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Modus operandi for a picture archiving and communication system.

The authors describe their 18 month experience operating an all digital nuclear medicine department, which led to the development of a system that allows multiple users to access patient reports and images through terminals located in various parts of the hospital. All user interactions are mediated through protocols that automatically acquire, process, display, and archive data. This approach offers distinct advantages over film and paper methods and is applicable in theory to any digital image format.

Computers↗

Picture archiving communication systems in the intensive care unit.

The authors review the potential of picture archiving and communication systems (PACS) in the critical care environment. Presented and analyzed is a university hospital intensive care unit (ICU) PACS project. The objective of the project is to provide reliable, timely viewing of bedside radiographs in selected ICUs within the hospital. Key features include redundancy of critical hardware components and viewing stations that are simple to operate. The system was implemented in phases with incremental evaluation of performance and goals. PACS subsystems are discussed. On the basis of this experience, ICU PACS specifications and general considerations are presented. Also presented is a comparison of 10-year cost projections for conventional screen-film radiography and an ICU PACS that incorporates computed radiography. The costs of the two alternatives are comparable. PACS is a viable alternative to conventional screen-film imaging in the ICU setting and offers improved and more efficient patient care.

Alabama↗

Diagnosis of Enterocytozoon bieneusi by the polymerase chain reaction in archival fixed duodenal tissue.

This study involved ninety five formalin-fixed paraffin-embedded duodenal biopsy specimens retrieved from hospital files that were microscopically observed for the presence of microsporidia. Eleven samples that revealed compatible organisms were analyzed by the polymerase chain reaction (PCR) with four different protocols for the detection of Enterocytozoon bieneusi. Amplicons of the right size were obtained by at least one method for nine samples, remaining two negative ones. We report a PCR methodology that allows the use of archival specimens obtained for traditional pathology.

Biopsy↗

Methylation profiling of archived non-small cell lung cancer: a promising prognostic system.

PURPOSE: Enhanced prognostication power is becoming more desirable in clinical oncology. In this study, we explored the prognostic potential of multigene hypermethylation profiling in non-small-cell lung cancer. EXPERIMENTAL DESIGN: We evaluated a panel of eight genes (p16, APC, ATM, hMLH1, MGMT, DAPK, ECAD, and RASSF1A) using methylation-specific PCR in 105 archived specimens of non-small-cell lung cancer representing all stages of the illness. We analyzed the effect of gene methylation status on outcome individually in a cumulative manner and in a combinatorial approach using recursive partitioning to identify methylation profiles, which affect overall survival. RESULTS: In this data set, tumors harboring promoter hypermethylation at two or more genes exhibit similar survival trends to others in the cohort. Using recursive partitioning, three genes (APC, ATM, and RASSF1A) emerged as determinants of prognostic groups. This designation retained its statistical significance even when disease stage and age were entered into a multivariate analysis. Using this approach, patients whose tumors were hypermethylated at APC and those hypermethylated at only ATM (not also at APC or RASSF1A) enjoyed substantially longer 1- and 2-year survival than patients in the remaining groups. In 32 adjacent histologically normal lung tissue specimens, we detected similar methylation abnormalities. CONCLUSION: Assessment of promoter hypermethylation aberrations may facilitate prognostic profiling of lung tumors, but validation in independent data sets is needed to verify these profiles. This system uses material that is abundantly available with linked outcome data and can be used to generate reliable epigenetic determinants.

Adaptor Proteins, Signal Transducing↗

Diagnosis of papillary thyroid carcinoma is facilitated by using an RT-PCR approach on laser-microdissected archival material to detect RET oncogene activation.

