bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mitochondrial Disorders. A Diagnostic Challenge in Clinical Chemistry
Authors: Bauer Matthias F. , Gempel Klaus , Hofmann Sabine , Jaksch Michaela , Philbrook Christine , Gerbitz Klaus-Dieter 
Published: October 2004

AbstractMitochondria play a pivotal role in cellular metabolism and in energy production in particular. Defects in structure or function of mitochondria, mainly involving the oxidative phosphorylation (OXPHOS), mitochondrial biogenesis and other metabolic pathways, have been shown to be associated with a wide spectrum of clinical phenotypes. The ubiquitous nature of mitochondria and their unique genetic features contribute to the clinical, biochemical and genetic heterogeneity of mitochondrial diseases. We will focus on the recent advances in the field of mitochondrial disorders and their consequences for an advanced clinical and genetic diagnostics. In addition, an overview on recently identified genetic defects and their pathogenic molecular mechanisms will be given.
Enrichment of Mutant Alleles by Chromatographic Removal of Wild Type Alleles: a New Principle for the Detection of Alleles with Unknown Point Mutations at Excess of Wild Type Alleles
Authors: Nollau Peter , Fischer Carsten , Tschentscher Peter , Wagener Christoph 
Published: October 2004

Abstract
In human carcinomas, mutations that alter tumour genes such as the KRAS, P53, or APC genes, are mostly point mutations. The detection of mutant alleles of tumour genes in specimens such as urine, pancreatic juice, sputum, and stool holds great promise for an early diagnosis of cancer. In addition, the detection of mutant tumour genes in tissue samples, such as lymph nodes or resection margins, may allow a sensitive diagnosis of residual malignant disease. However, the reliable detection of mutant alleles in excess of wild type alleles remains an unresolved analytical problem when the mutations are not known a priori. In the present communication, a new approach is described which makes possible the detection of unknown point mutations in tumour genes at excess of wild type alleles. The method is based on the removal of wild type alleles by hybridisation to immobilised complementary oligonucleotides. Using this approach, an enrichment of mutant KRAS, P53 and APC alleles of one mutant in up to 103 normal alleles has been achieved. Parallel miniaturised separation units with oligonucleotides complementary to defined sequences of a wild type allele should allow the detection of unknown point mutations as well as small insertions or deletions which occur in the sequence range covered by the oligonucleotides.
Quantitative Differences between Aberrant Transcripts Which Occur as Common Isoforms and due to Mutation-based Exon Skipping of the Mismatch Repair Gene hMLH1
Authors: Plaschke Jens , Bulitta Clemens , Saeger Hans D. , Schackert Hans K. 
Published: October 2004

Abstract
About one-third of hereditary non-polyposis colorectal cancer-related mutations in the mismatch repair gene hMLH1 result in the loss of entire exons from the wild type transcripts. Here we describe quantitative differences of hMLH1 transcripts without exon 15, exon 16 or exon 17 in several members of a family with hereditary non-polyposis colorectal cancer. The transcript lacking exon 15 is caused by a G to A transition affecting the last nucleotide of the respective exon and results in a truncated protein. The transcripts lacking exon 16 or exon 17, which are in-frame deletions, were also found in all tested samples of a normal population and represent common isoforms. Reverse transcription-polymerase chain reaction-based relative quantification revealed about 50 % signal intensity for the mutation-based transcript, but less than 10 % for the common isoforms, if compared to the wild type. All aberrant transcripts were detected from blood-derived cDNAs but not from samples of normal colon epithelium. Although the biological significance of the common isoforms is unknown, they might lead to false risk assessment in hereditary non-polyposis colorectal cancer cases.
Analytical and Clinical Performance of Two Cardiac Troponin I Immunoassays
Authors: Zaman Zahur , De Spiegeleer Sandy , Gerits Mathieu , Blanckaert Norbert 
Published: October 2004

