Diagnosis Of Malignant Disease Research Articles

General procedures in diagnosis of malignant diseases in dogs and cats

Tumors occur in all domestic and wild animals. They are most often diagnosed in dogs and cats, and their numbers increase from year to year. In the recent years, cancer is believed to be the most frequent cause of pet deaths. A speedy and reliable diagnosis is of paramount importance because it enables the veterinarian to begin therapy and make a prognosis. The objective of an early diagnosis is to enable the detection of neoplastic diseases before the tumor spreads throughout the organism, consequently enabling the timely administration of therapy and providing greater chances for curing the animal. A tumor is diagnosed on the grounds of the anamnesis, clinical picture, and special diagnostic procedures. The most frequently applied diagnostic procedures are laboratory diagnostics, cytology, biopsy and pathohistology, imaging diagnostics (roentgenography and roentgenoscopy, ultrasound diagnostics, endoscopy, computer tomography, magnetic resonance, and scintigraphy) and molecular diagnostics. Each of these methods has its advantages and faults in connection with costs, availability, sensitivity, specificity and quality of anatomic vs functional pictures. Every one of these techniques has its own field of implementation and each one provides different and additional information in connection with the nature and position of the primary lesion and the presence of metastases.

Congenital Bronchial Atresia: Diagnosis and Treatment

This study aimed to retrospectively summarize the clinical signs, diagnosis, and treatment of congenital bronchial atresia (CBA) in 12 patients. Chest radiographs and computed tomographic (CT) images of 12 patients with CBA treated in the Chinese People's Liberation Army General Hospital were reviewed. Analysis of chest radiographs revealed ten patients had hilar mass-like shadows and two had pneumonia-like shadows; most patients (n = 8) showed hyperlucency of the peripheral lung fields. CT revealed a mucocele in all the patients (n = 12); the mucoceles were round in four patients and club-like in eight. In 80% of the cases (n = 10), associated anomalies, including occlusions of the bronchus central to the mucocele, emphysematous changes of the peripheral lung fields, bronchogenic cyst, and anomalous branching of the bronchial tree and vascular structure were observed. CBA was detected in the right lobe in eight patients and the left lobe in the remaining four. No surgical intervention was performed in 5 CBA patients and the remaining 7 patients underwent surgery, including lobectomy in 5 patients and local resection in 2 patients. Among these 7 patients, 3 had a preoperative diagnosis of malignant disease, and the remaining 4 had severe clinical symptoms that could not be effectively treated by medicines. All patients were followed up, and none experienced obvious discomfort. CBA is a relatively rare and benign malformation disease. Chest CT is the procedure of choice for diagnosis. The presence of a bronchocele and surrounding emphysematous changes are typical radiologic findings in CBA. Surgery should be reserved only for patients with serious complications secondary to the atretic bronchus.

Functional protease profiling for diagnosis of malignant disease

Clinical proteomic profiling by mass spectrometry (MS) aims at uncovering specific alterations within mass profiles of clinical specimens that are of diagnostic value for the detection and classification of various diseases including cancer. However, despite substantial progress in the field, the clinical proteomic profiling approaches have not matured into routine diagnostic applications so far. Their limitations are mainly related to high-abundance proteins and their complex processing by a multitude of endogenous proteases thus making rigorous standardization difficult. MS is biased towards the detection of low-molecular-weight peptides. Specifically, in serum specimens, the particular fragments of proteolytically degraded proteins are amenable to MS analysis. Proteases are known to be involved in tumour progression and tumour-specific proteases are released into the blood stream presumably as a result of invasive progression and metastasis. Thus, the determination of protease activity in clinical specimens from patients with malignant disease can offer diagnostic and also therapeutic options. The identification of specific substrates for tumour proteases in complex biological samples is challenging, but proteomic screens for proteases/substrate interactions are currently experiencing impressive progress. Such proteomic screens include peptide-based libraries, differential isotope labelling in combination with MS, quantitative degradomic analysis of proteolytically generated neo-N-termini, monitoring the degradation of exogenous reporter peptides with MS, and activity-based protein profiling. In the present article, we summarize and discuss the current status of proteomic techniques to identify tumour-specific protease-substrate interactions for functional protease profiling. Thereby, we focus on the potential diagnostic use of the respective approaches.

