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1.
Haemophilia ; 21(1): 95-101, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25458735

ABSTRACT

UNLABELLED: Congenital factor XIII (FXIII) deficiency is a rare condition with substantial risk for life-threatening bleeding. Replacement of deficient FXIII with plasma-derived FXIII concentrate is a treatment option. The current 12-week study evaluated the steady-state pharmacokinetic (PK) and safety profile of prophylactic infusions of FXIII concentrate (human) in patients with congenital FXIII deficiency. Patients received FXIII concentrate (human) 40 IU kg(-1) on Days 0, 28, and 56. FXIII levels were assessed before and after each infusion; steady-state PK parameters were assessed up to 28 days after the infusion on Day 56. Treatment effectiveness in maintaining trough FXIII activity levels ≥ 5% over 28 days and safety parameters were also assessed. Fourteen patients received FXIII concentrate (human) and 13 completed the study. Post-infusion, FXIII activity levels increased to within the range found in patients without congenital FXIII deficiency without reaching supra-therapeutic levels. Non-baseline-adjusted trough FXIII activity levels were maintained at or above 10% at all post-baseline visits in all patients. Steady-state PK parameters were baseline-adjusted; maximum FXIII activity was 87.7% at 1.72 h post-infusion, subsequently declining to a minimum of 5.0%. The half-life was 6.6 days. FXIII concentrate (human) was generally well tolerated. Two patients had possibly treatment-related adverse events. There were no reports of thromboembolism, viral transmission, bleeding events or treatment-related hypersensitivity. These findings support use of FXIII concentrate (human) 40 IU kg(-1) every 28 days as an appropriate regimen for routine, long-term prophylaxis in children and adults with congenital FXIII deficiency. CLINICAL TRIAL REGISTRATION: www.clinicaltrials.gov/ct2/show/NCT00883090.


Subject(s)
Factor XIII Deficiency/congenital , Factor XIII/pharmacokinetics , Adolescent , Adult , Biomarkers, Pharmacological , Child , Child, Preschool , Factor XIII/therapeutic use , Factor XIII Deficiency/drug therapy , Female , Humans , Male , Young Adult
2.
Mar Biotechnol (NY) ; 13(1): 74-82, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20352272

ABSTRACT

The caudal fin represents a fundamental design feature of fishes and plays an important role in locomotor dynamics in fishes. The shape of caudal is an important parameter in traditional systematics. However, little is known about genes involved in the development of different forms of caudal fins. This study was conducted to identify and map quantitative trait loci (QTL) affecting the length of caudal fin and the ratio between tail length and standard body length in Asian seabass (Lates calcarifer). One F1 family containing 380 offspring was generated by crossing two unrelated individuals. One hundred and seventeen microsatellites almost evenly distributed along the whole genome were genotyped. Length of caudal fin at 90 days post-hatch was measured. QTL analysis detected six significant (genome-wide significant) and two suggestive (linkage-group-wide significant) QTL on seven linkage groups. The six significant QTL explained 5.5-16.6% of the phenotypic variance, suggesting these traits were controlled by multiple genes. Comparative genomics analysis identified several potential candidate genes for the length of caudal fin. The QTL for the length of caudal fin detected for the first time in marine fish may provide a starting point for the future identification of genes involved in the development of different forms of caudal fins in fishes.


Subject(s)
Animal Fins/anatomy & histology , Perciformes/genetics , Quantitative Trait Loci , Animal Fins/growth & development , Animal Fins/metabolism , Animals , Body Weights and Measures , Chromosome Mapping , Inheritance Patterns , Perciformes/anatomy & histology , Perciformes/growth & development , Phenotype
3.
Anim Genet ; 41(2): 208-12, 2010 Apr.
Article in English | MEDLINE | ID: mdl-19793264

ABSTRACT

Microsatellites are the most popular markers for parentage assignment and population genetic studies. To meet the demand for international comparability for genetic studies of Asian seabass, a standard panel of 28 microsatellites has been selected and characterized using the DNA of 24 individuals from Thailand, Malaysia, Indonesia and Australia. The average allele number of these markers was 10.82 +/- 0.71 (range: 6-19), and the expected heterozygosity averaged 0.76 +/- 0.02 (range: 0.63-1.00). All microsatellites showed Mendelian inheritance. In addition, eight standard size controls have been developed by cloning a set of microsatellite alleles into a pGEM-T vector to calibrate allele sizes determined by different laboratories, and are available upon request. Seven multiplex PCRs, each amplifying 3-5 markers, were optimized to accurately and rapidly genotype microsatellites. Parentage assignment using 10 microsatellites in two crosses (10 x 10 and 20 x 20) demonstrated a high power of these markers for revealing parent-sibling connections. This standard set of microsatellites will standardize genetic diversity studies of Asian seabass, and the multiplex PCR sets will facilitate parentage assignment.


