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PubMed2001

Caenorhabditis elegans homologues of the CLN3 gene, mutated in juvenile neuronal ceroid lipofuscinosis.

Neuronal ceroid lipofuscinoses (NCLs) are the most common hereditary neurodegenerative disorders of childhood. The first symptom of this heterogeneous group of devastating lysosomal storage diseases is progressive visual failure. The different forms of NCL can be distinguished by age of onset, clinical features and the characteristics of the accumulated materials. The juvenile form, Batten-Spielmeyer-Vogt disease which is caused by mutations in the CLN3 gene, is the most frequent form of the disease in which loss of vision becomes apparent around the age of 5-8 years. The gene was found to encode a novel integral membrane protein localizing to the lysosomes, confirming that the primary defect in NCL is in lysosomal function. The CLN3 protein function is still unknown, and is examined in several model organisms. We are studying the nematode Caenorhabditis elegans, and have identified three CLN3 homologues. In order to investigate the role of the CLN3 protein in C. elegans, Cecln-3 deletion mutants are being isolated from an ethyl methanesulphonate (EMS)-induced deletion mutant library. Examination of these mutants may provide us with information that will help in dissecting the processes in which the CLN3 protein is involved. In this library two mutated C. elegans Cln-3 loci have been identified, of which one mutant, NL748, was isolated. This mutant contains a deletion of the whole gene. The deletion mutant was characterized with regard to life expectancy, and showed no significant differences when compared with wild-type.

Animals↗

PubMed1999

Monte Carlo calculations to characterize the source for neutron therapy facilities.

Modern radiation treatment planning for photons includes full 3D modeling of the adsorbed dose distribution, accurate inclusion of the patient anatomy, and consideration of significant changes in material density and composition. Such efforts are founded in an accurate description of the radiation source and the beam delivery system. Modern fast neutron therapy facilities employ highly penetrating beams and isocentric beam delivery. Treatment planning is largely based on analytic models adapted from photon codes and interaction cross sections normalized to macroscopic attenuation. However, the recent PEREGRINE initiative at Lawrence Livermore Laboratory offers the possibility of fully stochastic modeling if the neutron source can be adequately described. In this article we report neutron source modeling of three high energy facilities. Neutron production is based on the intra-nuclear cascade model of the LAHET code while neutron transport through the beam delivery system is managed by MCNP using cross section libraries extended to 100 MeV neutron energy. PEREGRINE is then used to transport the neutron beam through typical phantoms. The resulting neutron sources are in excellent agreement with the limited experimental information and the measured phantom data are well described by the PEREGRINE transport using the LAHET/MCNP determined neutron sources.

Dose-Response Relationship, Radiation↗

PubMed1996

Chromosomal rearrangements detected by FISH and G-banding.

Fluorescence in situ hybridization (FISH) using chromosome-specific DNA libraries as painting probes, locus-specific unique sequence (cosmid) probes, and Y-specific repetitive sequences was applied in the analysis of eighteen cases of chromosomal rearrangements of undetermined nature. FISH clarified the origin of the extra or translocated chromosome segments in seventeen patients, one with 2q+, two with 4q+, one each with 6p+, 7p+, 9q+, 10p+, 11q+ and 12p+, two with 13q+, and one each with 15q+, 17p+, 18p+, 20p+, 21p+ and Yq+, as well as the nature of a de novo supernumerary chromosome marker in a previously reported case. By G-banding and molecular cytogenetic studies of the family members, six cases were determined to have unbalanced translocations inherited from the carrier parent. The extra translocated genetic material may cause specific trisomic syndromes, including partial 6p21.3-p23, 9q32-q34.3, 13q32-q34, 15q24-q26, and 17p11.2-p13 trisomies in those patients. A translocated 21q segment on 12p was shown by a painting probe in a patient with Down features. A patient with cat cry syndrome resulting from a loss of the terminal segment of the short arm of chromosome 5 was confirmed by a cosmid probe showing de novo reciprocal translocation between chromosomes 5 and 18:t(5;18) (p13.3;p11.31). With FISH, the extra material on the rearranged chromosome could also be identified as duplicated or translocated. The FISH technique thus provides a method for the analysis of extra structurally abnormal chromosomes (especially in de novo cases), recognizable syndromes (contiguous gene syndromes) caused by translocated deletion from parental balanced chromosome rearrangements, and supernumerary marker chromosomes. FISH subsequent to G-banding is also of great help in the confirmation of preliminary abnormal G-banded karyotypes after a modified destaining procedure. In conclusion, the combination of G-banding and FISH is very useful in the accurate diagnosis of chromosomal rearrangements.

