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Prostate cancer treatment planning is largely dependent upon examination of core-needle biopsies. The microscopic architecture of the prostate glands forms the basis for prognostic grading by pathologists. Interpretation of these convoluted three-dimensional (3D) glandular structures via visual inspection of a limited number of two-dimensional (2D) histology sections is often unreliable, which contributes to the under- and overtreatment of patients. To improve risk assessment and treatment decisions, we have developed a workflow for nondestructive 3D pathology and computational analysis of whole prostate biopsies labeled with a rapid and inexpensive fluorescent analogue of standard hematoxylin and eosin (H&E) staining. This analysis is based on interpretable glandular features and is facilitated by the development of image translation–assisted segmentation in 3D (ITAS3D). ITAS3D is a generalizable deep learning–based strategy that enables tissue microstructures to be volumetrically segmented in an annotation-free and objective (biomarker-based) manner without requiring immunolabeling. As a preliminary demonstration of the translational value of a computational 3D versus a computational 2D pathology approach, we imaged 300 ex vivo biopsies extracted from 50 archived radical prostatectomy specimens, of which, 118 biopsies contained cancer. The 3D glandular features in cancer biopsies were superior to corresponding 2D features for risk stratification of patients with low- to intermediate-risk prostate cancer based on their clinical biochemical recurrence outcomes. The results of this study support the use of computational 3D pathology for guiding the clinical management of prostate cancer. Significance: An end-to-end pipeline for deep learning–assisted computational 3D histology analysis of whole prostate biopsies shows that nondestructive 3D pathology has the potential to enable superior prognostic stratification of patients with prostate cancer.

Recent technological advances have demonstrated the feasibility of achieving high-throughput slide-free three-dimensional (3D) pathology of biopsy and surgical specimens. In comparison to conventional slide-based pathology, 3D pathology has the potential to provide a transformative improvement in diagnostic performance for a number of reasons: (1) vastly greater (multiple log orders) sampling of tissue specimens, (2) volumetric imaging of cell distributions and tissue structures that are prognostic and predictive, (3) nondestructive imaging, which allows valuable biopsy specimens to be used for downstream molecular assays, and (4) a simplified process with cost benefits for pathology laboratories. However, due to the immense size of these feature-rich datasets, new challenges exist in terms of data management and computer-aided interpretation. In this forward-looking Perspective, we first provide a brief overview of the imaging technologies that can enable nondestructive 3D pathology, including computational tools needed to support these 3D methods. We then provide a roadmap for how machine learning, which is already being developed within the context of 2D digital pathology, should be leveraged and refined for 3D pathology. Finally, we discuss future challenges and opportunities for the clinical validation, regulatory approval, and clinical adoption of this new paradigm for precision medicine, including for reducing health disparities across populations. This includes learning from, and integrating with, other diagnostic modalities such as radiology and genomics.

For breast cancer patients, the extent of regional lymph node (LN) metastasis influences the decision to remove all axillary LNs. Metastases are currently identified and classified with visual analysis of a few thin tissue sections with conventional histology that may underrepresent the extent of metastases.We sought to enable nondestructive three-dimensional (3D) pathology of human axillary LNs and to develop a practical workflow for LN staging with our method. We also sought to evaluate whether 3D pathology improves staging accuracy in comparison to two-dimensional (2D) histology.We developed a method to fluorescently stain and optically clear LN specimens for comprehensive imaging with multiresolution open-top light-sheet microscopy. We present an efficient imaging and data-processing workflow for rapid evaluation of HE-like datasets in 3D, with low-resolution screening to identify potential metastases followed by high-resolution localized imaging to confirm malignancy.We simulate LN staging with 3D and 2D pathology datasets from 10 metastatic nodes, showing that 2D pathology consistently underestimates metastasis size, including instances in which 3D pathology would lead to upstaging of the metastasis with important implications on clinical treatment.Our 3D pathology method may improve clinical management for breast cancer patients by improving staging accuracy of LN metastases.

Recent advances in optical clearing and light-sheet microscopy have provided unprecedented access to structural and molecular information from intact tissues. However, current light-sheet microscopes have imposed constraints on the size, shape, number of specimens, and compatibility with various clearing protocols. Here we present a multi-immersion open-top light-sheet microscope that enables simple mounting of multiple specimens processed with a variety of clearing protocols, which will facilitate wide adoption by preclinical researchers and clinical laboratories. In particular, the open-top geometry provides unsurpassed versatility to interface with a wide range of accessory technologies in the future., Light-sheet microscopes are increasingly used for imaging cleared tissues, but have imposed constraints on sample geometries and protocols. Here the authors present a multi-immersion open-top light-sheet microscope to overcome these limitations and enable high-throughput imaging of samples processed with various clearing protocols.