Subject(s)
Bass/genetics , Microsatellite Repeats , Animals , Asia, Southeastern , Australia , Genetic Variation , Genetics, Population
4.
Int J Biol Sci ; 4(5): 279-82, 2008 Aug 28.
Article in English | MEDLINE | ID: mdl-18781225

ABSTRACT

Self-cloning is quite rare in shrimp, lobsters, crayfish and crabs. Here we report the discovery of four natural clones of red swamp crayfish (Procambarus clarkii), each containing 2-6 genetically identical individuals, during the genotyping of 120 individuals with five microsatellites. The four clones were heterozygote at most of the five microsatellite loci. Phylogenetic analysis using microsatellite genotypes suggests recent origin of the four clones. Sequencing a part of the mitochondrial gene Cox I confirmed that the four clones were from the species Procambarus clarkii.


Subject(s)
Astacoidea/genetics , Microsatellite Repeats/genetics , Animals , Astacoidea/classification , Clone Cells/metabolism , Electron Transport Complex IV/genetics , Genotype , Phylogeny
6.
Anim Genet ; 39(1): 34-9, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18076743

ABSTRACT

Quantitative trait loci (QTL) affecting growth traits have previously been mapped in linkage groups (LG) 2, 3 and 23 of Barramundi (Lates carcalifer), but these QTL have not been verified in different genetic backgrounds and environments. Here, we report the identification and verification of QTL for growth traits on LG2, 3, 10 and 23 in F(1) families constructed using brooders from the Singapore Marine Aquaculture Center (MAC) and from wild stocks collected in Thailand (THAI). The previously detected QTL for body weight and length linked to marker Lca371 on LG2 were confirmed in both the MAC and THAI families, whereas other QTL previously mapped to LG3 and 23 were only detected in one of the two families. QTL for body weight and length were identified in the MAC family, but not in the THAI family, in a region where the insulin-like growth factor 2 (IGF2) and tyrosine hydroxylase 1 (TH1) genes are located on LG10. Significant epistatic interactions were identified between markers Lca287 on LG2 and IGF2 on LG10 for growth trait QTL in the MAC family, but not in the THAI family. Effects of the IGF2, TH1 and parvalbumin 1 candidate genes were family-specific. Our results indicate that some but not all QTL are family-specific in Barramundi.


Subject(s)
Perciformes/growth & development , Perciformes/genetics , Quantitative Trait Loci , Alleles , Animals , Animals, Wild/genetics , Animals, Wild/growth & development , Breeding , Chromosome Mapping , DNA/genetics , Epistasis, Genetic , Female , Fisheries , Genetic Variation , Insulin-Like Growth Factor II/genetics , Male , Microsatellite Repeats , Parvalbumins/genetics , Perciformes/classification , Phenotype , Singapore , Species Specificity , Thailand , Tyrosine 3-Monooxygenase/genetics
7.
Genome ; 49(8): 969-76, 2006 Aug.
Article in English | MEDLINE | ID: mdl-17036072

ABSTRACT

Barramundi (Lates calcarifer) is an important marine food fish species in Southeast Asia and Australia. Seventy-four novel microsatellites were isolated from a genomic DNA library enriched for CA repeats and were characterized in 24 unrelated individuals. Among the 74 microsatellites, 71 were polymorphic, with an average allele number of 7.0 +/- 3.6/locus. The average expected heterozygosity of these polymorphic markers was 0.66. Sixty-three of the 71 polymorphic microsatellites conformed to Hardy-Weinberg equilibrium. Linkage analyses were conducted in a reference family, leading to the assignment of 34 novel microsatellites and 16 published markers in 16 linkage groups. The novel microsatellites developed in this study will contribute significantly to the construction of a first-generation linkage map for mapping of quantitative trait loci in Barramundi, and supply a large choice of markers for studies on population genetics, stock management, and pedigree reconstruction.


