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Contains fulltext : 220546.pdf (Publisher’s version ) (Closed access) OBJECTIVE: To provide a contemporary update and recommendations for the diagnosis and management of low-grade non-muscle-invasive bladder cancer (BCa) based on current literature and expert consensus of the International Bladder Cancer Group. METHODS: We reviewed published trials, guidelines, meta-analyses and reviews (up to March 2019) and provide recommendations on baseline evaluations, treatment, endpoints, study design and surveillance protocols. RESULTS: Low-grade Ta BCa poses minimal risk to patients in terms of progression and disease-specific survival. Thus, to minimize patient morbidity, this entity should be managed appropriately. After initial diagnosis of low-grade Ta tumour, subsequent stable, low-grade-appearing recurrences can be managed conservatively with office cystoscopy and fulguration or even followed using an active surveillance protocol. Intravesical therapy other than single-dose peri-operative chemotherapy instillation should be used judiciously, and only after assigning appropriate risk points. Routine use of urinary cytology - other than at initial risk stratification, or for patients on active surveillance without therapy - is not recommended; and surveillance cystoscopy may be discontinued after 5 years. Clinical studies in this group of patients should focus on recurrence rates, and time to recurrence, rather than progression events. CONCLUSIONS: The International Bladder Cancer Group has developed formal recommendations regarding the diagnosis, treatment and surveillance of low-grade non-muscle-invasive BCa to minimize morbidity and encourage uniformity among studies in this disease.

To analyze outcomes and complication rates in an unselected cohort of men with unfavorable (NCCN intermediate and high-risk) PCa receiving combined-modality radiation treatment (CRT). Patients received androgen deprivation therapy for 1 year and combined-modality radiation treatment (CRT) consisting of external-beam radiotherapy (EBRT, 59.4 Gy, 33 fractions) and 125J seed-brachytherapy (S-BT, 100 Gy). Subgroups, including WHO group 3–5, and initial PSA (iPSA) 20 ng/ml were identified. Biochemical recurrence-free (BRFS), metastasis-free (MFS), cancer-specific (CSS) and overall survival (OS) were calculated at 5 and 10 years using the Kaplan–Meier method. Subgroups were compared using log-rank test and Cox proportional hazards regression. Urogenital and gastrointestinal side-effects were reported according to the CTCAE classification. After a median of 6.9 years (range 2–13) calculated 5- and 10-year rates for the whole cohort of 425 men were 92.8% and 82.5% for BRFS, 95.1%, and 88.8% for MFS, 98.2%, and 95.1 for CSS, and 95.4%, and 80.1% for OS, respectively. Univariate (UVA) and multivariate analysis (MV) identified a group with unfavorable outcome with iPSA > 20 ng/ml, comprising 24% of all patients, in which 55% of recurrences, 54% of metastases and 71% of cancer-specific deaths occurred. Side-effects were limited, with 20 ng/ml further, possibly systemic, treatment options should be identified.

Foodborne pathogens are responsible for hundreds of thousands of deaths around the world each year. Rapid screening of agricultural products for these pathogens is essential to reduce and/or prevent outbreaks and pinpoint contamination sources. Unfortunately, current detection methods are laborious, expensive, time-consuming and require a central laboratory. Therefore, a rapid, sensitive, and field-deployable pathogen-detection assay is needed. We previously developed a colorimetric sandwich immunoassay utilizing immuno-magnetic separation (IMS) and chlorophenol red-β-D-galactopyranoside for Salmonella detection on a paper-based analytical device (μPAD); however, the assay required many sample preparation steps prior to the μPAD as well as laboratory equipment, which decreased user-friendliness for future end-users. As a step towards overcoming these limitations in resource-limited settings, we demonstrate a reusable 3D-printed rotational manifold that couples with disposable μPAD layers for semi-automated reagent delivery, washing, and detection in 65 minutes. After IMS to clean the sample, the manifold performs pipette-free reagent delivery and washing steps in a sequential order with controlled volumes, followed by enzymatic amplification and colorimetric detection using automated image processing to quantify color change. Salmonella was used as the target pathogen in this project and was detected with the manifold in growth media and milk with detection limits of 4.4 × 102 and 6.4 × 102 CFU mL−1 respectively. The manifold increases user friendliness and simplifies immunoassays resulting in a practical product for in-field use and commercialization.

