Electrical impedance was recorded at 21 discrete frequencies (1 to 100 kHz) from 27 ex vivo human prostates. These electrical properties were measured by using custom designed Electrical Impedance Spectroscopy (EIS) sensing biopsy (Bx) needles. EIS-Bx needles gauge the electrical properties of tissue in tandem with the tissue extraction (used for histopathological assessment). The EIS-Bx probe has a signal-to-noise ratio (SNR) of 65 dB across the frequency range (1 kHz to 100 kHz). A total of 36 cancers and 288 benign regions were sampled from 27 human prostates. Mean resistance (R) of prostate decreased from 537.27 Ω to 126.74 Ω for benign tissues and 999.52 Ω to 340.67 Ω for malignant tissues across the 1 kHz - 100 kHz spectral range. Likewise, mean reactance (X) ranged from -391.41 Ω to -62.6 Ω for benign and -675.09 Ω to -162.28 Ω for cancer tissues over the same frequency range. Both R and X values are found to be significantly lower in normal prostate tissues than in malignant tissue (p<;0.001). Further testing to evaluate the clinical efficacy of this coupled device is underway.

Background: Osteoarthritis (OA) affects tens of millions of people worldwide. Twin studies and human GWAS have shown that there is a strong genetic component to the disease, however, to date relatively few genes have been linked to OA onset and progression. Using zebrafish, a genetically tractable organism, we aim to identify novel genes relevant to OA. Methods: We undertook a forward genetic screen of 600 zebrafish families, screening for cartilage and bone phenotypes by Alcian blue (cartilage) and Alizarin Red (bone) staining. We identified 5 families with phenotypes resembling OA, including progressive loss of joint mobility, cartilage matrix breakdown and osteophyte (bony spur) formation. We are mapping the causative genes using recombination distance based mapping techniques. Results: We have identified one of the causative genes as chst11 (also known as C4ST1) a gene involved in the metabolism of chondroitin sulphate. Chst11 has also been implicated in human OA. Loss of chst11 leads to misassembly of the cartilage matrix and changes to chondrocyte cell behaviour, such as altered cell proliferation and premature chondrocyte hypertrophy. Additionally loss of chst11 leads to increased osteoblast differentiation in vivo. Conclusions: Identification of a gene from a zebrafish forward genetic screen that has also been implicated in human OA pathogenesis acts as a proof of principle that zebrafish display sufficient similarities in their skeletal system to be a useful model in osteoarthritis. Zebrafish are not only genetically tractable, but are also transparent and use of fluorescent transgenic reporter lines allows us to track gene expression in the living fish in real time. We have generated numerous transgenic reporters marking chondrocytes at various stages of differentiation, as well as osteoblasts and important signalling molecules such as indian hedgehog, these allow us to track changes in the joint in real time at a level impossible in other model organisms. In conclusion zebrafish can form a useful complement to existing models of OA. Disclosure statement: The author has declared no conflicts of interest.