Subject(s)
Perciformes/genetics , Animals , Asia, Southeastern , Base Sequence , Chromosome Mapping , DNA/genetics , DNA Primers/genetics , Female , Gene Library , Genetic Linkage , Genetic Markers , Heterozygote , Male , Microsatellite Repeats , Polymorphism, Genetic
9.
Anim Genet ; 37(3): 266-8, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16734689

ABSTRACT

Parvalbumins are extremely abundant in fish muscle and play an important role in muscle relaxation. In this study, two parvalbumin genes (PVALB1 and PVALB2) were cloned from Asian seabass (Lates calcarifer). The cDNAs for PVALB1 and PVALB2 were 840 and 667 bp respectively. Both genes consisted of five exons and four introns, encoded 109 amino acids, and were of beta lineage. Using real-time polymerase chain reaction, expression of PVALB1 was detected in all 10 tissues tested, with expression in brain, kidney, muscle and small intestine being 15- to 322-fold higher than in the other tissues. Expression of PVALB2 was detected only in muscle, brain and intestine, and was up to 10-fold lower than PVALB1 expression. A (CT)(17) microsatellite in the 3'-untranslated region of PVALB1 and three single nucleotide polymorphisms (SNPs) in the third intron of PVALB2 were identified. The microsatellite in PVALB1 was significantly associated with body weight and body length at 90 days post-hatch (P < 0.01), whereas the SNPs in PVALVB2 were not associated with these traits.


Subject(s)
Bass/genetics , Fish Proteins/genetics , Parvalbumins/genetics , 3' Untranslated Regions , Animals , Bass/anatomy & histology , Bass/growth & development , Fish Proteins/metabolism , Fish Proteins/physiology , Haplotypes , Microsatellite Repeats , Parvalbumins/metabolism , Parvalbumins/physiology , Polymerase Chain Reaction , Polymorphism, Single Nucleotide
10.
Mar Biotechnol (NY) ; 8(1): 71-9, 2006.
Article in English | MEDLINE | ID: mdl-16228120

ABSTRACT

We determined the complete mtDNA nucleotide sequence of Lates calcarifer using the shotgun sequencing method. The mitochondrial DNA (mtDNA) was 16,535 base pairs (bp) in length, and contained 13 protein coding genes, 22 transfer RNAs, 2 ribosomal RNAs, and one major noncoding control region (CR). The CR was unusually short at only 768 bp. A striking feature of the mitochondrial genome was the high G+C content (46.1%), which is among the highest in fish. The gene order was identical to that of a typical vertebrate. Phylogenetic analyses using concatenated amino acid sequences of 12 protein-coding genes of 30 fish species representing 14 suborders clearly showed Lates calcarifer was located in the cluster of fish species from the order Perciformes, supporting the traditional systematic classification. We characterized single-nucleotide polymorphisms (SNPs) in the CR by sequencing the complete CR of 25 individuals obtained from Australia and Singapore. A total of 68 SNPs were detected. Eighteen SNPs were fixed with alternative nucleotides in Australian and Singapore seabass, and these SNPs could be used for differentiating fish from the two countries.


Subject(s)
DNA, Mitochondrial/chemistry , Genome/genetics , Locus Control Region/genetics , Mitochondria/genetics , Perciformes/genetics , Polymorphism, Single Nucleotide/genetics , Animals , Base Composition/genetics , Base Sequence/genetics , Biological Evolution , Gene Order/genetics , Genes, rRNA , Molecular Sequence Data , Perciformes/classification , Phylogeny , Polymerase Chain Reaction/veterinary , RNA, Transfer/genetics , Sequence Analysis, DNA/veterinary
11.
J Comb Chem ; 3(4): 341-5, 2001.
Article in English | MEDLINE | ID: mdl-11442390

ABSTRACT

Four polymer-supported benzotriazoles were prepared by linkage of 5-(hydroxymethyl)benzotriazole and benzotrizaole-5-carboxylic acid with Wang resin, Merrifield resin, and (monomethoxy)poly(ethylene glycol). The solid-phase and liquid-phase syntheses of tetrahydroquinolines were achieved by two-pair coupling reactions of aldehydes and aromatic amines using these polymer-supported benzotriazoles as the promoters. The ether-type benzotriazole prepared by loading 5-(hydroxymethyl)benzotriazole onto Merrifield resin turned out to be the catalyst of choice. Thus, a series of tetrahydroquinoline products were obtained in high purity by simple filtration, and the resin was recovered for reuse without loss of activity.