Diagnostische Bronchoskopie

The flexible bronchoscope is the instrument of choice for visualization of the trachea, and even peripheral, smaller airways. The majority of examinations can be carried out under sedation and local anesthesia. Bronchoscopy under general anesthesia, mostly performed with the rigid bronchoscope, is reserved for special situations and indications. In addition to inspection, bronchoscopy allows to gather materials with different techniques. Forceps biopsy supplemented by brushes, needles and various other techniques is the most common procedure. The removal of secretions as well as washing or lavage techniques play an additional role. This allows the diagnosis of malignant diseases, especially lung cancer and other thoracic malignomas as well as infectious and interstitial lung diseases. Bronchoscopy has a high sensitivity in chest diseases and also some extrathoracic illnesses, if these involve the chest. Bronchoscopy is a safe procedure and has a low complication rate as long as indications and contraindications are strictly followed, and if carried out by a trained physician.

Association of active pulmonary tuberculosis and malignant diseases: a clinical study

The association of pulmonary tuberculosis and bronchogenic carcinoma has been reported by many authors, however, there are rather few studies about the association of pulmonary tuberculosis and other malignant diseases and how the latter affects the outcome of the former. Between 1980 and 1993, we had in our hospital 104 patients who had both active pulmonary tuberculosis and malignant diseases. Pulmonary tuberculosis was diagnosed at the time or after the diagnosis of malignant diseases in 74 patients, of whom 92% (68 patients) were males and 42% (31 patients) were over the age of 70. There were 23 stomach cancer (31%), and 15 lung cancer (20%). In 11 patients tuberculosis developed after the initiation of radiation and/or chemotherapy. 67 patients could be followed up for more than 6 months after the initiation of chemotherapy for tuberculosis and the negative conversion rate was as high as 95.5% at 3 months. The fact indicates that the association of malignant diseases does not influence the course of tuberculosis and that these patients could be treated safely in general hospitals, provided the diagnosis is made properly without unnecessary delay.

Substrate optimization and clinical validation of reporter peptides for MS-based protease profiling in serum specimens: A new approach for diagnosis of malignant disease

The progression of many solid tumors is characterized by the release of tumor-associated proteases, such as cancer procoagulant, MMP2 and MMP7. Consequently, the detection of tumor-specific proteolytic activity in serum specimens has recently been proposed as a new diagnostic tool in oncology. However, tumor-associated proteases are highly diluted in serum specimens and it is challenging to identify substrates that are specifically cleaved. In this study, we describe the systematic optimization of a synthetic peptide substrate using a positional scanning synthetic combinatorial library (PS-SCL) approach. The initial reporter peptide (RP) comprises of the cleavage site, WKPYDAAD, that is part of the coagulation factor X, the natural substrate of the tumor-associated cysteine protease cancer procoagulant (EC 3.4.22.26). Specifically, the amino acid substitution of aspartatic acid (D) in position P1' against asparagine (N) improved the processing of respective RPs in serum specimens from patients with colorectal tumors compared to healthy controls. Proteolytic fragments of RPs accumulated during prolonged incubation with serum specimens and were quantified with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Finally, the optimized RP with the cleaved motif WKPYNAAD was combined with the RPs, VPLSLTMG and IPVSLRSG, that were cleaved by the tumor-associated proteases, MMP2 and MMP7, respectively. The diagnostic accuracy of MS-based protease profiling was evaluated for this triplex RP mix in a cohort of 50 serum specimens equally divided into colorectal cancer patients and healthy control individuals. Multiparametric analysis showed an AUC value of 0.90 for the receiver operating characteristic curve and was superior to the classification accuracy of the single markers. Our results demonstrate that RPs for MS-based protease profiling can systematically be optimized with a PS-SCL. Furthermore, the combination of different RPs can additionally increase the classification accuracy of functional protease profiling, and this in turn could lead to improved diagnosis, monitoring and prognosis of malignant disease.