Salmonella causes over a million foodborne illnesses per year in the United States resulting in more hospitalizations and deaths than any other foodborne bacterial pathogen. To help prevent outbreaks, a rapid, portable, sensitive, and reliable method for onsite detection of bacteria that can be used in different sample matrices would be beneficial. Herein, we present a colorimetric paper-based analytical device (PAD) combined with immunomagnetic separation (IMS) for detecting Salmonella typhimurium. IMS anti-Salmonella coated magnetic beads were applied to capture and separate bacteria from the sample matrix and preconcentrate it into small volumes before testing on paper. To directly detect S. typhimurium after IMS, a sandwich immunoassay was implemented into the procedure with β-galactosidase (β-gal) as the detection enzyme. Using the antibody/enzyme complex, we performed a colorimetric assay with chlorophenol red-β-d-galactopyranoside (CPRG) for bacteria quantification. The method was confirmed to be highly specific to S. typhimurium without interference from other pathogenic bacteria like Escherichia coli. Using this system, the limit of detection of S. typhimurium was found to be 10

Item does not contain fulltext Intravesical immunotherapy with live attenuated BCG remains the standard of care for patients with high-risk and intermediate-risk non-muscle-invasive bladder cancer (NMIBC). Most patients initially respond, but recurrence is frequent and progression to invasive cancer is a concern. No established and effective intravesical therapies are available for patients whose tumours recur after BCG, representing a clinically important unmet need. Development and discovery of treatment options for BCG-unresponsive NMIBC is a high priority in order to decrease the morbidity, burden of health-care expenditures, and mortality related to bladder cancer. This Review of treatment options after BCG failure focuses on principles of optimal management emerging therapies, thus enabling a synthesis of recommendations for management for such patients.

Purpose To compare a standard radio-oncological and a surgical biochemical failure definition after combined-modality radiation therapy (CRT) in men with intermediate- and high-risk prostate cancer. Methods 425 men were treated with external beam radiotherapy (59.4 Gy, 33 fractions) and 125J seed-brachytherapy (S-BT, 100 Gy). Biochemical recurrence (BR) was defined either as radio-oncologic (rBR), using a +2 ng/mL prostate-specific antigen (PSA) increase above a nadir value, or as surgical (sBR), using a 2-year posttreatment PSA of ≥0.2 ng/mL. Biochemical recurrence-free, metastasis-free, cancer-specific, and overall survival were calculated at 5 and 10 years using the Kaplan-Meier method. Standard validation tests were used to compare both thresholds. Results After a median of 7 years, overall recurrence rates were 10.4% and 31.5% for rBR and sBR definitions, respectively. Both failure definitions proved sensitive for the prediction of metastases and cancer-specific death, whereas the rBR definition was significantly more specific. The accuracies of a correct prediction of metastases and death of prostate cancer were 73.1% vs. 96.2% and 72.2% vs. 92.9% for sBR vs. rBR, respectively. The inferior validity results of the sBR definition were attributable to a PSA-bounce phenomenon occurring in 56% of patients with sBR. Still, using the less suitable sBR definition, the results of CRT compared favorably to BRFS rates of surgical interventions. Conclusion After CRT, the radio-oncological (aka Phoenix) failure definition is more reliable than a fixed surgical endpoint. Exclusively in high-risk patients, sBR offers a direct comparison across surgical and nonsurgical treatment options at 5 and 10 years.

Falsified and substandard antibiotics are a growing worldwide problem that leads to increased patient mortality and decreased trust in healthcare, and contributes to antimicrobial resistance. Monitoring falsified antibiotics is difficult because most falsified pharmaceuticals are most commonly found in developing countries, where detecting the active ingredient is difficult due to lack of access to complex instrumentation. Herein, we describe the development and optimization of a microfluidic paper-based analytical device (μPAD) to detect the active ingredient in the most falsified class of antibiotics, β-lactams. The assay is based on enzyme competition, making it the first demonstrated competitive enzyme assay reported in paper-based devices. The assay uses nitrocefin, a chromogenic substrate, to compete with β-lactam antibiotics in a reaction with β-lactamase. A yellow color indicates legitimate drugs, while a color change from yellow to red indicates falsified drugs. In addition to testing for the active ingredient, another section of the device was added to test the sample pH to further verify results and identify common falsified ingredients like aspirin or baking soda. Calibration curves for four different antibiotics, including cefazolin, have been generated making it the first paper-based device capable of detecting both cephalosporin and penicillin antibiotics. The μPAD has also been tested with common falsified ingredients and four antibiotics in tablet or injectable form, demonstrating its potential for in-field falsified antibiotic testing.

Purpose The aim of this study was to assess a non-endoscopic transoral versus extraoral technique in the open reduction and internal fixation of displaced or dislocated fractures of the condylar neck. Material and methods A total of 104 patients, treated from 2007 to 2012 with 114 class II or class IV fractures according to Spiessl and Schroll were included in this study. Facial nerve function, scarring, pain and functional clinical parameters, such as protrusion, mediotrusion and maximum interincisal distance, were judged clinically (at 21 ± 12.1 months); repositioning and reossification were measured upon preoperative, postoperative and follow-up (at 8.8 ± 7 months) radiographs. Patient satisfaction was evaluated using the OHIP-G 14 questionnaire. Results In all, 36 patients (35%) with 43 fractures (38%) presented for clinical follow-up. Both treatment groups showed clinically and radiologically comparable results. Scarring was obvious in all extraorally treated patients, and hypertrophic scars occurred in four class IV cases (24%). One class IV patient (6%) had a persistent facial nerve palsy; temporary pareses were more frequent ( n = 4; 24%). Conclusion The transoral approach did not jeopardize facial nerve function, and extraoral scars were avoided. Repositioning and fixation results and the frequency of revision operations were comparable. The transoral approach can be recommended generally in class II and class IV cases.

