Prediction of non-muscle invasive bladder cancer outcomes assessed by innovative multimarker prognostic models.

BACKGROUND: We adapted Bayesian statistical learning strategies to the prognosis field to investigate if genome-wide common SNP improve the prediction ability of clinico-pathological prognosticators and applied it to non-muscle invasive bladder cancer (NMIBC) patients. METHODS: Adapted Bayesian sequential threshold models in combination with LASSO were applied to consider the time-to-event and the censoring nature of data. We studied 822 NMIBC patients followed-up >10 years. The study outcomes were time-to-first-recurrence and time-to-progression. The predictive ability of the models including up to 171,304 SNP and/or 6 clinico-pathological prognosticators was evaluated using AUC-ROC and determination coefficient. RESULTS: Clinico-pathological prognosticators explained a larger proportion of the time-to-first-recurrence (3.1 %) and time-to-progression (5.4 %) phenotypic variances than SNPs (1 and 0.01 %, respectively). Adding SNPs to the clinico-pathological-parameters model slightly improved the prediction of time-to-first-recurrence (up to 4 %). The prediction of time-to-progression using both clinico-pathological prognosticators and SNP did not improve. Heritability (h (2)) of both outcomes was <1 % in NMIBC. CONCLUSIONS: We adapted a Bayesian statistical learning method to deal with a large number of parameters in prognostic studies. Common SNPs showed a limited role in predicting NMIBC outcomes yielding a very low heritability for both outcomes. We report for the first time a heritability estimate for a disease outcome. Our method can be extended to other disease models.

In vivo reflectance confocal microscopy characterization of silver deposits in localized cutaneous argyria.

Localized cutaneous argyria is a rare condition secondary to skin deposition of silver following exposure to substances containing this metal. The clinical appearance and dermoscopy findings require deep melanocytic lesions and particularly melanoma metastasis to be ruled out. Silver deposits are usually confirmed by scanning electron microscopy and/or energy-dispersive X-ray spectroscopy. Herein we describe the in vivo reflectance confocal microscopy (RCM) features observed in one case of localized cutaneous argyria. These features include the presence of a hyperrefractile network in the papillary dermis and a periadnexal dotted bright pattern. In vivo RCM might be a useful tool for an early diagnosis of this uncommon entity.

Clinical-pathological differences and smoking habit depending on gender in a cohort of patients with transitional cell carcinoma of the bladder: retrospective study.

OBJECTIVES: The increased incidence of transitional cell carcinoma (TCC) of the bladder in men is known, generally attributed to greater exposure to the effect carcinogenic products. Although it has been reported that cancer-specific outcome can be particularly adverse in women due to socioeconomic or biological factors, clinical-pathological differences of TCC at the time of diagnosis have not been sufficiently studied. The aim of this study is to analyze whether there are gender-related differences in grade and tumor stage in primary bladder TCC. METHODS: All consecutive primary bladder TCC diagnoses made in our institution between 1990 and 2009 have been included. We retrospectively analyzed age, smoking, symptoms at presentation, tumor grade (WHO 1973), tumor size and number, and TNM stage, comparing men and women. Statistical analysis was conducted using the Mann-Whitney U test as non-parametric test and Chi-squared test to compare frequencies. RESULTS: We analyzed 1196 patients (992 males and 204 females) with a 5:1 ratio. We found significant differences in age (69 years vs. 73 years), smoking (46.5% vs. 11.2%)and muscle-invasive stage (12.1% vs. 18.1%). Correcting by tobacco consumption, never-smoker women have larger and more aggressive tumors with a frequency of muscle-invasive disease three times higher than male never-smokers and equaling to male current-smokers. CONCLUSION: TCC of the bladder is more frequent in males than females. In this series, women are older at the time of diagnosis and most often affected by muscle-invasive disease particularly in never-smokers. We need studies to analyze the potential impact of passive smoking to justify these results.

A transcriptional signature associated with the onset of benign prostate hyperplasia in a canine model.

