Lymph node metastasis of squamous cell carcinoma from an unknown primary in the upper and middle neck: impact of 18F-fluorodeoxyglucose positron emission tomography/computed tomography

Purpose: The purpose of this study was to assess the potential of F-18-fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) imaging for detection of the primary tumor and its impact on treatment planning in patients presenting with cancer of unknown primary and squamous cell carcinoma (SCC)-positive cervical lymph nodes of the upper and middle neck. Methods: The study population consisted of 18 consecutive patients with biopsy-proven SCC involving lymph nodes of the upper and middle neck region and negative conventional diagnostic procedures with regard to the location of the primary. All patients underwent FDG-PET/CT according to a standard procedure in search for the primary, unidentified tumor. Results: In none of the patients FDG-PET/CT was able to indicate a primary tumor localization. Although FDGPET/CT did identify all sites of known lymph node involvement, neither additional sites of lymph node involvement nor sites of distant metastases were identified. Accordingly, FDG-PET/CT did not impact patient treatment planning. Conclusions: In this series, including patients suffering from lymph node metastases by an SCC of unknown primary in the upper and middle neck, FDG-PET/CT was unable to identify a primary tumor. In addition, FDGPET/CT did not modify the treatment planning in any of the patients studied

LC-MS characterization and cell-binding properties of chelate modified somatropin

Somatropin, a recombinant protein containing 191 amino acids, is derived from the endogenous human growth hormone, somatotropin. This protein is clinically used in children and adults with inadequate endogenous growth hormone to stimulate a normal bone and muscle growth. In addition, somatropin is recently being investigated for the diagnosis and radiotherapy of certain hormonal cancers. In some of these cancers, over-expression of the human growth hormone receptor (hGHR) is described. The modification of the protein with a chelating agent like NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) allows the inclusion of metals coupled to the protein. The NOTA unit is selectively introduced on a lysine side chain. As site-specific labelling is necessary to avoid active region interactions (1-16, 41-68, 103-119 and 167-175), characterization of the chelate-modified somatropin is indispensable. Therefore, we have applied an enzymatic digestion procedure using trypsin, chymotrypsin and a combination of both enzymes. The resulting peptides were then monitored using HPLC-MSn, allowing the investigation of the exact amino acid modifications. The use of a mixture of trypsin and chymotrypsin gave an enhanced information efficiency. Moreover, the intact protein, without enzymatic degradation, was analysed on a protein HPLC column using UV detection for quantification and ESI-MS/MS for characterization. Based upon the HPLC-MSn results of the digested somatropin, the chelating molecule is mainly bound to a specific lysine amino acid that is located away from the receptor binding site. Therefore, the cell-binding functionality of the characterized NOTA-somatropin is measured, using a HepG2 cell line

In vitro 2-deoxy-2-[18F]fluoro-D-glucose uptake: practical considerations

In oncology 2-deoxy-2-[F-18]fluoro-D-glucose ([F-18]-FDG), a glucose analogue, is the most used positron emission tomography (PET) tracer. There are however some limitations due to low metabolic activity or high surrounding physiological uptake in several tumors or regions. Investigating new tracers or methods is expensive and elaborative when animal experiments or phase I clinical trials are used. In vitro experiments can overcome these limitations. We analyzed the influence of incubation time, cell medium conditions, administered activity, and cell density on [F-18]-FDG uptake in six different cell cultures. Glucose transporter 1 (GLUT1)- and hexokinase 2 (HK2)-expression at high and low cell density was analyzed using immunocytochemistry. FDG-uptake increases over time and absence of glucose in the incubation medium increases uptake. By increasing the administered activity, uptake per protein also increases and tracer uptake per protein is lower at higher cell densities. Immunocytochemical analysis reveals a lower expression of both GLUT1 and HK2 at higher cell concentrations. All investigated parameters influenced FDG uptake and therefore we can conclude it is of utmost importance to keep administered activity, incubation medium, and time constant and to correct uptake when cell density changes due to environmental conditions, such as therapy

