Biomedical subjects

D Garza

Publications and source records attributed to D Garza.

At least 19 recordsLinked to original sources

PubMed2001

The 62E early-late puff of Drosophila contains D-spinophilin, an ecdysone-inducible PDZ-domain protein dynamically expressed during metamorphosis.

At the onset of metamorphosis in Drosophila melanogaster, the steroid hormone 20-OH ecdysone induces a small number of early and early-late puffs in the polytene chromosomes of the third-instar larval salivary gland whose activity is required for regulating the activity of a larger set of late puffs. Most of the corresponding early and early-late genes have been found to encode transcription factors that regulate a much larger set of late genes. In contrast, we describe here the identification of an ecdysone-regulated gene in the 62E early-late puff, denoted D-spinophilin, that encodes a protein similar to the mammalian protein spinophilin/neurabin II. The D-spinophilin protein is predicted to contain a highly conserved PP1-binding domain and adjacent PDZ domain, as well as a coiled-coil domain and SAM domain, and belongs to a family of related proteins from diverse organisms. Transcription of D-spinophilin is correlated with 62E puff activity during the early stages of metamorphosis and is ecdysone-dependent, making this the first member of this gene family shown to be regulated by a steroid hormone. Examination of the dynamic patterns of D-spinophilin expression during the early stages of metamorphosis are consistent with a role in central nervous system metamorphosis as well as a more general role in other tissues. As D-spinophilin appears to be the only member of this gene family in Drosophila, its study provides an excellent opportunity to elucidate the role of an important adaptor protein in a genetic model organism.

Amino Acid Sequence↗

PubMed2001

Self-antigen-presenting cells expressing diabetes-associated autoantigens exist in both thymus and peripheral lymphoid organs.

Recent reports indicate that genes with tissue-restricted expression, including those encoding the type 1 diabetes autoantigens insulin, glutamic acid decarboxylase (GAD), and the tyrosine-phosphatase-like protein IA-2 (or ICA512), are transcribed in the thymus. The reported modulation of diabetes susceptibility by genetically determined differences in thymic insulin levels and studies in transgenic mice provide correlative and functional evidence that thymic expression of peripheral proteins is crucial for immunological self-tolerance. However, there are no specific data about the existence, tissue distribution, phenotype, and function of those cells that express insulin and other self-antigens in the human thymus. We find that the human thymus harbors specialized cells synthesizing (pro)insulin, GAD, and IA-2, mainly localized in the medulla, and we demonstrate such cells also in peripheral lymphoid organs (spleen and lymph nodes). Phenotypic analysis qualifies these cells as antigen-presenting cells (APCs), including both dendritic cells and macrophages. These cells often appear surrounded by apoptotic lymphocytes, both in thymus and spleen, and may therefore be involved in the deletion of autoreactive lymphocytes. Our findings demonstrate the existence of, and define the tissue distribution and phenotype of, a novel subset of APCs expressing self-antigens in human lymphoid organs that appear to be involved in the regulation of self-tolerance throughout life.

Adolescent↗

PubMed2001

Differential splicing of the IA-2 mRNA in pancreas and lymphoid organs as a permissive genetic mechanism for autoimmunity against the IA-2 type 1 diabetes autoantigen.

Type 1 diabetes results from the autoimmune destruction of pancreatic beta-cells in genetically susceptible individuals. Growing evidence suggests that genetically determined variation in the expression of self-antigens in thymus may affect the shaping of the T-cell repertoire and susceptibility to autoimmunity. For example, both allelic variation and parent-of-origin effects influence the thymic expression of insulin (a known type 1 diabetes autoantigen), and insulin gene transcription levels in thymus inversely correlate with susceptibility in both humans and transgenic models. It is unclear why patients lose tolerance to IA-2 (insulinoma-associated tyrosine phosphatase-like protein, or islet cell antigen 512 [ICA512]), especially because IA-2 polymorphisms are not associated with type 1 diabetes. We report that alternative splicing determines differential IA-2 expression in islets compared with thymus and spleen. Islets express full-length mRNA and two alternatively spliced transcripts, whereas thymus and spleen exclusively express an alternatively spliced transcript lacking exon 13. This encodes for the transmembrane (TM) and juxta-membrane (JM) domains that comprise several type 1 diabetes target epitopes, supporting the concept that tolerance to IA-2 epitopes not expressed in lymphoid organs may not be achieved. We propose differential splicing as a regulatory mechanism of gene expression playing a permissive role in the development of autoimmune responses to IA-2. Our findings also show that candidate gene expression studies can help in dissecting the complex genetic determinants of a multifactorial disease such as type 1 diabetes.

