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1.
Blood Cells Mol Dis ; 68: 203-208, 2018 02.
Article in English | MEDLINE | ID: mdl-28274788

ABSTRACT

Gaucher Disease type 1 (GD1) is a lysosomal disorder that affects many systems. Therapy improves the principal manifestations of the condition and, as a consequence, many patients show a modified phenotype which reflects manifestations of their disease that are refractory to treatment. More generally, it is increasingly recognised that information as to how a patient feels and functions [obtained by patient- reported outcome measurements (PROMs)] is critical to any comprehensive evaluation of treatment. A new set of management goals for GD1 in which both trends are reflected is needed. To this end, a modified Delphi procedure among 25 experts was performed. Based on a literature review and with input from patients, 65 potential goals were formulated as statements. Consensus was considered to be reached when ≥75% of the participants agreed to include that specific statement in the management goals. There was agreement on 42 statements. In addition to the traditional goals concerning haematological, visceral and bone manifestations, improvement in quality of life, fatigue and social participation, as well as early detection of long-term complications or associated diseases were included. When applying this set of goals in medical practice, the clinical status of the individual patient should be taken into account.


Subject(s)
Gaucher Disease/complications , Gaucher Disease/therapy , Quality of Life , Consensus , Disease Management , Europe/epidemiology , Gaucher Disease/epidemiology , Gaucher Disease/psychology , Humans
2.
Mol Ecol ; 26(3): 740-751, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27891694

ABSTRACT

Many aspects of blue whale biology are poorly understood. Some of the gaps in our knowledge, such as those regarding their basic taxonomy and seasonal movements, directly affect our ability to monitor and manage blue whale populations. As a step towards filling in some of these gaps, microsatellite and mtDNA sequence analyses were conducted on blue whale samples from the Southern Hemisphere, the eastern tropical Pacific (ETP) and the northeast Pacific. The results indicate that the ETP is differentially used by blue whales from the northern and southern eastern Pacific, with the former showing stronger affinity to the region off Central America known as the Costa Rican Dome, and the latter favouring the waters of Peru and Ecuador. Although the pattern of genetic variation throughout the Southern Hemisphere is compatible with the recently proposed subspecies status of Chilean blue whales, some discrepancies remain between catch lengths and lengths from aerial photography, and not all blue whales in Chilean waters can be assumed to be of this type. Also, the range of the proposed Chilean subspecies, which extends to the Galapagos region of the ETP, at least seasonally, perhaps should include the Costa Rican Dome and the eastern North Pacific as well.


Subject(s)
Balaenoptera/genetics , Genetic Variation , Genetics, Population , Animal Migration , Animals , Central America , Chile , DNA, Mitochondrial/genetics , Ecuador , Microsatellite Repeats , Pacific Ocean , Peru
3.
Orphanet J Rare Dis ; 11(1): 53, 2016 04 29.
Article in English | MEDLINE | ID: mdl-27129405

ABSTRACT

BACKGROUND: Gaucher disease (GD) is a rare lysosomal storage disease caused by deficiency in the enzyme beta-glucocerebrosidase. Along with visceral, hematologic, and bone manifestations, patients may experience chronic fatigue resulting in functional disability and reduced quality of life. Management of the disease includes therapeutic intervention, supportive therapies, and regular monitoring of all clinically relevant disease signs and symptoms. However, current practice guidelines do not include measurement of fatigue or therapeutic goals for fatigue. OBJECTIVE: To provide insight regarding key considerations for fatigue in GD. METHODS: We conducted a systematic PubMed literature search and an exploratory, hypothesis-generating survey regarding fatigue in GD. RESULTS: Our literature search resulted in 19 publications. Of these, 6 were identified that assessed fatigue, including 2 that used specific fatigue assessment instruments. In our survey involving 14 patients with Type 1 GD and 19 physicians, patients ascribed greater importance to fatigue than other disease parameters, while physicians placed more emphasis on objective measures of visceral and hematologic disease manifestations. CONCLUSIONS: Collectively, the results of our literature analysis and survey underscore the need for further investigation and in-office evaluation of fatigue in patients with GD, which will require a reliable, validated, and disease-specific instrument. Criteria for clinically significant fatigue in patients with GD should be established along with the development of a fatigue scale specifically designed for this patient population to provide a more objective means to potentially incorporate fatigue assessment into routine monitoring practices.