Abstract
Cardiac troponin I assays for Axsym (Abbott Diagnostics, Abbott Park, IL, USA) and Immuno 1 (Bayer Corporation, Tarrytown, NY, USA) analysers were evaluated. Heparin plasma or serum could be used for both assays. Samples were stable for 24 h at ambient temperature, 3 days at 4–8 ℃ and 3 months at −20 °C. After 10 months' storage at −80 °C, the recoveries were well above 100 % by both assays. Total coefficients of variation for Axsym assay were 9.0 %, 5.8 % and 5.3 % at concentrations of 2.6 μg/l, 9.83 μg/l and 34.3 μg/l respectively; for Immuno 1 these were 4.4 %, 1.6 % and 1.8 % at 2.3 μg/l,6.27 μg/l and 44.35 μg/l respectively. It was ≥ 20 % at concentration of ≤ 0.5 μg/l for Axysm assay and ≤ 0.15 μg/l for Immuno 1 assay. Recoveries were ≤ 90 % at ≤ 0.22 μg/l on Axsym and at ≤ 1.47 μg/l on Immuno 1. Neither method showed significant interference with haemoglobin, bilirubin, triglycerides or rheumatoid factor. Correlation between the two methods was excellent (r = 0.997, Y (Axsym) = 4.2X (Immuno 1) + 3.2). The highest concentrations detected in 50 healthy subjects were 0.3 μg/l and 0.1 μg/l by Axsym and Immuno 1 methods, respectively. Twelve out of 43 renal failure patients had troponin I 0.13–0.9 μg/l using Axsym method and 4 had levels of 0.07–0.13 μg/l using Immuno 1. In muscle trauma patients, troponin I was undetectable.
Development of an Automated Immunoturbidimetric Ferritin Assay
Authors: Borque Luis , Rus Antonio , Bellod Laura , Seco Ma Luisa 
Published: October 2004

Abstract
We have developed a new procedure for turbidimetric measurement of ferritin concentration in human serum, based on latex microparticle agglutination technology. The procedure has been automated using the Falcor 300 analyzer. Carboxilated latex particles (336 nm in diameter) were covalently coupled with immunopurified F(ab′)2 fragments of anti-ferritin IgG antibodies. Coated microparticles were automatically mixed with undiluted sample and the resulting absorbance due to agglutination was measured at 550 nm. The procedure generated a calibration curve with a measuring range of 0 to 558 μg/l, showing a day-today imprecision lower than 5.7 %. The detection limit was 4 μg/l. There were no interferences from bilirubin, hemoglobin or rheumatoid factors. Turbid and lipemic samples caused an important interference which could be avoided by pretreating those samples prior to measurement. A prozone effect was provisionally obtained with ferritin concentrations over 1800 μg/l. The results suggested a hook-like effect due to a rapid microparticle precipitation in the reaction media, that could be avoided by increasing the reaction medium density by adding sucrose to the buffer, up to 150 g/l concentration. This sucrose addition resulted in a displacement of the Heidelberger curve with a prozone phenomenon occuring at concentration higher than 3000 μg/l of ferritin. Results obtained with the present procedure correlated well with those obtained by a nephelometric procedure and with those obtained by an RIA. We conclude that this latex turbidimetric immunochemical procedure is simple, rapid, has a good analytical and operational performance on the Falcor 300 analyzer and is well suited for the measurement of ferritin concentration in human serum.
Does the Uncertainty of Commonly Performed Glucose Measurements Allow Identification of Individuals at High Risk for Diabetes?
Authors: Kallner Anders , Waldenström Johan 
Published: October 2004

Abstract
Revised recommendations for diagnosis of diabetes introduce the intermediary risk group of impaired fasting glucose (IFG), defined as individuals with a fasting blood-glucose concentration between 5.6 and 6.0 mmol/l. We apply the concept of uncertainty to identifiable steps of sampling and measuring blood-glucose. Since many instruments in primary health care measure plasma-glucose and report results as blood-glucose and vice versa, factors affecting the transformation are also considered. The study identifies the measurement procedure as the major source of uncertainty, closely followed by preanalytical sources. The estimated uncertainties indicate that the presently available procedures do not allow identification of IFG by a single investigation. The approach to establish an uncertainty budget can be used to evaluate the clinical usefulness of measurements.
Autoantibodies against Oxidized LDL in the First Phase of Life
Authors: Steinerová Alexandra , Racek Jaroslav , Stožický František , Tatzber Franz , Lapin Alexander 
Published: October 2004

