Advanced chondrosarcoma of the pelvis: a rare case of urinary obstruction

Chondrosarcoma is the second most common malignant tumor of the bone with an incidence of 1 in 200.000 per year. Axial skeleton is frequently involved showing poorer oncological outcomes than appendicular one: human pelvis is a site predilection. It is rarely associated to urinary obstruction but according to its localization, it can be frequently linked to compression of pelvic organs as bladder, prostate or bowel. We describe the case of a 52 years old caucasian male with history of advanced pelvic chondrosarcoma and severe hydronephrosis due to total bladder dislocation

“Urinalysis, urinary proteome and metabolome of zoo-housed giraffes (Giraffa camelopardalis) through noninvasive sampling method”.

The study of non-domestic animals withholds more difficulties compared to the domestic counterpart, thus using noninvasive techniques to collect biological samples might play an important role in assessing the health status of wild animals. 1 The present study established the reliability of urine sampling from the ground. A preliminary study was run with 10 urine samples of 10 cows (Bos taurus) housed in a dairy farming in Northern Italy. Urine samples, collected both in sterile cups and from the ground, were analyzed and compared. Results revealed no statistical differences in the variables investigated (p > 0.05, dipstick parameters and USG, protein quantification and UPC and protein electrophoresis), which proved the reliability of this noninvasive sampling method. This method was used for sampling 103 urine samples from 44 zoo-housed giraffes (Giraffa camelopardalis) of four Italian zoos. Urine samples were used to establish the urinalysis reference values in this species and to study the urinary proteome for the first time. The urinary reference values reported as median (lower and upper limit) were: urine specific gravity (USG), 1.030 (1006 - 1.049); urine total proteins (uTP), 17.58 (4.54 – 35.31) mg/dL; urine creatinine (uCr), 154.62 (39.59 – 357.95) mg/dL; urine protein: creatinine ratio (UPC), 0.11 (0.07 – 0.16). In giraffes, most urinary proteins had a low molecular mass (MM) and were present in low quantities. Proteomics disclosed fifteen different proteins, which were involved in the defense against microbes and in the ability of giraffes to concentrate urine. Albumin, lysozyme C, and ubiquitin were the most represented urinary proteins in giraffes. In addition, to define the urinary metabolome profile, 35 urine samples from 35 zoo-housed giraffes of five Italian zoos were used. Metabolomics allowed to identify and quantify 39 molecules and the most represented metabolites were hippurate, creatinine and phenylacetylglycine. This analysis provided information on physiological adaptations of giraffes. Besides, urinary metabolites were influenced by sex: urinary metabolome profile of female showed higher level of acetate, succinate, and lactate, conversely hippurate, phenylacetylglycine, and thymine were more concentrated in male urines. Similarly, the age affected the concentration of three urinary metabolites, namely formate, alanine, and valerate

Prevalence of p53 dysregulations in feline oral squamous cell carcinoma and non-neoplastic oral mucosa

Squamous cell carcinoma is the most common malignant oral tumor in cats. The late presentation is one of the factors contributing to the detrimental prognosis of this disease. The immunohistochemical expression of the p53 tumor suppressor protein has been reported in 24% to 65% of feline oral squamous cell carcinomas, but no study has systematically evaluated in this tumor the presence of p53 encoding gene (TP53) mutations. The aim of this retrospective study was to determine whether p53 immunohistochemistry accurately reflects the mutational status of the TP53 gene in feline oral squamous cell carcinoma. Additionally, the prevalence of p53 dysregulation in feline oral squamous cell carcinoma was compared with that of feline non-neoplastic oral mucosa, in order to investigate the relevance of these dysregulations in cancer development. The association between p53 dysregulations and exposure to environmental tobacco smoke and tumor characteristics was further assessed. Twenty-six incisional biopsies of oral squamous cell carcinomas and 10 cases each of lingual eosinophilic granuloma, chronic gingivostomatitis and normal oral mucosa were included in the study. Eighteen squamous cell carcinomas (69%) expressed p53 and 18 had mutations in exons 5\u20138 of TP53. The agreement between immunohistochemistry and mutation analysis was 77%. None of non-neoplastic oral mucosa samples had a positive immunohistochemical staining, while one case each of eosinophilic granuloma and chronic gingivostomatitis harbored TP53 mutations. Unlike previously hypothesized, p53 dysregulations were not associated with exposure to environmental tobacco smoke. These results suggest an important role of p53 in feline oral tumorigenesis. Additionally, the immunohistochemical detection of p53 expression appears to reflect the presence of TP53 mutations in the majority of cases. It remains to be determined if the screening for p53 dysregulations, alone or in association with other markers, can eventually contribute to the early detection of this devastating disease