OBJECTIVE: The purpose of this study was to investigate the value of the expression of the RET oncogene (rearranged during transfection) in papillary thyroid carcinomas (PTC) and its variants in the differential diagnosis of thyroid neoplasias. According to the literature RET oncogene activation by chromosomal rearrangements has been exclusively implicated in PTCs. METHODS: To establish the incidence of RET activation in PTCs we used 5- to 10-microm sections from archival paraffin blocks. Either parts of the tissue slices were manually dissected or a few distinct cells were microdissected by laser-mediated manipulation with the Robot-MicroBeam system. RNA was extracted from paraffin-embedded thyroid tumors and the corresponding normal tissue. RT and nested PCR were performed using primers for RET/PTC1, PTC2 and PTC3, or for RET exons 12 and 13. PCR products were resolved by gel electrophoresis. RESULTS: We detected RET transcription in approximately 85% of the PTCs including follicular variants and in isolated cells of the same tissues, but not in nonmalignant thyroid tissue. CONCLUSIONS: Our method may serve as an additional diagnostic tool to characterize ambiguous neoplasias and to identify especially nonpapillary, i.e. follicular tumors, as papillary carcinomas. Additionally, this study has demonstrated that expressed genes can be analyzed from routine histopathological tissue slides or pooled single cells. Large retrospective studies can also be performed with this method.

Adult↗

Fluorescent in situ hybridization on archival G-banded slides.

We report a simple procedure that allows the application of chromosomal in situ suppression hybridization to standard G-banded and Entellan-mounted archival preparations that had been stored for 4-8 y. The procedure includes removing the cover slips in xylene, refixation in buffered formaldehyde-acetone mixture, and washing in 2 x SSC, Tween 20.

Chromosome Banding↗

Point mutation analysis of archived cytogenetic slide DNA.

Archived Giemsa-stained cytogenetic slide repositories represent valuable DNA resources for medical, scientific, and forensic studies. Sequencing readily identified a Charcot-Marie-Tooth disease point mutation in a 209-bp PCR amplified product. With optimal PCR primers and amplification conditions, our protocol quickly and reliably isolated sufficient DNA for at least 12 independent PCR amplification reactions for forensic and medical applications from single slides up to 5 years old.

Alleles↗

Reverse transcriptase polymerase chain reaction for detecting Ewing's sarcoma in archival fine needle aspiration biopsies.

OBJECTIVE: Previous studies on pediatric soft tissue sarcomas have demonstrated a chromosomal 11;22 (q24;q12) translocation in Ewing's sarcoma (ES) and peripheral primitive neuroectodermal tumors that appears to be a unique oncogenic marker. To investigate the usefulness of reverse transcriptase polymerase chain reaction (RT-PCR) as an ancillary method for cytodiagnosis, we tested archival aspirates derived from ES patients to establish whether any beta-actin RNA expression or tumor-specific EWS/FLI-1 gene translocation had occurred. STUDY DESIGN: Sixteen skeletal specimens aspirated 10-15 years earlier from patients with cytologic and histologic diagnoses of ES were prepared for PCR. The amplification products were sequenced. RESULTS: Amplifiable RNA was detected in smears from 12 patients by beta-actin RT-PCR. Seven aspirates from beta-actin-positive patients showed ES-specific genomic translocation (58%). Sequence analysis of the resulting PCR fragments revealed EWS/FLI-1 fusion transcriptions of varying length. CONCLUSION: When RNA was retrievable, RT-PCR applied to routinely stained aspirate smears was a highly specific method in differential diagnosis of ES.

Base Sequence↗

Associations between dimensions of religious commitment and mental health reported in the American Journal of Psychiatry and Archives of General Psychiatry: 1978-1989.

The authors assessed all measures of religious commitment (N = 139) reported in research studies published in the American Journal of Psychiatry and Archives of General Psychiatry in 1978 through 1989 (N less than 35). For nearly two-thirds of the measures, the studies either made no hypotheses or reported no results concerning the relationship of religious commitment to mental health status. For the great majority of the measures assessed, the studies reported a positive relationship between religious commitment and mental health.

Humans↗

Inflammatory response to chronic otitis media in DiGeorge syndrome: a case study using immunohistochemistry on archival temporal bone sections.