Significance: Processing and diagnosing a set of 12 prostate biopsies using conventional histology methods typically take at least one day. A rapid and accurate process performed while the patient is still on-site could significantly improve the patient’s quality of life. Aim: We develop and assess the feasibility of a one-hour-to-diagnosis (1Hr2Dx) method for processing and providing a preliminary diagnosis of a set of 12 prostate biopsies. Approach: We developed a fluorescence staining, optical clearing, and 3D open-top light-sheet microscopy workflow to enable 12 prostate needle core biopsies to be processed and diagnosed within an hour of receipt. We analyzed 44 biopsies by the 1Hr2Dx method, which does not consume tissue. The biopsies were then processed for routine, slide-based 2D histology. Three pathologists independently evaluated the 3D 1Hr2Dx and 2D slide-based datasets in a blinded, randomized fashion. Turnaround times were recorded, and the accuracy of our method was compared with gold-standard slide-based histology. Results: The average turnaround time for tissue processing, imaging, and diagnosis was 44.5 min. The sensitivity and specificity of 1Hr2Dx in diagnosing cancer were both >90%. Conclusions: The 1Hr2Dx method has the potential to improve patient care by providing an accurate preliminary diagnosis within an hour of biopsy.

Intraoperative identification of carcinoma at lumpectomy margins would enable reduced re-excision rates, which are currently as high as 20% to 50%. Although imaging of disease-associated biomarkers can identify malignancies with high specificity, multiplexed imaging of such biomarkers is necessary to detect molecularly heterogeneous carcinomas with high sensitivity. We have developed a Raman-encoded molecular imaging (REMI) technique in which targeted nanoparticles are topically applied on excised tissues to enable rapid visualization of a multiplexed panel of cell surface biomarkers at surgical margin surfaces. A first-ever clinical study was performed in which 57 fresh specimens were imaged with REMI to simultaneously quantify the expression of four biomarkers HER2, ER, EGFR, and CD44. Combined detection of these biomarkers enabled REMI to achieve 89.3% sensitivity and 92.1% specificity for the detection of breast carcinoma. These results highlight the sensitivity and specificity of REMI to detect biomarkers in freshly resected tissue, which has the potential to reduce the rate of re-excision procedures in cancer patients. Cancer Res; 77(16); 4506–16. ©2017 AACR.

Slide-free digital pathology techniques, including nondestructive 3D microscopy, are gaining interest as alternatives to traditional slide-based histology. In order to facilitate clinical adoption of these fluorescence-based techniques, software methods have been developed to convert grayscale fluorescence images into color images that mimic the appearance of standard absorptive chromogens such as hematoxylin and eosin (H&E). However, these false-coloring algorithms often require manual and iterative adjustment of parameters, with results that can be inconsistent in the presence of intensity nonuniformities within an image and/or between specimens (intra- and inter-specimen variability). Here, we present an open-source (Python-based) rapid intensity-leveling and digital-staining package that is specifically designed to render two-channel fluorescence images (i.e. a fluorescent analog of H&E) to the traditional H&E color space for 2D and 3D microscopy datasets. However, this method can be easily tailored for other false-coloring needs. Our package offers (1) automated and uniform false coloring in spite of uneven staining within a large thick specimen, (2) consistent color-space representations that are robust to variations in staining and imaging conditions between different specimens, and (3) GPU-accelerated data processing to allow these methods to scale to large datasets. We demonstrate this platform by generating H&E-like images from cleared tissues that are fluorescently imaged in 3D with open-top light-sheet (OTLS) microscopy, and quantitatively characterizing the results in comparison to traditional slide-based H&E histology.

Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful method for rapid and optically efficient 3D microscopy. Initial LSFM designs utilized a static sheet of light, termed selective plane illumination microscopy (SPIM), which exhibited shadowing artifacts and deteriorated contrast due to light scattering. These issues have been addressed, in part, by multidirectional selective plane illumination microscopy (mSPIM), in which rotation of the light sheet is used to mitigate shadowing artifacts, and digital scanned light-sheet microscopy (DSLM), in which confocal line detection is used to reject scattered light. Here we present a simple passive multidirectional digital scanned light-sheet microscopy (mDSLM) architecture that combines the benefits of mSPIM and DSLM. By utilizing an elliptical Gaussian beam with increased angular diversity in the imaging plane, mDSLM provides shadow-free contrast-enhanced imaging of fluorescently labeled samples.One Sentence SummaryGlaser et al. describe a light-sheet microscopy architecture that enables passive multidirectional illumination with confocal line detection to enable both uniform fluorescence excitation and contrast-enhanced imaging of fluorescently labeled samples.

Megavoltage radiation beams used in External Beam Radiotherapy (EBRT) generate Cherenkov light emission in tissues and equivalent phantoms. This optical emission was utilized to excite an oxygen-sensitive phosphorescent probe, PtG4, which has been developed specifically for NIR lifetime-based sensing of the partial pressure of oxygen (pO2). Phosphorescence emission, at different time points with respect to the excitation pulse, was acquired by an intensifier-gated CCD camera synchronized with radiation pulses delivered by a medical linear accelerator. The pO2 distribution was tomographically recovered in a tissue-equivalent phantom during EBRT with multiple beams targeted from different angles at a tumor-like anomaly. The reconstructions were tested in two different phantoms that have fully oxygenated background, to compare a fully oxygenated and a fully deoxygenated inclusion. To simulate a realistic situation of EBRT, where the size and location of the tumor is well known, spatial information of a prescribed region was utilized in the recovery estimation. The phantom results show that region-averaged pO2 values were recovered successfully, differentiating aerated and deoxygenated inclusions. Finally, a simulation study was performed showing that pO2 in human brain tumors can be measured to within 15 mmHg for edge depths less than 10-20 mm using the Cherenkov Excited Phosphorescence Oxygen imaging (CEPhOx) method and PtG4 as a probe. This technique could allow non-invasive monitoring of pO2 in tumors during the normal process of EBRT, where beams are generally delivered from multiple angles or arcs during each treatment fraction.