12.
Chirality ; 13(5): 266-71, 2001 May 15.
Article in English | MEDLINE | ID: mdl-11317348

ABSTRACT

The absolute configuration of beta-hydroxy-alpha-amino acids was studied by CD exciton chirality method using 7-diethylaminocoumarin-3-carboxylate as a red-shifted chromophore. The CD spectra of bischromophoric derivatives of (S)-serine and (2S,3R)-threonine methyl esters (2 and 7) were compared with those of acyclic vic-aminoalcohols and diols (3--6 and 8--9). This study indicates that the polar carboxylate group of beta-hydroxy-alpha-amino acids makes them a unique subclass of vic-aminoalcohols. By combining the data of CD and NMR coupling constants, we are able to correlate their preferred conformer B and positive CD to the corresponding absolute configuration.


Subject(s)
Amino Acids/chemistry , Circular Dichroism , Magnetic Resonance Spectroscopy , Molecular Structure , Spectrophotometry, Ultraviolet , Stereoisomerism
13.
Bioorg Med Chem Lett ; 9(5): 709-12, 1999 Mar 08.
Article in English | MEDLINE | ID: mdl-10201833

ABSTRACT

A facile preparation of triacetylated derivative of 2-phenylthioglycoside of N-acetylneuraminic acid (4) was achieved by treatment with lipase PS in vinyl acetate. The major product 4 has a free hydroxyl group at C-7. Results of time-course HPLC analysis indicate that the reactivity of the hydroxyl groups under this condition is in the following order; C-9 > C-4 > C-8 > C-7.


Subject(s)
Lipase/metabolism , N-Acetylneuraminic Acid/metabolism , Acetylation , Catalysis , Chromatography, High Pressure Liquid , N-Acetylneuraminic Acid/analogs & derivatives , N-Acetylneuraminic Acid/chemical synthesis , Pseudomonas/enzymology , Time Factors , Vinyl Compounds/metabolism
14.
Science ; 275(5302): 945-8, 1997 Feb 14.
Article in English | MEDLINE | ID: mdl-9020070

ABSTRACT

For the past decade the immune system has been exploited as a rich source of de novo catalysts. Catalytic antibodies have been shown to have chemoselectivity, enantioselectivity, large rate accelerations, and even an ability to reroute chemical reactions. In many instances catalysts have been made for reactions for which there are no known natural or man-made enzymes. Yet, the full power of this combinatorial system can only be exploited if there was a system that allows for the direct selection of a particular function. A method that allows for the direct chemical selection for catalysis from antibody libraries was so devised, whereby the positive aspects of hybridoma technology were preserved and re-formatted in the filamentous phage system to allow direct selection of catalysis. This methodology is based on a purely chemical selection process, making it more general than biologically based selection systems because it is not limited to reaction products that perturb cellular machinery.


Subject(s)
Antibodies, Catalytic , Peptide Library , Animals , Antibodies, Catalytic/genetics , Antibodies, Catalytic/metabolism , Catalysis , Cloning, Molecular , Coliphages , Dithiothreitol , Enzyme-Linked Immunosorbent Assay , Escherichia coli/genetics , Escherichia coli/metabolism , Galactosides/metabolism , Haptens , Hybridomas , Immunoglobulin Fab Fragments/genetics , Immunoglobulin Fab Fragments/metabolism , Indoles/metabolism , Isopropyl Thiogalactoside/metabolism , Mice , Nitrophenylgalactosides/metabolism , Polymerase Chain Reaction , Serum Albumin, Bovine , Transformation, Bacterial , beta-Galactosidase/metabolism
16.
Biochemistry ; 34(31): 9893-6, 1995 Aug 08.
Article in English | MEDLINE | ID: mdl-7632688