The development of transparency-based electrochemical and paper-based colorimetric analytic detection platforms is presented as complementary methods for food and waterborne bacteria detection from a single assay. Escherichia coli and Enterococcus species, both indicators of fecal contamination, were detected using substrates specific to enzymes produced by each species. β-galactosidase (β-gal) and β-glucuronidase (β-glucur) are both produced by E. coli, while β-glucosidase (β-gluco) is produced by Enterococcus spp. Substrates used produced either p-nitrophenol (PNP), o-nitrophenol (ONP), or p-aminophenol (PAP) as products. Electrochemical detection using stencil-printed carbon electrodes (SPCEs) was found to provide optimal performance on inexpensive and disposable transparency film platforms. Using SPCEs, detection limits for electrochemically active substrates, PNP, ONP, and PAP were determined to be 1.1, 2.8, and 0.5 μM, respectively. A colorimetric paper-based well plate system was developed from a simple cardboard box and smart phone for the detection of PNP and ONP. Colorimetric detection limits were determined to be 81 μM and 119 μM for ONP and PNP respectively. While colorimetric detection methods gave higher detection limits than electrochemical detection, both methods provided similar times to positive bacteria detection. Low concentrations (10

Context: The European Association of Urology (EAU) Guideline Group for urothelial cell carcinoma of the upper urinary tract (UUT-UCC) has prepared new guidelines to aid clinicians in assessing the current evidence-based management of UUT-UCC and to incorporate present recommendations into daily clinical practice. Objective: This paper provides a brief overview of the EAU guidelines on UUT-UCC as an aid to clinicians in their daily practice. Evidence acquisition: The recommendations provided in the current guidelines are based on a thorough review of available UUT-UCC guidelines and papers identified using a systematic search of Medline. Data on urothelial malignancies and UUT-UCCs in the literature were searched using Medline with the following keywords: urinary tract cancer, urothelial carcinomas, upper urinary tract, carcinoma, transitional cell, renal pelvis, ureter, bladder cancer, chemotherapy, nephroureterectomy, adjuvant treatment, neoadjuvant treatment, recurrence, risk factors, and survival. A panel of experts weighted the references. Evidence synthesis: There is a lack of data in the current literature to provide strong recommendations due to the rarity of the disease. A number of recent multicentre studies are now available, whereas earlier publications were based only on limited populations. However, most of these studies have been retrospective analyses. The TNM classification 2009 is recommended. Recommendations are given for diagnosis as well as for radical and conservative treatment; prognostic factors are also discussed. Recommendations are provided for patient follow-up after different therapeutic options. Conclusions: These guidelines contain information for the diagnosis and treatment of individual patients according to a current standardised approach. When determining the optimal treatment regimen, physicians must take into account each individual patient's specific clinical characteristics with regard to renal function including medical comorbidities; tumour location, grade and stage; and molecular marker status. (C) 2011 European Association of Urology. Published by Elsevier B. V. All rights reserved.

Resumen Contexto El Grupo de Guias Clinicas sobre el carcinoma de celulas uroteliales de las vias urinarias superiores (CCU-VUS) de La Asociacion Europea de Urologia (EAU) ha elaborado una nueva guia clinica para ayudar a los medicos a evaluar el tratamiento del CCU-VUS basado en los datos cientificos, y a incorporar las presentes recomendaciones a la practica clinica diaria. Objetivo Este documento ofrece una breve vision general de la guia clinica de la EAU sobre el CCU-VUS como una ayuda para los medicos en su practica diaria. Adquisicion de datos cientificos Las recomendaciones proporcionadas en la guia actual se basan en una meticulosa revision de las guias y documentos sobre CCU-VUS disponibles, identificados mediante una busqueda sistematica en Medline. Los datos sobre neoplasias uroteliales malignas y CCU-VUS en la literatura se buscaron mediante Medline con las siguientes palabras clave: cancer del tracto urinario, carcinomas uroteliales, tracto urinario superior, carcinoma, celulas de transicion, pelvis renal, ureter, cancer vesical, quimioterapia, nefroureterectomia, tratamiento adyuvante, tratamiento neoadyuvante, recidiva, factores de riesgo y supervivencia. Un equipo de expertos sopeso las referencias. Sintesis de los datos cientificos Hay una falta de datos en la literatura actual para proporcionar recomendaciones consistentes, debido a la rareza de la enfermedad. Una serie de recientes estudios multicentricos ya estan disponibles, mientras que las publicaciones anteriores se basaban solo en poblaciones limitadas. Sin embargo, la mayoria de estos estudios han sido analisis retrospectivos. Se recomienda la clasificacion TNM2009. Se hacen recomendaciones para el diagnostico, asi como para el tratamiento radical y conservador; tambien se tratan los factores pronosticos. Se proporcionan recomendaciones para el seguimiento del paciente despues de diferentes opciones terapeuticas. Conclusiones Esta guia contiene informacion para el diagnostico y tratamiento de los pacientes individuales de acuerdo a un enfoque estandar actual. Al determinar el regimen de tratamiento optimo, los medicos deben tener en cuenta las caracteristicas clinicas especificas de cada paciente con respecto a la funcion renal, incluyendo comorbilidades medicas, localizacion del tumor, grado y estadio y el estado de los marcadores moleculares.