Enriched sera protein profiling for detection of non-small cell lung cancer biomarkers

<p>Abstract</p> <p>Background</p> <p>Non Small Cell Lung Cancer (NSCLC) is the major cause of cancer related-death. Many patients receive diagnosis at advanced stage leading to a poor prognosis. At present, no satisfactory screening tests are available in clinical practice and the discovery and validation of new biomarkers is mandatory. Surface Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-ToF-MS) is a recent high-throughput technique used to detect new tumour markers. In this study we performed SELDI-ToF-MS analysis on serum samples treated with the ProteoMiner™ kit, a combinatorial library of hexapeptide ligands coupled to beads, to reduce the wide dynamic range of protein concentration in the sample. Serum from 44 NSCLC patients and 19 healthy controls were analyzed with IMAC30-Cu and H50 ProteinChip Arrays.</p> <p>Results</p> <p>Comparing SELDI-ToF-MS protein profiles of NSCLC patients and healthy controls, 28 protein peaks were found significantly different (p < 0.05), and were used as predictors to build decision classification trees. This statistical analysis selected 10 protein peaks in the low-mass range (2-24 kDa) and 6 in the high-mass range (40-80 kDa). The classification models for the low-mass range had a sensitivity and specificity of 70.45% (31/44) and 68.42% (13/19) for IMAC30-Cu, and 72.73% (32/44) and 73.68% (14/19) for H50 ProteinChip Arrays.</p> <p>Conclusions</p> <p>These preliminary results suggest that SELDI-ToF-MS protein profiling of serum samples pretreated with ProteoMiner™ can improve the discovery of protein peaks differentially expressed between NSCLC patients and healthy subjects, useful to build classification algorithms with high sensitivity and specificity. However, identification of the significantly different protein peaks needs further study in order to provide a better understanding of the biological nature of these potential biomarkers and their role in the underlying disease process.</p

Inflammation: an important parameter in the search of prostate cancer biomarkers

Background A more specific and early diagnostics for prostate cancer (PCa) is highly desirable. In this study, being inflammation the focus of our effort, serum protein profiles were analyzed in order to investigate if this parameter could interfere with the search of discriminating proteins between PCa and benign prostatic hyperplasia (BPH). Methods Patients with clinical suspect of PCa and candidates for trans-rectal ultrasound guided prostate biopsy (TRUS) were enrolled. Histological specimens were examined in order to grade and classify the tumor, identify BPH and detect inflammation. Surface Enhanced Laser Desorption/Ionization-Time of Flight-Mass Spectrometry (SELDI-ToF-MS) and two-dimensional gel electrophoresis (2-DE) coupled with Liquid Chromatography-MS/MS (LC-MS/MS) were used to analyze immuno-depleted serum samples from patients with PCa and BPH. Results The comparison between PCa (with and without inflammation) and BPH (with and without inflammation) serum samples by SELDI-ToF-MS analysis did not show differences in protein expression, while changes were only observed when the concomitant presence of inflammation was taken into consideration. In fact, when samples with histological sign of inflammation were excluded, 20 significantly different protein peaks were detected. Subsequent comparisons (PCa with inflammation vs PCa without inflammation, and BPH with inflammation vs BPH without inflammation) showed that 16 proteins appeared to be modified in the presence of inflammation, while 4 protein peaks were not modified. With 2-DE analysis, comparing PCa without inflammation vs PCa with inflammation, and BPH without inflammation vs the same condition in the presence of inflammation, were identified 29 and 25 differentially expressed protein spots, respectively. Excluding samples with inflammation the comparison between PCa vs BPH showed 9 unique PCa proteins, 4 of which overlapped with those previously identified in the presence of inflammation, while other 2 were new proteins, not identified in our previous comparisons. Conclusions The present study indicates that inflammation might be a confounding parameter during the proteomic research of candidate biomarkers of PCa. These results indicate that some possible biomarker-candidate proteins are strongly influenced by the presence of inflammation, hence only a well-selected protein pattern should be considered for potential marker of PCa