Subject(s)
Fatigue/etiology , Gaucher Disease/complications , Fatigue/pathology , Gaucher Disease/pathology , Humans
4.
Mol Ecol ; 23(24): 5998-6010, 2014 12.
Article in English | MEDLINE | ID: mdl-25492593

ABSTRACT

Blue whales (Balaenoptera musculus) were among the most intensively exploited species of whales in the world. As a consequence of this intense exploitation, blue whale sightings off the coast of Chile were uncommon by the end of the 20th century. In 2004, a feeding and nursing ground was reported in southern Chile (SCh). With the aim to investigate the genetic identity and relationship of these Chilean blue whales to those in other Southern Hemisphere areas, 60 biopsy samples were collected from blue whales in SCh between 2003 and 2009. These samples were genotyped at seven microsatellite loci and the mitochondrial control region was sequenced, allowing us to identify 52 individuals. To investigate the genetic identity of this suspected remnant population, we compared these 52 individuals to blue whales from Antarctica (ANT, n = 96), Northern Chile (NCh, n = 19) and the eastern tropical Pacific (ETP, n = 31). No significant differentiation in haplotype frequencies (mtDNA) or among genotypes (nDNA) was found between SCh, NCh and ETP, while significant differences were found between those three areas and Antarctica for both the mitochondrial and microsatellite analyses. Our results suggest at least two breeding population units or subspecies exist, which is also supported by other lines of evidence such as morphometrics and acoustics. The lack of differences detected between SCh/NCh/ETP areas supports the hypothesis that eastern South Pacific blue whales are using the ETP area as a possible breeding area. Considering the small population sizes previously reported for the SCh area, additional conservation measures and monitoring of this population should be developed and prioritized.


Subject(s)
Balaenoptera/genetics , Genetics, Population , Animal Migration , Animals , Antarctic Regions , Bayes Theorem , Cell Nucleus/genetics , Chile , Cluster Analysis , DNA, Mitochondrial/genetics , Female , Gene Flow , Genetic Variation , Genotype , Haplotypes , Male , Microsatellite Repeats , Pacific Ocean , Sequence Analysis, DNA
5.
Blood Cells Mol Dis ; 50(3): 206-11, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23199589

ABSTRACT

Taliglucerase alfa (Protalix Biotherapeutics, Israel) is a carrot-cell-expressed recombinant human beta-glucocerebrosidase recently approved in the United States for the treatment of type 1 Gaucher disease (GD). As bone disease is one of the most debilitating features of GD, quantification of bone marrow involvement is important for monitoring the response to treatment. Therefore, bone marrow fat fraction (Ff) measured by quantitative chemical shift imaging (QCSI) was included as exploratory parameter to evaluate bone marrow response in treatment naïve GD patients participating in a double-blind, randomized phase III study. Eight GD patients with intact spleens were treated with 30 or 60U/kg biweekly. Ff results were compared to outcomes in 15 untreated Dutch GD patients with a follow-up interval of 1year. Five taliglucerase alfa treated patients had a Ff below the threshold that relates to complication risk (<0.23) at baseline (median (n=8) 0.19, range 0.11-0.35). Ff significantly increased compared to baseline (p=0.012) and compared to untreated patients (p=0.005), already after 1year of follow-up with further improvement up to 36months. In four patients with the lowest Ff, the higher dose resulted in increases above 0.23 within 1year. All patients had sustained improvements in all other parameters. There was no influence of antibodies on response parameters. Treatment with taliglucerase alfa results in significant increases in lumbar spine fat fractions, which indicates clearance of Gaucher cells from the bone marrow.


