cells, especially when the tumor content is low. The minimum percentage of tumor cells required to detect genetic abnormalities is a major variable. Information on tumor cell percentage is essential for a correct interpretation of the result. In daily practice, the percentage of tumor cells is estimated by pathologists on hematoxylin and eosin (H&E)-stained slides, the reliability of which has been questioned. This study aimed to determine the reliability of estimated tumor cell percentages in tissue samples by pathologists. On 47 H&E-stained slides of lung tumors a tumor area was marked. The percentage of tumor cells within this area was estimated independently by nine pathologists, using categories of 0-5%, 6-10%, 11-20%, 21-30%, and so on, until 91-100%. As gold standard, the percentage of tumor cells was counted manually. On average, the range between the lowest and the highest estimate per sample was 6.3 categories. In 33% of estimates, the deviation from the gold standard was at least three categories. The mean absolute deviation was 2.0 categories (range between observers 1.5-3.1 categories). There was a significant difference between the observers (P<0.001). If 20% of tumor cells were considered the lower limit to detect a mutation, samples with an insufficient tumor cell percentage (<20%) would have been estimated to contain enough tumor cells in 27/72 (38%) observations, possibly causing false negative results. In conclusion, estimates of tumor cell percentages on H&E-stained slides are not accurate, which could result in misinterpretation of test results. Reliability could possibly be improved by using a training set with feedback.

Background: In lung cancer minimally invasive staging of the mediastinum with endobronchial ultrasonography with transbronchial needle aspiration (EBUS-TBNA) has become an important alternative to the gold standard of mediastinoscopy. Aims: First: To determine the diagnostic yield of EBUS-TBNA and calculate the reduction in number of mediastinoscopies that can be achieved when this technique is used as initial modality for mediastinal staging in lung cancer. Second: Calculate the reduction in health care costs when EBUS-TBNA is used in this setting. Methods: In a retrospective cohort study all patients in our hospital in whom EBUS-TBNA was performed for mediastinal staging in lung cancer from September 2008 until January 2011 were identified and the results of EBUS-TBNA were analysed. If metastatic tumour cells were found there was no indication for additional mediastinoscopy. Diagnostic yield of EBUS-TBNA and the number of mediastinoscopies that were avoided were calculated, as well as the achieved cost reduction. Results: EBUS-TBNA was performed on 77 patients for mediastinal staging: 47 male and 30 female, average age 62.1 years (extremes 39-81). In 51% of patients (39/77) mediastinal lymph node metastasis were found and mediastinoscopy could be avoided. Sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy were 91%, 100%, 100%, 80% and 93% respectively. The achieved cost reduction was € 321 per patient (31%). Conclusion: Mediastinoscopy can be avoided in more that 50% of lung cancer patients when EBUS-TBNA is used as initial staging modality for mediastinal staging, leading to a significant reduction of health care costs.

Background Frequencies of EGFR and KRAS mutations in non-small cell lung cancer (NSCLC) have predominantly been determined in East Asian and North American populations, showing large differences between these populations. The aim of the present study was to determine the frequency of EGFR and KRAS mutations in NSCLC in the West European Dutch population in primary carcinomas and different metastatic locations.Methods EGFR (exons 19, 20 and 21) and KRAS (exons 2 and 3) mutation test results of NSCLC samples of patients in 13 hospitals were collected. The tests were performed on paraffin-embedded tissue or cytological material of primary and metastatic lung carcinomas.Results EGFR mutations were detected in 71/778 (9.1 %) tested patients; in 66/620 (10.6 %) adenocarcinomas. EGFR mutations were significantly more often detected in female than in male patients (13.4 % vs. 5.5 %, p<0.001). KRAS mutations were found in 277 out of 832 (33.3 %) tested patients; in 244/662 (36.9 %) adenocarcinomas. A significantly increased frequency of EGFR mutations was observed in patients with malignant pleural/pericardial effusions (26.5 %; odds ratio (OR) 2.80, 95 % confidence interval (CI) 1.22-6.41), whereas the frequency of KRAS mutations was significantly decreased (18.8 %; OR 0.35, 95 % CI 0.14-0.86).Conclusions In the investigated Dutch cohort, patients with malignant pleural/pericardial effusion of lung adenocarcinoma have an increased frequency of EGFR mutations. The overall frequency of EGFR mutations in lung adenocarcinomas in this West European population is within the frequency range of North American and South European populations, whereas KRAS mutation frequency is higher than in any population described to date.

We aimed to determine the role of HPV in the pathogenesis and outcome of oropharyngeal squamous cell carcinoma (OSCC) in lifelong nonsmoking and nondrinking patients. A case-case analysis was performed to compare the presence of HPV-DNA in tumor cells of 16 nonsmoking and nondrinking with 16 matched smoking and drinking patients (matching criteria: age at incidence, gender, tumor sublocation, tumor stage). HPV was detected using 2 PCR tests, FISH analysis, and p16(INK4A) immunostaining. Nonsmoking and nondrinking patients had more HPV-positive tumors than smoking and drinking patients (n = 12; 75% versus n = 2; 13%; P < 0.001). All HPV-positive tumors showed p16(INK4A) overexpression, and 1 HPV-negative tumor had p16(INK4A) overexpression, (P < 0.001). Overall survival and disease-specific survival were higher for HPV-positive compared to HPV-negative cases (P = 0.027, P = 0.039, resp.). In conclusion, HPV is strongly associated with OSCC of nonsmoking and nondrinking patients. Specific diagnostic and therapeutic actions should be considered for these patients to achieve a better prognosis.