Adult↗

PubMed2001

Alaska Natives assessing the health of their environment.

The changes in Alaska's ecosystems caused by pollution, contaminants and global climate change are negatively impacting Alaska Natives and rural residents who rely on natural resources for food, culture and community identity. While Alaska commerce has contributed little to these global changes and impacts, Alaska and its resources are nonetheless affected by the changes. While Alaska Natives have historically relied on Alaska's land, water and animals for survival and cultural identity, today their faith in the safety and quality of these resources has decreased. Alaska Natives no longer believe that these wild resources are the best and many are turning to alternative store-bought foods. Such a change in diet and activity may be contributing to a decline in traditional activities and a decline in general health. Contaminants are showing up in the animals, fish and waters that Alaska Natives use. Efforts need to be expanded to empower Alaska Native Tribes to collect and analyze local wild foods for various contaminants. In addition existing information on contaminants and pollution should be made readily available to Alaska residents. Armed with this type of information Alaska Native residents will be better prepared to make informed decisions on using wild foods and materials.

Alaska↗

PubMed2000

The E23 early gene of Drosophila encodes an ecdysone-inducible ATP-binding cassette transporter capable of repressing ecdysone-mediated gene activation.

At the onset of Drosophila metamorphosis, the steroid hormone 20-OH ecdysone directly induces a small number of early puffs in the polytene chromosomes of the larval salivary gland. Proteins encoded by the early genes corresponding to these transcriptional puffs then regulate the activity of both the early puffs themselves and a much larger set of late puffs. Three of these early genes encode transcription factors that play critical regulatory roles during metamorphosis. Here we report the cloning, DNA sequence, genomic structure, ecdysone inducibility, and temporal expression of an early gene residing in the 23E early puff and denoted E23 (Early gene at 23). In contrast to other early genes, E23 encodes a protein with similarity to ATP-binding cassette transporters. Using heat shock-inducible transgenes, we found that E23 overexpression not only produces phenotypic abnormalities and lethality, but also interferes with ecdysone-mediated gene activation, demonstrating that E23 is capable of modulating the ecdysone response. Our results suggest the existence of a previously unrecognized regulatory mechanism for modulating steroid hormone signaling in Drosophila.

ATP-Binding Cassette Transporters↗

PubMed2000

The L63 gene is necessary for the ecdysone-induced 63E late puff and encodes CDK proteins required for Drosophila development.

The pulse of ecdysone that triggers Drosophila metamorphosis activates six early genes in a primary response made visible by polytene chromosome puffs. The secondary response is detected by the induction of over 100 late puffs, only a few of which have been subject to molecular genetic analysis. We present a molecular and mutational analysis of the L63 gene responsible for the late puff at 63E. This gene contains overlapping L63A, B, and C transcription units of which the A unit encodes two isoforms and the B unit three. The C unit, which exhibits little activity, encodes one of the B isoforms. Evidence that L63B, but not L63A, transcription is ecdysone responsive derives from their developmental transcription profiles and from P-element mutagenesis showing that ecdysone induction of the 63E puff requires sequences adjacent to the 5' end of L63B but not those adjacent to the 5' end of L63A. L63-specific lethal mutations showed that L63 is required not only for metamorphosis, but also maternally and for embryonic and larval development. The L63 proteins contain a common C-terminal 294-aa sequence that is 71% identical to the CDK sequence of the murine PFTAIRE protein. In vivo tests of L63 proteins altered by site-directed mutagenesis showed that they exhibit CDK functions. L63 proteins are widely distributed among late larval and prepupal tissues and are unlikely to be involved in cell cycle functions.

Amino Acid Sequence↗

PubMed2000

Construction and transposition of a 100-kilobase extended P element in Drosophila.

We have used P element deletion derivatives at defined locations in the Drosophila genome to construct a 100-kb extended P element more than twice the size of any previously available. We demonstrate that this prototypical extended P element is capable of transposition to new sites in the genome. The structural and functional integrity of a transposed extended P element was confirmed using molecular, genetic, and cytogenetic criteria. This is the first method shown to be capable of producing large, unlinked transpositional duplications in Drosophila. The ability to produce functional transposable elements from half-elements is novel and has many potential applications for the functional analysis of complex genomes.

Animals↗

PubMed1999

The 82F late puff contains the L82 gene, an essential member of a novel gene family.