HLA-G Expression in Cancers: Potential Role in Diagnosis, Prognosis and Therapy

Human leukocyte antigen G (HLA-G) is a non-classic major histocompatibility complex (MHC) class I molecule that functions as an immune suppressive molecule. HLA-G has direct inhibitory effects on natural killer cells(NK), dendritic cells (DC), T cells and has long-term tolerogenic indirect effects by inducing regulator T cells (Treg). HLA-G has been reported to be involved in various physio-pathological conditions such as reproduction, transplantation, autoimmunity, infectious and malignant diseases. In this context, aberrant expression of HLA-G in malignant diseases including hematological and solid tumors has been extensively investigated and its relevance to clinicoparameters and potential significance in diagnosis, prognosis and immune target therapy has been postulated. We here summarized the HLA-G expression in malignancies and emphasis its clinical relevance to malignant disease diagnosis, prognosis, and its potential in target-based immunotherapy was also discussed.

Obstructive Jaundice- Aetiological Spectrum, Clinical, Biochemical And Radiological Evaluation At A Tertiary Care Teaching Hospital.

Objective–This study was done to assess the aetiological spectrum of obstructive jaundice as well as common clinical findings and relevance of laboratory and radiological imaging investigations.Methods: A prospective study of 110 patients with obstructive jaundice confirmed by thorough history and physical examination, biochemical tests and radiological investigations was done with final confirmation of diagnosis on histopathology. The data was analyzed using SOFA analytical software ver 0.9.24.Results: Out of 110 patients 62 (56%) were male and 48 (44%) were female. The mean age of the study population was 50.4 years (range 3–85 years). Malignant obstructive was more common than benign (62.73% Vs 47.27%). Abdominal pain and clay coloured stools were more frequent in patients with malignant disease. Carcinoma (Ca) of the head of pancreas was commonest aetiology 37/110 (33.63%) followed by Choledocholithiasis 32/110 (29%), Ca gall bladder 20/110 (18.18%), periampullary carcinoma 6/110 (5.45%), cholangiocarcinoma 4/110 (3.64%), CBD stricture 3/110 (2.73%), acute pancreatitis 3/110 (2.73%) and choledochal cyst 3/110(2.73%). and HCC(1.8%). Regarding etiology of the obstruction, the accuracy of ultrasound, CT scan, MRCP and ERCP was 87.3%, 92.7%, 90% and 100%, respectively. The sensitivities of USG, CT, MRCP and ERCP in the diagnosis of benign disease were 85.3%, 84.6%, 92.3% and 100%, respectively, whereas specificities were 88.4%, 94.2%, 86% and 100%, respectively. Sensitivities for diagnosis of malignant disease were 88.4 %, 94.2 %, 86 % and 100% for USG, CT, MRCP and ERCP respectively whereas specificities were 85.3%, 85%, 92% and 100% respectively.Conclusion: Malignant obstructive jaundice is predominant in males compared to females. Benign obstruction is seen at a comparatively younger age group compared to malignant (Mean age 38.6yrsVs 58.7yrs). Carcinoma of head of pancreas and choledocholithiais were the commonest malignant and benign etiology respectively. Ultrasound and CT have high diagnostic accuracy, sensitivities and specificities and along with MRCP have largely confined the role of invasive cholangiography (ERCP/PTC) to therapeutic/palliative procedures in biliary obstruction.

Abstract 3997: Introducing the GPDE: A biological proteomics database to identify biomarker candidates