Intraoperative assessment of breast surgical margins will be of value for reducing the rate of re-excision surgeries for lumpectomy patients. While frozen-section histology is used for intraoperative guidance of certain cancers, it provides limited sampling of the margin surface (typically

A method was developed utilizing Cherenkov imaging for rapid and thorough determination of the two gantry angles that produce the most uniform treatment plane during dual-field total skin electron beam therapy (TSET).Cherenkov imaging was implemented to gather 2D measurements of relative surface dose from 6 MeV electron beams on a white polyethylene sheet. An intensified charge-coupled device camera time-gated to the Linac was used for Cherenkov emission imaging at sixty-two different gantry angles (1° increments, from 239.5° to 300.5°). Following a modified Stanford TSET technique, which uses two fields per patient position for full body coverage, composite images were created as the sum of two beam images on the sheet; each angle pair was evaluated for minimum variation across the patient region of interest. Cherenkov versus dose correlation was verified with ionization chamber measurements. The process was repeated at source to surface distance (SSD) = 441, 370.5, and 300 cm to determine optimal angle spread for varying room geometries. In addition, three patients receiving TSET using a modified Stanford six-dual field technique with 6 MeV electron beams at SSD = 441 cm were imaged during treatment.As in previous studies, Cherenkov intensity was shown to directly correlate with dose for homogenous flat phantoms (R(2) = 0.93), making Cherenkov imaging an appropriate candidate to assess and optimize TSET setup geometry. This method provided dense 2D images allowing 1891 possible treatment geometries to be comprehensively analyzed from one data set of 62 single images. Gantry angles historically used for TSET at their institution were 255.5° and 284.5° at SSD = 441 cm; however, the angles optimized for maximum homogeneity were found to be 252.5° and 287.5° (+6° increase in angle spread). Ionization chamber measurements confirmed improvement in dose homogeneity across the treatment field from a range of 24.4% at the initial angles, to only 9.8% with the angles optimized. A linear relationship between angle spread and SSD was observed, ranging from 35° at 441 cm, to 39° at 300 cm, with no significant variation in percent-depth dose at midline (R(2) = 0.998). For patient studies, factors influencing in vivo correlation between Cherenkov intensity and measured surface dose are still being investigated.Cherenkov intensity correlates to relative dose measured at depth of maximum dose in a uniform, flat phantom. Imaging of phantoms can thus be used to analyze and optimize TSET treatment geometry more extensively and rapidly than thermoluminescent dosimeters or ionization chambers. This work suggests that there could be an expanded role for Cherenkov imaging as a tool to efficiently improve treatment protocols and as a potential verification tool for routine monitoring of unique patient treatments.

This study's goal was to determine how Cherenkov radiation emission observed in radiotherapy is affected by predictable factors expected in patient imaging. Factors such as tissue optical properties, radiation beam properties, thickness of tissues, entrance/exit geometry, curved surface effects, curvature and imaging angles were investigated through Monte Carlo simulations. The largest physical cause of variation of the correlation ratio between of Cherenkov emission and dose was the entrance/exit geometry (˜50%). The largest human tissue effect was from different optical properties (˜45%). Beyond these, clinical beam energy varies the correlation ratio significantly (˜20% for X-ray beams), followed by curved surfaces (˜15% for X-ray beams and ˜8% for electron beams), and finally, the effect of field size (˜5% for X-ray beams). Other investigated factors which caused variations less than 5% were tissue thicknesses and source to surface distance. The effect of non-Lambertian emission was negligible for imaging angles smaller than 60 degrees. The spectrum of Cherenkov emission tends to blue-shift along the curved surface. A simple normalization approach based on the reflectance image was experimentally validated by imaging a range of tissue phantoms, as a first order correction for different tissue optical properties.

Summary Background Outbreaks of infections with multidrug-resistant bacteria in neonatal intensive care units (NICUs) pose a major threat, especially to extremely preterm infants. This study describes a 35-day outbreak of multidrug-resistant Escherichia coli ( E. coli ) in a tertiary-level NICU in Germany. Aim To underline the importance of surveillance policies in the particularly vulnerable cohort of preterm infants and to describe the efficacy of outbreak control strategies. Methods Data were collected retrospectively from medical reports. Infants and environment were tested for E. coli . Findings The outbreak affected a total of 13 infants between 25 +1 and 35 +0 weeks of gestation with seven infants showing signs of infection. The outbreak strain was identified as E. coli sequence type 131. Environmental screening provided no evidence for an environmental source. Through colonization surveillance and immediate and adequate treatment of potentially infected preterm infants, no fatalities occurred. Outbreak control was achieved by strict contact precautions, enhanced screening and temporary relocation of the NICU. Relocation and reconstruction improved the NICU's structural layout, focusing on isolation capacities. Follow-up indicated carriage for several months in some infants. Conclusion Routine surveillance allowed early detection of the outbreak. The identification of carriers of the outbreak strain was successfully used to direct antibiotic treatment in case of infection. Enhanced hygienic measures and ward relocation were instrumental in controlling the outbreak.