Proteomic analysis of urine in medication-overuse headache patients: possible relation with renal damages

Medication-overuse headache (MOH) is a chronic disorder associated with overuse of analgesic drugs, triptans, non-steroidal anti-inflammatory drugs (NSAIDs) or other acute headache compounds. Various epidemiologic investigations proved that different drug types could cause nephrotoxicity, particularly in chronic patients. The aim of the present work was to analyze, by a proteomic approach, the urinary protein profiles of MOH patients focusing on daily use of NSAIDs, mixtures and triptans that could reasonably be related to potential renal damage. We selected 43 MOH patients overusing triptans (n = 18), NSAIDs (n = 11), and mixtures (n = 14), for 2–30 years with a mean daily analgesic intake of 1.5 ± 0.9 doses, and a control group composed of 16 healthy volunteers. Urine proteins were analyzed by mono-dimensional gel electrophoresis and identified by mass spectrometry analysis. Comparing the proteomic profiles of patients and controls, we found a significantly different protein expression, especially in the NSAIDs group, in which seven proteins resulted over-secreted from kidney (OR = 49, 95% CI 2.53–948.67 vs. controls; OR = 11.6, 95% CI 0.92–147.57 vs. triptans and mixtures groups). Six of these proteins (uromodulin, α-1-microglobulin, zinc-α-2-glycoprotein, cystatin C, Ig-kappa-chain, and inter-α-trypsin heavy chain H4) were strongly correlated with various forms of kidney disorders. Otherwise, in mixtures and in triptans abusers, only three proteins were potentially associated to pathological conditions (OR = 4.2, 95% CI 0.33–53.12, vs. controls). In conclusion, this preliminary proteomic study allowed us to define the urinary protein pattern of MOH patients that is related to the abused drug. According with the obtained results, we believe that the risk of nephrotoxicity should be considered particularly in MOH patients who abuse of NSAIDs

Ischaemic priapism tamsulosin-induced in chronic renal failure: may this represent a risk factor?

Introduction: Ischaemic priapism following adrenergic alpha-antagonist is a rare emergency but some cases have been reported in literature. While opportunely describeing a case report of priapism following tamsulosin ingestion which presented to us, we reviewed the literature using PubMed library and selected all the articles about the casual relationship between those two entities: furthermore we looked for any other cases where priapism a-blockers-induced and chronic renal failure were both present in order to evaluate if this last condition may represent a risk factor. Patient and Methods: A 52 years old caucasian male with a clinical history of transplanted kidney presented to us for lower urinary tract symptoms related to a BPH condition. His prostate was firm and enlarged on digito-rectal examination. PSA and urine analysis were normal while blood examination revealed a chronic renal failure (creatinine 2,0 mg/dl). Empirical treatment with tamsulosin was initiated. Two days after the first dose of drug he developed a painfull persistent erection according to priapism diagnosis. After performing a distal shunt of the corpora and intracavernosal injection of etilephrine, full penile detumescence was obtained. Discussion: The proposed mechanism responsible for priapism following A1-antagonist is an alpha adrenergic blockade which directly inhibits the sympathetic impulse of detumescence expecially if a higher dose has been taken or its concentration is increased by metabolism inhibition. Although more common in non uroselective agents, few cases related to tamsulosin have been also described. In a systematic review of the literature 14 articles about priapism following a-blockers were found but to our knowledge this is the first case related to chronic renal function condition.. Conclusions: Adrenergic a-blockers are safe and effective drugs but, although rare, association with priapism is documented. Patient should be informed of the risk especially whenever affected by renal failure in order to not to delay medical care and earlier treat this urological emergenc