Abstract
The study presents a comparison of data concerning lipid metabolism and lipid oxidation (oxidative stress) in children at the time of their birth and 3 months later, as well as of their mothers at the time of delivery, compared to a control group of non-pregnant women of the same age. The data confirm that labour represents an oxidative stress for both mother and child; it is expressed as a significant increase of malondialdehyde concentration in mothers immediately after delivery in comparison with non-pregnant women (p < 0.001). Its concentration in newborns was even higher than in their mothers (p < 0.005). Concentration of antibodies against oxidized LDL (oxLDLAb) was comparable in mothers and newborns due to their transplacental transport. During the first three months of life these autoantibodies increased almost two-fold. The importance of this unique observation is discussed with respect to possible early atherogenesis.
Approaches to Handling, Breeding, Strain Preservation, Genotyping, and Drug Administration for Mouse Models of Cancer
Authors: Boggess Dawnalyn , Silva Kathleen A. , Landel Carlisle P. , Mobraaten Larry , Sundberg John P. 
Published: October 2004

Abstract
Mouse models are extremely valuable biomedical tools for cancer research. Their value to individual investigators depends upon the investigator's ability to manage a colony and manipulate the mice. Basic genotyping to maintain a colony of genetically engineered mice is a critical technique. Understanding the differences between inbred and outbred (nonsegregating vs. segregating backgrounds) determines the reproducibility of models. Preserving valuable colonies for future use through cryopreservation of embryos, sperm, or just the constructs used to create the models enables future work to be done. This chapter provides a broad overview of many of the methods commonly used to perform these tasks as well as how and where to find more specific information.
Pathology of Mouse Models of Cancer
Authors: Cardiff Robert D. 
Published: October 2004

Abstract
Analysis of the changes in structure (gross and histopathology) is essential for the validation of mouse models of human cancer. However, many investigators do not have immediate access to qualified pathologists who understand the unique features of genetically engineered mouse pathology. Further, the gross examination of the mouse and the preparation of tissues are usually done by the laboratory personnel and, most frequently, by relatively untrained graduate students or fellows. Some laboratories have the good fortune of having experienced laboratory technologists who perform the services and teach the incoming students and fellows the correct techniques. However, many laboratories do not have this advantage and frequently relegate the necropsy to the least experienced animal technician or the newest graduate student. Unfortunately, this often leads to disastrous results that cannot be overcome. Therefore, this chapter is not about how to be a pathologist but rather is about how to prepare a mouse for pathologic analysis and is meant to be a primer for the neophyte who needs basic help.
Intercurrent Infections in Genetically Engineered Mice
Authors: Barthold Stephen W. 
Published: October 2004

AbstractThe laboratory mouse was domesticated from fancy mice that arose from geographically diverse genetic origins. The diverse origins of the mouse also contributed to a lengthy list of infectious agents that are carried by mice. Despite years of microbial quality control, the burgeoning use of mice for research is causing a resurgence of infectious disease among laboratory mice. Infections can be introduced through mice, but also through biologic products such as serum, hybridomas, and transplantable tumors and through iatrogenic introduction by scientists. Retroviruses represent an important group of agents with significant effects upon mouse research, but the transmissible exogenous viruses are no longer an issue among laboratory mice. The diversity of other infectious agents induce a wide variety of effects upon cancer research, including immunomodulation, tumor graft rejection, misinterpretation of phenotype, modified carcinogenesis, and zoonotic risk. Prudent research practices require that microbial status of research mice does not necessarily have to be standardized among laboratories, but it must be defined so that research results are reproducible and valid.
Germline Modification Strategies
Authors: Grisendi Silvia , Pandolfi Pier Paolo 
Published: October 2004

AbstractModeling human disease in the mouse has become an extremely important source of knowledge in biomedical research by deepening our understanding of human tumorigenesis and generating experimental systems for the evaluation of novel therapeutic approaches. Various strategies exist for engineering gain and loss‐of‐function mutant alleles by means of conventional and conditional germline modification methodologies. We describe classical transgenic and gene‐targeting approaches as well as their more advanced conditional versions, providing specific examples that illustrate both their utility and their disadvantages in modeling human cancer in the mouse.
Somatic Cell Gene Transfer
Authors: Uhrbom Lene , Holland Eric C. 
Published: October 2004