To identify achievable camera performance and hardware needs in a clinical Cherenkov imaging system for real-time, in vivo monitoring of the surface beam profile on patients, as novel visual information, documentation, and possible treatment verification for clinicians.Complementary metal-oxide-semiconductor (CMOS), charge-coupled device (CCD), intensified charge-coupled device (ICCD), and electron multiplying-intensified charge coupled device (EM-ICCD) cameras were investigated to determine Cherenkov imaging performance in a clinical radiotherapy setting, with one emphasis on the maximum supportable frame rate. Where possible, the image intensifier was synchronized using a pulse signal from the Linac in order to image with room lighting conditions comparable to patient treatment scenarios. A solid water phantom irradiated with a 6 MV photon beam was imaged by the cameras to evaluate the maximum frame rate for adequate Cherenkov detection. Adequate detection was defined as an average electron count in the background-subtracted Cherenkov image region of interest in excess of 0.5% (327 counts) of the 16-bit maximum electron count value. Additionally, an ICCD and an EM-ICCD were each used clinically to image two patients undergoing whole-breast radiotherapy to compare clinical advantages and limitations of each system.Intensifier-coupled cameras were required for imaging Cherenkov emission on the phantom surface with ambient room lighting; standalone CMOS and CCD cameras were not viable. The EM-ICCD was able to collect images from a single Linac pulse delivering less than 0.05 cGy of dose at 30 frames/s (fps) and pixel resolution of 512 × 512, compared to an ICCD which was limited to 4.7 fps at 1024 × 1024 resolution. An intensifier with higher quantum efficiency at the entrance photocathode in the red wavelengths [30% quantum efficiency (QE) vs previous 19%] promises at least 8.6 fps at a resolution of 1024 × 1024 and lower monetary cost than the EM-ICCD.The ICCD with an intensifier better optimized for red wavelengths was found to provide the best potential for real-time display (at least 8.6 fps) of radiation dose on the skin during treatment at a resolution of 1024 × 1024.

Introduction: Ureaplasma spp. have been implicated in the pathogenesis of both preterm labor and neonatal morbidity including pneumonia and sepsis and the development of chronic lung disease of prematurity. Data on Ureaplasma meningitis are limited and partly controversially discussed. Patient: We report the unique case of a 9-month-old infant with progressive internal hydrocephalus of unknown origin and developmental delay due to a history of>200 days of inflammation of the central nervous system. The female extremely low birth weight infant had been referred to our hospital for ventriculoperitoneal shunt implantation. She had been born at 26+3 weeks of gestation with a birth weight of 940 g. With the exception of a moderate respiratory distress syndrome, postnatal period had been reported uneventful. However, internal hydrocephalus had become manifest at 4 weeks of postnatal age. Intraventricular hemorrhage had not been documented by cranial ultrasound and magnetic resonance imaging. Cerebrospinal fluid (CSF) analysis had repetitively revealed pronounced inflammation reflected by pleocytosis (50–86 leukocytes/μL, 60% lymphocytes), CSF protein levels of 578–1 026 mg/dL and undetectable CSF glucose. Although suggesting bacterial meningitis, microbial diagnostics had not been indicative, and empirical antibiotics had not affected the CSF findings. On admission to our hospital, CSF analysis still documented significant inflammation (125 leukocytes/μL, CSF protein 565 mg/dL, CSF glucose Results: Due to a prenatal history of cerclage, we initiated microbial diagnostics on Ureaplasma spp. and Mycoplasma hominis. U. parvum was detected in CSF by culture and PCR, no other pathogens were isolated. On intravenous treatment with chloramphenicol, CSF profile continuously normalized, and cultures and PCR became negative. Treatment was continued for 3 weeks, and the infant was discharged after uncomplicated ventriculoperitoneal shunt placement. During a 12-month observation period she has shown encouraging recovery. Conclusion: In preterm infants, in particular, internal hydrocephalus of unknown origin and sustained CSF inflammation are highly suggestive of Ureaplasma meningitis. Our case highlights that infection may escape detection if not explicitly considered, since microbial diagnosis requires complex media and PCR.