Immunohistochemical analyses were conducted on archival celloidin-embedded human temporal bone sections from an 8-month-old boy with chronic otitis media and DiGeorge syndrome. We employed antigen retrieval methods with saturated sodium hydroxide-methanol solution, microwave incubation, and proteolytic treatment to demonstrate the distribution of T-lymphocytes, B-lymphocytes, macrophages, and intercellular adhesion molecule 1 (ICAM-1) expression in the middle ear. B-lymphocytes and macrophages were observed predominantly within the middle ear mucosa. T-lymphocytes were rare. Further, ICAM-1 was expressed in the vascular endothelium of the lamina propria, as well as infiltrating mononuclear cells. This suggests that the expression of ICAM-1 can be induced in the middle ear with otitis media, even if T-lymphocytes are depressed in a cell-mediated immunodeficiency disorder such as DiGeorge syndrome.

Antibodies, Monoclonal↗

Dual Y-chromosome painting and immunofluorescence staining of archival human liver transplant biopsies.

Combining fluorescence in situ hybridization (FISH) and indirect immunofluorescence staining of protein markers provides a highly specific method for identifying chromosomes in phenotypically defined cells and tissues. We developed a technique enabling dual chromosome painting and immunofluorescence staining of archival formalin-fixed, paraffin-embedded material, and used this to phenotype chimeric cells in female-to-male human liver transplants.

Chromosome Painting↗

Immunofluorescence and confocal laser scanning microscopy of chronic myeloproliferative disorders on archival formaldehyde-fixed bone marrow.

Spatial analysis of the histoarchitecture and photographic documentation at high resolution are the principal advantages of confocal laser scanning microscopy (CLSM) over conventional fluorescence microscopy (CFM) if combined with appropriate software. Restrictions for the use of CFM and CLSM, on the other hand, include nonspecific background fluorescence, fading of photolabile fluorochromes, and both tissue-specific and fixation-induced autofluorescence. Most of those shortcomings can now be avoided. Autofluorescence, the most limiting factor of high-resolution CLSM, was recently controlled also for paraffin sections of archival formaldehyde-fixed tissues. This allowed the present study on cytoskeletal fibers and extracellular matrix proteins in both neoplastic cells of myeloproliferative disorders and in medullary stromal cells using CLSM under proper autofluorescence control. By multiple fluorescence labeling, we found that the intracellular smooth muscle alpha-actin (SMA) fibers and the two extracellular adhesive matrix proteins tenascin and fibronectin vary in their presence in stromal and/or myeloid cells according to the degree of bone marrow fibrosis in chronic myeloproliferative disorders (CMPDs). CLSM offers further insight in our attempts to understand a complex interplay between the two cellular compartments.

Bone Marrow↗

An archival study of depression before and after age 55.

In this archival study, 58 patients with unipolar depression who were 55 years of age or older were compared to 155 depressed control patients age 54 years or less. The older patients with unipolar depression had a significantly worse outcome, with only 51.7% experiencing a period of full remission during follow-up. Medical disorders were common, and older depressed patients showed a clear excess of cardiovascular disorders. Clinical symptoms did not separate the two groups, with the exception of psychomotor retardation and agitation. Electroconvulsive therapy was associated with less continuous hospitalization and an overall superior outcome.

Adolescent↗

Establishment of proliferative cell nuclear antigen gene as an internal reference gene for polymerase chain reaction of a wide range of archival and fresh mammalian tissues.

Polymerase chain reaction (PCR) from paraffin-embedded tissues provides a powerful tool to amplify DNA from a variety of recent and archival material. Because DNA from paraffin-embedded samples is more degraded than from fresh material, the amplification of reference genes is essential to exclude false-negative results. This study describes the use of the proliferative cell nuclear antigen (PCNA) gene as a reference gene in a range of animal species and in humans. The PCNA-PCR to amplify a fragment extending from exon 5 through exon 6 and including the intervening intron 6 gave a reproducible pattern, with a 280-base pair (bp) band from canine, equine, bovine, ovine, and caprine samples showing high sequence homology. Porcine, guinea pig, tiger, and lion samples, however, gave an additional fragment of approximately 197 bp. The whole intron 6 from these fragments is missing, possibly representing a pseudogene. In feline samples only the 197-bp fragment could be detected. This study shows that the PCNA gene is highly conserved across a broad range of animal species and is well suited as an internal control for PCR analysis in veterinary medicine.