BACKGROUND: Benign prostatic hyperplasia (BPH) represents the most frequent proliferative abnormality of the human prostate. In spite of the well-characterized architectural development of BPH, little is known about the cellular and molecular events that contribute to it. METHODS: We have developed an animal model to evaluate the follow-up of hormone-induced BPH and the analysis of the gene expression associated with BPH. Immunohistochemistry on human patient samples validated the BPH-related molecular alterations. RESULTS: Canine specific Affymetrix microarray analysis performed on sequential biopsies obtained from a beagle dog dynamic model characterized a number of genes altered during the onset of BPH. In addition to the genes involved in calcification, matrix remodeling, detoxification, cell movement, and mucosa protection (MGP, MMP2, TIMP2, ITIH3, GST, MT2A, SULT1A1, FKBP1B, MUC1, STRBP, TFF3), the up-regulation of TGFB3 and CLU indicated a complete adjustment of the transdifferentiation, senescence and apoptosis programs. The up-regulation of Clusterin was validated by RT-qPCR and immunohistochemistry, both in the dog dynamic model and in human samples, further confirming the suitability of the animal model for the study of the molecular alterations associated with BPH. CONCLUSIONS: Transcriptome analysis performed on a dynamic animal model that accurately mimicked the human clinic, allowed us to characterize a gene expression pattern associated with the onset of BPH.

Occupation and bladder cancer in a hospital-based case-control study in Spain.

OBJECTIVES: We investigated the association between occupation and bladder cancer in a hospital-based case-control study conducted in Spain. METHODS: 1219 patients with transitional cell carcinoma of the urinary bladder and 1271 controls selected from 18 hospitals in Spain between June 1998 and September 2000 provided detailed information on life-time occupational history, smoking habits, medical history, and other factors. We used unconditional logistic regression to calculate odds ratios (OR) and 95% confidence intervals (CI) for each occupation and industry, adjusting for age, hospital region, smoking duration, and employment in a high-risk occupation for bladder cancer. RESULTS: Statistically significant increased risks were observed among men employed as machine operators in the printing industry (OR 5.4; 95% CI 1.6 to 17.7), among men employed in the transportation equipment industry (OR 1.6; 95% CI 1.1 to 2.6) and among those who had worked for >/=10 years in the electrical/gas/sanitary services (OR 3.9; 95% CI 1.5 to 10.4) and in hotels and other lodgings (OR 3.1; 95% CI 1.3 to 7.3). Men who worked as miscellaneous mechanics and repairers (OR 2.0; 95% CI 1.1 to 3.6) and as supervisors in production occupations (OR 2.1; 95% CI 1.2 to 3.6) also had excess risks for bladder cancer. Male farmers and those who worked in crop and livestock production had decreased risks for bladder cancer. We found no significant associations between occupation or industry and bladder cancer risk among women. CONCLUSIONS: We did not observe excess bladder cancer risk for many of the occupations identified as being a priori at high risk. Examination of more detailed job exposure information should help clarify these associations.

Primitive neuroectodermal tumor of the central nervous system with glial differentiation: a FISH study of an adult case.

Primitive neuroectodermal tumors (PNETs) of the central nervous system (CNS), a rare occurrence in adults, may show glial differentiation and can be misinterpreted as pure astrocytic neoplasms. Few fluorescence in situ hybridization (FISH) studies have been carried out on these tumors; isochromosome 17q was found to be the major chromosomal abnormality. We present the case of an adult in which we performed a FISH study of both the glial and neuronal components. A complex array of FISH changes, not including an isochromosome 17q were identified.

Expression of androgen, oestrogen alpha and beta, and progesterone receptors in the canine prostate: differences between normal, inflamed, hyperplastic and neoplastic glands.

The expression of receptor for androgen (AR), oestrogen alpha and beta (ERalpha and ERbeta) and progesterone (PR) was examined immunohistochemically in canine prostate specimens (normal, hyperplastic, inflamed [prostatitis] or neoplastic). AR immunolabelling was seen in 100% of epithelial cells of normal and hyperplastic tissue, the corresponding figures for inflamed and carcinomatous tissue being 74% and 65%, respectively. ERalpha labelling was seen in 85% of epithelial cells in normal prostate glands, the corresponding figures for hyperplastic, inflamed and neoplastic glands being 35%, 22% and 12%, respectively. ERbeta labelling was seen in 85% of epithelial cells of normal glands and in about 70% of such cells in glands showing pathological changes. On the other hand, PR expression (weak) in normal glands was observed in fewer epithelial cells (44%) than in hyperplastic (70%), inflamed (62%) or neoplastic (64%) glands.