Evidence in humans and rodents suggests that normal circadian rhythmicity is important for supporting reproductive function. A molecular clock underlies circadian rhythmicity. Impaired fertility is observed in some genetically altered mice with deficiencies in genes of the molecular clock, suggesting a critical role for these genes in reproduction. Here we systematically characterize the reproductive phenotype of females deficient in the clock gene Bmal1. Bmal1(-/-) females are infertile. They exhibit progression through the estrous cycle, although these cycles are prolonged. Normal follicular development occurs in Bmal1(-/-) females, and healthy embryos of the expected developmental stage are found in the reproductive tract of Bmal1(-/-) females 3.5 d after mating to wild-type males. However, serum progesterone levels are significantly lower in Bmal1(-/-) vs. Bmal1(+/+/-) females on d 3.5 of gestation. Low progesterone levels in Bmal1(-/-) females are accompanied by decreased expression of steroidogenic acute regulatory protein in corpora lutea of Bmal1(-/-) vs. Bmal1(+/+/-) females. Whereas implantation of embryos is not observed in untreated or vehicle-treated Bmal1(-/-) females, exogenous administration of progesterone to Bmal1(-/-) females is able to reinstitute implantation. These data suggest that implantation failure due to impaired steroidogenesis causes infertility of Bmal1(-/-) females.

The administration of endostatin, a potent anti-angiogenic agent, will be required for extended periods of time as a cancer treatment. The aim of the present study was to induce endogenous endostatin secretion in a continuous fashion, based on retroviral gene transfer. The tumor response was evaluated in an orthotopic murine tumor model of human lung cancer. Human non-small-cell lung cancer cells (KNS 62) were retrovirally transduced with the human endostatin gene. An orthotopic murine xenotransplant model was used to investigate tumor growth, metastases and survival. After 4 weeks of subcutaneous growth, endostatin expression was measured by immunoblot analysis in tumor lysates. The growth of the subcutaneous tumors was significantly delayed, and orthotopic tumor growth and pleural metastases were significantly reduced in endostatin-transduced KNS 62 tumors. Prolongation of survival subsequent to orthotopic tumor induction was demonstrated. Strong endostatin expression was found in subcutaneous tumors after 4 weeks. Retroviral transduction of the human endostatin gene is capable of achieving long-term endostatin expression. Endostatin transduction provides significant anti-tumor effects with regard to local tumor growth, metastases and survival.

Summary Human induced pluripotent stem cells (iPSCs) from a healthy 86-year-old male were differentiated into neural stem cells and grafted into adult immunodeficient rats after spinal cord injury. Three months after C5 lateral hemisections, iPSCs survived and differentiated into neurons and glia and extended tens of thousands of axons from the lesion site over virtually the entire length of the rat CNS. These iPSC-derived axons extended through adult white matter of the injured spinal cord, frequently penetrating gray matter and forming synapses with rat neurons. In turn, host supraspinal motor axons penetrated human iPSC grafts and formed synapses. These findings indicate that intrinsic neuronal mechanisms readily overcome the inhibitory milieu of the adult injured spinal cord to extend many axons over very long distances; these capabilities persist even in neurons reprogrammed from very aged human cells. Video Abstract

Background . Combretastatin A-4 prodrug (CA-4PD) has been identified as a potent antivascular agent in various rodent tumor models. The aim of this study was to investigate the effect of CA-4PD on human non–small cell lung cancer (NSCLC). Methods . Cytostatic and cytotoxic effects of CA-4PD on selected NSCLC cells, Colo-699 and KNS-62, were studied in vitro. After subcutaneous xenotransplantation the effect of systemically administrated CA-4PD on tumor growth was investigated in vivo. A newly established orthotopic xenotransplant model was employed to estimate prolongation of survival after intrapulmonary tumor induction with secondary metastatic disease. Results . In vitro, CA-4PD displayed a time and dose dependent antiproliferative effect on human lung cancer cells. In vivo, CA-4PD significantly delayed growth of subcutaneously induced lung cancer. This growth delay was translated into a prolongation of survival in the metastasizing orthotopic xenotransplant model. Conclusions . In vitro CA-4PD inhibits proliferation of NSCLC cells, most likely by disruption of microtubule assembly. In vivo, systemic treatment inhibits growth of subcutaneously xenotransplanted tumors by an antivascular effect. In the case of metastasizing human lung cancer this translated into a prolongation of survival.