Background Apoptotic cells (ACs) induce murine B cells to produce IL-10 and in this way inhibit collagen-induced arthritis [1]. This led us to question whether regulatory B cells also play a role in the classical autoimmune disease SLE. Objectives We aimed to demonstrated a role for AC in the induction of regulatory B cells in SLE. Methods 14 healthy donors and 14 SLE patients volunteered for the study. Prior ethical approval was granted. Written consent was obtained. Venous blood was collected. B cells were isolated by density gradients and magnetic bead sorts or flow sorts. These were then incubated with pre-prepared apoptotic cells and 10ng/ml IL-4. These were then cultured for 72 hours at 37°C with 5% CO2. Supernatants were harvested and tested for IL-10 by sandwich ELISA. Results Healthy human B cells produced significantly more IL-10 (3.27 fold, p<0.004) when co-cultured with ACs than B cells, or ACs alone. Flow sorted memory, marginal zone B (MZB) cells and naive B cells were cultured with ACs. Memory and MZB cells produced significantly more IL-10 than naive B cells. B cell IL-10 induction by ACs was significantly lower in SLE patients with high anti-dsDNA titres (p<0.0031), and there was an inverse logarithmic relationship between anti-dsDNA titre and B cell IL-10 production (p<0.0011). There was a trend towards a significantly higher level of AC induced B cell IL-10 in patients with quiescent disease compared to active disease (P<0.28), and a weak inverse relationship between ESR and B cell IL-10 production (p<0.312). Conclusions Others have shown that the rate of apoptosis may be increased in SLE [2], however an increased rate of phagocytosis has also been demonstrated in the condition [3]. These events may be related and not be directly causing autoimmunity. Indeed high apoptosis rates are ordinarily found in organs such as the thymus without the development of autoimmunity. The current data suggests that a defect in the appropriate immunoregulatory recognition of apoptotic cells in SLE may be an alternative explanation of the evolution of this condition. We have recently published evidence to support this in both murine and human models. We have demonstrated that CD27+ B cells are the sub-population most able to produce IL-10 in response to apoptotic cells [4]. The kinetics of this has yet to be confirmed. It may be that the generation of apoptotic cells acts as a natural immunosuppressant to dampen exaggerated immune responses, with MZB cell dampening innate responses, and memory cells dampening adaptive responses. References Gray M, et al. Apoptotic cells protect mice from autoimmune inflammation by the induction of regulatory B cells. Proc. Natl. Acad. Sci. U. S. A. 2007;104:14080-14085. Emlen W, et al. Accelerated in vitro apoptosis of lymphocytes from patients with systemic lupus erythematosis. Journal of Immunology 1994; 152:3685-3692. Sarmiento L, et al. Opsonization by anti-dsDNA antibodies of apoptotic cells in systemic lupus erythematosis. Autoimmunity 2007; 40:337-339. Miles K, et al. A tolerogenic role for Toll-like receptor 9 is revealed by B-cell interaction with DNA complexes expressed on apoptotic cells. Proc. Natl. Acad. Sci. U. S. A. 2012; 109:887-892. Disclosure of Interest None Declared

A novel imaging method to detect prostate cancer and the clinical findings is presented in this study. The dual modal imaging system incorporates a transrectal ultrasound system and an electrical impedance tomography (EIT) system. The reconstructed electrical property image (EPI) is correlated with semi-quantitative pathological findings. An example of reconstructed EPI is also presented in this study. Additionally, at lower frequencies of 0.4 kHz, 3.2 kHz and 25.6 kHz, statistical tests suggest that the conductivity of cancerous regions is significantly greater than that of normal region (p<;0.0304). The preliminary clinical findings point to potential electrical impedance measuring applications for prostate cancer detecting.

Diagnostic confirmation of cancer in solid organs is based on biopsy findings. In a standard 12-core prostate biopsy protocol, conventional biopsy needles sample only 0.95% (∼0.228 cm³) of a typical 24-cm³ prostate gland. The primary objective of this study was to enhance the sensitivity of standard biopsy protocol by gauging electrical properties of tissue simultaneously with tissue extraction for histopathology analysis. A conventional biopsy (Bx) needle was instrumented with an electrical impedance spectroscopy (EIS) sensor to interrogate the tissue volume surrounding the needle tip. The EIS-Bx device was evaluated in a series of saline bath and ex vivo porcine experiments. It was found to sense a volume of 0.286 cm³ of tissue around the needle tip. EIS measurements were recorded from three ex vivo human prostates using the device, and the extracted biopsy cores were histologically assessed. Prostate conductivity σ ranged from 0.179 to 0.3310 S/m for benign tissues and 0.0746 to 0.0837 S/m for malignant tissues at frequencies ranging from 1 to 100 kHz. Relative permittivity ϵ(r) ranged from 2.10×10⁶ to 2.9 × 10⁴ for benign and 6.63×10⁵ to 5.3 × 10³ for cancer tissues over the same frequency range. Both are found to be significantly higher in normal prostate tissues than in malignant tissue (p < 0.00001).

