Ronald S. Weinstein

Inside-Out Red Cell Membrane Vesicles: Preparation and Puirification

Plasma membranes purified from human red cells were Converted into small vesicleS by disruption in alkaline buffer of low ionic strength. Most of these vesicles were inside-out. The presence of divalent cations prevented this inversion. The inside-out vesicles were separatcd from right-side-out vesicles by centrifugration to equilibrium in dextran density gradients.

Overview of telepathology, virtual microscopy, and whole slide imaging: prospects for the future ☆

Telepathology, the practice of pathology at a long distance, has advanced continuously since 1986. Today, fourth-generation telepathology systems, so-called virtual slide telepathology systems, are being used for education applications. Both conventional and innovative surgical pathology diagnostic services are being designed and implemented as well. The technology has been commercialized by more than 30 companies in Asia, the United States, and Europe. Early adopters of telepathology have been laboratories with special challenges in providing anatomic pathology services, ranging from the need to provide anatomic pathology services at great distances to the use of the technology to increase efficiency of services between hospitals less than a mile apart. As to what often happens in medicine, early adopters of new technologies are professionals who create model programs that are successful and then stimulate the creation of infrastructure (ie, reimbursement, telecommunications, information technologies, and so on) that forms the platforms for entry of later, mainstream, adopters. The trend at medical schools, in the United States, is to go entirely digital for their pathology courses, discarding their student light microscopes, and building virtual slide laboratories. This may create a generation of pathology trainees who prefer digital pathology imaging over the traditional hands-on light microscopy. The creation of standards for virtual slide telepathology is early in its development but accelerating. The field of telepathology has now reached a tipping point at which major corporations now investing in the technology will insist that standards be created for pathology digital imaging as a value added business proposition. A key to success in teleradiology, already a growth industry, has been the implementation of standards for digital radiology imaging. Telepathology is already the enabling technology for new, innovative laboratory services. Examples include STAT QA surgical pathology second opinions at a distance and a telehealth-enabled rapid breast care service. The innovative bundling of telemammography, telepathology, and teleoncology services may represent a new paradigm in breast care that helps address the serious issue of fragmentation of breast cancer care in the United States and elsewhere. Legal and regulatory issues in telepathology are being addressed and are regarded as a potential catalyst for the next wave of telepathology advances, applications, and implementations.

The Structure and Function of Intercellular Junctions in Cancer

Publisher Summary Intercellular junctions are a set of structurally complex membrane components that are incorporated into the general plasma membrane at the sites of close cell-to-cell apposition. The primary function of some types of junctions, such as gap junctions, remains obscure; this is unfortunate because a considerable body of information on the occurrence, biochemical ultrastructure, and physical properties of junctions suggests that they probably do play a central role in important biological phenomena. The quantitative evaluations of the occurrence of junctions are subjective in most of the reports, although there are a few reports in which the data were obtained by quantitative electron microscopy techniques. There are a number of ways in which genes of neoplastic transformation may influence intercellular junction structure and function. Additional research is required to define the contribution of tumor gene products to the pathogenesis of junctional abnormalities in tumors and to elucidate the roles, if any, of these cell membrane defects in malignant growth.

P-glycoproteins in pathology: The multidrug resistance gene family in humans

Many cancers do not respond to chemotherapy on primary exposure to drugs, thus manifesting intrinsic drug resistance. Other cancers that do initially respond subsequently become resistant to the same drugs and simultaneously to other drugs to which the patient has had no previous exposure. This is a form of acquired drug resistance. There is a pressing need to better understand the mechanisms of drug resistance and to use this information to develop strategies for the chemosensitization of drug-resistant tumors. A goal of the pathology laboratory is to offer chemosensitivity tests that identify intrinsic or acquired resistance of tumors to specific drugs or classes of drugs to enable the clinician to tailor therapy to the biology of cancers in individual patients. Multidrug resistance is one type of drug resistance. It can be present in either an intrinsic or acquired form. The human gene that confers human multidrug resistance, the MDR1 gene, has been cloned and classified as a member of the MDR gene family. Its encoded protein, called Mdr1, is an energy-driven membrane efflux transporter that maintains intracellular concentrations of certain chemotherapeutic drugs at nontoxic levels. Useful model systems for studying multidrug resistance have been developed in several research laboratories. Applying selection pressure by exposing cultured cancer cells to escalating doses of natural product anti-cancer drugs allows cross-resistant cell lines to be produced which share patterns of drug resistance with human cancers. A common feature of these drug-resistant lines is the expression of Mdr1. Using techniques of genetic engineering, molecular probes have been developed that can be used to measure MDR1 mRNA and MDR1 gene amplification. Mdr can be measured by immunochemistry methods. Currently, such measurements are being used to stratify patients in clinical trials designed to determine if chemosensitization by inhibition of the pump function of Mdr is a clinically useful therapeutic strategy. If successful, Mdr/MDR1 mRNA laboratory testing might significantly increase the clinical laboratorys role in cancer patient management.

Cytogenetics of 158 patients with regional or disseminated melanoma. Subset analysis of near-diploid and simple karyotypes.