Asthma is among the most frequent chronic diseases in childhood. Although numerous environmental risk factors have already been identified, the basis for familial occurrence of asthma remains unclear. Previous genome screens for atopy in British/Australian families and for asthma in different American populations showed inconsistent results. We report a sib pair study of a sample of 97 families, including 415 persons and 156 sib pairs. Following an extensive clinical evaluation, all participants were genotyped for 351 polymorphic dinucleotide markers. Linkage analysis for asthma identified four chromosomal regions that could to be linked to asthma: chromosome 2 (at marker D2S2298,P= 0.007), chromosome 6 (around D6S291, lowestP= 0.008), chromosome 9 (proximal to D9S1784,P= 0.007), and chromosome 12 (D12S351,P= 0.010). These linkage regions could be reproduced for all loci by analysis of total or specific immunoglobulin E (minimumPvalues at these regions were 0.003, 0.001, 0.010, and 0.015, respectively).

Eight DSM-III-R female schizophrenics were investigated by magneto-encephalographically (MEG) detected long latency (N100m) auditory evoked fields (AEFs). In contrast to controls, in schizophrenics a lack of interhemispheric asymmetry was present, which was found to be due to alterations of the right hemisphere.

Purpose The combination of gemcitabine plus cisplatin (GC) is a standard regimen in patients with locally advanced or metastatic urothelial cancer. A phase I/II study suggested that a three-drug regimen that included paclitaxel had greater antitumor activity and might improve survival. Patients and Methods We conducted a randomized phase III study to compare paclitaxel/cisplatin/gemcitabine (PCG) with GC in patients with locally advanced or metastatic urothelial carcinoma. Primary outcome was overall survival (OS). Secondary outcomes were progression-free survival (PFS), overall response rate, and toxicity. Results From 2001 to 2004, 626 patients were randomly assigned; 312 patients were assigned to PCG, and 314 patients were assigned to GC. After a median follow-up of 4.6 years, the median OS was 15.8 months on PCG versus 12.7 months on GC (hazard ratio [HR], 0.85; P = .075). OS in the subgroup of all eligible patients was significantly longer on PCG (3.2 months; HR, 0.82; P = .03), as was the case in patients with bladder primary tumors. PFS was not significantly longer on PCG (HR, 0.87; P = .11). Overall response rate was 55.5% on PCG and 43.6% on GC (P = .0031). Both treatments were well tolerated, with more thrombocytopenia and bleeding on GC than PCG (11.4% v 6.8%, respectively; P = .05) and more febrile neutropenia on PCG than GC (13.2% v 4.3%, respectively; P < .001). Conclusion The addition of paclitaxel to GC provides a higher response rate and a 3.1-month survival benefit that did not reach statistical significance. Novel approaches will be required to obtain major improvements in survival of incurable urothelial cancer.

Herr and Dalbagni aptly noted that comparisons between therapies for BCG failure patients have been hampered by the lack of standard definitions for BCG failure and BCG-refractory transitional cell carcinoma (TCC) [2]. Some series have defined BCG failure after a single induction course of BCG [3, 4], others after two courses [5]. In addition, the methods of reporting the results have been inconsistent. Most studies have included all patients who received one or more courses of BCG [6, 7, 8]. Investigators have often combined patients with persistent disease (non-responders) and patients with recurrent disease after an initial response [3, 4, 5] and a few studies have combined patients who were non-responders to BCG and patients who could not complete BCG therapy because of toxicity (BCG intolerant) [5, 6]. Finally, most studies did not indicate the disease-free interval after the last BCG course. These inconsistencies have led to comparisons of outcome in a very heterogeneous population. In the most general sense any recurrent disease after initiation of BCG therapy can be referred to as “BCG failure.” However, to provide more uniformity in reporting, the following alternative descriptive terms for specific types of BCG failure should be used whenever possible: BCG refractory Failure to achieve a disease-free state by 6 months after initial BCG therapy with either maintenance or re-treatment at 3 months due to either persistent or rapidly recurrent disease. Also includes any progression in stage, grade, or disease extent by 3 months after first cycle of BCG, i.e., non-improving or worsening disease despite BCG. BCG resistant Recurrence or persistence of disease at 3 months after induction cycle but of lesser degree, stage or grade which subsequently is no longer present at 6 months from BCG re-treatment ± TUR, i.e., disease improves then resolves with further BCG. BCG relapsing Recurrence of disease after achieving a disease-free status by 6 months, i.e., disease resolves after BCG then returns. Relapse is further defined by time of recurrence: early (within 12 months); intermediate (12–24 months); late (>24 months). Caution: relapsing disease while on active maintenance (within 3 months) may qualify as BCG refractory. BCG intolerant Disease recurrence after a less than adequate course of therapy is applied due to a serious adverse event or symptomatic intolerance that mandates discontinuation of further BCG, i.e., recurrent disease in setting of inadequate BCG treatment from drug toxicity.