Abstract The discovery of novel biomarkers is essential for the early diagnosis of malignant diseases as well as for patient stratification to improve oncological therapies. Nevertheless, most biomarkers discovered so far have not satisfied the hopes laid upon them. Following a novel strategy we have started to systematically investigate different types of cells in different functional states to create a “protein-atlas”. We believe that only if we know the “normal” proteome of a cell at different functional states we will be able to efficiently deduct specific pathological changes. The associated proteome alterations may thus qualify as biomarkers. Based on this cell centered approach we created a novel proteomics database: the Griss Proteomics Database (GPDE). The GPDE is free, open-source software available at http://gpde.sourceforge.net. The GPDE is based on a standardized data format for proteomics experiments, the Proteomics Idenitification Database (PRIDE) data format, and thus allows the integration of proteomics experiments from different research groups. Here we present a new set of tools that was specifically developed to support the identification of biomarker candidates. The main feature of these new tools is the automated interpretation of any given protein list with respect to cells already represented in the GDPE. To ensure correct comparison results protein accessions are converted to the IPI accession system, updated to the currently suggested accession and cleared of duplicate entries caused by redundant accessions. In addition, biological knowledge is integrated into the GPDE and presented to the user to aid the interpretation of the results. With this tool it is, e.g., possible to analyze the proteome profile of hepatocytes isolated from a hepatocellular carcinoma (HCC) against other types of hepatocytes represented in the GPDE. By this comparison to multiple references the function now returns three main conclusions: (1) A quality assessment regarding reference protein profiles of this cell type. A wide coverage of proteins known to be present in the investigated cell type is an indicator for the reliability of the experiment. (2) Identification of specifically expressed proteins implies the presence of functional cell states in the investigated sample. Thus, functional states such as proliferation, occurrence of apoptosis, inflammatory activation or establishment of drug resistance can immediately be identified. (3) The most likely and plausible biomarkers are promptly highlighted by the automated identification of atypical, unexpected alterations which may be specific for the investigated disease. With the GPDE any researcher gets the possibility to quickly and easily assess, interpret and further use huge amounts of complex proteomics data. Thus the often experienced gap between the abstract results of basic research and their biological relevance is maybe a little bit closed. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3997.

Risk Factors and Consequences of Incidental Parathyroidectomy during Thyroidectomy

The objectives of this retrospective study were to investigate the incidence, risk factors, and clinical significance of incidental parathyroidectomy during thyroidectomy. In this study, there were 315 patients who underwent thyroidectomy between 1996 and 2006. All the operations were performed by a single experienced surgeon (J.S.). Pathology reports on all specimens were reviewed and information regarding patient demographics, diagnosis, operative details, and postoperative complications were collected. A total of 315 thyroidectomies were performed: 101 total thyroidectomies (32.1%) and 214 subtotal thyroidectomies (67.9%). Two hundred and eighty patients were operated on for benign disease (88.9%) and 35 for malignant disease (11.1%). Incidental parathyroidectomy was identified in 32 cases (10.2%). Preoperative diagnosis of malignant disease (P = 0.009) and duration of the disease (P = 0.001) were significant predictors of incidental parathyroidectomy. Incidental parathyroidectomy was significantly correlated with postoperative hypoparathyroidism (P = 0.03). Transient postoperative hypoparathyroidism occurred in 18 cases (6%) and permanent in three cases (1%). The duration of the disease was the sole significant predictor of postoperative hypoparathyroidism (P < 0.001). Incidental parathyroidectomy seems to be the result of the use of extensive surgical procedures. The preoperative diagnosis of malignant disease and the delay of the operation, which leads to a more advanced thyroid disease, make the use of extensive surgical procedures necessary.

Sialyl Lewis X-i (SLX) in the bronchoalveolar lavage fluid from patients with lung cancer

The measurement of serum SLX is thought to be a useful aid in the diagnosis of malignant diseases, particularly adenocarcinoma of the lung. In the present investigation, we measured and compared SLX values in BALF from affected and normal bronchi, obtained from 83 patients. They consisted of 64 males and 19 females, with mean age of 60 years, consisting of 8 normal controls, 19 cases of benign lung disease, and 56 cases of primary lung cancer. SLX value in BALF from normal bronchi was significantly higher in patients with lung cancer than in normal controls, but there was no significant difference in SLX value between lung cancer and benign lung disease. On the other hand, SLX value from affected bronchi was significantly higher in patients with lung cancer than in normal controls and patients with benign lung disease. The rate of elevated SLX in BALF from affected bronchi was significantly higher in patients with lung cancer than in those with benign lung disease. These results suggest that measurement of SLX levels in BALF from affected bronchi may be a useful method for differential diagnosis of primary lung cancer.