Erythrokeratoderma variabilis-like ichthyosis in Chanarin-Dorfman syndrome.

Neutral lipid storage disease (Chanarin-Dorfman syndrome) is an autosomal recessive metabolic disorder associated with congenital ichthyosis and a multisystemic accumulation of neutral lipids (lipid droplets) in various types of cells. The clinical presentation has been reported to correspond to that of nonbullous congenital ichthyosiform erythroderma. We report a 4-year-old boy presenting a generalized ichthyosiform disorder manifested by migrating scaly plaques alternating with areas of normal-looking skin, showing erythematous borders with sharp margins, clinically suggestive of erythrokeratoderma variabilis (EKV). A peripheral blood smear revealed cytoplasmic vacuoles in most granulocytes. Genetic studies from the patient and his parents revealed that the patient carried two different and novel mutations of the ABHD5 gene: a nonsense mutation in exon 6 (transmitted by the father) and an insertion/deletion in exon 4 (transmitted by the mother). Our observation demonstrates the clinical heterogeneity of the ichthyosiform dermatoses observed in Chanarin-Dorfman syndrome and widens the clinical range of conditions presenting migrating scaly plaques mimicking EKV.

Alendronate and etidronate do not regulate interleukin 6 and 11 synthesis in normal human osteoblasts in culture.

Bisphosphonates exert a potent inhibitory effect on bone resorption. Several studies have been performed, with contradictory results, to ascertain whether the effect of bisphosphonates on osteoclasts could be produced, at least in part, by modulation of the synthesis of resorption-promoting factors by osteoblasts. The aim of this study was to evaluate the effect of etidronate (10-4-10-9 M) and alendronate (10-7-10-12 M) on the production of IL-6 and IL-11 using human osteoblast cultures. Cytokines were quantified by ELISA, and mRNA expression was tested. Treatment with alendronate and etidronate had no effect on the synthesis of IL-6 or IL-11, and IL-6 and IL-11 mRNA levels. These results were obtained both in nonstimulated cultures and in cultures stimulated by means of TNF-a, IL-1b, and TNF-a+IL-1b, with or without FCS. In conclusion, a possible indirect osteoclast-mediated effect of alendronate and etidronate on bone resorption would not be exerted through reduction in osteoblastic synthesis of IL-6 and IL-11.

[Fiber-optic bronchoscopic biopsy of bronchial smooth muscle. Efficacy of the technique in individuals with normal lung function and patients with COPD]. / Biopsia de músculo liso bronquial mediante fibrobroncoscopia. Eficacia de la técnica en individuos con función pulmonar normal y en pacientes con EPOC.

BACKGROUND AND OBJECTIVES: The epithelium and airway smooth muscles of patients with chronic obstructive pulmonary disease (COPD) or bronchial asthma undergo certain structural changes that are probably related to increased expression of inflammatory molecules and cell growth factors. Studying the relation between disease and changes in bronchial smooth muscle is difficult if investigation is restricted to samples from autopsies or thoracotomies. This study was designed to evaluate the probability of obtaining bronchial smooth muscle by endoscopic bronchial biopsy in patients with COPD and from individuals with normal lung function, the relation of disease to bronchial epithelial histology, and the potential usefulness of studying airway muscle remodeling events. METHODS: Forty-two patients undergoing diagnostic fiberoptic bronchoscopy were enrolled. Bronchial biopsies were taken systematically from the lobar and segmental dividing ridges. The epithelial structure was analyzed by conventional histology. The smooth muscle was identified by immunohistochemistry (anti-desmin antibody assay) and Western-blot analysis (anti-desmin, actin and myosin antibodies). RESULTS: Sixty-nine percent of the biopsies contained bronchial smooth muscle. The probability of obtaining smooth muscle was higher in segmental than in lobar biopsies (72 vs 30%, p < 0.05). This probability was unrelated to the presence of COPD or to signs of epithelial inflammation. The fragments allowed us to use electrophoresis to identify protein structures (myosin, actin, desmin) involved in muscle remodeling processes. CONCLUSIONS: Endoscopic biopsy of the bronchi allows us to obtain bronchial smooth muscle samples in a large percentage of patients, particularly when performed on segmental bronchi. The technique may be useful for future studies examining the processes of airway smooth muscle remodeling.