Subject(s)
Enzyme Replacement Therapy , Gaucher Disease/drug therapy , Glucosylceramidase/therapeutic use , Adipose Tissue/metabolism , Adult , Aged , Antibodies/immunology , Antibodies, Neutralizing/immunology , Bone Marrow/drug effects , Bone Marrow/metabolism , Enzyme Replacement Therapy/adverse effects , Female , Glucosylceramidase/administration & dosage , Glucosylceramidase/immunology , Humans , Male , Middle Aged , Treatment Outcome , Young Adult
6.
Heredity (Edinb) ; 107(4): 349-61, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21427750

ABSTRACT

Genetic analyses of population structure can be placed in explicit environmental contexts if appropriate environmental data are available. Here, we use high-coverage and high-resolution oceanographic and genetic sequence data to assess population structure patterns and their potential environmental influences for humpback dolphins in the Western Indian Ocean. We analyzed mitochondrial DNA data from 94 dolphins from the coasts of South Africa, Mozambique, Tanzania and Oman, employing frequency-based and maximum-likelihood algorithms to assess population structure and migration patterns. The genetic data were combined with 13 years of remote sensing oceanographic data of variables known to influence cetacean dispersal and population structure. Our analyses show strong and highly significant genetic structure between all putative populations, except for those in South Africa and Mozambique. Interestingly, the oceanographic data display marked environmental heterogeneity between all sampling areas and a degree of overlap between South Africa and Mozambique. Our combined analyses therefore suggest the occurrence of genetically isolated populations of humpback dolphins in areas that are environmentally distinct. This study highlights the utility of molecular tools in combination with high-resolution and high-coverage environmental data to address questions not only pertaining to genetic population structure, but also to relevant ecological processes in marine species.


Subject(s)
Dolphins/genetics , Ecosystem , Animals , DNA, Mitochondrial/genetics , Dolphins/classification , Environment , Genetic Variation , Genetics, Population , Indian Ocean , Marine Biology , Phylogeny , Population Dynamics , Remote Sensing Technology
7.
N Engl J Med ; 361(17): 1651-61, 2009 Oct 22.
Article in English | MEDLINE | ID: mdl-19846850

ABSTRACT

BACKGROUND: Recent studies indicate an increased frequency of mutations in the gene encoding glucocerebrosidase (GBA), a deficiency of which causes Gaucher's disease, among patients with Parkinson's disease. We aimed to ascertain the frequency of GBA mutations in an ethnically diverse group of patients with Parkinson's disease. METHODS: Sixteen centers participated in our international, collaborative study: five from the Americas, six from Europe, two from Israel, and three from Asia. Each center genotyped a standard DNA panel to permit comparison of the genotyping results across centers. Genotypes and phenotypic data from a total of 5691 patients with Parkinson's disease (780 Ashkenazi Jews) and 4898 controls (387 Ashkenazi Jews) were analyzed, with multivariate logistic-regression models and the Mantel-Haenszel procedure used to estimate odds ratios across centers. RESULTS: All 16 centers could detect two GBA mutations, L444P and N370S. Among Ashkenazi Jewish subjects, either mutation was found in 15% of patients and 3% of controls, and among non-Ashkenazi Jewish subjects, either mutation was found in 3% of patients and less than 1% of controls. GBA was fully sequenced for 1883 non-Ashkenazi Jewish patients, and mutations were identified in 7%, showing that limited mutation screening can miss half the mutant alleles. The odds ratio for any GBA mutation in patients versus controls was 5.43 across centers. As compared with patients who did not carry a GBA mutation, those with a GBA mutation presented earlier with the disease, were more likely to have affected relatives, and were more likely to have atypical clinical manifestations. CONCLUSIONS: Data collected from 16 centers demonstrate that there is a strong association between GBA mutations and Parkinson's disease.