Chromosome Aberrations↗

PubMed2000

Nitinol - its use in vascular surgery and other applications.

OBJECTIVES: to describe the physical properties of shape-memory alloys and the surgical, scientific and commercial applications of nitinol, in particular. DESIGN AND METHODS: a Medline, Internet and library search with contributions from commerce to describe the alloy's structure, behaviour and biocompatibility, and design for devices constructed from nitinol. RESULTS: nitinol has the properties of thermal shape memory and superelasticity that make it ideal for many vascular and general surgical prostheses and disposables, and for various commercial applications. CONCLUSIONS: further research into shape-memory alloys from scientific and commercial groups should widen their use in vascular and endovascular surgery.

Alloys↗

PubMed2005

Juan Comas's summary history of the American association of physical anthropologists (1928-1968).

This translation of Juan Comas's Summary History of the American Association of Physical Anthropologists was originally published in Spanish by the Instituto Nacional de Antropología e Historia, Mexico, in 1969 (Departamento de Investigaciones Antropológicas, Publication 22). Physical anthropologists from North America and members of the American Association of Physical Anthropologists owe Juan Comas a debt of gratitude for having labored to produce this Summary History of the AAPA. There is much useful and interesting material in this document: extensive endnotes that are helpful to the historian of the profession; an appendix of the Journal issues where the proceedings of annual meetings can be found; a detailed listing of contributors of papers to annual meetings from 1930-1968; a warm acknowledgment and history of the contributions of the Wenner-Gren Foundation to biological anthropology; a history of the Yearbook of Physical Anthropology; and comments on the short-lived PA Newsletter. In addition, there are appendices with the founding AAPA Constitution and By-Laws from 1930 and as they existed in 1968. All of this synoptic information saves the reader with interests in the history of the AAPA considerable effort, especially when few university and college libraries have the full (old and new) series of the AJPA on their shelves. We have tried to provide a translation of Comas's history that is faithful to the original Spanish-language publication. In a few cases, we shortened sentences and applied a slightly more modern usage than was popular in the late 1960s.

Anthropology, Physical↗

PubMed2000

Purification, cloning and autoproteolytic processing of an aspartic proteinase from Centaurea calcitrapa.

Plant aspartic proteinases (APs) have been isolated from several seed and leaf sources but the only well characterized enzymes from flowers are cardosins and cyprosins from cardoon, Cynara cardunculus L. Here we report a full-length cDNA clone encoding an AP named cenprosin from the flowers of Centaurea calcitrapa L., a thistle related to cardoon. As found for all eukaryotic APs, the deduced primary sequence consists of a signal sequence, a propart and a mature enzyme. In addition, an internal sequence region of 104 residues typical only of plant APs (a plant-specific insert) is present in the primary structure. Northern analysis revealed that the strongest expression is in fresh flowers. The enzyme is also expressed in fairly high amounts in seeds and in leaves, a feature not detected for cardoon APs. The corresponding enzyme was purified in its precursor form from fresh flowers using ammonium-sulfate precipitation followed by ion-exchange and hydrophobic-interaction chromatography. The processing of the precursor into its mature form was studied in vitro. The enzyme underwent autocatalytic processing at pH 3.0 resulting in two chains of 16 and 30 kDa. When dried flowers were used as a starting material for purification, only 16- and 30-kDa chains were obtained, suggesting that autoproteolytic activation of procenprosin in vivo occurs mainly during drying of the flowers. This may indicate a specific degradative role for the enzyme during senescence of the flowers.