Introduction: Incidence rates of bladder cancer are notably higher in men than women. While there is evidence that reproductive and hormonal risk factors may influence risk of bladder cancer, data are inconclusive.Materials and methods: We examined reproductive, menstrual and hormonal use history in our population-based case-control study of bladder cancer in New Hampshire (NH), USA (n = 207 women cases and n = 463 women controls). Additionally, we performed a meta-analysis of the published literature. We used unconditional logistic regression analysis to compute adjusted odds ratios associated with each risk factor in the NH study. We combined these estimates with those from the published literature using inverse variance effects models.Results: In the NH study, a slightly decreased odds ratio was found among women who had ever had a birth compared to nulliparous women and an elevated odds ratio among women who underwent surgical menopause (bilateral oophorectomy), especially at an early age. No overall associations were found with oral contraceptive use or hormone replacement therapy. These findings were generally in agreement with the meta-analytic results for which the combined relative risk (RR) estimate was reduced among ever parous women (combined RR estimate for ever parous versus nulliparous = 0.66, 95% confidence intervals [95% CI] 0.55-0.79) and elevated among those undergoing an early menopause (combined RR estimate for early versus late menopause = 1.59, 95% CI 1.31-1.92). No consistent risk was observed for the other factors.Discussion: Some reproductive and menstrual factors appear to be related to the incidence of bladder cancer among women; but whether effects are due to female hormones is uncertain. (C) 2010 Elsevier Ltd. All rights reserved.

Electrical properties of benign and malignant prostate tissues have been suggested for use in cancer detection. Estimation of spectral parameters that model the dispersive nature of these properties has the potential to provide significant contrast between different prostate tissues. Electrical conductivity and permittivity were gauged at 31 frequencies between 100 Hz and 100 kHz from 800 prostate tissue specimens. The spectral parameters (σ ∞ , σ0, f c , α) best fitting a Cole-based parameterization of the recorded electrical property spectrum were extracted for each specimen probed. The regions probed were histologically assessed in order to assign the quantity of different tissue types making up the region probed. The primary tissue types probed consisted of prostate cancer (CaP), benign prostatic hyperplasia (BPH), nonhyperplastic glandular (Gl), and stroma (Str) tissues. Of the four spectral parameters, significant (p<0.05) positive linear correlations were observed between σ ∞  and f c and cancer content and between σ0 and α and stroma content. The mean σ ∞  and f c in cancer were significantly larger (p<0.0001) than in benign tissues. Of the spectral parameters, f c provided the maximum contrast, being 4.4 times larger in cancer than in benign tissues. Spectral electrical properties provide sufficient levels of cancer discrimination and may potentially enhance disease localization once prostate cancer is suspected. Development of novel technologies gauging these properties in vivo has the potential to provide new tissue characterizing tools for prostate cancer detection and identification.

Use of immunosuppressive drugs post organ transplantation, and prolonged use of glucorticoids for other conditions have been associated with subsequent risk of certain malignancies, that is, skin cancers and lymphoma. There is evidence that the incidence of bladder cancer is also elevated among organ transplant recipients, however, it is unknown whether other groups of patients, that is, those taking oral glucocorticoids, likewise are at an increased risk. In a population-based case–control study in New Hampshire, USA, we compared the use of glucocorticoids in 786 bladder cancer cases and in 1083 controls. We used unconditional logistic regression analysis to compute adjusted odds ratios (ORs) associated with oral glucocorticoid use. In our analysis, the risk of bladder cancer was related to a history of prolonged oral glucocorticoid use (OR=1.85, 95% CI=1.24–2.76, adjusted for age, gender and smoking). Associations with oral glucocorticoid use were stronger for invasive tumours (OR=2.12, 95% CI=1.17–3.85) and tumours with high (3+) p53 staining intensity (OR=2.35, 95% CI=1.26–4.36). Our results raise the possibility of an increased risk of bladder cancer from systemic use of glucocorticoids, and a potential role of immune surveillance in bladder cancer aetiology.

