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PubMed2006

Protein expression in a transformed trabecular meshwork cell line: proteome analysis.

PURPOSE: Characterization of the human trabecular meshwork (TM) proteome is hindered by the small mass of intact tissue and the slow growth of cultured cell strains. We have previously characterized a transformed TM cell strain (GTM3) that demonstrates many of the same protein expression and cell signaling systems of nontransformed cell strains. The aim of this study was to initiate a proteomic survey of GTM3 cells as the initial step toward characterization of the complete human TM proteome. METHODS: GTM3 cells were cultured to confluence, harvested and solubilized in urea/Nonidet. The protein extract (600 mug) was focused in immobilized isoelectric focusing (IEF) strips, separated by 10% SDS PAGE, and visualized with colloidal Coomassie Blue. Spots of interest were excised, destained, and the contained proteins subjected to in-gel reduction, derivatization, and tryptic digestion. Tryptic peptides were extracted and analyzed by electrospray LC/MS/MS. Protein identification was made using the TurboSequest search algorithm and a recent version of the nonredundant human protein database downloaded from the National Center for Biotechnology Information (NCBI). RESULTS: Eighty-seven (87) primary proteins and 93 variants of these proteins were identified. A website was created (TM proteome) that combines data such as graphic spot location within the gel, peptide sequence, apparent and calculated pI, apparent and calculated mass, percentage of coverage, and protein informatic website links. CONCLUSIONS: Proteomic analysis of a transformed human TM cell line has been initiated combining preparative two-dimensional PAGE separation, LC/MS/MS analysis of major proteins, and bioinformatic cataloging of the data. Further investigation of data from the transformed cell strain will be used in a comparative fashion for spot identification of analytical proteomic gels of human TM tissue and cultured normal cells. These initial data will form the base from which the characterization of protein expression in the normal and glaucomatous TM can be accomplished.

Cell Line, Transformed↗
PubMed2006

Why innovation in health care is so hard.

Health care in the United States--and in most other developed countries--is ailing. Medical treatment has made astonishing advances, but the packaging and delivery of health care are often inefficient, ineffective, and user unfriendly. Problems ranging from costs to medical errors beg for ingenious solutions-and indeed, enormous investments have been made in innovation. But too many efforts fail. To find out why, it's necessary to break down the problem, look at the different types of innovation, and examine the forces that affect them. Three kinds of innovation can make health care better and cheaper: One changes the ways consumers buy and use health care, another taps into technology, and the third generates new business models. The health care system erects an array of barriers to each type of innovation. More often than not, organizations can overcome the barriers by managing the six forces that have an impact on health care innovation: players, the friends and foes who can bolster or destroy;funding, the revenue-generation and capital-acquisition processes, which differ from those in other industries; policy, the regulations that pervade the industry; technology, the foundation for innovations that can make health care delivery more efficient and convenient; customers, the empowered and engaged consumers of health care; and accountability, the demand from consumers, payers, and regulators that innovations be safe, effective, and cost-effective. Companies can often turn these six forces to their advantage. The analytical framework the author describes can also be used to examine other industries. Cataloging the innovation types and identifying the forces that aid or undermine them can reveal insights on how to treat chronic innovation ills- prescriptions that will make any industry healthier.

Delivery of Health Care↗
PubMed1996

Digitized media--we used to call it clip art.

One major problem when dealing with thousands of images is finding the exact image that you want to use. All of the packages reviewed in this article come with a printed catalog of thumbnail sketches of the various individual images, as well as an electronic image browser that will call up on the screen selected images that fulfill specific search criteria.

Computers↗
PubMed2006

Discovering cultural aspects of nurse-patient relationships.

The purpose of this article is to illuminate modes of inquiry that unconceal cultural aspects of the meaningful life-world of individuals. To present strategies for acquiring cultural awareness, sensitivity, and competence based on insights gained from these modes of inquiry. Nurses can acquire and implement culturally competent patient care by inquiring into individual's personal interpretations of their life's world experiences rather than relying on catalogs of cultural attributes or by adhering to popularly held opinions. By following the procedures for augmenting culturally competent nursing outlined in this article nurses reported being able to provide a more culturally competent, higher quality of patient care.