Metamorphosis in Drosophila results from a hierarchy of ecdysone-induced gene expression initiated at the end of the third larval instar. A now classical model of this hierarchy was proposed based on observations of the activity of polytene chromosome "puffs" which distinguished "early" puffs as those directly induced by ecdysone and "late" puffs as those which become active as a secondary response to the hormone. We report here the isolation and characterization of the L82 gene corresponding to the extensively characterized late puff at 82F. L82 is a complex gene that spans at least 50 kb of genomic DNA, produces at least seven different nested mRNAs, and has homology to a novel gene family. In contrast to most previously characterized puff genes, the broad developmental expression pattern of L82 suggests that it is controlled by both ecdysone-dependent and ecdysone-independent regulatory mechanisms. L82 mutations were identified by transgene rescue of developmental delay and eclosion lethal phenotypes.

Amino Acid Sequence↗

PubMed1999

Process mapping in screening mammography.

Successful screening mammography programs aim to screen large numbers of women efficiently and inexpensively. Development of an effective screening mammography program requires skilled personnel, solid infrastructure, and a robust computer system. A group of physicians, technologists, computer support personnel, and administrators carefully analyzed a growing screening mammography program as a series of steps, starting with the request for the examination and ending with the receipt of a hard-copy consultation. The analysis involved a detailed examination of every step and every possible outcome in the screening process. The information gained through process mapping may be used for identification of systemic and personnel problems, allocation of resources, modification of workplace architecture, and design of computer networks. Process mapping is helpful for those involved in designing and improving screening mammography programs. Viewing a process (i.e., obtaining a screening mammogram) as a series of steps may allow for the identification of inefficient components that may limit growth.

Adult↗

PubMed1997

The clinical value of serum ferritin tests in endurance athletes.

OBJECTIVE: It is common practice to measure serum ferritin levels in endurance athletes because of the belief that low iron stores may compromise performance. The direct relationship between endurance performance and iron deficiency anemia is well known, but there are theoretical reasons to believe that endurance performance may be adversely affected by low iron stores even in the absence of frank anemia. The purpose of this article is to provide a critical review of the scientific evidence relating low iron stores to endurance performance. DATA SOURCES: Medline was searched using MeSH for articles related to ferritin and endurance published since 1985. Additional references were reviewed from the bibliographies of the retrieved articles. STUDY SELECTION: All clinical study designs were reviewed as well as relevant animal studies. Conclusions regarding endurance performance in humans were limited to data from clinical studies. DATA EXTRACTION AND SYNTHESIS: In reviewing the literature, the relative strengths of the study designs were examined carefully. Particular attention of the effectiveness of each study in isolating ferritin as the key independent variable. Dependent measures of endurance capacity were also evaluated. MAIN RESULTS: Eight studies isolated serum ferritin as the experimental variable. Only one study reported a significant improvement in endurance performance (time to exhaustion) in subjects with low ferritin levels treated with oral iron, but this finding may have been magnified by an unexplained decrease in time to exhaustion in the control group. Iron dosages differed in the studies reviewed. Two additional studies that reported increases in performance parameters following increases in ferritin were confounded by concomitant increases in hemoglobin levels. CONCLUSIONS: Iron supplementation can raise serum ferritin levels, but increases in ferritin concentration, unaccompanied by increases in hemoglobin concentration, have not been shown to increase endurance performance. Of concern to the clinician is that athletes with low ferritin levels but hemoglobin in the low-normal range may have iron deficiency anemia responsive to iron supplementation. Low ferritin with hemoglobin in the mid- to upper normal range is at best a relative indication for iron supplementation: low ferritin with hemoglobin in the low normal range is a stronger, yet still relative, indication for iron supplementation in athletes.

Anemia, Iron-Deficiency↗

PubMed1997

Detection of coronary collaterals using dipyridamole PET myocardial perfusion imaging with rubidium-82.

UNLABELLED: This study evaluated the ability of dipyridamole PET myocardial perfusion imaging to detect coronary collaterals. A previous study showed an association between dipyridamole-induced coronary steal on PET imaging and the presence of coronary collaterals on angiography. METHODS: Dipyridamole PET myocardial perfusion imaging using 82Rb was performed in 45 patients who had recent coronary angiography. The stress/rest count ratio (rubidium activity with stress divided by activity at rest)-was used to express the change in regional tracer uptake with dipyridamole and was calculated manually and automatically. The accuracy of the stress/rest count ratio for detecting coronary collaterals was determined. RESULTS: A manual stress/rest count ratio < or = 0.80 identified coronary collaterals with 81% sensitivity, 92% specificity and 90% accuracy (p < 0.0001). An automated ratio < or = 0.80 had 90% sensitivity, 88% specificity and 90% accuracy (p < 0.0001). Vascular beds incorrectly identified by PET as having collaterals had an increased frequency of severe stenoses and abnormal wall motion. CONCLUSION: PET perfusion imaging using the stress/rest count ratio can serve as a unique imaging method to identify coronary collaterals noninvasively.