Tissue electrical impedance is a function of its architecture and has been used to differentiate normal and cancer tissues in a variety of organs including breast, cervix, skin, and bladder. This paper investigates the possibility of differentiating normal and malignant prostate tissue using bioimpedance spectra. A probe was designed to measure impedance spectra over the range of 10 kHz to 1 MHz. The probe was fully characterized using discrete loads and saline solutions of different concentrations. Impedance spectra of five ex vivo prostates were measured in the operating room immediately following radical prostatectomy. Wilcoxon signed-rank tests were used to compare the normal and malignant findings. The impedance probe had a signal-to-noise ratio (SNR) > 84 dB across the entire spectrum and measured a tissue volume of approximately 46 mm(3). At 10 kHz, prostate conductivity (or) ranged from 0.232 S/m to 0.310 S/m for tumor and from 0.238 S/m to 0.901 S/m for normal tissue. At 1 MHz the ranges were 0.301 S/m to 0.488 S/m for tumor and 0.337 S/m to 1.149 S/m for normal. Prostate permittivity (epsilonr) ranged from 6.64 x10(4) to 1.25 x 10(5) for tumor and from 9.08 x 10(4) to 4.49 x 10(5) for normal tissues at 10 kHz. And, at 1 MHz the er ranges were 9.23 x 10(2) to 1.88 x 10(3) for tumor and 1.16 x 10(3) to 2.18 x 10(3) for normal tissue. Both sigma and epsilonr of tumor tissue were found to be significantly lower than that of normal tissue (P < 0.0001). Conductivity and permittivity are both higher in normal prostate tissues than they are in malignant tissue making them suitable parameters for tissue differentiation. This is in agreement with trends observed in other tissues reported in much of the literature. Expanded studies are needed to further validate this finding and to explore the biological mechanism responsible for generating the results.

Rinderpest, or cattle plague, is caused by Rinderpest virus (RPV), which is related most closely to human Measles virus (MV), both being members of the genus Morbillivirus , a group of viruses known to have strong immunosuppressive effects in vitro and in vivo . Here, it was shown that peripheral blood mononuclear cells (PBMCs) isolated from cattle experimentally infected with either wild-type or vaccine strains of RPV impaired the proliferation of PBMCs derived from uninfected animals; however, in contrast to either mild or virulent strains of wild-type virus, the inhibition induced by the vaccine was both weak and transient. Flow-cytometric analysis of PBMCs obtained from cattle infected with different strains of RPV showed that the proportion of infected cells was virus dose-dependent and correlated with lymphoproliferative suppression.

ABSTRACT Immune suppression associated with morbillivirus infections may influence the mortality rate by allowing secondary bacterial infections that are lethal to the host to flourish. Using an in vitro proliferation assay, we have shown that all members of the genus Morbillivirus inhibit the proliferation of a human B-lymphoblast cell line (BJAB). Proliferation of freshly isolated, stimulated bovine and caprine peripheral blood lymphocytes is also inhibited by UV-inactivated rinderpest (RPV) and peste-des-petits ruminants viruses. As for measles virus, coexpression of both the fusion and the hemagglutinin proteins of RPV is necessary and sufficient to induce immune suppression in vitro.

Activated B cells and T cells express CD154/CD40 ligand in vitro. The in vivo expression and function of B cell CD154 remain unclear and therefore were examined. Tonsillar B and T cells expressed CD154 at a similar density both in situ and immediately ex vivo, whereas a significantly higher percentage of the former expressed CD154. CD154-expressing B cells were most frequent in the CD38positiveIgD+ pre-germinal center (GC)/GC founder, CD38positive GC and CD38-IgD- memory populations, and were also found in the CD38-IgD+ naive and CD38brightIgD+ plasmablast subsets, but not in the CD38brightIgD- plasma cell subset. B cell expression of CD154 was induced by engaging surface Ig or CD40 by signals that predominantly involved activation of AP-1/NF-AT and NF-kappaB, respectively. The functional importance of CD154-mediated homotypic B cell interactions in vivo was indicated by the finding that mAb to CD154 inhibited differentiation of CD38positiveIgD- GC B cells to CD38-IgD- memory cells. In addition, mAb to CD154 inhibited proliferation induced by engaging sIg or CD40, indicating the role of up-regulation of this molecule in facilitating B cell responsiveness. Of note, CD154 itself not only functioned as a ligand but also as a direct signaling molecule as anti-CD154-conjugated Sepharose beads costimulated B cell responses induced by engaging surface Ig. These results indicate that CD154 is expressed by human B cells in vivo and plays an important role in mediating B cell responses.