Amino Acid Sequence↗

PubMed2004

Consent for regional anaesthesia in the United Kingdom: what is material risk?

Legal principles that apply to the process of informed consent have changed in recent years. Patients should now be given the information that they wish to receive, not the information that health professionals may consider reasonable for them. In obstetric practice informed consent is especially important as young, fit patients may request and receive non-essential but potentially life-threatening interventions. The quantity and detail of information parturients desire do not remain static. They vary over time and from country to country. Our paper examines current opinion amongst parturients in the United Kingdom. We asked 100 obstetric patients to choose the complications of regional anaesthesia that they would like to learn about during informed consent. Nearly all women (82-94%) wished to know about common, less severe side effects. A substantial majority (70-77%) also wished to know about rarer but more severe complications, such as permanent neurological deficit, meningitis and high spinal block. Despite the availability of information for patients from sources such as the Obstetric Anaesthetists' Association and the National Electronic Library for Health, there remains little consensus amongst anaesthetists about what information to provide. Frequently some complications that patients would consider important are not discussed. Changing legal and public expectations demand that we adapt our current practice and improve the accuracy and timing of information provided.

Adolescent↗

PubMed2004

Purification and cDNA cloning of the ovigerous-hair stripping substance (OHSS) contained in the hatch water of an estuarine crab Sesarma haematocheir.

The egg attachment system of an estuarine crab Sesarma haematocheir is formed on the maternal ovigerous hairs just after egg laying, and slips off these hairs just after hatching. The stripping is caused by an active factor that we call OHSS (ovigerous-hair stripping substance), which is released by the embryo upon hatching. OHSS was purified, and its active form had a molecular mass of 25 kDa. The cDNA of OHSS cloned from an embryonic cDNA library was 1759 bp long, encoding 492 amino acids in a single open reading frame (ORF). The C-terminal part of the predicted protein was composed of a trypsin-like serine protease domain, with homology to counterparts in other animals of 33-38%. The predicted protein (54.7 kDa) secreted as a zymogen may be cleaved post-translationally, separating the C-terminal from the N-terminal region. The OHSS gene was expressed in the embryo at least 2 weeks before hatching. Expression was also detected in the zoea larva 1 day after hatching and in the brain of the female. However, it was not detected in the muscle, hepatopancreas or ovigerous seta of the female. Ultrastructural analysis indicated that the material investing maternal ovigerous hair, i.e. the outermost layer (E1) of the egg case, is attached at the special sites (attachment sites) arranged at intervals of 130-160 nm on the hair. It is suggested that OHSS acts specifically at these sites, lysing the bond with the coat, thus disposing of the embryo attachment system. This enables the female to prepare the next clutch of embryos without ecdysis.

Amino Acid Sequence↗

PubMed1983

Bibliotherapy: an adjunct to care of patients with problems of living.

Bibliotherapy, or the selected use of reading as an adjunct to the treatment of medical and psychological problems, has a long history in the library science literature. However, the use of bibliotherapy by practicing physicians has not been significant. Many patients who see family physicians are candidates for bibliotherapy as adjunctive treatment. In the past five years numerous well-written books that address various patient problems of living have been made available. The authors surveyed a variety of health care specialists in the Denver area to develop a reading list for people with life problems. These problems have been categorized under the following headings: dealing with life crises and transitions, parents and children, parenting, coping with illness and disability, death and dying, lifestyle modification, sexuality, and coping with feelings. The resulting annotated bibliography may be used as a guide for practicing physicians who feel that appropriate reading material may be beneficial to their patients with such problems.

Adjustment Disorders↗

PubMed1969

Thesaural problems in an on-line system.