Invasive pulmonary aspergillosis (IPA) is a life-threatening infection in immunocompromised patients. Mortality rates of cerebrally disseminated IPA approach 100%. We report on a case of a 9-year-old girl with acute myeloid leukemia, who acquired cerebrally disseminated IPA during chemotherapy-induced leukopenia. Longtime survival was achieved by left pneumonectomy and neurosurgical resection of the intracerebral lesion combined with systemic application of itraconazole and liposomal amphotericin B. A review of literature revealed 7 other cases of cerebrally disseminated IPA with survival of more than 12 months.

Objectives: On behalf of the European Association of Urology (EAU), guidelines for the diagnosis, therapy and follow-up of patients with urothelial carcinoma in situ (CIS) have been established. Method: The recommendations in these guidelines are based on a recent comprehensive overview and meta-analysis in which two panel members have been involved (RS and AVDM). A systematic literature search was conducted using Medline, the US Physicians' Data Query (PDQ), the Cochrane Central Register of Controlled Trials, and reference lists in trial publications and review articles. Results: Recommendations are provided for the diagnosis, conservative and radical surgical treatment, and follow-up of patients with CIS. Levels of evidence are influenced by the lack of large randomized trials in the treatment of CIS.

The Ki-67 protein is a nuclear and nucleolar protein that is strictly associated with cell proliferation. Recently it has been suggested to play a role in the control of the higher order chromatin structure (1). Because the protein is produced only in dividing cells, the anti Ki-67 antibody MIB-1 has been widely used in histopathologic studies to estimate the growth fraction of human neoplastic tissue samples in situ. For a variety of human tumors, including bladder carcinomas, the Ki-67 labeling index has been shown to be of prognostic value for tumor recurrence and for patient survival (2)(3)(4). We investigated whether Ki-67 RNA in total urine of bladder tumor patients is correlated with the Ki-67 labeling index of the corresponding tumor tissue. Spontaneously voided clean-catch urines from 68 patients were collected with informed consent, stored at 4 °C, and processed within 4 h of collection. Approval was obtained from the Ethikkommission of the University of Luebeck. Three groups of patients were included: ( a ) healthy donors (n = 14); ( b ) patients with urinary tract infection as detected by analysis of urine sediment (n = 28); and ( c ) patients with bladder carcinoma (n = 26). All urine samples from patients with tumors were checked for significant bacteriuria or leukocyturia (>10/μL) as well as for the presence of erythrocytes (>10/μL). The specific weights of all urine samples were in the range 1.010–1.015 kg/L, showing that there were no major differences in urine concentration. After routine diagnostics were completed, 1 mL of urine was mixed 1:1 with lysis buffer [5.64 mol/L guanidinium thiocyanate, 5 g/L sarcosyl, 50 mmol/L sodium acetate (pH 6.5), 1 mmol/L β-mercaptoethanol], the pH was adjusted to 7 by addition of 1.5 mol/L HEPES (pH 8.0), and samples were frozen at −80 °C until RNA extraction was performed. …

This study assesses the chemotherapeutic drug gemcitabine in the human non-small cell lung cancer (NSCLC) cell line KNS62 in relation to the CD95-induced apoptotic pathway, and the role of the anti-apoptotic protein Bcl-xL in vitro and in vivo. Apoptosis was determined by JAM assay and DAPI staining analysis. Activation of key apoptotic proteins, including caspases 3, 8 and 9 and BID, as well as cytochrome c release and mitochondrial transmembrane potential (MTP), were measured. The impact of the caspase inhibitor zVAD on gemcitabine-induced apoptosis was quantified. The in vitro results were verified in vivo in an orthotopic murine xenotransplantation model. Gemcitabine treatment, as well as stimulation of CD95, resulted in cleavage of effector caspase 3 as well as its substrate PARP and caspase 9, followed by DNA fragmentation. Cleavage of caspase 8 was demonstrated after CD95 activation but not after the application of gemcitabine. In KNS62-Bcl-xL clones, release of cytochrome c and loss of mitochondrial transmembrane potential were suppressed. Consequently, apoptosis after gemcitabine therapy, as well as CD95-induced apoptosis, were significantly inhibited. Caspase inhibitor zVAD only partly reversed gemcitabine-induced DNA fragmentation. In vivo, there was a significant reduction in tumour volume under gemcitabine therapy. Bcl-xL over-expressing tumours were completely resistant to gemcitabine therapy. In NSCLC cell line KNS62 gemcitabine activated the mitochondrial apoptotic pathway downstream of mitochondria without activation of initiator caspases. Bcl-xL over-expression induced significant resistance to gemcitabine. In vivo, the anti-apoptotic effect of Bcl-xL was more pronounced than in vitro. Gemcitabine also induced caspase-independent DNA fragmentation in KNS62 cells.