Subject(s)
Glucosylceramidase/genetics , Mutation , Parkinson Disease/genetics , Aged , Case-Control Studies , Genotype , Humans , Jews/genetics , Logistic Models , Middle Aged , Multivariate Analysis , Odds Ratio
8.
Proc Biol Sci ; 272(1562): 533-42, 2005 Mar 07.
Article in English | MEDLINE | ID: mdl-15846869

ABSTRACT

Right whales (genus: Eubalaena) are among the most endangered mammals, yet their taxonomy and phylogeny have been questioned. A phylogenetic hypothesis based on mitochondrial DNA (mtDNA) variation recently prompted a taxonomic revision, increasing the number of right whale species to three. We critically evaluated this hypothesis using sequence data from 13 nuclear DNA (nuDNA) loci as well as the mtDNA control region. Fixed diagnostic characters among the nuclear markers strongly support the hypothesis of three genetically distinct species, despite lack of any diagnostic morphological characters. A phylogenetics analysis of all data produced a strict consensus cladogram with strong support at nodes that define each right whale species as well as relationships among species. Results showed very little conflict among the individual partitions as well as congruence between the mtDNA and nuDNA datasets. These data clearly demonstrate the strength of using numerous independent genetic markers during a phylogenetics analysis of closely related species. In evaluating phylogenetic support contributed by individual loci, 11 of the 14 loci provided support for at least one of the nodes of interest to this study. Only a single marker (mtDNA control region) provided support at all four nodes. A study using any single nuclear marker would have failed to support the proposed phylogeny, and a strong phylogenetic hypothesis was only revealed by the simultaneous analysis of many nuclear loci. In addition, nu DNA and mtDNA data provided complementary levels of support at nodes of different evolutionary depth indicating that the combined use of mtDNA and nuDNA data is both practical and desirable.


Subject(s)
Genetic Variation , Phylogeny , Whales/genetics , Animals , Base Sequence , Cell Nucleus/genetics , DNA, Mitochondrial/genetics , Molecular Sequence Data , Sequence Analysis, DNA , Species Specificity , Whales/classification
9.
Mol Genet Metab ; 79(2): 104-9, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12809640

ABSTRACT

Among the phenotypes associated with Gaucher disease, the deficiency of glucocerebrosidase, are rare patients with early onset, treatment-refractory parkinsonism. Sequencing of glucocerebrosidase in 17 such patients revealed 12 different genotypes. Fourteen patients had the common "non-neuronopathic" N370S mutation, including five N370S homozygotes. While brain glucosylsphingosine levels were not elevated, Lewy bodies were seen in the four brains available for study. The shared clinical and neuropathologic findings in this subgroup suggest that the deficiency in glucocerebrosidase may contribute to a vulnerability to parkinsonism.


Subject(s)
Gaucher Disease/genetics , Genetic Predisposition to Disease , Glucosylceramidase/deficiency , Glucosylceramidase/genetics , Parkinson Disease/etiology , Sphingosine/analogs & derivatives , Adult , Blotting, Southern , Brain/metabolism , Brain/pathology , Deoxyribonucleases, Type II Site-Specific/genetics , Female , Gaucher Disease/complications , Gaucher Disease/metabolism , Gaucher Disease/pathology , Homozygote , Humans , Levodopa/therapeutic use , Male , Middle Aged , Mutation , Parkinson Disease/drug therapy , Parkinson Disease/metabolism , Parkinson Disease/pathology , Psychosine/analogs & derivatives , Sphingosine/metabolism
10.
J Hered ; 93(6): 389-99, 2002.
Article in English | MEDLINE | ID: mdl-12642638

ABSTRACT

To examine whether demographic and life-history traits are correlated with genetic structure, we contrasted mtDNA lineages of individual humpback whales (Megaptera novaeangliae) with sighting and reproductive histories of female humpback whales between 1979 and 1995. Maternal lineage haplotypes were obtained for 323 whales, either from direct sequencing of the mtDNA control region (n = 159) or inferred from known relationships along matrilines from the sequenced sample of individuals (n = 164). Sequence variation in the 550 bp of the control region defined a total of 19 maternal lineage haplotypes that formed two main clades. Fecundity increased significantly over the study period among females of several lineages among the two clades. Individual maternal lineages and other clades were characterized by significant variation in fecundity. The detected heterogeneity of reproductive success has the potential to substantially affect the frequency and distribution of maternal lineages found in this population over time. There were significant yearly effects on adult resighting rates and calf survivorship based on examination of sighting histories with varying capture-recapture probability models. These results indicate that population structure can be influenced by interactions or associations between reproductive success, genetic structure, and environmental factors in a natural population of long-lived mammals.