Attitude of Health Personnel↗
PubMed2006

[The database and its data sharing of neuropathic images].

This paper introduces the basic principle and method of establishing a database for neuropathic images, and discusses its significance and data sharing. The database is composed of three data volumes about basic knowledge for images, cranial sectional anatomy and neuropathic images. The data sharing is achieved by using a method of linking the dynamic network of neuropathic images with database of neuropathic images. There are three searching approaches: catalog searching, key words searching and code searching.

Databases, Bibliographic↗
PubMed2006

Chronic mountain sickness: the reaction of physical disorders to chronic hypoxia.

Chronic mountain sickness (CMS) is a condition in which hematocrit is increased above the normal level in residents at high altitude. In this article we take issue with the "Consensus Statement On Chronic And Subacute High Altitude Diseases" of 2005 on two essential points: using a questionnaire to evaluate the symptoms of CMS to use the term "loss of adaptation" as opposed to "adaptation to disease in the hypoxic environment". We opine that CMS is rather an adaptive reaction to an underlying malfunction of some organs and no specific symptoms could be quantified. To substantiate our line of reasoning we reviewed 240 CMS cases seen at the High Altitude Pathology Institute in La Paz. Patients who had a high hematocrit (<58%) underwent pulmonary function studies in search for the cause of hypoxia: hypoventilation, diffusion alteration, shunts, and uneven ventilation-perfusion. The tests included arterial blood gas tests, chest x-rays, spirometry, hyperoxic tests, flow-volume curves, ventilation studies at rest and during exercise, ECG, exercise testing and doppler color echocardiography to assess heart structure and function. When correlated with clinical history these results revealed that CMS is practically always secondary to some type of anomaly in cardio-respiratory or renal function. Therefore, a questionnaire that tries to catalog symptoms common to many types of diseases that lead to hypoxia is flawed because it leads to incomplete diagnosis and inappropriate treatment. CMS, once again, was shown to be an adaptation of the blood transport system to a deficient organs' function due to diverse disease processes; the adaptation aimed at sustaining normoxia at the cellular level in the hypoxic environment at high altitude.

Adaptation, Physiological↗
PubMed2006

Identifying protein complexes in high-throughput protein interaction screens using an infinite latent feature model.

We propose a Bayesian approach to identify protein complexes and their constituents from high-throughput protein-protein interaction screens. An infinite latent feature model that allows for multi-complex membership by individual proteins is coupled with a graph diffusion kernel that evaluates the likelihood of two proteins belonging to the same complex. Gibbs sampling is then used to infer a catalog of protein complexes from the interaction screen data. An advantage of this model is that it places no prior constraints on the number of complexes and automatically infers the number of significant complexes from the data. Validation results using affinity purification/mass spectrometry experimental data from yeast RNA-processing complexes indicate that our method is capable of partitioning the data in a biologically meaningful way. A supplementary web site containing larger versions of the figures is available at http://public.kgi.edu/wild/PSBO6/index.html.

Algorithms↗
PubMed1991

Identification and characterization of rat intestinal keratins. Molecular cloning of cDNAs encoding cytokeratins 8, 19, and a new 49-kDa type I cytokeratin (cytokeratin 21) expressed by differentiated intestinal epithelial cells.

In the previous paper (Quaroni, A., Calnek, D., Quaroni, E., and Chandler, J.S. (1991) J. Biol. Chem. 266, 11923-11931) we describe the use of a panel of "antikeratin" monoclonal antibodies to study cytokeratin distribution in rat intestinal epithelium. In the present paper we describe the use of three antikeratin monoclonal antibodies to identify and recovery cDNA clones expressing immunologically specific fusion proteins from a rat intestinal cDNA library. DNA sequence analysis identified each cDNA encoded epitope including the carboxyl-terminal portions of cytokeratins 8 and 19 (as cataloged by Moll, R., Franke, W.W., and Schiller, D.L. (1982) Cell 31, 11-24) recognized by antibodies RK4 and RK7, respectively. In addition, antibody RK5 was used to recover a cDNA clone (pRK5) encoding a portion of a 48-kDa keratin-related protein with unique tissue and cellular distribution, designated cytokeratin 21. Translation of cDNA-selected mRNAs yielded individual proteins which could be resolved and identified by their specific immunoreactivities. The pRK5 cDNA was used to recover a larger (approximately 1.3 kilobase pairs) cDNA clone (KB2) from an independent cDNA library for DNA sequence analysis and for the recovery of additional overlapping cDNA clones. The resulting cDNA sequence (1519 base pairs) contains the complete coding region of cytokeratin 21 (49,387 daltons). The predicted amino acid sequence of cytokeratin 21 confirms its identity as a novel type I cytokeratin expressed predominantly in the intestinal epithelium.