Collateral Circulation↗

PubMed1996

Detection of bony metastases of androgen-independent prostate cancer by PET-FDG.

Fourteen F-18 fluorodeoxyglucose (FDG) studies were carried out in 13 patients known to have bony metastases from carcinoma of the prostate. One patient was newly diagnosed. The remaining patients had various types of therapy and were considered hormonally resistant. The average age was 67. All patients had extensive bony metastases shown on the conventional Tc99m-MDP bone scans. Only about 18% of bony lesions apparent on the conventional bone scans showed corresponding increase of FDG uptake. Anatomical correlation was performed by using co-registered images of SPECT and PET in the same area. The positive FDG uptake was not related to the duration of illness, level of PSA, previous therapy, and magnitude of disease involvement. It appears that only a small percentage of bony metastases is associated with increased glycolysis. It is possible that other metabolic processes are more important than glycolysis for providing prostate cancer with a source of energy and nutrients.

Aged↗

PubMed1995

Production of antigen-specific human antibodies from mice engineered with human heavy and light chain YACs.

Our paper describes the introduction of large fragments of both the human heavy and light chain Ig genes into the mouse germline to create a mouse strain capable of producing a broad repertoire of antigen-specific, fully human antibodies. The human immunoglobulin gene sequences were functional in the context of the mouse machinery for antibody recombination and expression, either in the presence or absence of functional endogenous genes. This was demonstrated by their ability to undergo diverse rearrangement, to be expressed at significant levels, and to exclude expression of mouse immunoglobulins irrespective of their copy number or site of integration. The decrease in susceptibility to influence by adjacent genomic sequences may reflect the greater size, variable gene content, or structural integrity of the human Ig YACs and/or the presence of unidentified but important regulatory elements needed for optimal expression of the human immunoglobulin genes and their correct regulation. Our results show that mouse B cells coexpressing human heavy and kappa chains, upon immunization, can produce antigen-specific, fully human antibodies. Furthermore, the human heavy and kappa chain YACs induced differentiation and maturation of the growth-arrested B-cell lineage in mice with inactivated endogenous Ig genes, leading to the production of a diverse repertoire of fully human antibodies at levels approaching those in normal serum. These results suggest the potential value of these mice as a source of fully human antibodies for human therapy. Furthermore, it is expected that such mice would lack immunological tolerance to and thus readily yield antibodies to human proteins, which may constitute an important class of targets for monoclonal antibody therapy. Our findings suggest that the introduction of even larger portions of the human heavy and light chain loci, which should be achievable with the ES cell-yeast spheroplast fusion technology described, will result in strains of mice ultimately capable of recapitulating the full antibody repertoire characteristic of the human humoral response to infection and immunization. The present and future mouse strains may prove to be valuable tools for studying the molecular mechanisms and regulatory sequences influencing the programmed assembly and expression of human antibodies in the normal immune response, as well as the abnormal response characteristic of autoimmune disease and other disorders. The strategy we have described for the introduction of large segments of the human genome into mice in conjunction with the inactivation of the corresponding mouse loci may also have broad applicability to the investigation of other complex or uncharacterized loci.

Animals↗

PubMed1995

Analysis of the structural integrity of YACs comprising human immunoglobulin genes in yeast and in embryonic stem cells.

With the goal of creating a strain of mice capable of producing human antibodies, we are cloning and reconstructing the human immunoglobulin germline repertoire in yeast artificial chromosomes (YACs). We describe the identification of YACs containing variable and constant region sequences from the human heavy chain (IgH) and kappa light chain (IgK) loci and the characterization of their integrity in yeast and in mouse embryonic stem (ES) cells. The IgH locus-derived YAC contains five variable (VH) genes, the major diversity (D) gene cluster, the joining (JH) genes, the intronic enhancer (EH), and the constant region genes, mu (C mu) and delta (C delta). Two IgK locus-derived YACs each contain three variable (V kappa) genes, the joining (J kappa) region, the intronic enhancer (E kappa), the constant gene (C kappa), and the kappa deleting element (kde). The IgH YAC was unstable in yeast, generating a variety of deletion derivatives, whereas both IgK YACs were stable. YACs encoding heavy chain and kappa light chain, retrofitted with the mammalian selectable marker, hypoxanthine phosphoribosyltransferase (HPRT), were each introduced into HPRT-deficient mouse ES cells. Analysis of YAC integrity in ES cell lines revealed that the majority of DNA inserts were integrated in substantially intact form.