Subject(s)
DNA, Mitochondrial/genetics , Whales/genetics , Whales/physiology , Animals , Base Sequence , Female , Fertility , Genetic Variation , Genetics, Population , Haplotypes , Maine , Population Density , Reproduction , Seawater , Whales/growth & development
11.
Appl Environ Microbiol ; 67(9): 4206-14, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11526025

ABSTRACT

Total DNA of a population of uncultured organisms was extracted from soil samples, and by using PCR methods, the genes encoding two different 2,5-diketo-D-gluconic acid reductases (DKGRs) were recovered. Degenerate PCR primers based on published sequence information gave internal gene fragments homologous to known DKGRs. Nested primers specific for the internal fragments were combined with random primers to amplify flanking gene fragments from the environmental DNA, and two hypothetical full-length genes were predicted from the combined sequences. Based on these predictions, specific primers were used to amplify the two complete genes in single PCRs. These genes were cloned and expressed in Escherichia coli. The purified gene products catalyzed the reduction of 2,5-diketo-D-gluconic acid to 2-keto-L-gulonic acid. Compared to previously described DKGRs isolated from Corynebacterium spp., these environmental reductases possessed some valuable properties. Both exhibited greater than 20-fold-higher kcat/Km values than those previously determined, primarily as a result of better binding of substrate. The Km values for the two new reductases were 57 and 67 microM, versus 2 and 13 mM for the Corynebacterium enzymes. Both environmental DKGRs accepted NADH as well as NADPH as a cosubstrate; other DKGRs and most related aldo-keto reductases use only NADPH. In addition, one of the new reductases was more thermostable than known DKGRs.


Subject(s)
Bacteria/genetics , Cloning, Molecular , DNA, Bacterial/genetics , Polymerase Chain Reaction/methods , Soil Microbiology , Sugar Alcohol Dehydrogenases/genetics , Amino Acid Sequence , Bacteria/enzymology , Base Sequence , DNA Primers , Kinetics , Molecular Sequence Data , Sequence Analysis, DNA , Sugar Alcohol Dehydrogenases/chemistry , Sugar Alcohol Dehydrogenases/isolation & purification , Sugar Alcohol Dehydrogenases/metabolism
12.
Mol Ecol ; 9(11): 1793-802, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11091315

ABSTRACT

Few studies have examined systematic relationships of right whales (Eubalaena spp.) since the original species descriptions, even though they are one of the most endangered large whales. Little morphological evidence exists to support the current species designations for Eubalaena glacialis in the northern hemisphere and E. australis in the southern hemisphere. Differences in migratory behaviour or antitropical distribution between right whales in each hemisphere are considered a barrier to gene flow and maintain the current species distinctions and geographical populations. However, these distinctions between populations have remained controversial and no study has included an analysis of all right whales from the three major ocean basins. To address issues of genetic differentiation and relationships among right whales, we have compiled a database of mitochondrial DNA control region sequences from right whales representing populations in all three ocean basins that consist of: western North Atlantic E. glacialis, multiple geographically distributed populations of E. australis and the first molecular analysis of historical and recent samples of E. glacialis from the western and eastern North Pacific Ocean. Diagnostic characters, as well as phylogenetic and phylogeographic analyses, support the possibility that three distinct maternal lineages exist in right whales, with North Pacific E. glacialis being more closely related to E. australis than to North Atlantic E. glacialis. Our genetic results provide unequivocal character support for the two usually recognized species and a third distinct genetic lineage in the North Pacific under the Phylogenetic Species Concept, as well as levels of genetic diversity among right whales world-wide.