PURPOSE:This phase II study was designed to evaluate effectiveness and toxicity of a combined chemoradiotherapy program with selective bladder preservation in the management of patients with invasive bladder cancer.PATIENTS AND METHODS:Ninety-one eligible patients with invasive bladder cancer stages T2M0 to T4AM0 suitable for radical cystectomy received two courses of methotrexate, cisplatin, and vinblastine (MCV regimen) followed by radiotherapy with 39.6 Gy and concurrent cisplatin. After complete urologic evaluation, operable patients who achieved complete response were selected for bladder preservation and treated with consolidation cisplatin-radiotherapy.RESULTS:Of 91 eligible patients, 85 underwent complete urologic evaluation and 68 (75%; 95% confidence interval [CI], 59% to 84%) had documented complete responses. Fourteen operable patients with residual tumor underwent immediate cystectomy. Of 70 patients treated with consolidation cisplatin-radiotherapy, 36 subsequently developed bladder recurrences, 23 of which were invasive. Patients with invasive recurrence (n = 16), extensive noninvasive recurrence (n = 6), or severe treatment complications (n = 1) underwent salvage cystectomy. Thus, a total of 37 of 91 patients (40%) required cystectomy. The 4-year cumulative risk of invasive local failure (which includes induction failures) was 43% (95% CI, 33% to 53%). The 4-year actuarial risk of distant metastasis was 22% (95% CI, 13% to 31%). The 4-year actuarial survival rate of the entire group was 62% (95% CI, 52% to 72%). The 4-year actuarial rate of survival with bladder intact was 44% (95% CI, 34% to 54%).CONCLUSION:Initial results of this combined chemoradiotherapy program show that bladder preservation can be achieved in the majority of patients, and that overall survival is similar to that reported with aggressive surgical approaches. Long-term survival and quality-of-life assessments require longer follow-up study.

PURPOSE:This Phase II study was designed to test the tolerance and effectiveness of concurrent cisplatin-radiotherapy in the treatment of invasive bladder cancer. Objectives were to determine toxicity, complete response rate, bladder preservation rate, and survival.METHODS AND MATERIALS:Patients with invasive bladder cancer, clinical Stages T2-4, NO-2 or NX, MO were treated with pelvic radiotherapy 40 Gy in 4 weeks and cisplatin 100 mg/m2 on days 1 and 22. Complete responders were given an additional 24 Gy bladder boost plus a third dose of cisplatin; patients with residual tumor after 40 Gy were assigned radical cystectomy.RESULTS:The complete remission rate following cisplatin and 40 Gy for evaluable cases was 31/47 (66%). Acute toxicity was acceptable with only two patients not completing induction therapy. Patients with poorly differentiated tumors were more likely to achieve complete remission. Of fully evaluable patients, 28/42 (67%) achieved complete remission with induction therapy, 11 remain continuously in remission, and eight have relapsed with bladder as the only site of failure. Five of these eight cases relapsed with noninvasive tumor. Of the 14 patients who failed to achieve complete remission, only three remain disease-free. Median survival is not reached, with 17/42 (19/48) deaths reported. Actuarial survival is 64% at 3 years.CONCLUSION:This combined cisplatin-radiotherapy regimen was moderately well-tolerated and associated with tumor clearance in 66% of patients treated. Isolated bladder recurrences with invasive carcinoma are infrequent. Better definition of pretreatment selection criteria is needed if combined modality treatment is to achieve disease control and organ preservation for patients with bladder cancer.

Primary amyloidosis of the lower urinary tract is a rare condition with an excellent prognosis in most cases. Three patients with this condition are described. In the cases of localized amyloidosis of the urethra and urinary bladder, the clinical presentation mimicked cancer of the respective sites. This was also true in the case of primary systemic amyloidosis involving the bladder. If significant associated systemic or local disease can be excluded, management is symptomatic and expectant.