Background . Therapy failures have been reported for retroviral gene transfer of herpes simplex virus thymidine kinase (HSV-TK) gene followed by systemic ganciclovir application in human lung cancer. Use of the HSV-TK mutant TK 30 in combination with a VSV-G pseudotyped retroviral vector was found to enhance the efficacy of prodrug therapy. The present study evaluated this therapeutic strategy in human non-small cell lung cancer cell lines in a preclinical murine xenotransplant model. Methods . Intrathoracally tumors induced by HSV-TK 30 transduced non-small cell lung cancer cell lines Colo 699 (adenocarcinoma) and KNS 62 (squamous cell carcinoma) were monitored for local tumor growth, survival, and metastases. So-called bystander effects were investigated in tumors consisting of as little as 25% TK 30 transfected cells and by analysis of gap junctional protein connexin-43 expression. Results . Survival was significantly prolonged, and tumor growth and pleural metastases were reduced in HSV-TK 30 -positive tumors of both cell lines. A significant therapeutic effect in bystander experiments was observed in squamous cell carcinoma. This was correlated with higher expression of connexin-43. Conclusions . Delivery of HSV-TK 30 in a VSV-G pseudotyped retroviral vector and subsequent ganciclovir application provided therapeutic efficacy. Despite of low transduction rates achievable in gene transfer in situ, prodrug therapy appears to be feasible in tumor cells with efficient bystander effects.

Background and aims. Recently we demonstrated that phosphatidylinositol 3-kinase (PI3K) is overexpressed in human lung cancer. This study evaluated whether the PI3K inhibiting agent wortmannin affects proliferation of human lung cancer cells in vitro and in vivo. Methods. Effects of exposure of human non-small-cell lung cancer (NSCLC) cells (KNS-62, Colo-699) to wortmannin were investigated in vitro by proliferation, cytotoxicity, and DNA fragmentation assays. In vivo we examined the effects of blocking PI3K by wortmannin prior to xenotransplantation of human NSCLC cells into SCID-bg mice and the effect of systemic wortmannin administration following intrapulmonary xenotransplantation of human NSCLC. Results. Exposure of KNS-62 and Colo-699 lung cancer cells to wortmannin inhibited proliferation in correlation to concentration in vitro. In vivo the blocking of PI3K by wortmannin prior to xenotransplantation caused a significant delay in the growth of subcutaneously induced tumors. Systemic wortmannin administration increased mean survival after intrapulmonary xenotransplantation of human NSCLC significantly by 38% and 47%. Conclusions. These data suggest inhibition of PI3K activity as a potential target for treatment of human NSCLC. Systemic toxicity of wortmannin requires development of improved PI3K inhibitors with favorable pharmacological properties.

Overall prognosis in human NSCLC remains poor. Antiangiogenic treatment has become a promising concept for the treatment of solid malignancies. Our purpose was to evaluate the efficacy of recombinant HSENDO for the treatment of human NSCLC in an orthotopic murine xenotransplantation model. The efficacy of HSENDO was tested in vitro in cell-proliferation, cell-migration and tube-formation assays. In vivo, the effect of HSENDO on tumor growth was tested in s.c. xenotransplanted human NSCLC and on intrapulmonary induced human NSCLC. In vitro, HSENDO inhibited both human and rodent endothelial cell proliferation in a time- and dose-dependent fashion. Endothelial cell migration was inhibited by 97%. Tube formation of murine endothelial cells was inhibited and preexisting tubes degenerated after HSENDO exposure. In vivo, HSENDO delayed growth of s.c. xenotransplanted tumors. Immunohistochemic staining demonstrated no change in microvessel density but a significant reduction of proliferating tumor cells and an increase in bFGF and VEGF expression, reflecting the antiangiogenic effect of HSENDO. Intrapulmonary tumor induction caused death subsequent to metastatic disease. Systemic HSENDO application extended survival significantly. HSENDO was demonstrated to inhibit endothelial cell proliferation, migration and tube formation effectively. In vivo growth of s.c. transplanted tumors was delayed and survival extended by 32% and 69%, respectively, after intrapulmonary NSCLC induction.