We report on the cytogenetic analyses of 158 cases of metastatic malignant melanoma, comprised of 63 cases with regional disease (RD) and 95 cases with distant (metastatic) disease (DD). Clonal structural abnormalities were identified in 126 (80%) cases and were significantly increased ( < 0.01 after adjusting for multiple comparisons) on chromosomes (in order of frequency of involvement) 1, 6, 7, 11, 9, and 3. Clustering of breakpoints occurred at 1p36, 1p22-q21, 6p11-q21, 9p, 11q23-qter, 13p (especially for cases with DD), and 19q13. The most common clonal numerical abnormalities, in a subset of 49 near-diploid cases were -10, -22, -9, +7, -19, and -Y. Analysis of chromosome segment gains and losses (CSRP) showed frequent loss of chromosomes 6 and 10, followed by equal rates of involvement of chromosomes 1, 7, and 9. Whole or segmental losses of chromosome 9 (especially 9p) correlate well with recent molecular genetic studies identifying putative suppressor genes, and are also likely important genetic abnormalities. However, based on the frequency of abnormalities in this large series of metastatic melanomas, it is likely that structural abnormalities of 1 and 6, and 10 are important in the pathogenesis of sporadic advanced melanoma.

Advances in Flow Cytometry for Diagnostic Pathology

Only 3 years have passed since Lovett et al. (147) first reviewed applications of flow cytometry (FCM) to diagnostic pathology in Laboratory Investigation. Progress in this field has been rapid, however, and the importance of having timely, practical knowledge of this technology has increased significantly for pathologists. Lovett’s lucid explanation of the basic function of the instrument would still be difficult to improve upon. Therefore, this review is not intended to duplicate or replace the earlier substantial monograph. Rather, we will attempt to update the reader on what we regard to be important recent developments in the field. We will also consider some older work which seems in retrospect, to have been insufficiently covered in Lovett’ review. Readers with little or no background in FCM may benefit from referring to the earlier review before starting this one, or else consulting other more comprehensive works, especially Shapiro’s monograph (204). For the many pathologists who have been following the development of FCM, we hope this review will aid in bringing advances in the state-of-the-art into perspective.

Toward the development of miniaturized imaging systems for detection of pre-cancer

In this paper, we describe the progress toward the development of miniaturized imaging systems with applications in medical imaging, and specifically, detection of pre-cancer. The focus of the article is a miniature, optical-sectioning, fluorescence microscope. The miniature microscope is constructed from lithographically printed optics and assembled using a bulk micro-machined silicon microoptical table. Optical elements have been printed in a negative tone hybrid glass to a maximum depth of 59 /spl mu/m and an rms surface roughness between 10-45 nm, fulfilling the requirements of the miniature microscope. Test optical elements have been assembled using silicon-spring equipped mounting slots. The design of silicon springs is presented in this paper. Optical elements can be assembled within the tolerances of an NA=0.4 miniature microscope objective, confirming the concept of simple, zero-alignment assembly.

The Empirical Foundations of Telemedicine Interventions for Chronic Disease Management

The telemedicine intervention in chronic disease management promises to involve patients in their own care, provides continuous monitoring by their healthcare providers, identifies early symptoms, and responds promptly to exacerbations in their illnesses. This review set out to establish the evidence from the available literature on the impact of telemedicine for the management of three chronic diseases: congestive heart failure, stroke, and chronic obstructive pulmonary disease. By design, the review focuses on a limited set of representative chronic diseases because of their current and increasing importance relative to their prevalence, associated morbidity, mortality, and cost. Furthermore, these three diseases are amenable to timely interventions and secondary prevention through telemonitoring. The preponderance of evidence from studies using rigorous research methods points to beneficial results from telemonitoring in its various manifestations, albeit with a few exceptions. Generally, the benefits include reductions in use of service: hospital admissions/re-admissions, length of hospital stay, and emergency department visits typically declined. It is important that there often were reductions in mortality. Few studies reported neutral or mixed findings.

Amplification of 19q13.1-q13.2 sequences in ovarian cancer: G-band, FISH, and molecular studies

In this study of ovarian carcinoma, we extended previous findings by performing FISH using chromosome 19 paint and microFISH probes and patient samples with and without abnormalities of chromosome 19 identified by G-banding. Karyotype interpretations of der(19) were confirmed, while additional 19 translocations were also detected by FISH with 19WCP in some cases. Similar FISH studies of ovarian carcinoma cell lines found chromosome 19 abnormalities even after extensive in vitro culture. MicroFISH probes were generated by chromosome microdissection from two cases with hsr(19) and mapped to 19q13.2 and 19q13.1-.2, respectively. FISH with these microFISH probes alone or in combination with a 19WCP probe to four patient samples and seven cell lines showed that 65% of chromosome 19 structural abnormalities contained 19q13.1-q13.2 sequences, sometimes as large hsrs. Ovarian cancer cell lines showed amplification and overexpression of the AKT2 putative oncogene, but not the ERCC-2 DNA repair gene in this chromosomal region. In addition to AKT2, amplification and overexpression of other yet-unidentified genes in the 19q13.1-q13.2 region may contribute to ovarian carcinoma pathogenesis or progression.

Blood group-related antigens as markers of malignant potential and heterogeneity in human carcinomas‡

The expression of BGR-Ags is often aberrant in human carcinomas. The observation that BGR-Ag expression in human bladder carcinomas correlates with prognosis for patients with these tumors is especially interesting in light of the numerous reports of correlations between cell surface glycosylation and malignant phenotype in experimental animal tumors. Many observations suggest how this relation might be mediated. It seems reasonable to anticipate that the study of the BGR-Ags and their expression in carcinoma may emerge from its current predominantly descriptive phase and become an important part of the investigation of human tumor biology.