PurposeCurrent assessment of lymph node metastasis in patients with head and neck squamous cell carcinoma is not accurate enough to prevent overtreatment. The aim of this study was validation of a gene expression signature for distinguishing metastasizing (N+) from nonmetastasizing (N0) squamous cell carcinoma of the oral cavity (OSCC) and oropharynx (OPSCC) in a large multicenter cohort, using a diagnostic DNA microarray in a Clinical Laboratory Improvement Amendments/International Organization for Standardization-approved laboratory.MethodsA multigene signature, previously reported as predictive for the presence of lymph node metastases in OSCC and OPSCC, was first re-evaluated and trained on 94 samples using generic, whole-genome, DNA microarrays. Signature genes were then transferred to a dedicated diagnostic microarray using the same technology platform. Additional samples (n = 222) were collected from all head and neck oncologic centers in the Netherlands and analyzed with the diagnostic microarray. Human papillomavirus status was determined by real-time quantitative polymerase chain reaction.ResultsThe negative predictive value (NPV) of the diagnostic signature on the entire validation cohort (n = 222) was 72%. The signature performed well on the most relevant subset of early-stage (cT1-T2N0) OSCC (n = 101), with an NPV of 89%.ConclusionCombining current clinical assessment with the expression signature would decrease the rate of undetected nodal metastases from 28% to 11% in early-stage OSCC. This should be sufficient to enable clinicians to refrain from elective neck treatment. A new clinical decision model that incorporates the expression signature is therefore proposed for testing in a prospective study, which could substantially improve treatment for this group of patients.

CD36 is the receptor for long chain fatty acids (LCFA), and is expressed in lingual taste cells from rodents. In these animals, CD36 has been proposed to play an important role in oral detection of LCFA, and subsequently, determines their dietary fat preference. Humans also seem to detect LCFA in the oral cavity, however, information on the molecular mechanism of this human orosensory LCFA recognition is currently lacking. The aim of our study was to investigate whether CD36 is also expressed in lingual human and porcine taste buds cells. Using fluorescence immunohistochemistry, apical CD36 expression was revealed in human and porcine taste bud cells from circumvallate and foliate papillae. These data suggest CD36 as the putative orosensory receptor for dietary LCFA in human, and, therefore, may be involved in our preference for fatty foods.

Background: The intervertebral disc (IVD) is dependent on nutrient provision through a cartilage layer with underlying subchondral bone, analogous to joint cartilage. In the joint, subchondral bone remodeling has been associated with osteoarthritis (OA) progression due to compromised nutrient and gas diffusion and reduced structural support of the overlaying cartilage. However, subchondral bone changes in IVD degeneration have never been quantified before.Objective: The aim of this study is to determine the subchondral bone changes at different stages of IVD degeneration by micro-CT.Methods: Twenty-seven IVDs including the adjacent vertebral endplates were obtained at autopsy. Midsagittal slices, graded according the Thompson score, were scanned. Per scan 12 standardized cylindrical volumes of interest (VOI) were selected. Six VOIs contained the bony endplate and trabeculae (endplate VOIs) and six accompanying VOIs only contained trabecular bone (vertebral VOIs). Bone volume as percentage of the total volume (BV/TV) of the VOI, trabecular thickness (TrTh) and connectivity density (CD) were determined.Results: An increase in BV/TV and TrTh was found in endplate VOIs of IVDs with higher Thompson score whereas these values remained stable or decreased in the vertebral VOIs.Conclusion: The increase in bone volume combined with the increase in TrTh in endplate VOIs strongly suggest that the subchondral endplate condenses to a more dense structure in degenerated IVDs. This may negatively influence the diffusion and nutrition of the IVD. The endplate differences between intact and mild degenerative IVDs (grade II) indicate an early association of subchondral endplate changes with IVD degeneration. (C) 2010 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Granzyme-mediated cell death is the major pathway for cytotoxic lymphocytes to kill virus-infected and tumor cells. In humans, five different granzymes (i.e. GrA, GrB, GrH, GrK, and GrM) are known that all induce cell death. Expression of intracellular serine protease inhibitors (serpins) is one of the mechanisms by which tumor cells evade cytotoxic lymphocyte-mediated killing. Intracellular expression of SERPINB9 by tumor cells renders them resistant to GrB-induced apoptosis. In contrast to GrB, however, no physiological intracellular inhibitors are known for the other four human granzymes. In the present study, we show that SERPINB4 formed a typical serpin-protease SDS-stable complex with both recombinant and native human GrM. Mutation of the P2-P1-P1' triplet in the SERPINB4 reactive center loop completely abolished complex formation with GrM and N-terminal sequencing revealed that GrM cleaves SERPINB4 after P1-Leu. SERPINB4 inhibited GrM activity with a stoichiometry of inhibition of 1.6 and an apparent second order rate constant of 1.3×10(4) M(-1) s(-1). SERPINB4 abolished cleavage of the macromolecular GrM substrates α-tubulin and nucleophosmin. Overexpression of SERPINB4 in tumor cells inhibited recombinant GrM-induced as well as NK cell-mediated cell death and this inhibition depended on the reactive center loop of the serpin. As SERPINB4 is highly expressed by squamous cell carcinomas, our results may represent a novel mechanism by which these tumor cells evade cytotoxic lymphocyte-induced GrM-mediated cell death.