OBJECTIVE: To compare the efficacy and safety of sertraline and amitriptyline in a German outpatient population. METHODS: Patients with Major Depression (DSM-III-R) and HAM-D (21 items) > or = 21 in 19 German centers received double-blind treatment with sertraline (initial dose 50 mg, titration up to 100 mg) or amitriptyline (75 mg, up to 150 mg) over 6 weeks. HAM-D (21 items), HAM-D Bech, CGI, DSI and SDS were evaluated for the efficacy analysis. FSUCL (Fischer Somatic and Undesired Effects Check List) and spontaneously reported adverse events were used for safety analysis. RESULTS: Of the 240 patients enrolled in the study, 205 (100 sertraline; 105 amitriptyline) were evaluable for efficacy. No statistically significant differences were detected between the two groups in the ITT and ATP efficacy analyses. Response, defined as score 1 (very much improved) or 2 (much improved) of the CGI improvement score, was 76% in the sertraline and 81% in the amitriptyline group (efficacy evaluable patients = ATP population). In the structured FSUCL, the side-effect burden (FSUCL score >2 for drug related symptoms) was significantly higher in the amitriptyline group at all follow up visits (p

Purpose To determine whether Cherenkov light imaging can visualize radiation therapy in real time during breast radiation therapy. Methods and Materials An intensified charge-coupled device (CCD) camera was synchronized to the 3.25-μs radiation pulses of the clinical linear accelerator with the intensifier set × 100. Cherenkov images were acquired continuously (2.8 frames/s) during fractionated whole breast irradiation with each frame an accumulation of 100 radiation pulses (approximately 5 monitor units). Results The first patient images ever created are used to illustrate that Cherenkov emission can be visualized as a video during conditions typical for breast radiation therapy, even with complex treatment plans, mixed energies, and modulated treatment fields. Images were generated correlating to the superficial dose received by the patient and potentially the location of the resulting skin reactions. Major blood vessels are visible in the image, providing the potential to use these as biological landmarks for improved geometric accuracy. The potential for this system to detect radiation therapy misadministrations, which can result from hardware malfunction or patient positioning setup errors during individual fractions, is shown. Conclusions Cherenkoscopy is a unique method for visualizing surface dose resulting in real-time quality control. We propose that this system could detect radiation therapy errors in everyday clinical practice at a time when these errors can be corrected to result in improved safety and quality of radiation therapy.

Čerenkov radiation emission occurs in all tissue, when charged particles (either primary or secondary) travel at velocity above the threshold for the Čerenkov effect (about 220 KeV in tissue for electrons). This study presents the first examination of optical Čerenkov emission as a surrogate for the absorbed superficial dose for MV x-ray beams.In this study, Monte Carlo simulations of flat and curved surfaces were studied to analyze the energy spectra of charged particles produced in different regions near the surfaces when irradiated by MV x-ray beams. Čerenkov emission intensity and radiation dose were directly simulated in voxelized flat and cylindrical phantoms. The sampling region of superficial dosimetry based on Čerenkov radiation was simulated in layered skin models. Angular distributions of optical emission from the surfaces were investigated. Tissue mimicking phantoms with flat and curved surfaces were imaged with a time domain gating system. The beam field sizes (50 × 50-200 × 200 mm(2)), incident angles (0°-70°) and imaging regions were all varied.The entrance or exit region of the tissue has nearly homogeneous energy spectra across the beam, such that their Čerenkov emission is proportional to dose. Directly simulated local intensity of Čerenkov and radiation dose in voxelized flat and cylindrical phantoms further validate that this signal is proportional to radiation dose with absolute average discrepancy within 2%, and the largest within 5% typically at the beam edges. The effective sampling depth could be tuned from near 0 up to 6 mm by spectral filtering. The angular profiles near the theoretical Lambertian emission distribution for a perfect diffusive medium, suggesting that angular correction of Čerenkov images may not be required even for curved surface. The acquisition speed and signal to noise ratio of the time domain gating system were investigated for different acquisition procedures, and the results show there is good potential for real-time superficial dose monitoring. Dose imaging under normal ambient room lighting was validated, using gated detection and a breast phantom.This study indicates that Čerenkov emission imaging might provide a valuable way to superficial dosimetry imaging in real time for external beam radiotherapy with megavoltage x-ray beams.

Radiation from a linear accelerator induces Cherenkov emission in tissue, which has recently been shown to produce biochemical spectral signatures that can be interpreted to estimate tissue hemoglobin and oxygen saturation or molecular fluorescence from reporters. The Cherenkov optical light levels are in the range of 10(-6) to 10(-9) W/cm2, which limits the practical utility of the signal in routine radiation therapy monitoring. However, due to the fact that the radiation is pulsed, gated-acquisition of the signal allows detection in the presence of ambient lighting, as is demonstrated here. This observation has the potential to significantly increase the value of Cherenkov emission spectroscopy during radiation therapy to monitor tissue molecular events.

The frequency of primary systemic small vessel vasculitides (PSV) varies among different geographic regions and age categories. We studied PSV in patients from middle-eastern Crete (Greece), and compared clinical characteristics in younger (65 years) versus older (or = 65 years) adult patients.The records of 67 patients (33 younger, 34 older adults) diagnosed with PSV during 1995-2003 who were referred to a mixed secondary/tertiary care University Hospital in Crete were reviewed. Data on clinical manifestations, diagnosis, therapy, and adverse outcomes (end stage renal disease, death) during a median follow-up of 6 (range 0-12) years were recorded. Multivariate regression analysis was applied to identify independent predictors for adverse outcomes.The overall annual incidence of PSV was 19.5/million (95% confidence interval [CI] 15.7-23.4), 48.9/million (95% CI 33.8-63.9) in older and 12.4/million (95% CI 7.7-17) in younger adults. Microscopic polyangiitis was more prevalent in older patients (65%) and Wegener's Granulomatosis in younger patients (52%). Thirty-one percent of older patients developed end-stage renal disease as compared to 11% of younger patients (p=0.103). Mortality rates were 60% in older patients and 19% in younger patients (p=0.001). In multivariate regression analysis age (Beta=0.33 per 1-year, p=0.005), serum creatinine (Beta=0.29 per 1-mg/dL, p=0.011), and lung involvement (Beta=0.36, p=0.002) at the time of diagnosis were independent predictors for end stage renal disease and/or death.This study documents increased frequency and significant mortality of PSV among older people in Crete, with MPA being the most prevalent type. Age, serum creatinine, and lung involvement are important predictors for adverse outcome in these patients.