Ranking soccer teams on the basis of their current strength : a comparison of maximum likelihood approaches

We present 10 different strength-based statistical models that we use to model soccer match outcomes with the aim of producing a new ranking. The models are of four main types: Thurstone-Mosteller, Bradley-Terry, independent Poisson and bivariate Poisson, and their common aspect is that the parameters are estimated via weighted maximum likelihood, the weights being a match importance factor and a time depreciation factor giving less weight to matches that are played a long time ago. Since our goal is to build a ranking reflecting the teams' current strengths, we compare the 10 models on the basis of their predictive performance via the Rank Probability Score at the level of both domestic leagues and national teams. We find that the best models are the bivariate and independent Poisson models. We then illustrate the versatility and usefulness of our new rankings by means of three examples where the existing rankings fail to provide enough information or lead to peculiar results

HPLC-MS characterisation of chelate modified somatropin

Somatropin is a recombinant human growth hormone, consisting of 191 amino acids. This protein is clinically used in children and adults with inadequate endogenous growth hormone to stimulate a normal bone and muscle growth. In addition, somatropin is currently being investigated for the diagnosis and radiotherapy of certain hormonal cancers. The modification of the protein with the chelating agent NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) allows the inclusion of metals coupled to the protein for diagnostic (e.g. 68Ga) or therapeutic (e.g. 90Y) purposes. The NOTA unit is selectively introduced on a lysine side chain. This yields 9 possible labelling sites for somatropin: FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYSFLQNPQTSLCFSESIPTPSNREETQQKSNLELLRISLLLIQSWLEPVQFLRSVFANSLVYGASDSNVYDLLKDLEEGIQTLMGRLEDGSPRTGQIFKQTYSKFDTNSHNDDALLKNYGLLYCFRKDMDKVETFLRIVQCRSVEGSCGF We have applied an enzymatic digestion procedure for the characterisation of the modified somatropin, using trypsin, chymotrypsin and Staphylococcus aureus V-8 proteases. The resulting peptides were then monitored using HPLC-MS2, allowing the characterisation of the modified protein

Dual time-point FDG PET/CT for differentiating benign from malignant solitary pulmonary nodules in a TB endemic area

Objective. Fluorodeoxyglucose (FDG)-positron emission tomography (PET) is an accurate non-invasive imaging test for differentiating benign from malignant solitary pulmonary nodules (SPNs). We aimed to assess its diagnostic accuracy for differentiating benign from malignant SPNs in a tuberculosis (TB)-endemic area. Methods. Thirty patients, 22 men and 8 women, mean age 60 years, underwent dual time point FDG-PET/computed tomography (CT) imaging, followed by histological examination of the SPN. Maximum standard uptake values (SUVmax) with the greatest uptake in the lesion were calculated for two time points (SUV1 and SUV2), and the percentage change over time per lesion was calculated (%DSUV). Routine histological findings served as the gold standard. Results. Histological examination showed that 14 lesions were malignant and 16 benign, 12 of which were TB. SUVmax for benign and malignant lesions were 11.02 (standard deviation (SD) 6.6) v. 10.86 (SD 8.9); however, when tuberculomas were excluded from the analysis, a significant difference in mean SUV1max values between benign and malignant lesions was observed (p=0.0059). Using an SUVmax cut-off value of 2.5, a sensitivity of 85.7% and a specificity of 25% was obtained. Omitting the TB patients from analysis resulted in a sensitivity of 85.7% and a specificity of 100%. Mean %DSUV of benign lesions did not differ significantly from mean %DSUV of malignant lesions (17.1% (SD 16.3%) v. 19.4% (SD 23.7%)). Using a cut-off of %DSUV >10% as indicative of malignancy, a sensitivity of 85.7% and a specificity of 50% was obtained. Omitting the TB patients from the analysis yielded a sensitivity of 85.7% and a specificity of 75%. Conclusion. Our findings suggest that FDG-PET cannot distinguish malignancy from tuberculoma and therefore cannot reliably be used to reduce futile biopsy/thoracotomy