Amino Acid Sequence↗
PubMed2006

[Mutations of hemochromatosis gene in volunteer blood donors and Chilean porphyria cutanea tarda patients].

In patients with porphyria cutanea tarda (PCT), hepatic iron accumulation associated to hereditary hemochromatosis (HH) could play a role in the etiology and in the clinical expression of the disease. The H63D and C282Y mutations of the HFE gene frequency were studied in a PCT group of patients and compared with the frequency observed in a group of volunteer blood donors. PCT patients were cataloged as hereditary or acquired PCT carriers, whether or not they presented uroporphyrinogen decarboxilase gene mutations. Fifty percent of PCT patients were carriers of the disease's genetic type. Such percentage is significantly higher than what other authors have previously informed. H63D and C282Y mutations were present in 23% and 2.4% of the volunteer blood donors, respectively. Similar frequencies were informed by others authors in Chilean white ethnic populations, and also in Spaniard and Argentinean populations, but significantly higher than that observed in Chile's Araucanean aboriginal population. Probably the frequency of H63D and C283Y mutations are related to the Spaniard ascendancy dominance of Chile's white ethnic population. The frequency of HFE gene mutations in PCT patients was not different than what was observed in volunteer blood donors. Similarly, there was no statistical difference in the frequency of these mutations among patients with acquired or genetic PCT disease. With the obtained results, it is not possible postulate an association between PCT and the hereditary hemochromatosis of HFE gene mutations carrier conditions.

Blood Donors↗
PubMed2006

[Dynamics of genetic diversity in winter common wheat Triticum aestivum L. cultivars released in Russia from 1929 to 2005].

Genealogical analysis was used to study the dynamics of genetic diversity in Russian cultivars of winter common wheat from 1929 to 2005. The Shannon diversity index of the total set of released cultivars remained almost unchanged, although the number of original ancestors (landraces and genetic lines) increased almost tenfold in the period under study. This was explained in terms of the dependence of the modified Shannon diversity index on two parameters, the number of original ancestors and the mean coefficient of parentage. Significant direct effects were revealed: a positive effect of the former parameter and a negative of the latter. As a result, the increase in the number of original ancestors was compensated by the increase in relatedness of cultivars. Genetic erosion of realized diversity was observed, as a half of Russian landraces were lost. Although the mean coefficient of parentage did not reach its critical value (R = 0.25), cultivars of some regions (Central and Volga-Vyatka) proved to be closely related. A favorable gradual decrease in the mean coefficient of parentage was observed in the past 15 years. A set of modem winter wheat cultivars, which were introduced in the Russian State Catalog from 2002 to 2005, displayed a cluster structure. The overwhelming majority of cultivars formed two clusters originating from Bezostaya 1 (67% of cultivars) and Mironovskaya 808 (31%).

Genetic Variation↗
PubMed1991

[The use of monoclonal antibodies to cytokeratin peptides for the diagnosis of basal cell carcinoma of the skin].

25 skin basaliomas were studied immunohistochemically using 6 different monoclonal antibodies (MAB) to the low-molecular cytokeratins N 8, 18 (according to Moll's catalog) as well as to the cytokeratins N 1, 2, 9, 10, 11 and N 17. Prekeratin N 17 was found in all tumours while cytokeratin N 18 was found in no tumour. Prekeratin N 8 was expressed in 24% of tumours. MAB EE 21-06 to the cytokeratin N 1, 2, 9, 10, 11 are reactive with tumour cells in 20% of cases. Basalioma cells (apart from trichobasalioma) did not react with MAB G 36-19. These results suggest a dissimilar origin of basaliomas and may help in the differential diagnosis of the skin malignant tumours.