Procion orange tracer dye technique vs. identification of intrafibrillar fibronectin in the assessment of sarcolemmal damage.

BACKGROUND: The use of Procion orange dye (POD) is one of the most widely accepted techniques to assess sarcolemmal damage. This phenomenon has been related to functional adaptation in skeletal muscles. The POD method includes intravenous injection of this colorant in vivo, enabling its identification inside those fibres with membrane leaks (fluorescence). However, the safety of the use of POD has not been proven. AIM: This study was designed to compare POD with a safer alternative, involving the identification of intracellular fibronectin using specific antibodies. METHOD: Eight Swiss mice were submitted to electrical stimulation of the lower limbs at different frequencies (10-80 Hz). Subsequently, the POD solution was infused, and samples from the vastus medialis muscle were obtained 24 h later. Samples were processed and serial sections were analysed using immunohistochemistry (monoclonal antibodies against fibronectin) and epifluorescence microscopy. RESULTS: Ninety-eight per cent of the fibres were equally classified by both techniques, which in addition showed good correlation (percentages of damaged fibres, r = 0.998, P < 0.001) and concordance (R1 = 0.82) in quantitative terms. CONCLUSIONS: Although the two techniques compared here are based on different principles, both are comparable in assessing sarcolemmal damage. This would facilitate comparisons between human and experimental studies. In addition, the fibronectin technique appears to be a suitable alternative for long-term studies including repeated biopsies.

Renal cell carcinoma with syncytial giant cell component.

We report a case of clear cell renal cell carcinoma in which a prominent multinucleated giant cell component was intermingled with clear, granular, and spindle cells. Histological, ultrastructural, cytometric, and cytogenetic features of giant cells were similar to those of mononucleated cells in the tumor, and therefore they were not from stromal or osteoclast derivation. These giant cells had homogeneous, finely granular, abundant cytoplasm, often with scalloped cell borders, and contained from 5 to more than 50 nuclei, all of them very similar in size and shape, with prominent central nucleoli. Occasionally, surrounding inflammatory cells were also engulfed in the cytoplasm. This syncytial appearance was more similar to that of some giant cell carcinomas from the lung than to the pleomorphic giant cells often encountered in high grade renal cell tumors. Although the patient is alive and free of disease 6 years after diagnosis, a longer follow-up will be required to assess the potential prognostic influence of this peculiar histological appearance.

Injury of the human diaphragm associated with exertion and chronic obstructive pulmonary disease.

Injury of the diaphragm may have clinical relevance having been reported in cases of sudden infant death syndrome or fatal asthma. However, examination of diaphragm injury after acute inspiratory loading has not been reported. The purpose of this study was to determine whether an acute inspiratory overload induces injury of the human diaphragm and to determine if diaphragm from chronic obstructive pulmonary disease (COPD) is more susceptible to injury. Eighteen patients with COPD and 11 control patients with normal pulmonary function (62 +/- 10 yr) undergoing thoracotomy or laparotomy were studied. A threshold inspiratory loading test was performed prior to surgery in a subset of seven patients with COPD and five control patients. Samples of the costal diaphragm were obtained during surgery and processed for electron microscopy analysis. Signs of sarcomere disruption were found in all diaphragm samples. The range of values of sarcomere disruption was wide (density: 2-45 abnormal areas/100 microm(2); area fractions: 1.3-17.3%), significantly higher in diaphragm from patients with COPD (p < 0.05) and with the greatest injury after inspiratory loading. We conclude that sarcomere disruption is common in the human diaphragm, is more evident in patients with COPD, and is higher after inspiratory loading, especially in the diaphragm of those with COPD.