Animals↗

Ethical issues in the documentary data analysis of Internet posts and archives.

The documentary analysis of e-mail posts and archives for qualitative research has been outlined elsewhere. Although there is an increase in the number of studies being conducted on listserv and newsgroup material in health research, this has not always been accompanied by a careful, in-depth consideration of the concomitant ethical issues. Therefore, this article outlines the ethical considerations surrounding this form of research, including issues of accessing voices, consent, privacy, anonymity, interpretation, and ownership and authorship of research material.

Communication↗

Evaluation of a hospital picture archiving and communication system.

OBJECTIVES: To establish the net costs to the hospital and the broad range of benefits associated with a hospital-wide picture archiving and communication system (PACS) that comprised digital acquisition, storage and transmission of radiological images via a hospital-wide network to 150 workstations. METHODS: 'Before and after' comparisons and time series analyses at Hammersmith Hospital (London, UK), and comparison with five other British hospitals where PACS was not being installed. The cost analysis considered implementation costs and changes in key elements of hospital running costs, including the impact of changes in the length of inpatient stays. A range of benefit measures were investigated, including image availability, avoidance of repeat imaging, avoidance of exposure to radiation, patient turn-round speed, time from examination to image availability in intensive care, avoidance of diagnostic 'errors' by casualty doctors, the additional diagnostic value of PACS-based images and clinician satisfaction. RESULTS: The annual equivalent capital cost of the PACS was 1.7 million Pounds (annual equivalent replacement cost: 0.8 million Pound). Overall, the PACS substantially increased running costs. No convincing evidence of a PACS-induced change in length of inpatient stay was found. PACS was associated with some improvements in the performance of the radiology department: improved image availability (97.7% versus 86.9%), lower repeat imaging rate (7.3% versus 9.9%) and 20% lower total radiation doses for examinations of the lateral lumbar spine. No improvements were identified in the quality of the radiology reporting service. Benefits outside radiology included shorter time from examination to image availability for routine uses in intensive care (19 versus 37 minutes), and a lower rate of diagnostic 'errors' in casualty (0.65% versus 1.51%). High levels of satisfaction with PACS were found amongst both providers and clinical users. CONCLUSIONS: PACS was almost universally preferred by users and brought many operational and clinical benefits. However, these advantages came at a significant capital and net running cost.

Cost-Benefit Analysis↗

In situ hybridization with human papillomavirus using biotinylated DNA probes on archival cervical smears.

We report a method of in situ hybridization (ISH) of 10-year-old archival cervical smears with a cocktail of nick-translated human papillomavirus (HPV) DNA types 6, 11, 16, 18, and 31. The method, which does not require destaining, results in excellent preservation of morphological detail with only 2% cell loss. Methods of smear treatment and detection of the biotinylated probe with a multistep avidin-biotin-immunoperoxidase method are described. Biotinylated PBR 322 plasmid and biotinylated human DNA were used as negative and positive controls in each run. Twenty-nine of 50 smears (58%) showing changes consistent with CIN I-II were positive for HPV. Fourteen corresponding cervical biopsies were also studied by ISH, seven corresponding to HPV-positive smears and seven to HPV-negative smears. HPV DNA was demonstrated in six of seven biopsies (87%) from the positive group but none could be demonstrated in the negative group. We conclude that retrospective study can be performed on routine alcohol-fixed, Papanicolaou-stained cervical smears with biotinylated HPV probes with excellent cell preservation, minimal cell loss, and high degrees of specificity.

Biotin↗