Antibodies, Monoclonal↗
PubMed2006

Gene expression profile of the adult human retinal ganglion cell layer.

PURPOSE: Pathophysiological events in the retinal ganglion cell layer (GCL) are a prominent feature of several optic neuropathies including glaucoma. The purpose of this study was to identify and catalog genes whose expression in the human retina is restricted to the GCL. METHODS: Laser capture microdissection (LCM) technology was used to isolate tissue from the perimacular retina of three human donors without retinal or optic nerve disease. RNA was isolated from the (1) retinal GCL and (2) the inner and outer nuclear layers of the same retina, and the gene expression profiles of both fractions were determined using Affymetrix Hu133Plus 2.0 GeneChips. Data were analyzed to identify those genes whose expression is substantially more prevalent in the GCL when compared to the outer retinal layers. Differential expression of selected genes was confirmed by real-time PCR and immunohistochemistry. RESULTS: The results show that mRNA levels of previously described ganglion cell markers, e.g. the neurofilament genes (NEFH, NEF3, or NEFL) and the Brn3a transcription factor (POU4F1), were highly enriched in the isolated GCL fraction. In contrast, transcripts for genes associated with phototransduction (RHO), photoreceptor development (NR2E3), or interphotoreceptor matrix constituents (IMPG1) were nearly absent from the GCL fraction. Using bioinformatics approaches over 80 genes were identified whose expression in the human retina appears to be limited to the GCL. CONCLUSIONS: We have successfully used LCM technology to generate gene expression profiles of highly enriched GCL fractions of the normal human retina. These data not only provide clues to the normal function of retinal ganglion cells but also serve as a resource in the development of ganglion cell specific markers or transfection vectors, and the identification of candidate genes for hereditary forms of glaucoma.

Aged↗
PubMed1976

Cystic diseases of the kidney in children.

Many genetically determined cystic kidney disorders occur in children. Presenting manifestations related to the kidney include failure to thrive, abdominal enlargement or flank masses, progressive renal failure, or urinary tract infection. Radiological techniques often yield information helpful in establishing the type and extent of renal lesions. Cataloging the cystic renal disease in terms of genetic patterns, the involvement of liver or another organs, and the nature of the disorder of renal architecture is helpful in establishing the proper handling of affected children, and in genetic counseling.

Abnormalities, Multiple↗
PubMed1991

The establishment of an academic health sciences library in a developing country: a case study.

The development of a Faculty of Medical Sciences (FMS) and an academic health sciences library for the University of the West Indies (UWI) has proven to be a polemical and political issue due to the depressed economy of the country. Although FMS is still shrouded in politics and controversy after its inaugural year, the Medical Sciences Library (MSL) has expanded its dimensions and is actively developing a biomedical information network within the country. This will result in better dissemination and control of biomedical information. The library now participates in joint projects with other health sciences libraries in the country with the goal of joint automated listings of holdings and shared cataloging projects. This paper examines the development of the library and explains the difficulties experienced in its developmental stages due to politics, the delay in appointment of a medical sciences librarian, and the financial decline in the local economy.

Developing Countries↗
PubMed1990

The biologic actions of extracellular adenosine triphosphate.

The study of the biologic effects of extracellular ATP is rapidly progressing from cataloging the integrated biologic responses of isolated tissues to characterizing the biochemical basis of signal transduction in isolated cells. Clearly, many fundamental issues remain; these include the classification of receptor subtypes, identification of defined or new signal-transduction pathways, generation of specific high-affinity ligands for the various ATP receptor subtypes, and ultimately, the development of protocols for the identification, isolation, cloning, and physicochemical characterization of the receptor moities per se. Such work will be essential for developing selective ATP-receptor antagonists. Such antagonists, in turn, will facilitate evaluation of the role of these receptors in physiological and/or pathological processes. A possible therapeutic role for drugs selective for these receptors is suggested by the critical pathophysiological roles of the human cell types (phagocytic leukocytes, vascular endothelial cells) known to express such receptors. The earlier, seminal work on the biologic actions of extracellular ATP was largely performed by smooth-muscle physiologists and pharmacologists. With the expanding range of, and interest in, this topic, recent studies have been carried out largely by diverse groups of pharmacologists, neurobiologists, physiologists, and biochemists. Not surprisingly, these various scientists often investigate certain aspects of extracellular ATP action from different perspectives, using the specialized tools and methods of their respective disciplines. Hopefully, by integrating these interdisciplinary perspectives on the biologic actions of extracellular ATP, there will be rapid progress in this field during the next few years.