Subject(s)
Whales/genetics , Animals , Atlantic Ocean , Base Sequence , DNA, Mitochondrial/genetics , Databases, Factual , Genetic Variation , Genetics, Population , Pacific Ocean , Phylogeny , Species Specificity
13.
Isr Med Assoc J ; 2(1): 22-4, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10892366

ABSTRACT

BACKGROUND: The etiology of bone marrow failure, a prominent feature of paroxysmal nocturnal hemoglobulinuria, is presently unknown. OBJECTIVES: To evaluate the possible influence of cellular immune mechanisms in the bone marrow failure of PNH. METHODS: We studied marrow erythroid colony formation in a patient with paroxysmal nocturnal hemoglobinuria without hypoplastic/aplastic marrow complications. RESULTS: In vitro assays revealed a pronounced inhibition of primitive erythroid (BFU-E) progenitor cell growth by marrow T lymphocytes. Removal of T cells prior to culture resulted in a 4.5-fold enhancement of BFU-E numbers. Reevaluation of in vitro erythropoiesis during steroid administration indicated a persistent, albeit less prominent, T cell inhibitory effect. CONCLUSION: Our findings provide the first direct evidence for a cellular immune inhibitory phenomenon accompanying PNH.


Subject(s)
Erythroid Precursor Cells/metabolism , Erythropoiesis/immunology , Hemoglobinuria, Paroxysmal/immunology , T-Lymphocytes/immunology , Adult , Anemia, Refractory , Case-Control Studies , Female , Humans , Immunity, Cellular , In Vitro Techniques , Pregnancy , Pregnancy Complications, Hematologic
14.
Leuk Res ; 23(4): 323-30, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10229317

ABSTRACT

The skin involvement of the myelodysplastic syndrome (MDS) can take the form of either a neoplastic infiltration or various non specific lesions. The occurrence of these lesions may be the presenting feature of the disease (MDS) or may herald its progression to acute leukemia. Recognition and early diagnosis have therapeutic and prognostic significance.


Subject(s)
Myelodysplastic Syndromes/pathology , Skin/pathology , Adolescent , Aged , Aged, 80 and over , Anemia, Refractory, with Excess of Blasts/etiology , Anemia, Refractory, with Excess of Blasts/pathology , Female , Humans , Leukemia, Myelomonocytic, Chronic/pathology , Leukemic Infiltration/pathology , Male , Middle Aged , Myelodysplastic Syndromes/complications , Pyoderma Gangrenosum/etiology , Pyoderma Gangrenosum/pathology , Sweet Syndrome/etiology , Sweet Syndrome/pathology
16.
Mol Ecol ; 7(6): 695-707, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9640650

ABSTRACT

The population structure of variation in a nuclear actin intron and the control region of mitochondrial DNA is described for humpback whales from eight regions in the North Pacific Ocean: central California, Baja Peninsula, nearshore Mexico (Bahia Banderas), offshore Mexico (Socorro Island), southeastern Alaska, central Alaska (Prince Williams Sound), Hawaii and Japan (Ogasawara Islands). Primary mtDNA haplotypes and intron alleles were identified using selected restriction fragment length polymorphisms of target sequences amplified by the polymerase chain reaction (PCR-RFLP). There was little evidence of heterogeneity in the frequencies of mtDNA haplotypes or actin intron alleles due to the year or sex composition of the sample. However, frequencies of four mtDNA haplotypes showed marked regional differences in their distributions (phi ST = 0.277; P < 0.001; n = 205 individuals) while the two alleles showed significant, but less marked, regional differences (phi ST = 0.033; P < 0.013; n = 400 chromosomes). An hierarchical analysis of variance in frequencies of haplotypes and alleles supported the grouping of six regions into a central and eastern stock with further partitioning of variance among regions within stocks for haplotypes but not for alleles. Based on available genetic and demographic evidence, the southeastern Alaska and central California feeding grounds were selected for additional analyses of nuclear differentiation using allelic variation at four microsatellite loci. All four loci showed significant differences in allele frequencies (overall FST = 0.043; P < 0.001; average n = 139 chromosomes per locus), indicating at least partial reproductive isolation between the two regions as well as the segregation of mtDNA lineages. Although the two feeding grounds were not panmictic for nuclear or mitochondrial loci, estimates of long-term migration rates suggested that male-mediated gene flow was several-fold greater than female gene flow. These results include and extend the range and sample size of previously published work, providing additional evidence for the significance of genetic management units within oceanic populations of humpback whales.