Background . For effective palliation of patients with malignant pleural effusion due to advanced neoplastic disease, any proposed treatment should have low procedure-related mortality and morbidity. Methods . The clinical outcome of 119 thoracoscopies in 101 patients (56 women, 45 men), from 42 to 91 years of age (mean, 68 ± 9 years) with malignant pleural effusions was evaluated in a retrospective study. Video-assisted thoracoscopy (VATS) talc pleurodesis was done in 105 instances, and a pleuroperitoneal shunt was performed 14 times as an alternative when complete expansion of the lung could not be achieved due to tumor implants on the visceral pleura. Results . The VATS talc pleurodesis resulted in clinically significant improvement of dyspnea in 92.2% of the patients. Thirty-day mortality was 2.8% and morbidity was 2.8%. The mean duration of postoperative survival was 6.7 months. Recurrent pleural effusion occurred in 5.7% of patients after a mean interval of 6 months. Clinical relief of dyspnea was obtained in 73% of the patients treated with pleuroperitoneal shunts. Thirty-day mortality in this group was 21% and morbidity was 14.3%. The mean duration of survival was 4.2 months. Conclusions . The VATS talc pleurodesis is appropriate for palliation of patients with malignant pleural effusions and should be performed once the diagnosis has been confirmed. Patients with lungs trapped by visceral carcinomatosis may benefit from placement of a pleuroperitoneal shunt as an alternative.

Background . Overall prognosis in human lung cancer is still poor. A highly reproducible, easy to perform in vivo model, which closely resembles the clinical features of advanced human lung cancer, is required for the evaluation of novel therapies. Methods . Tumor cells, originated from a human adenocarcinoma, a squamous cell carcinoma, and an undifferentiated large cell carcinoma, were xenotransplanted heterotopically by subcutaneous and intravenous injection and compared with orthotopic intrapleural and intrapulmonary xenotransplantation by a facilitated engraftment procedure into SCID bg mice. Results . Subcutaneous injection of tumor cells resulted in a 100% engraftment rate with establishment of solid tumors without clinically relevant metastases. Intravenous injection had poor engraftment rates by hematogenous spread. Depending on the cell line, a 80% to 100% engraftment rate in orthotopic xenotransplantation was achieved, resulting in a consistent pattern of mediastinal and bilateral pulmonary metastases. Conclusions . The facilitated orthotopic xenotransplantation of human lung cancer is easy to perform and results in a reproducible in vivo model that closely resembles the clinical features of advanced human lung cancer. Consequently, this model appears suitable for in vivo evaluation of novel cancer therapies in preclinical tests.

Functional and structural changes in 10 DSM-III-R male schizophrenics and 10 healthy volunteers were investigated using magnetoencephalographically (MEG) detected long-latency (N100 m) auditory evoked fields (AEFs) and magnetic resonance imaging (MRI). The AEFs were characterized by single moving equivalent dipoles, which were superimposed on MRIs. There were significant differences in dipole orientations and in AEF latencies in the left hemisphere of schizophrenics, when compared to the controls. The MEG-detected alterations were found to be associated with a bilateral volume reduction of the posterior superior temporal gyrus (pSTG), which was more pronounced in the left hemisphere. Separate analysis of white and gray matter has shown that the pSTG volume reduction resulted from decreased gray matter volumes without white matter changes. Both the functional and the morphological data indicate a left-hemispheric disturbance in our patients.

4685 Background: Based on the results of our monocenter phase I study of intravesical administration of gemcitabine (Gemzar) immediately after transurethral resection of superficial bladder cancer ...

Two chimpanzees were born to parents with chronic non-A, non-B hepatitis and remained with their mothers until 12 and 18 months, respectively. The infants were followed from 7 to 8 weeks of age with biweekly or monthly blood samples and with monthly liver biopsies from 4 to 7 months after birth. Another chimpanzee, along with both of its parents, was held throughout the parents' acute infection with non-A, non-B hepatitis; at this time the infant was 14-16 months of age, and it was followed with bi-weekly blood samples and monthly biopsies from the time of potential exposure for 20 months. No abnormalities indicative of non-A, non-B hepatitis were detected in these animals. During the 29 to 35 months of follow-up, alanine aminotransferases and gamma glutamyl-transferases (GGPT) levels remained well within normal range for animals held in the same facility. Histologic and electron microscopic examination of liver tissue revealed no abnormalities.

We analyzed the outer cell membranes of normal kidney cells, hypernephroma cells, nude mouse transplants of hypernephroma tissue, and cell lines derived from hypernephroma using horseradish peroxidase-coupled lectins of different specificity, in order to investigate their binding to cryostat sections or the cell surfaces of suspended single cells. We showed that hypernephroma cells express higher amounts of receptors for Robinia pseudoacacia lectin, soy bean lectin and Phaseolus vulgaris lectin compared to those prepared from normal kidneys. In contrast, Helix pomatia lectin is bound to connective tissue elements of both malignant and non-malignant tissue exclusively. L-fucose and D-galactose residues are detectable on malignant cells, but not on their non-malignant counterparts. However, the expression of the latter varies considerably among cells from different hypernephromas. No significant difference could be demonstrated between hypernephroma, hypernephroma cell lines and hypernephroma transplants in regard to the carbohydrate profile on the cell surfaces.