ABSTRACT

The specific, high affinity binding of plant-derived digitalis glycosides by the mammalian sodium and potassium transporting adenosine triphosphatase (Na,K-ATPase, or sodium pump), a plasma membrane enzyme with critical physiological importance in mammalian tissues, has raised the possibility that a mammalian analog of digitalis might exist. We previously isolated and structurally characterized from bovine hypothalamus a novel isomer of the plant glycoside, ouabain, which differs structurally only in the attachment site and/or the stereochemistry of the steroid moiety [Tymiak et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 8189-8193]. Hamlyn and co-workers reported a molecule purified from human plasma which by mass spectrometry could not be distinguished from plant ouabain [Hamlyn et al. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 6259-6263]. Since rhamnoside cardiotonic steroids are not known as natural products from mammalian sources, it became important to compare these two pure isolates to determine if the same or structurally distinct compounds has been found. Our results indicate that the human and bovine Na,K-ATPase-inhibitors are identical, but different from plant ouabain. This supports the notion that the human sodium pump may be under specific physiological regulation by a mammalian analog of the digitalis glycosides.


Subject(s)
Enzyme Inhibitors/blood , Enzyme Inhibitors/chemistry , Hypothalamus/chemistry , Ouabain/analogs & derivatives , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitors , Animals , Cattle , Chromatography, High Pressure Liquid , Circular Dichroism , Humans
17.
J Neurosci ; 14(11 Pt 1): 6370-83, 1994 Nov.
Article in English | MEDLINE | ID: mdl-7965042

ABSTRACT

Recently, the Dlx family of homeobox genes have been identified as candidates for regulating patterning and differentiation of the forebrain. We have made a polyclonal antiserum to the protein product of the Dlx-2 gene. Using this antiserum, we have characterized the spatial and temporal pattern of DLX-2 protein expression during murine development and in the adult mouse brain. These studies demonstrate that, like the mRNA from the Dlx-2 gene, DLX-2 protein is expressed in mouse embryonic forebrain, limbs, tail, genital tubercle, and branchial arches. Within the embryonic forebrain, DLX-2 protein is expressed within specific transverse and longitudinal domains. Analysis of expression within the wall of the forebrain shows that DLX-2 is expressed in proliferative regions including the ventricular and subventricular zones. DLX-2 is expressed in the same cells as MASH-1, a marker of relatively undifferentiated cells, but in a reciprocal fashion to MAP-2, a marker of terminal neuronal differentiation. A number of DLX-2-expressing cells, but not all, can be labeled with bromodeoxyuridine (BrdU). Using the patterns of DLX-2, MASH-1, MAP-2 expression, and bromodeoxyuridine incorporation, we identify four molecularly distinct populations of cells that may correspond to different stages of neuronal differentiation in the mouse basal forebrain, in which DLX-2 is expressed at the transition from proliferation to terminal differentiation.


Subject(s)
Bromodeoxyuridine/metabolism , Embryo, Mammalian/physiology , Gene Expression , Genes, Homeobox , Pheromones , Prosencephalon/embryology , Animals , Basic Helix-Loop-Helix Transcription Factors , DNA-Binding Proteins/metabolism , Embryo, Mammalian/cytology , Fungal Proteins , Ganglia/embryology , Immunohistochemistry , Median Eminence/embryology , Mice/embryology , Mice/genetics , Mitosis , Protein Sorting Signals/metabolism , Transcription Factors/metabolism
18.
Acta Chem Scand (Cph) ; 48(3): 240-51, 1994 Mar.
Article in English | MEDLINE | ID: mdl-8155432

ABSTRACT

In screening for antifungal metabolites, a novel compound, malonofungin, exhibiting growth inhibitory activity against Botrytis cinerea (grey mould), has been isolated from fermentations of Phaeoramularia fusimaculans CBS 616.87. Its structure is established as (E)-(3R,4S,5S)-5-acetoxy-2-amino-2-carboxy-3,4-dihydroxy-14-oxoicos++ +-6-enoic acid, representing an addition to the rare class of naturally occurring aminomalonic acids. 1H NMR data and extensive use of CD spectroscopy have been utilized to establish the absolute stereochemistry of malonofungin. The structural and biological relationship of malonofungin to previously reported fungal metabolites is discussed.