Subject(s)
Genetic Variation , Genetics, Population , Whales/genetics , Alleles , Animals , DNA, Mitochondrial/genetics , Feeding Behavior , Female , Geography , Haplotypes , Introns , Male , Microsatellite Repeats , Pacific Ocean , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Seasons
17.
J Biol Chem ; 273(13): 7657-67, 1998 Mar 27.
Article in English | MEDLINE | ID: mdl-9516471

ABSTRACT

Granulocyte-macrophage colony-stimulating factor (GM-CSF) activity is mediated by a cellular receptor (GM-CSFR) that is comprised of an alpha-chain (GM-CSFRalpha), which specifically binds GM-CSF, and a beta-chain (betac), shared with the interleukin-3 and interleukin-5 receptors. GM-CSFRalpha exists in both a transmembrane (tmGM-CSFRalpha) and a soluble form (sGM-CSFRalpha). We designed an sGM-CSFRalpha-Fc fusion protein to study GM-CSF interactions with the GM-CSFRalpha. The construct was prepared by fusing the coding region of the sGM-CSFRalpha with the CH2-CH3 regions of murine IgG2a. Purified sGM-CSFRalpha-Fc ran as a monomer of 60 kDa on reducing SDS-polyacrylamide gel electrophoresis but formed a trimer of 160-200 kDa under nonreducing conditions. The sGM-CSFRalpha-Fc bound specifically to GM-CSF as demonstrated by standard and competitive immunoassays, as well as by radioligand assay with 125I-GM-CSF. The sGM-CSFRalpha-Fc also inhibited GM-CSF-dependent cell growth and therein is a functional antagonist. Kinetics of sGM-CSFRalpha-Fc binding to GM-CSF were evaluated using an IAsys biosensor (Affinity Sensors, Paramus, NJ) with two assay systems. In the first, the sGM-CSFRalpha-Fc was bound to immobilized staphylococcal protein A on the biosensor surface, and binding kinetics of GM-CSF in solution were determined. This revealed a rapid koff of 2.43 x 10(-2)/s. A second set of experiments was performed with GM-CSF immobilized to the sensor surface and the sGM-CSFRalpha-Fc in solution. The dissociation rate constant (koff) for the sGM-CSFRalpha-Fc trimer from GM-CSF was 1.57 x 10(-3)/s, attributable to the higher avidity of binding in this assay. These data indicate rapid dissociation of GM-CSF from the sGM-CSFRalpha-Fc and suggest that in vivo, sGM-CSFRalpha may need to be present in the local environment of a responsive cell to exert its antagonist activity.


Subject(s)
Immunoglobulin Fc Fragments/metabolism , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Recombinant Fusion Proteins/metabolism , Animals , Biosensing Techniques , Cell Line , Cloning, Molecular , Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Immunoglobulin Fc Fragments/genetics , Kinetics , Mice , Protein Binding , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/isolation & purification , Solubility
18.
Ann Pharmacother ; 32(1): 60-2, 1998 Jan.
Article in English | MEDLINE | ID: mdl-9475823