Endoprotease Profiling with Double-Tagged Peptide Substrates: A New Diagnostic Approach in Oncology

The measurement of disease-related proteolytic activity in complex biological matrices like serum is of emerging interest to improve the diagnosis of malignant diseases. We developed a mass spectrometry (MS)-based functional proteomic profiling approach that tracks degradation of artificial endoprotease substrates in serum specimens. The synthetic reporter peptides that are cleaved by tumor-associated endopeptidases were systematically optimized with regard to flanking affinity tags, linkers, and stabilizing elements. Serum specimens were incubated with reporter peptides under standardized conditions and the peptides subsequently extracted with affinity chromatography before MS. In a pilot study an optimized reporter peptide with the cleavage motif WKPYDAADL was added to serum specimens from colorectal tumor patients (n = 50) and healthy controls (n = 50). This reporter peptide comprised a known cleavage site for the cysteine-endopeptidase "cancer procoagulant." Serial affinity chromatography using biotin- and 6xHis tags was superior to the single affinity enrichment using only 6xHis tags. Furthermore, protease-resistant stop elements ensured signal accumulation after prolonged incubation. In contrast, signals from reporter peptides without stop elements vanished completely after prolonged incubation owing to their total degradation. Reporter-peptide spiking showed good reproducibility, and the difference in proteolytic activity between serum specimens from cancer patients and controls was highly significant (P < 0.001). The introduction of a few structural key elements (affinity tags, linkers, d-amino acids) into synthetic reporter peptides increases the diagnostic sensitivity for MS-based protease profiling of serum specimens. This new approach might lead to functional MS-based protease profiling for improved disease classification.

Circulating tumor cells (CTCs) and endothelial cells (CECs) in the diagnosis of malignant pleural mesothelioma (MPM): A single institutional prospective study

e22107 Background: Circulating tumor cell (CTC), a surrogate of distant metastasis, and circulating endothelia cell (CEC), a surrogate of angiogenesis, are potentially useful in the diagnosis of malignant tumors. Following a promising result of our preliminary study showing the diagnostic value of CTC/CEC in malignant pleural mesothelioma (MPM)(Tanaka F, et al. ASCO 2008), we conducted a prospective study. Methods: Patients (pts), who presented at our institute with suspicion or diagnosis of MPM, were eligible in the study. CTCs and CECs in peripheral blood (7.5mL and 4.0mL, respectively) were captured and quantitatively evaluated with the “CellSearch” system without knowledge of clinical characteristics of patients. Results: A total of 92 pts were enrolled into the study, and the final diagnosis was MPM in 68, other malignant tumors in 7, and non-malignant diseases in 17 pts ( Table ). CTC was positive (CTC-count, 1 or more per 7.5mL of the peripheral blood) in 35%(24/68) of MPM pts (range of CTC-count, 0–27 cells/7.5 mL). Among non-malignant pts, 3 pts (18%) showed a positive-CTC, but no patient showed 2 or more CTCs in 7.5mL of the peripheral blood. The mean CEC-count (/4.0mL) was significantly higher in MPM pts than in non-malignant pts (105.1 versus 40.2; p=0.047). When the cut-off value of CEC-count for the diagnosis of MPM was defined as 50(cells/4.0mL), the sensitivity, specificity, positive predictive value, and negative predictive value for the diagnosis of malignant diseases were 66%, 70%, 93%, and 26%, respectively. There was no correlation between CTC-positivity and clinical stage of MPM pts, but was a trend of increase in the mean CEC-count along with tumor progression (mean CEC-count for stage I, II, III, and IV pts: 63.0, 82.4, 95.6, and 116.7, respectively). Conclusions: CTC and CEC are useful clinical markers in the diagnosis of MPM. [Table: see text] No significant financial relationships to disclose.

178. Microrna Profiling in Pheochromocytoma

Pheochromocytomas are neuroendocrine tumors of chromaffin cell origin, arising from the adrenal medulla and less commonly from extra-adrenal sympathetic paraganglia within the abdomen, thorax and neck. In approximately 10% of cases pheochromocytoma will be malignant. The definitive diagnosis of malignant disease is often only made in retrospect once metastasis has occurred as there are currently no reliable prognostic markers to predict whether a tumor will be malignant. Malignant pheochromocytoma responds poorly to chemotherapy and radiotherapy and therefore patients with malignant pheochromocytoma generally have a poor outcome with an overall survival of 50% at five years.