The management of patients with a laparostoma due to peritonitis is a challenge for every surgeon and intensivist. The goal of this study was to compare the different treatment strategies for the open abdomen: Abdominal Dressing (AD), the classic V.A.C. therapy (CV) and conventional open therapy (CT).Between 2001 and 2005 we identified 62 patients in 4 surgical departments in Austria who had to be treated with a laparostoma due to peritonitis. 27 patients were conventionally treated, 16 with the Classic V.A.C. therapy and 19 patients with V.A.C. abdominal dressing.The mortality was 3/16 (14 %) in the AD group vs. 4/12 (21 %) patients in the CV group and 18/9 (59 %) in conventional therapy. There was no significant difference for survivors in the length of ICU stay: 26.6 +/- 23.0 days in the CT group, 34.6 +/- 30.2 days in the CV group and 38.9 +/- 27.2 days in the AD group. Apache II Score and Mannheimer Peritonitis Score showed no difference between the groups.We found a reduction of mortality in the V.A.C. Abdominal Dressing group by approximately 40 % (AD: 14 %, CT: 59 %). Although we could identify a difference in age in our retrospective study we believe that V.A.C. Abdominal Dressing is the important factor for the different clinical outcome. These first results indicate the need for further prospective evaluation of the V.A.C. Abdominal Dressing therapy, to prove if a new standard in the therapy of the open abdomen is created.

A previously healthy patient became comatose 8 h after osteosynthesis of a fractured femur and humerus. After an initially normal cranial computed tomography, extended lesions were found 10 h later in the control scan as an indication of an embolization in the basilar artery. After neurosurgeons assessed the situation to be unfavorable, no other interventions were conducted and 6 days later brain death was confirmed. During the subsequent heart transplantation the suspicion of a patent foramen ovale as the reason for the paradoxical cerebral embolization was confirmed. This tragic course raises the question whether a patent foramen ovale should be excluded or closed by interventional measures in similar situations.

This study reports on the implementation of a real-time MR acquisition, reconstruction, and display system on standard hardware for 3D contrast-enhanced MR abdominal angiography. The system allows for dynamic imaging of the contrast passage with good spatial resolution in a single breathhold. It is capable of synchronously acquiring and processing multiple coil data. Bolus arrival can be detected confidently from the display of MIP images from sagittal 3D slabs with moderate spatial resolution. Upon detection of the contrast agent, the patient is asked to hold their breath and a novel 3D acquisition is started for a coronal volume. Temporal imaging within the breathhold is achieved through applying a modified time-resolved imaging of contrast kinetics (TRICKS) technique with elliptical centric view ordering. The technique displays contrast passage from the arterial phase through enhancement of the hepatic venous system. It also provides the ability to quantify motion of the diaphragm over a long breathhold.

We have developed a method of spraying assay reagents onto a target gel in the Micro-Arrayed Compound Screening ( micro ARCS) format. After application of target gels to compound sheets, subsequent reagents can be applied by spraying onto the target gel. The spraying method conserves on assay reagents by up to 10-fold, eliminates the need for casting additional agarose gels, and increases the throughput of a screen by 3-fold. To demonstrate the efficacy of applying the spraying method to micro ARCS, we screened over 600,000 compounds for inhibitors of histone deacetylase (HDAC). Commercially available HDAC substrate and reaction developer were sprayed directly onto the gel to initiate the reaction and to amplify the signal, respectively. Picks in the primary screen were retested at a density of 384 compounds per sheet in the micro ARCS format. IC(50) values for active compounds were confirmed in a 96-well plate assay. The screen identified several small molecule inhibitors of the enzyme, including members of several classes of known HDAC inhibitors. The combination of the high-density format of micro ARCS, the efficiency of the spraying method, and a timed sequence of adding assay reagents permitted a screening throughput of 200,000 tests an hour. We present the details of the screening format and the analysis of the hits from the screen.

Rectus sheath hematoma (RSH) is a rare entity that can mimic an acute abdomen. Therefore, we designed a study to analyze the etiology, frequency, diagnosis using ultrasound, and treatment of RSH.A total of 1,257 patients admitted for abdominal ultrasound for acute abdominal pain or unclear acute abdominal disorders were evaluated.In 23 (1.8%) patients, an RSH was diagnosed; three of them were not diagnosed preoperatively by ultrasound. Of 13 men and 10 women (mean age, 57 +/- 23 years), 13 developed RSH after local trauma, three after severe coughing, two after defecation, and five spontaneously. Fifteen had nonsurgical therapy, and eight underwent surgery. The use of anticoagulants was accompanied by a larger diameter of the RSH (p.012), and surgical therapy was more frequently required in these patients. In the surgically treated group, more intraabdominal free fluid could be detected by ultrasound (p.0005), patients required less analgesics (p.001), and the mean hospital stay was shorter (p.001).RSH is a rare condition that is usually associated with abdominal trauma and/or anticoagulation therapy. Ultrasound is a good screening technique. Nonsurgical therapy is appropriate but leads to a greater need for analgesics. Surgery should be restricted to cases with a large hematoma or free intraabdominal rupture.