High-abundance proteins depletion for serum proteomic analysis: concomitant removal of non-targeted proteins.

In clinical and pharmaceutical proteomics, serum and plasma are frequently used for detection of early diagnostic biomarkers for therapeutic targets. Although obtaining these body fluid samples is non-invasive and easy, they contain some abundant proteins that mask other protein components present at low concentrations. The challenge in identifying serum biomarkers is to remove the abundant proteins, uncovering and enriching at the same time the low-abundance ones. The depletion strategies, however, could lead to the concomitant removal of some non-targeted proteins that may be of potential interest. In this study, we compared three different methods aimed to deplete high-abundance proteins from human serum, focusing on the identification of non-specifically bound proteins which might be eventually removed. A Cibacron blue-dye-based method for albumin removal, an albumin and IgG immunodepletion method and an immunoaffinity column (Multiple Affinity Removal System) that simultaneously removes a total of six high-abundance proteins, were investigated. The bound proteins were eluted, separated by two-dimensional gel electrophoresis and identified by Nano LC-CHIP-MS system. Flow-through fractions and bound fractions were also analysed with the ProteinChip technology SELDI-TOF-MS. Our results showed that the methods tested removed not only the targeted proteins with high efficiency, but also some non-targeted proteins. We found that the Multiple Affinity Removal Column improved the intensity of low-abundance proteins, displayed new protein spots and increased resolution. Notably, the column showed the lowest removal of untargeted proteins, proved to be the most promising depletion approach and a reliable method for serum preparation prior to proteomic studies

Research of Prostate Cancer Urinary Diagnostic Biomarkers by Proteomics: The Noteworthy Influence of Inflammation

Nowadays, in the case of suspected prostate cancer (PCa), tissue needle biopsy remains the benchmark for diagnosis despite its invasiveness and poor tolerability, as serum prostate-specific antigen (PSA) is limited by low specificity. The aim of this proteomic study was to identify new diagnostic biomarkers in urine, an easily and non-invasively available sample, able to selectively discriminate cancer from benign prostatic hyperplasia (BPH), evaluating whether the presence of inflammation may be a confounding parameter. The analysis was performed by two-dimensional gel electrophoresis (2-DE), mass spectrometry (LC-MS/MS) and Enzyme-Linked Immunosorbent Assay (ELISA) on urine samples from PCa and BPH patients, divided into subgroups based on the presence or absence of inflammation. Significant quantitative and qualitative differences were found in the urinary proteomic profile of PCa and BPH groups. Of the nine differentially expressed proteins, only five can properly be considered potential biomarkers of PCa able to discriminate the two diseases, as they were not affected by the inflammatory process. Therefore, the proteomic research of novel and reliable urinary biomarkers of PCa should be conducted considering the presence of inflammation as a realistic interfering element, as it could hinder the detection of important protein targets

Schistosomiasis infection mimicking a bladder cancer

Schistosomiasis is an acute and chronic disease caused by parasitic worms: it’s trasmitted by contact with water contaminated with parasites. Farmers, fishermen and people using unclean water, especially in endemic areas, can be easily infected. Urogenital, intestinal and hepatic localization are common. Furthermore, it’s estimated that at least 92% of patient requiring treatment for schistosomiasis live in Africa. It’s a very uncommon disease in Italy, but immigration, especially from North Africa, is changing its prevalence in our country. We describe a case report of a schistosomiasis infection mimicking a bladder cancer in a young black male coming from Africa