Risk Factors for Febrile Morbidity After Abdominal Hysterectomy in a University Hospital in Thailand

Febrile morbidity is the most commonly reported adverse event after hysterectomy. Its incidence ranges from 9.1 to 37.4%. Risk factors reported in the literature include prolonged operative time, history of previous surgery, higher parity, greater blood loss, abdominal approach, and no antibiotic prophylaxis. Antibiotic prophylaxis in abdominal hysterectomy may provide protection against febrile morbidity and reduce its incidence, but does not eliminate it. Limited data are available evaluating risk factors for febrile morbidity after hysterectomy. This historical cohort study assessed the incidence of febrile morbidity and associated risk factors in 1980 Thai women, who had an abdominal hysterectomy between 1998 and 2005. Of the 1980 women, febrile morbidity occurred in 517. This represents an overall incidence of 26.1%. Univariate analysis of potential risk factors showed that the incidence of febrile morbidity was higher in patients with lower preoperative hematocrit, more extensive surgery, longer operative time, greater intraoperative blood loss and malignant disease. Median intraoperative blood loss was lower in the nonfebrile group than the febrile group (400 versus 500 ml, P ≤ .0005) and median operative time was shorter (135 versus 150 minute, P ≤ .0005). When both pre and postoperative variables were included in the logistic regression model, the only two risk factors independently associated with febrile morbidity were intraoperative blood loss of ≥750 ml (odds ratio [OR],1.52; 95% confidence interval [CI], 1.08–2.13; P = .036) and a diagnosis of malignant disease (OR 1.86; 95% CI 1.45–2.13, P < .0005).

Molecular basis of tumor formation. Significance for the diagnosis of malignant diseases

Molecular basis of tumor formation. Significance for the diagnosis of malignant diseases

Risk Factors for Febrile Morbidity after Abdominal Hysterectomy in a University Hospital in Thailand

Aim: To identify the risk factors for febrile morbidity after hysterectomy. Methods: This was a historical cohort study of 1,980 women who had undergone hysterectomy between October 1998 and December 2005. Multiple logistic regression was used to identify risk factors for febrile morbidity. Results: Among the cohort, 1,463 women were nonfebrile and 517 febrile – an incidence of febrile morbidity of 26.1%. The incidence of febrile morbidity was higher in patients with lower preoperative hematocrit, more extensive surgery, longer operative time, greater blood loss and malignant disease. The median intraoperative blood loss was 500 ml in the febrile group and 400 ml in the non-febrile group (p < 0.0005). Median operative time was 150 min in the febrile group and 135 min in the non-febrile group (p < 0.0005). Two variables were identified as independent risk factors for febrile morbidity: intraoperative blood loss of ≧750 ml (compared with <250 ml OR 1.52; 95% CI 1.08–2.13; p = 0.036) and a diagnosis of malignant disease (OR 1.86; 95% CI 1.45–2.13; p < 0.0005). Conclusions: Independent risk factors for febrile morbidity were an intraoperative blood loss of ≧750 ml and malignant disease.

Long‐term survival rate of pediatric patients after blood transfusion

Studies in adults report posttransfusion survival rate (PTSR) at 1 to 10 years of 41 to 67 percent. There are no published studies specifically addressing PTSR in pediatric patients. The objectives of this study were to evaluate PTSR and risk factors associated with death in children receiving transfusions. A database of all patients receiving their first transfusion between 1990 and 1992 at Sainte-Justine Hospital (SJH) was used. Patients' demographic data, primary diagnosis, surgical procedures, and information on all labile components transfused were collected. Death was confirmed by the SJH database, RAMQ (universal health care provider for Québec residents), and the Quebec Commission for Access to Information (death certificate). The study population consisted of 1100 children. Median age was 16 months (range, 0-204 months). The most frequent primary diagnoses were cardiac disease (22%), prematurity (21.5%), malignant disease (11%), and hematologic disease (9.5%). Patients received a median of three transfusions (range, 1-126). PTSR was 86.9 percent at 1 year and 82.3 percent at 10 years. For nonsurvivors, median survival time was 22 days. Multivariate analysis showed that significant risk factors for death are age of less than 1 month, a diagnosis of malignant disease, more than 20 transfusions, and administration of more than one type of blood component. PTSR in children receiving their first transfusion between 1990 and 1992 was considerably higher than the PTSR reported in adults. Death occurred early after the first transfusion; survival rate remained relatively stable thereafter.