AbstractExperimentally induced, virally mediated oncogenesis has been used to study cancer for nearly 100 years. During this time, many of the fundamentals of cancer biology have been established. The technology differs from germline modification strategies in that the viral vectors that induce cancer in experimental animals achieve their goal by transferring genes into a limited number of somatic cells, usually after birth. New advances in these systems now support experimental designs aimed at assessing the effects of mixtures of genetic alterations, identification of the contribution of specific alterations on the histology of tumor cells, and the identification of cell‐of‐origin for certain cancers. These somatic cell gene transfer strategies are complementary to germline modification technologies in unraveling the complexities of cancer biology.
Lung Cancer
Authors: Kurie Jonathan M. , Minoo Parviz , Hecht Stephen S. , Nikitin Alexander , Demayo Franco J. 
Published: October 2004

AbstractLung cancer most often stems from a single environmental cause but has diverse genetic, biochemical, and histologic characteristics. This diversity has made the basic causes of lung cancer difficult to surmise. Current mouse models support genetic alterations in lung epithelial cell transformation, but these models do not recapitulate the diversity of histologic appearance, site of origin, or natural history of lung cancer in humans. Advances in mouse model technology provide hope that the significance of the myriad genetic and biochemical alterations in human lung cancer will eventually be understood and that future models will better represent their human counterpart. Progress in this area will enhance both the basic understanding of lung tumorigenesis and the speed with which the anticancer efficacy of investigational drugs can be investigated in preclinical trials.
Mammary Gland Cancer
Authors: Barkan Dalit , Montagna Cristina , Ried Thomas , Green Jeffrey E. 
Published: October 2004

AbstractIt has been 20 years since the first genetically engineered mouse model of mammary cancer was published (Stewart et al.,) using transgenic technology. Since the first report there have been well over 100 mouse models of mammary cancer developed which have provided critical insights into the functions of numerous genes involved in mammary oncogenesis. Advances in manipulating the mouse genome have provided the technology to induce mammary cancers in mice that arise from genetic mutations that have been identified in human cancer, thus generating models based upon molecular lesions relevant to the human disease. Recent work has focused on validating these models as systems which represent, at least in part, various aspects of the human disease and which may be used for preclinical testing of novel therapies. The challenge remains to establish the predictive value of such models for chemoprevention and therapeutic response for treating human breast cancer.
Prostate Cancer
Authors: Ellwood‐Yen Katharine , Sawyers Charles 
Published: October 2004

AbstractProstate cancer is one of the leading causes of cancer death among men in the United States. Development of model systems to study the disease is imperative. Both transgenic and knockout mice have been generated and offer great hope for future studies. While this type of research is still in its infancy, several promising models have been generated which share similar features to the human disease. Many experiments have been conducted using these models to identify proteins and pathways important to the initiation and progression of prostate cancer. Perhaps the most significant are microarray experiments performed on mouse tissues that have identified expression changes in several genes such asNkx3.1,Pim‐1, and prostate stem cell antigen (PSCA). These genes have already been implicated in human prostate cancer and therefore strengthen the argument for use of mouse model systems to tease out the molecular mechanisms of disease progression. It is only a matter of time before researchers are able to interpret data from the mouse and translate the results to human therapeutics.
Skin Cancer
Authors: Bosenberg Marcus 
Published: October 2004

Abstract
Skin cancer is the most common form of human cancer and causes significant morbidity and mortality. A wide variety of techniques have been used to model skin cancer in mice, including genetically engineered mice, exposure to chemical carcinogens or ultraviolet light, and xenograft studies of human skin and human skin reconstructs. These models have been used to validate human cancer genetics, identify new oncogenes and tumor suppressors, and to develop novel therapies for use in human skin cancer.
Ovarian Cancer
Authors: Orsulic Sandra 
Published: October 2004