Partial posterior fundoplication improves esophageal peristalsis in patients with gastroesophageal reflux disease (GERD) associated with poor esophageal body function. The aim of this study was to investigate whether postoperative administration of cisapride enhances the effect of surgery on esophageal peristalsis. Laparoscopic partial posterior fundoplication was performed on 34 consecutive GERD patients with poor esophageal body motility. These patients were randomized in groups without and with postoperative treatment with cisapride 20 mg twice daily for six months. Esophageal manometry was performed preoperatively and six months following surgery. Esophageal body function improved significantly following partial posterior fundoplication without or with postoperative treatment with cisapride. However, this effect was more pronounced in the group of patients receiving cisapride. Partial posterior fundoplication combined with postoperative treatment with cisapride should be the therapy of choice in GERD patients with poor esophageal body motility.

Gastroesophageal reflux disease (GERD) is the most common disease of the upper gastrointestinal tract. With the introduction of proton pump inhibitors medical treatment of GERD has been significantly improved. However, the development of laparoscopic antireflux surgery resulted in an increasing interest of surgeons in this disease. An interactive meeting was organized in order to develop an agreement between gastoenterologists and surgeons regarding therapeutic decisions and this is the main topic of this paper.

Retrospective analysis was done of three cases with severe liver trauma and excessive serum bilirubin levels caused by a traumatic biliovenous fistula. The literature is reviewed.Diagnostic measures included laboratory findings, computed tomography, ultrasonography, and endoscopic retrograde cholangiopancreatography (ERCP).The biliovenous fistula was detected by ERCP in two cases. In one case a left hemihepatectomy was carried out, and the patient was cured. The other patient received drainage of a huge necrotic cavity in the center of the liver. Ten months later the patient underwent reoperation, and left hepatic resection was performed. The patient died of liver function failure on postoperative day 7. In the third case the fistula subsided spontaneously.An excessively high serum level of direct bilirubin and only moderately elevated liver enzymes indicate bilhemia in trauma patients. ERCP is most reliable in localizing the fistula; computed tomography/ultrasonography are valuable in detecting the extension and localization of the parenchymal destruction. Conservative therapy is justified if the patient is in good condition or if the localization of the fistula is unclear. Spontaneous closure of the fistula may occur. Surgical treatment options are partial liver resection and suture of the fistula and T-tube drainage of the common bile duct and drainage of the rupture site.

From January 1983 to January 1991, radical surgical treatment for carcinoma of the rectum was performed upon 154 patients in our department. In 30 instances, local treatment failure occurred and patients were treated either conservatively or operatively. Survival times of these 30 patients were compared to evaluate if operation, even in instances in which only palliative resection is possible, can prolong survival and if early diagnosis of recurrence leads to a higher rate of radical resections and subsequent cure.Patients were divided into three groups. Group 1 consisted of patients not undergoing an operation, patients in group 2 had a palliative resection and patients in group 3 had radical resections. The median survival time was estimated for each group.The median survival period was six months for group 1, 17 months for group 2, and 35.5 months for group 3. Four patients who underwent reoperation for cure are still alive: one with recurrent tumor after 28 months, and three without evidence of disease after 32, 42 and 43 months. The most valuable diagnostic mean in the detection of local recurrence was endosonography.Surgical treatment for recurrent carcinoma of the rectum is justified not only in cases having radical resection but also as a palliative approach. Compared to other investigative methods, endosonography seems to detect recurrences earlier, at a time when curative retreatment is still possible.

Endoscopic sphincterotomy (EST) is regarded as an alternative therapeutic approach to the surgical treatment of choledocholithiasis. To clarify the indication for each of these two methods, a prospective study has been performed comparing patients with calculi of the common bile duct who had undergone endoscopic or surgical treatment. Mortality, morbidity and stone clearance were used as criteria for the evaluation of the dependence of age on results achieved by the two modalities. Of 306 patients with choledocholithiasis, 199 underwent surgical exploration of the duct and 107 underwent endoscopic sphincterotomy. Patients who were more than 60 years of age and had previously undergone cholecystectomy underwent endoscopic sphincterotomy. All other patients had surgical treatment. There were no significant differences among the two groups with regard to mortality rates. The incidence of relevant complications as well as the incidence of surgical revisions related to postoperative complications in the operation group was, however, significantly higher (p less than 0.05). In contrast with this, the incidence of retained stones was significantly lower (p less than 0.01) in the group that underwent surgical treatment (2.5 percent) than in the group that had endoscopy (11.2 percent). When assessing the results in terms of stone clearance, complications and mortality rates, findings seem to indicate that a safe limit for the application of surgical treatment would be 60 years of age, whereas older patients should be primarily considered as candidates for EST.