Subject(s)
Antifungal Agents/pharmacology , Fatty Acids, Unsaturated/pharmacology , Malonates/pharmacology , Mitosporic Fungi/chemistry , Acetylation , Antifungal Agents/chemistry , Antifungal Agents/isolation & purification , Circular Dichroism , Fatty Acids, Unsaturated/chemistry , Fatty Acids, Unsaturated/isolation & purification , Magnetic Resonance Spectroscopy , Malonates/chemistry , Malonates/isolation & purification , Microbial Sensitivity Tests , Mitosporic Fungi/drug effects , Molecular Conformation , Stereoisomerism
19.
Cell ; 75(3): 463-76, 1993 Nov 05.
Article in English | MEDLINE | ID: mdl-8221886

ABSTRACT

The mouse Mash-1 gene, like its Drosophila homologs of the achaete-scute complex (AS-C), encodes a transcription factor expressed in neural precursors. We created a null allele of this gene by homologous recombination in embryonic stem cells. Mice homozygous for the mutation die at birth with apparent breathing and feeding defects. The brain and spinal cord of the mutants appear normal, but their olfactory epithelium and sympathetic, parasympathetic, and enteric ganglia are severely affected. In the olfactory epithelium, neuronal progenitors die at an early stage, whereas the nonneuronal supporting cells are present. In sympathetic ganglia, the mutation arrests the development of neuronal precursors, preventing the generation of sympathetic neurons, but does not affect glial precursor cells. These observations suggest that Mash-1, like its Drosophila homologs of the AS-C, controls a basic operation in development of neuronal progenitors in distinct neural lineages.


Subject(s)
Autonomic Nervous System/embryology , DNA-Binding Proteins/genetics , Olfactory Mucosa/embryology , Transcription Factors/genetics , Adrenal Medulla/innervation , Animals , Animals, Newborn , Autonomic Nervous System/abnormalities , Base Sequence , Basic Helix-Loop-Helix Transcription Factors , Chromaffin System/embryology , Epithelium/embryology , Ganglia/abnormalities , Ganglia/embryology , Genes, Lethal , Mice , Mice, Mutant Strains , Molecular Sequence Data , Mutagenesis , Neural Crest/embryology , Neuroglia , Neurons, Afferent , Olfactory Mucosa/abnormalities , Olfactory Mucosa/innervation , Phenotype , Stem Cells , Time Factors
20.
Proc Natl Acad Sci U S A ; 90(17): 8189-93, 1993 Sep 01.
Article in English | MEDLINE | ID: mdl-8396262

ABSTRACT

Recent reports have shown the presence of a ouabain-like inhibitor of Na+/K(+)-ATPase in humans. We have purified a bovine hypothalamic Na+/K(+)-ATPase inhibitory factor (HIF) by using affinity chromatography combined with HPLC. This inhibitor has a molecular weight of 584 as determined by ion-spray mass spectrometry, making it isobaric with ouabain. Glycosidase treatment or acid hydrolysis of HIF released only L-rhamnose, the hexose isomer found in ouabain, as detected by chiral GC/MS. Additionally, enzymatically generated desrhamnosyl HIF was found to have a molecular weight of 438, as does ouabagenin, the aglycone of ouabain. HIF and its aglycone were indistinguishable from ouabain and ouabagenin, respectively, by reversed-phase HPLC retention times. However, derivatization with naphthoylimidazole followed by HPLC revealed different retention times for naphthoylation products of HIF and ouabain. Subsequent CD spectroscopy on isolated naphthoylation products of HIF and ouabain confirmed that they were different. This study provides chromatographic and spectroscopic evidence that ouabain and HIF are isomeric cardenolides. The structural difference is presumed to account for the significant differences in biological properties observed for HIF and ouabain.


Subject(s)
Hypothalamus/chemistry , Ouabain/chemistry , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitors , Animals , Cattle , Chromatography, Affinity , Chromatography, High Pressure Liquid , Gas Chromatography-Mass Spectrometry , Glycoside Hydrolases , Hydrolysis , Isomerism , Mass Spectrometry , Molecular Structure , Multienzyme Complexes , Ouabain/isolation & purification , Spectrometry, Mass, Fast Atom Bombardment , beta-Glucosidase
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