Abstract
Mice have been used in ovarian cancer research mainly as hosts for cell lines derived from human ovarian tumors and ascites. Such models provided valuable information into the nature of metastatic ovarian cancer and possible treatment strategies. However, the complexity of genetic aberrations in human ovarian cancer cell lines precluded understanding of the initiating events responsible for ovarian cancer induction. Since the majority of ovarian cancer patients present at an advanced stage of the disease, it has been difficult to identify the precursor lesions that could be used to study the early morphologic and genetic changes in ovarian cancer. It is thought that the development of animal models in which ovarian cancer can be induced and studied during its early stages will enable better understanding of early ovarian cancer lesions and elucidate molecular events that support ovarian cancer progression. The difficulties in generating such models include the lack of an adequate ovary‐specific promoter, the uncertainty about the tissue of origin for different histologic types of ovarian cancer, and the deficiency in understanding the genetic aberrations responsible for ovarian cancer induction. In spite of these difficulties, the first advances toward generating mouse models for ovarian cancer have been made.
Peripheral Nervous System Tumors
Authors: Gutmann David H. , Perry Arie , Rangwala Reshma , Sherman Larry S. 
Published: October 2004

Abstract

Tumors of the peripheral nervous system arise from Schwann cells and include neurofibromas, schwannomas, and malignant peripheral nerve sheath tumors (MPNSTs). Individuals with the inherited tumor predisposition syndromes, neurofibromatosis 1 (NF1) and neurofibromatosis 2 (NF2), develop neurofibromas and schwannomas, respectively, suggesting that the
NF1
and
NF2
tumor suppressor genes are critical regulators of Schwann cell growth. Mouse models of these tumors have focused on the
Nf1
and
Nf2
genes, while MPNSTs in mice and humans require additional cooperating genetic alterations.
Central Nervous System Tumors
Authors: Lassman Andrew B. , Holland Eric C. 
Published: October 2004

AbstractWithin the last several decades, a plethora of mutations and signaling abnormalities in human brain tumors have been discovered. This represents an exciting development that holds the potential for new therapies. However, conquering such tumors will also require the successful differentiation of oncogenic abnormalities from epiphenomena. Modeling of brain tumors in mice is an important technique in this endeavor; transgenic, knockout, conditional tissue expression and somatic cell gene transfer methods have all been used to create these models, both separately and in combination. In addition, use of these models will become an important modality to test newly developed small‐molecule pathway inhibitors. Developed models include those of the most common primary brain tumors: the gliomas and medulloblastomas. These mouse tumors have substantial similarities to their human counterparts with respect to both histologic appearance and noninvasive imaging. In this chapter, the pathways important in driving both gliomagenesis and medulloblastomagenesis are described, along with the key features of individual tumor models presently in use.
Myeloid Malignancies
Authors: Kogan Scott C. 
Published: October 2004

AbstractA wealth of information is available regarding genetic abnormalities that contribute to human myeloid malignancies. Modeling these mutations in mice has been facilitated by the ability to manipulate the hematopoietic system not only through germline modification but also by retroviral transduction of bone marrow. Chimeric fusion proteins resulting from balanced chromosomal aberrations as well as intragenic mutations have been introduced into mice to generate models of many types of human myeloid leukemias. These models recapitulate important features of the human diseases and have been utilized to elucidate mechanisms through which particular genetic changes contribute to myeloid neoplasia.The study of human myeloid neoplasms has led to many advances in our understanding of these disorders and of other human malignancies. The association of specific chromosomal changes in neoplastic cells with particular malignancies was begun by the identification of the Philadelphia chromosome in chronic myelogenous leukemia (CML) (Nowell,). CML has remained a leading source of insight into the pathogenesis and treatment of human neoplasms: from identifying that the Philadelphia chromosome creates an oncogenic BCR/ABL fusion protein to the development of imatinib mesylate as a molecularly targeted therapeutic agent. In addition to the t(9;22) of CML, numerous recurrent balanced chromosomal aberrations have been identified in human myeloid neoplasms. In fact, the genes altered by all of the major (and many of the minor) balanced aberrations of myeloid leukemia have been identified. Most of these aberrations result in the production of abnormal fusion proteins, while others result in dysregulated gene expression. In addition, a substantial number of recurrent point mutations and small intragenic changes have also been associated with human leukemias and related disorders. Because there is a wealth of information regarding the genetic changes that are present in these human diseases, it has been possible to model numerous such changes in mice.