The histological slides of 39 cases of cancer of the pancreatic head were analysed using an interactive image analyser system. Some 14 cases were classified as periampullary, 25 as ductal pancreatic cancer. All cases had undergone radical tumor resection according to Whipple's procedure. Morphometric data, tumor size and metastatic nodal involvement were correlated to prognosis. Univariate statistical analysis showed that the classical differentiation between ductal and periampullary cancer was a weaker prognosticator than morphometric variables. In fact, multivariant statistical analysis showed that the morphometric variable irregularity was the best prognosticator (p = 0.0001). No other variable added significant prognostic information. Irregularity is a newly developed variable describing the nuclear shape corrected for roundness. We conclude that morphometry can be of essential prognostic information for the clinician in cancer of the pancreatic head.

During the period from 1984-1988 at the University Clinic, Innsbruck (in cooperation with the Department of Radiotherapy and Surgery) 40 patients with pancreas carcinoma were treated with intraoperative radiotherapy. Based upon the preoperative CT's, the operation reports and the patho-anatomic judgement of the operative specimen for 19 patients the quality of the performed IORT (with curative intention combined with partial pancreaticoduodenectomy) was examined. This examination was based upon a standardized concept. In 53% of the cases the chosen irradiation field was too small and, in most cases the applied single dose was from radiooncological point of view not sufficient. These results emphasize the need for preoperative treatment planning depending from the surgical intention, the need for individually shaped cones and direct field control of the applied dose.

Sigmoid volvulus is an emergency which requires rapid diagnosis and immediate adequate treatment. Diagnosis is primarily radiological. Emergency laparotomy carries a high mortality and should therefore not be the immediate treatment. In our opinion, detorsion by colonoscopy under radiological control is a promising form of treatment. Selective resection of the sigmoid, thereby removing the cause of the abnormality, should not be forgotten, since this prevents recurrences. For this method, mortality after adequate gut preparation is only 5.4%. An exception to the above is gangrene of the bowel, when an emergency laparotomy cannot be avoided.

The nuclear DNA distribution pattern of the neoplastic parenchymal cells of 100 conventionally formalin-fixed and paraffin-embedded specimens from pancreatic adenocarcinomas and from 8 specimens of chronic pancreatitis was assessed by means of image cytometry. All material originated from pancreatic restrictions. Evaluable DNA histograms could be obtained for 77 carcinomas, and clinical data were available for 71 of these. In these 71 specimens, the nuclear DNA ploidy pattern was also investigated by means of flow cytometry. In 76 of the 77 cases, the image-cytometric DNA ploidy pattern obtained showed a "nondiploid" distribution with modal values as high as 8.5 c. In 21 cases, the neoplastic cells showed modal values in the "triploid" region. The analogous 71 flow-cytometric DNA histograms could only be evaluated in 50 cases because of excessively high amounts of background and/or excessively broad peaks. In 47 cases, the nuclear DNA histogram was nondiploid according to both techniques. The patients with carcinomas whose cell nuclei showed a triploid DNA distribution showed a significantly shorter survival time than those with tumor cell populations of nontriploid DNA distribution patterns. In the 8 specimens of chronic pancreatitis, the parenchymal cells were all equipped with nuclei showing diploid DNA distribution patterns.

Intraoperative radiotherapy (IORT) was administered to 13 patients with carcinoma of the pancreas, using high-velocity electrons of a LINAC accelerator in a single dose of maximally 25 Gy applied directly to the tumour or tumour bed with the abdomen open. There were no disorders of wound healing or other serious early complications which could be ascribed to the radiation. Stomach and intestines were excluded from the radiation field to avoid late radiation damage, such as stenoses or ulcers. The method is particularly indicated for locally non-resectable, non-metastasized pancreatic carcinoma, possibly also as tumour-bed radiation during radical surgery. Patients with distant metastases were not given radiation. First results point to IORT providing lasting improvement of pancreatic pain. It is too early to know whether survival time is favourable influenced.

According to the literature synchronous adenocarcinomas of both, the colon and the gallbladder or the colon and the appendix are extremely unusual. To our knowledge the simultaneous occurrence of all three tumors has never been published before. Preoperative diagnosis of early stage tumors of the gallbladder and the vermiform appendix is also made very seldom. The following case report records an instance of synchronous adenocarcinomas of the gallbladder, the transverse colon, in a colonic polyp and the appendix. Owing to the patient's advanced age simultane cancer occurrence may be mere coincidence. Nevertheless, this extremely rare entity could also be ascribed to a genetic defect or an unknown carcinogenic agent.

Thirteen patients suffering from adenocarcinomas of the pancreas were submitted to an intraoperative fast electron "boost" therapy with or without percutaneous photon irradiation. A duodeno-cephalo-pancreatectomy with subsequent irradiation of the tumor bed could be performed in three patients. Ten patients were inoperable because of advanced tumors and formation of metastases. The average survival is 6.5 months, at present six patients are alive without major troubles. An analgetic effect was obtained in ten patients. The first results are encouraging with respect to local control, the little acute and chronic morbidity, and palliation achieved in advanced stages.