ABSTRACT

BACKGROUND: Amiodarone hydrochloride is classified as a Vaughan Williams class III antiarrhythmic agent, although class I, II, and IV effects may contribute to its favorable antiarrhythmic profile. It is associated with a wide variety of adverse effects, such as hypothyroidism, hyperthyroidism, interstitial pulmonary disease, hepatitis, coagulation disorders, skin photosensitivity, corneal microdeposits, alopecia, peripheral neuropathy, and cardiovascular arrhythmias. SUBJECTS: Bone marrow aspirations and biopsies were performed on two patients treated with amiodarone, on the first during a follow-up for myelofibrosis and on the second for a suspected lymphoproliferative disorder. Several bone marrow granulomas were found in both patients. The bone marrow specimens for tuberculosis and fungal stains were negative. CONCLUSIONS: The temporal relationship between the amoidarone therapy and the development of two cases of asymptomatic bone marrow granuloma suggest the possibility that this antiarrhythmic agent is involved in the etiology of these granulomas.


Subject(s)
Amiodarone/adverse effects , Anti-Arrhythmia Agents/adverse effects , Bone Marrow Diseases/chemically induced , Granuloma/chemically induced , Aged , Amiodarone/therapeutic use , Anti-Arrhythmia Agents/therapeutic use , Female , Humans , Lymphoproliferative Disorders/drug therapy , Middle Aged , Primary Myelofibrosis/drug therapy
19.
DNA Seq ; 8(5): 317-27, 1998.
Article in English | MEDLINE | ID: mdl-10993602

ABSTRACT

In numerous population genetic and disease association studies decisions about the ancestry of polymorphic alleles are often made based on the relative frequency of the alleles in the extant populations with the most frequent allele being deemed as ancestral. However, the frequency of an allele in a population is generally not a perfect indicator of its ancestral status. A more accurate method to assess ancestral/derived status of polymorphic alleles involves identification of shared alleles between species. We used this strategy to examine genomic regions homologous to several human polymorphisms in four species of non-human primates. Cross species polymerase chain reaction (CS-PCR), with primers designed from human sequence, was used to investigate regions of interest. Nineteen polymorphisms at six loci (DRD2, HOXB@, PAH, D4S10, RBP3, and RET) were examined either by restriction fragment length analysis of PCR products (PCR-RFLP) or by direct sequencing. At seventeen of the eighteen PCR-RFLPs, non-human primates were monomorphic and identical to each other for either lack of restriction enzyme site or presence of the site. Thus, at these seventeen polymorphic sites the shared alleles are most likely to be the ancestral ones in humans. In several cases we have used sequence data to further demonstrate that the nucleotide at the site of the polymorphism is conserved between species confirming the hypothesis of a single ancestral allele. However, not all human alleles can be simply resolved into ancestral and derived; sequence data from one PCR-RFLP (in an intron of the PAH locus) and a single strand conformational polymorphism (SSCP) in the 3' untranslated region (UTR) of the DRD2 gene illustrate this point.


Subject(s)
Alleles , Evolution, Molecular , Polymorphism, Genetic , Animals , Gene Frequency , Gorilla gorilla/genetics , Humans , Pan paniscus/genetics , Pan troglodytes/genetics , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , Polymorphism, Single-Stranded Conformational , Pongo pygmaeus/genetics , Sequence Analysis, DNA/methods , Species Specificity
20.
Mol Ecol ; 6(7): 677-81, 1997 Jul.
Article in English | MEDLINE | ID: mdl-9226948

ABSTRACT

DNA was isolated from an early twentieth century museum specimen of northern right whale baleen. A system of stringent controls and a novel set of cetacean specific primers eliminated contamination from external sources and ensured the authenticity of the results. Sequence analysis revealed that there were informative nucleotide positions between the museum specimen and extant members of the population and closely related species. The results indicate that museum specimens of baleen can be used to assess historical genetic population structure of the great whales.


Subject(s)
DNA, Mitochondrial/genetics , Whales/genetics , Animals , Base Sequence , DNA Primers , DNA, Mitochondrial/chemistry , DNA, Mitochondrial/isolation & purification , Genetics, Population , Molecular Sequence Data , Museums , Polymerase Chain Reaction , Sequence Alignment , Sequence Homology, Nucleic Acid , Whales/classification
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