Animals↗

PubMed1995

Serial myocardial perfusion imaging with dipyridamole and rubidium-82 to assess restenosis after angioplasty.

UNLABELLED: The purpose of this study was to determine whether patients at high risk for clinical restenosis, following coronary angioplasty, could be identified by myocardial perfusion imaging performed with dipyridamole- 82Rb PET. METHODS: Forty-five patients (34 men, 11 women; mean age 58.5 yr) who had successful single-vessel angioplasty and were asymptomatic had dipyridamole-82Rb PET at 1 and 3 mo after the procedure. Abnormal flow reserve in the distribution of the angioplasty artery on PET was considered to be a decrease of > or = 1 perfusion grade in response to dipyridamole (assessed qualitatively from tomographic images and polar coordinate maps). Follow-up was performed for 6 mo postangioplasty. Clinical restenosis was defined as recurrent angina similar to that occurring before angioplasty and/or > or = 50% stenosis at the angioplasty site documented angiographically. We analyzed abnormal flow reserve in the distribution of the angioplasty vessel to identify which patients were at high risk for clinical restenosis. RESULTS: Fourteen patients developed clinical restenosis between 1 and 6 mo postangioplasty. Abnormal relative flow reserve in the distribution of the angioplasty vessel was present prior to the development of symptoms in 13 of 14 patients with clinical restenosis and in 8 of 31 patients without clinical restenosis (sensitivity 93%, specificity 74%, p < 0.0001). PET imaging successfully separated postangioplasty patients into groups with high (62%) and low (4%) risk of clinical restenosis. CONCLUSION: Abnormal relative flow reserve in the distribution of the angioplasty vessel on dipyridamole PET identifies asymptomatic postangioplasty patients at risk for clinical restenosis.

Angioplasty, Balloon, Coronary↗

PubMed1995

Silent myocardial ischemia during PET.

A resting perfusion defect detected during radioisotope myocardial perfusion imaging is generally thought to represent myocardial infarction. The administration of nitroglycerin during cardiac PET imaging results in improved resting defects, which suggests transiently abnormal resting perfusion with silent myocardial ischemia.

Aged↗

PubMed1994

Antigen-specific human monoclonal antibodies from mice engineered with human Ig heavy and light chain YACs.

We describe a strategy for producing human monoclonal antibodies in mice by introducing large segments of the human heavy and kappa light chain loci contained on yeast artificial chromosomes into the mouse germline. Such mice produce a diverse repertoire of human heavy and light chains, and upon immunization with tetanus toxin have been used to derive antigen-specific, fully human monoclonal antibodies. Breeding such animals with mice engineered by gene targeting to be deficient in mouse immunoglobulin (Ig) production has led to a mouse strain in which high levels of antibodies are produced, mostly comprised of both human heavy and light chains. These strains should provide insight into the adoptive human antibody response and permit the development of fully human monoclonal antibodies with therapeutic potential.

Adult↗

PubMed1992

3 alpha-Androstanediol and 3 alpha-androstanediol glucuronide. Maternal and umbilical cord plasma concentrations in normal pregnancy.

Concentrations of 3 alpha-androstanediol (A) and 3 alpha-androstanediol glucuronide (AG) were measured with a radioimmunoassay in pregnancy plasma samples collected from the mother in the first trimester (first, n = 18) and second trimester (second, n = 20) and at term (n = 14). At term, along with the maternal samples, matching cord plasma samples were also obtained (n = 14). Throughout pregnancy the levels of A and AG were significantly higher than in normally menstruating women. Although the total concentration of A and AG did not change significantly throughout pregnancy, the levels of A in the second and at term were significantly higher than in the first. In contrast, AG concentrations in the second were significantly lower than in the first and at term. As compared to the maternal concentrations at term, the cord levels of AG and A plus AG were not significantly different. However, the levels of A in the cord were significantly lower than those in the maternal circulation at term. Significant correlations were observed between the term maternal and cord levels of AG and A plus AG. No sex-related differences in the plasma levels of A and AG were noted. The results show that levels of A and AG increase significantly in the first trimester and remain elevated throughout pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

Androstane-3,17-diol↗