Adenosine Triphosphate↗
PubMed1990

Microcephaly: general considerations and aids to nosology.

Microcephaly is defined as an occipito-frontal head circumference (OFC) 2 or more standard deviations below the mean for age and sex using the new Roche et al. [Pediatrics 1987;79:706-712] charts, and corrected for parental OFC by the method of Weaver and Christian [J Pediatr 1980;96:990-994]. "Relative" microcephaly, i.e., a small head on a small child, may be associated with a much better intellectual prognosis than absolute microcephaly, although the average IQ of children with absolute microcephaly ascertained in a normal school system is normal when compared with that of appropriate control children. "Primary" microcephaly means an abnormal OFC at birth (corrected for gestational age and length), and "secondary" microcephaly a normal birth OFC with later, acquired microcephaly due to deceleration of brain growth reflecting infection, trauma, intoxication, metabolic disease, the Rett syndrome, or a true CNS degenerative disease. Some cases of syndromal microcephaly may be associated with normal intelligence including some "primordial dwarfs," children with Dubowitz syndrome, FAS, mild SC-Roberts syndrome, and an occasional Brachmann-de Lange individual. The nosology of (syndromal) microcephaly is extraordinarily complex and requires the assistance of special library resources and information retrieval expertise. At a minimum, it requires McKusick's Catalog of Mendelian Inheritance in Man (MIM); however, we find that our work is greatly enhanced by recently developed electronic databases such as MIM-online (OMIM), POSSUM, SYNDROME, and MEDLINE, as well. Three groups of syndromal and non-syndromal microcephaly are discussed selectively in order to illustrate the marvels of pleiotropy in human development and its abnormalities and the difficulties encountered in splitting and lumping entities with overlapping manifestations.

Abnormalities, Multiple↗
PubMed1990

Regional health library service in northern Ireland.

The regional medical library service provided to physicians, hospitals, nurses, social workers, and health care administrators throughout Northern Ireland by the Queen's University of Belfast is described. A brief outline of the National Health Service in the United Kingdom is given, and the library service is described in terms of collections, cataloging, interlibrary loan, and reference.

Capital Financing↗
PubMed1989

Neutral sphingomyelinase from human urine. Purification and preparation of monospecific antibodies.

A neutral sphingomyelinase which cleaves phosphorylcholine from sphingomyelin at a pH optima of 7.4 was purified 440-fold to apparent homogeneity from normal human urine concentrate employing Sephadex G-75 column chromatography, preparative isoelectric focusing, and sphingosylphospholcholine CH-Sepharose column chromatography. The enzyme is composed of a single polypeptide whose apparent molecular weight is 92,000. Analytical isoelectric focusing revealed that the pI of this enzyme is 6.5. Purified neutral sphingomyelinase was devoid of beta-galactosidase and beta-N-acetylglucosaminidase activity originally present in the urine concentrate. The purified neutral sphingomyelinase (N-SMase) had low levels of phospholipase A1 and A2 activity when phosphatidylcholine was used as a substrate and detergents were included in the assay mixture. However, it had no phospholipase activity toward phosphatidylglycerol and sphingomyelin at pH 4.5 irrespective of the presence or absence of detergents. Monospecific polyclonal antibodies raised against N-SMase immunoprecipitated approximately 70% of N-SMase activity from urine, human kidney proximal tubular cells, and partially purified membrane-bound N-SMase from these cells. Western immunoblot assays revealed that the monospecific polyclonal antibody against urinary N-SMase recognized both the urinary N-SMase and the membrane-bound N-SMase. Because this enzyme is distinct biochemically and immunologically as compared to acid sphingomyelinase (EC 3.1.4.12), we would like to assign it an enzyme catalog number of EC 3.1.4.13. The availability of N-SMase and corresponding antibody will be useful in studying various aspects of this enzyme in biological systems.

Acetylglucosaminidase↗