This paper describes the construction of a synonym thesaurus or entry vocabulary for the SUNY Biomedical Communication Network, which will permit the user greater ease of access to the MeSH-indexed material without previously consulting a printed list of indexing terms. In order to discover the actual terminology used by a researcher, words were extracted from titles of articles appearing in Index Medicus, and compared with the subject heading under which they appeared. As well as strict synonyms, grammatical variants were also included. Work is continuing on relating other indexing vocabularies, such as Excerpta Medica and Current Medical Terminology, used in the biomedical world to MeSH terms.

Computers↗

PubMed1995

Cloning and heterologous expression of the Candida albicans gene PMI 1 encoding phosphomannose isomerase.

Using a DNA fragment derived from the Saccharomyces cerevisiae phosphomannose isomerase (PMI) structural gene as a probe against a random ordered array library of genomic DNA from the pathogenic fungus Candida albicans, we have cloned the C. albicans PMI 1 gene. This gene, which is unique in the C. albicans genome, can functionally complement PMI-deficient mutants of both S. cerevisiae and Escherichia coli. The DNA sequence of the PMI 1 gene predicts a protein with 64.1% identity to PMI from S. cerevisiae. Sequential gene disruption of PMI 1 produces a strain with an auxotrophic requirement for D-mannose. The heterologous expression of the PMI 1 gene at levels up to 45% of total cell protein in E. coli leads to partitioning of the enzyme between the soluble and particulate fractions. The protein produced in the soluble fraction is indistinguishable in kinetic properties from the material isolated from C. albicans cells.

Amino Acid Sequence↗

PubMed2005

Production and purification of self-assembling peptides in Ralstonia eutropha.

Self-assembling peptides have emerged as an attractive scaffold material for tissue engineering, yet the expense associated with solid phase chemical synthesis has limited their broad use. In addition, the fidelity of chemical synthesis constrains the length of polypeptides that can be produced homogeneously by this method. Template-derived biosynthesis by recombinant DNA technology may overcome both of these problems. However, recovery of polypeptides from recombinant protein expression systems typically involves multi-step purification schemes. In this study, we report an integrated approach to recombinantly produce and purify self-assembling peptides from the recently developed expression host Ralstonia eutropha. The purification is based on the specific affinity of carbohydrate binding modules (CBMs) to cellulose. In a first step, we identified CBMs that express well in R. eutropha by assembling a fusion library of green fluorescent protein (GFP) and CBMs and determining the fluorescence of cell-free extracts. Three GFP::CBM fusions were found to express at levels similar to GFP alone, of which two CBMs were able to mediate cellulose binding of the GFP::CBM fusion. These two CBMs were then fused to multiple repeats of the self-assembling peptide RAD16-I::E (N-RADARADARADARADAE-C). The fusion protein CBM::E::(RAD16-I::E)4 was expressed in R. eutropha and purified using the CBM's affinity for cellulose. Subsequent proteolytic cleavage with endoproteinase GluC liberated RAD16-I::E peptide monomers with similar properties to the chemically synthesized counterpart RAD16-I.

Carrier Proteins↗

PubMed2004

A new compact disc format of high density array synthesis applied to peptide nucleic acids and in situ MALDI analysis.

A fully automated synthesizer was constructed and designed to perform high speed miniaturized syntheses of compound libraries using the SPOT technique. Utilizing magnetically controlled drop-on-demand ink jet nozzles, an r/phi array format of 2500 spots can be simultaneously dispensed from up to 24 separate reagent valves onto a rotating disc as the solid phase in less than three minutes. In addition, a complete wash station is on board allowing for fully programmable combinatorial syntheses without manual attention. A new carbon black/polypropylene composite solid phase disc was developed and tested for its functionalisation/loading, spot detection, durability and MALDI-TOF target capabilities. The carbon black/polypropylene composite was then successfully employed jointly as the solid phase in the syntheses of short peptide and PNA oligomers and as the target probe holder for MALDI-TOF measurement without transfer of the material. Several protocols for PNA syntheses were also investigated and an optimised PNA methodology for the carbon black/polypropylene composite is reported.