Einsatz bildgebender Verfahren bei der Diagnostik der Osteoporose Interpretation der Röntgenbilder und Messprotokolle der Knochendichte

According to the recently published BoneEVA study, 7.8 million Germans (6.5 million women) are affected by osteoporosis. Of them, 4.3% experienced at least one clinical fracture. Only 21.7% were treated with an anti-osteoporotic drug, whereby only 10% received a bisphosphonate and 17% given calcium and vitamin D. On the other hand, as osteoporosis may be associated with severe pain in 90% of patients, analgesics are prescribed. The total direct costs attributable to osteoporosis amounted to Euro 5.4 billion in 2003. One out of three postmenopausal women and one out of five men over the age of 50 years will experience osteoporotic fractures unless preventive measures are undertaken. According to the German guidelines for diagnosis and treatment of osteoporosis, bone densitometry using dual energy x-ray absorptiometry (DXA) together with other clinical risk factors (previous low trauma fracture, use of nicotine, low body weight [BMI<20 kg/m2], immobilisation, and more than two falls during the last six months) are recommended for diagnosis. Using typical cases out of clinical practice, this article delineates frequent mistakes in the interpretation of DXA measurements. Furthermore, the present paper clarifies the role of classical x-rays, which still represent the predominant procedure for the identification of fractures and especially vertebral fractures. In comparison to x-rays, CT or MRI are more important in differential diagnosis of malignant disease and bone metastases. Essentially a reduction of vertebral height without evidence of central endplate fracture in postmenopausal women may be unrelated to osteoporosis. Quantitative morphometry should not be used alone for the assessment of vertebral fracture in clinical decision-making. Therefore, we recommend differential diagnosis of morphometric vertebral deformities by an expert reader to rule out deformities related to degenerative disease and norm variants of which we will present several examples to train the view of the reader.

Measurement of Cyclin E Genomic Copy Number and Strand Length in Cell-Free DNA Distinguish Malignant versus Benign Effusions

Previous studies have shown that the concentration of cell-free DNA was higher and its strand length longer in body fluids obtained from patients with cancer as compared to patients with benign diseases. We hypothesized that analysis of both DNA copy number and strand length of cell-free DNA from an amplified chromosomal region could improve the diagnosis of malignant diseases in body fluids. To test this hypothesis, we used ovarian cancer effusion as an example and applied a quantitative real-time PCR to measure the relative copy number and strand length of DNA fragments from one of the most frequently amplified genes, cyclin E, in ovarian serous carcinomas. As compared with nonamplified chromosomal loci, including beta-actin, p53, 2p24.1, and 4p15.31, measurement of cyclin E DNA copy number (100 bp) had the best performance in distinguishing malignant (n = 88) from benign (n = 70) effusions after normalization to effusion volume or Line-1 DNA with areas under the receiver operating characteristics curve (AUC) of 0.832 and 0.847, respectively. Different DNA lengths of the cyclin E locus were further analyzed and we found that the AUC was highest by measuring the 400-bp cyclin E locus (AUC = 0.896). The AUC was improved to 0.936 when it was combined with the length integrity index as defined by the relative abundance of 400 bp cyclin E to 100 bp p53 loci. Cyclin E real-time PCR assay had a higher sensitivity (95.6%) than routine cytology examination (73.9%) and was able to diagnose false-negative cytology cases in this study. The above findings indicate that measurement of the DNA copy number and strand length of the cyclin E locus is a useful cancer diagnostic tool.