Fine-needle aspiration of chromophobe renal-cell carcinoma metastatic to the thyroid gland.

Chromophobe renal cell carcinoma is an unusual variant of renal carcinoma that has less aggressive behavior than clear cell carcinomas. There are few documented cases of metastases, none of which occurred in the thyroid gland. A case is presented of chromophobe renal cell carcinoma metastatic to the thyroid eight years after right nephrectomy, suspected by FNA-biopsy and confirmed histologically. Although metastases of chromophobe renal cell carcinoma are rare, they may also present in thyroid, even many years after primary tumor diagnosis, just like clear cell carcinomas. Even though the FNA cytology of chromophobe renal cell carcinoma has distinctive features, in the context of the thyroid, it can be mistaken for a primary tumor of that organ. In our case, the history of a previous renal tumor was essential in suggesting a metastatic lesion, and histologic and ultrastructural features allowed its precise identification.

Bone metaplasia in a case of bilateral renal cell carcinoma.

We present a case of synchronous, bilateral renal cell carcinoma with osseous metaplasia. In renal cell carcinoma, bone can originate through two different processes: osseous differentiation or osseous metaplasia. The case we report here represents the second process. We discuss pathological differential diagnosis and prognosis of the infrequent tumor.

Mucins as differentiation markers in bronchial epithelium. Squamous cell carcinoma and adenocarcinoma display similar expression patterns.

UNLABELLED: Highly glycosylated apomucins are important to maintain the viscoelastic properties of the mucus. Changes in their expression are frequently associated with inflammatory and neoplastic conditions. We analyzed the expression of apomucins in normal respiratory tract (n = 8) and compared it with distal, peritumoral, and tumoral epithelia from patients with squamous cell carcinoma (n = 20), adenocarcinoma (n = 13), and small cell carcinoma (n = 12). Squamous metaplasia (n = 16) was also analyzed. MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC8 apomucins were detected by immunohistochemistry, and mucin transcripts by in situ hybridization and reverse transcriptase polymerase chain reaction. Bronchial epithelium from normal individuals and distal epithelium from cancer patients showed a similar expression pattern: MUC1, MUC4, and MUC8 were always present, MUC2 and MUC5AC showed more variability, and MUC6 was focally detected. MUC5AC was downregulated in peritumoral epithelium and in squamous metaplasia, and MUC6 was upregulated in peritumoral epithelium. A reduced expression of MUC4, MUC5AC, and MUC8 was observed in non-small cell carcinomas, regardless of their histologic subtype. In small cell tumors, only MUC1 was consistently expressed. CONCLUSIONS: (1) peritumoral epithelium and squamous metaplasia show an abnormal pattern of mucin expression; (2) squamous cell carcinomas and adenocarcinomas display a similar pattern of mucin gene expression, supporting the concept of a common cellular origin.

Malignant pigmented clear cell epithelioid tumor of the kidney: clear cell ("sugar") tumor versus malignant melanoma.

A 73-year-old woman presented with an hemorrhagic kidney tumor initially interpreted as a renal cell carcinoma (RCC). A retroperitoneal recurrence infiltrating the duodenal wall was made up of clear cells, some of which contained Fontana-Masson positive pigment, immunopositive for HMB-45, S-100 protein, actin, and vimentin. The same immunohistochemical profile was retrospectively reproduced in the kidney tumor, where melanosomes were also found ultrastructurally. Lipomatous differentiation was not observed. There was no history of malignant melanoma (MM), or stigmata of tuberous sclerosis. The patient died of disease 5 years after the initial diagnosis. This neoplasm can be considered as a malignant, pigmented, clear-cell epithelioid variant of angiomyolipoma, or "sugar" tumor of the kidney, with the peculiarity of having a previously unreported component of pigmented cells visible on light microscopy. This finding raises the possibility that the exceptional cases of MM reported in renal parenchyma may be pigmented variants of epithelioid angiomyolipoma rather than true MM.