Amino Acid Sequence↗

PubMed1992

Cloning of highly polymorphic microsatellites in the horse.

We have isolated equine microsatellites by screening a genomic library with (TG)n and (TC)n probes. TG microsatellites were found to be more abundant than TC repeats, with an estimated frequency of one per 100,000bp. Sequence analysis of eight TG-positive clones revealed varying structures of the repeat regions; perfect stretches of TG repeats, imperfect stretches of TG repeats and compound regions of TG and TC repeats. Five loci were analysed by PCR and showed extensive polymorphism; three to seven alleles and heterozygosities of 0.40-0.76 were observed when screening 20-30 unrelated individuals. The high degree of polymorphism, their abundance and the possibility of automating the typing procedure make these loci ideal for standardized paternity testing in the horse. Furthermore, we demonstrate that single hairs can be used as starting material for the PCR analysis.

Animals↗

PubMed2007

High-throughput synthesis of conopeptides: a safety-catch linker approach enabling disulfide formation in 96-well format.

Conotoxins exhibit a high degree of selectivity and potency for a range of pharmacologically relevant targets. The rapid access to libraries of conotoxin analogues, containing multiple intramolecular disulfide bridges for use in drug development, can be a very labor intensive, multi-step task. This work describes a high-throughput method for the synthesis of cystine-bridged conopeptides. Peptides were assembled on a peptide synthesizer employing the Fmoc solid-phase strategy using a safety-catch amide linker (SCAL). Side-chain protecting groups were removed on solid phase before SCAL activation with ammonium iodide in TFA, finally releasing the peptide into the TFA solution. Disulfide bond formation was performed in the cleavage mixture employing DMSO. This improved method allows mixtures of oxidized peptides to be obtained in parallel directly from a peptide synthesizer. A single HPLC purification of the resulting crude oxidized material produced peptides of > 95% purity.

Acrylic Resins↗

PubMed2003

[Triple mounting of the Ernst Mach memorial tablet in Brno-Chirlice].

On the occasion of his 100th birthday, the outstanding Austrian physicist and philosopher Ernst Mach (1838-1916) was honored in a ceremony on 13 February 1938 with a commemorative plaque that was placed on the front side of the house of his birth in Chirlice/Chirlitz which belongs to Brno today. The bronze plaque is derived from a design created by the sculptor Karl Korschann who was a professor at the German Technical University in Brno. It is 140 x 90 cm and in the central field (59 x 71 cm) bears a portrait of Mach in half-relief, and to the right and left of it inscriptions in German and Czech. This commemorative plaque endured a history filled with vicissitudes that are here - insofar as it is possible - sketched in on the basis of archival materials and the testimony of witnesses. After the ceremonial unveiling in 1938, the plaque was preserved in its original condition for only a short time: during the occupation of Czechoslovakia by the Germans the word 'ĉeskoslovenský' was painted over in the Czech text, and in 1943 a massive campaign to collect metal led to the removal of the plaque, but not to its being melted down. The place where it was kept for the next five years could not be determined. By decision of the town council the plaque was again put up in 1948 in a low-key event. As early as 1950 it was again removed - this time secretly in a nighttime operation and undoubtedly for political reasons -, and transported to the municipal library and later stored in the basement of the nearby little castle. There, boys who were playing found the plaque at the end of the fifties and cut out an approximately 40 x 30 cm piece in order to sell the metal. Soon after that the damaged plaque was no longer in its place. Since then there has been no trace of its whereabouts, it probably ended up in a smelting furnace in 1965. The approaching 150th birthday of Mach led in the eighties to renewed interest in the plaque. A massive search was unsuccessful, which led to the commissioning of a simple granite commemorative plaque, the unveiling of which occurred on 14 May 1988 during the celebration of Ernst Mach days in Brno: the house of Mach's birth again had a commemorative plaque after a long period of anonymity. - Should Korschann's model of the bronze commemorative plaque appear again, then a new cast of the original plaque was to be made: then the image of Mach could again (for the third time) be put up - and Goethe's "You have to say it three times" (Faust I, line 1531) would thereby be fulfilled.

Austria↗

PubMed1995

A multimedia-based histology laboratory course: elimination of the traditional microscope laboratory.

UNLABELLED: We have developed a multimedia-based laboratory course which has enabled us to eliminate the microscope and traditional microscope laboratory that have been mainstays of our histology course and histology courses at almost all institutions where histology is taught. The multimedia laboratory uses a library of histology images (approximately 24,000) stored on videodisc ( HISTOLOGY: A Photographic Atlas, by S. Downing) as its microscope slide collection and accesses those images through barcode and computer interfaces. The laboratory workstations consist of a videodisc player, videodisc monitor, computer, and computer monitor. One workstation is available for every 4-students, and our students are encouraged to work together in groups of four or five. In our current set-up, the students are introduced to and instructed in the basic principles of histology using a computer program that interfaces with the videodisc images. The computer program is divided into 19 chapters (the chapters are typical of the chapters found in a normal histology textbook) and has: (1) a laboratory component that covers the material traditionally covered in the microscope laboratory, and (2) a lecture component that enables the students to evaluate their understanding of the lecture material in a non-punishing way. The laboratory section of each chapter is divided into a "MicroLab" section, an "InFo Time" section, and a "Quiz Time" section. Each of these sections interfaces with histological images stored on the videodisc. The students are encouraged to work through the "MicroLab" section of each chapter before moving on to the "InFo Time" and "Quiz Time" sections. The "MicroLab" sections introduce the students to the various tissues and organs of the body and is interfaced with the videodisc player and the histology images stored on the videodisc. These sections describe the basic histological features of the various tissues and organs and give the students access to multiple examples of what they are studying. The "InFo Time" sections bring up specific images and ask the students to think about the images. Information about the images being observed is available if the students want it and the students can flag those images that they found difficult. The Quiz Time section of the program is also interfaced with the videodisc player and provides access to a large number of histology images stored on the videodisc. The "Quiz Time" sections provide non-punishing review questions that the students can study after she has worked her way through the "MicroLab" and "InFo Time" sections. In addition to the use of a computer program to access the histology images stored on videodisc, we use barcodes that address specific images on the histology videodisc in a variety of ways to augment the students' laboratory and lecture experience. The benefits of using multimedia in place of the traditional microscope and microscope slide collection are numerous and include the speed at which specific histological images can be accessed and reviewed (when compared to finding a structure on a glass slide), a significant reduction in the amount of laboratory time needed by the student to learn the same amount of information, the ease of tutoring on a large monitor screen (when compared to trying to discuss a histological structure with a student through the eyepiece of a microscope), the encouragement of group study (which is difficult to do when a student is working 1-on-with a microscope), and the reduction of the number of faculty necessary to cover a typical histology laboratory session. The use of barcodes that address specific videodisc histology images has greatly changed our examination procedures and has significantly expanded the usefulness of the traditional lecture note handouts given to our students.

Computer-Assisted Instruction↗

PubMed1997

Meeting clinician information needs by integrating access to the medical record and knowledge resources via the Web.

MINDscape is a web based integrated interface to diverse sources of clinical information including both patient specific information (electronic medical record) as well as medical knowledge (the "digital library") to provide "just in time" information at the point of care. It was developed at the University of Washington to meet clinical information needs both as identified locally and by a review of the literature. Beta testing by over 600 clinicians is in progress and medical centers wide access scheduled for Fall 1997. We describe the information needs we sought to meet and the ongoing evaluation approach we are taking to ensure the information needs of a diverse group of clinicians are met. The iterative evolution of the interface from prototype, to alpha to large scale beta testing is reported. Integration of information occurs at three levels: integration of information by patient, integration of information by provider, and integration of patient specific information with medical reference material and decision support tools.

Computer Communication Networks↗