ABSTRACT
The turbot is a flatfish species of great relevance to marine aquaculture in Europe. Only a limited number of microsatellites have been isolated to date in this species. To increase the number of potentially useful mapping markers, we screened simple sequence repeat (SSR)--enriched genomic libraries obtained from several di-, tri-, and tetranucleotide tandem repeat motifs. A total of 248 new polymorphic microsatellites were successfully optimized. The efficiency of the protocol applied (6.4%) was higher than that in other studies of fish that used the same method. Dinucleotide and perfect microsatellites were predominant in this species; the (AC)n motif was the most frequent class of repeat. Polymorphism and structural properties at these loci, together with 30 variable loci previously reported in turbot, were evaluated in 6 wild individuals. The number of alleles per locus ranged from 2 to 10, with an average of 4.046. The microsatellite markers characterized in this study will contribute to the development of the turbot genetic map, which can be used for quantitative trait locus (QTL) identification, marker-assisted selection programs, and other applications to improve its culture.
Subject(s)
Flatfishes/genetics , Microsatellite Repeats , Animals , Chromosome Mapping , Dinucleotide Repeats , Europe , Genomic Library , Polymorphism, Genetic , Quantitative Trait Loci , SoftwareABSTRACT
Reaction of the title ligands (HPyTSC and HS(S)PPh2, respectively) with R2SnO (R = Me, Et, Bu) in ethanol (EtOH) afforded the complexes [SnMe2(PyTSC) (S2PPh2)].EtOH (1) and [SnR2(PyTSC) (S2PPh2)] (R = Et (2), Bu (3)). The structures of 1 and 2 were determined by single-crystal X-ray diffractometry. In both these complexes the tin atom is coordinated to an N,N,S-dentate thiosemicarbazonate ligand, an anisobidentate dithiophosphinato ligand and the two R groups. The coordination polyhedrons can be described as distorted pentagonal bipyramids. A comparative study of the IR spectra of 1, 2 and 3 indicates that the butyl complex has a similar structure. Multinuclear (1H, 13C, 31P and 119Sn) NMR data suggest that the structures of 1 and 2 probably remain in CDCl3 (or DMSO-d6) solution but compound 3 partially decomposes in these media. Preliminary results on the effects of the complexes on the proliferation and differentiation of FLC, CEM, U937, K562 and TOM-1 leukaemia cells, and on the clonogenic activity of K562 cells are also described.
Subject(s)
Organotin Compounds/chemistry , Organotin Compounds/pharmacology , Cell Differentiation/drug effects , Cell Division/drug effects , Crystallography, X-Ray , Humans , K562 Cells , Ligands , Magnetic Resonance Spectroscopy , Molecular Structure , Organotin Compounds/chemical synthesis , Spectrophotometry, InfraredABSTRACT
The complexes [SnR2(L)] (R = Me, Et, Bu, Ph; H2L = pyridoxal thiosemicarbazone) have been prepared and characterized. In the light of the spectral properties of the complexes in the solid state (IR, mass, Mössbauer) the bideprotonated thiosemicarbazonato anion is O(phenolic)-, N(3)-, S-bonded to the tin atom which probably has trigonal bipyramidal coordination with N(3) atom and R groups occupying equatorial positions. NMR ( 1H, 13C and 119Sn) data in CDCl3 or DMSO-d6 suggest that this coordinative picture remains in these solutions. The ethyl, butyl and phenyl derivatives suppress proliferation of Friend erithroleukaemia cells (FLC). Of the pyridoxal thiosemicarbazone complexes so far evaluated. [SnBu2(L)] and [SnPh2(L)] showed the lowest thresholds for inhibition of FLC proliferation. The effects of these compounds on DMSO-induced differentiation of FLC, DNA synthesis and reverse transcriptase were also assayed.
Subject(s)
Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/pharmacology , Organotin Compounds/chemical synthesis , Organotin Compounds/pharmacology , Pyridoxal/analogs & derivatives , Thiosemicarbazones/chemical synthesis , Thiosemicarbazones/pharmacology , Animals , Antineoplastic Agents/chemistry , Cell Differentiation/drug effects , Cell Division/drug effects , DNA, Neoplasm/biosynthesis , Dimethyl Sulfoxide/pharmacology , Friend murine leukemia virus , Leukemia, Erythroblastic, Acute/drug therapy , Leukemia, Erythroblastic, Acute/metabolism , Leukemia, Erythroblastic, Acute/pathology , Magnetic Resonance Spectroscopy , Mass Spectrometry , Mice , Organotin Compounds/chemistry , Pyridoxal/chemical synthesis , Pyridoxal/chemistry , Pyridoxal/pharmacology , Reverse Transcriptase Inhibitors/chemical synthesis , Reverse Transcriptase Inhibitors/chemistry , Reverse Transcriptase Inhibitors/pharmacology , Spectroscopy, Mossbauer , Thiosemicarbazones/chemistry , Tumor Cells, CulturedABSTRACT
The synthesis, X-ray structure, behavior in solution, and biological properties of the complex [SnMe2(PyTSC)(OAc)].HOAc (HPyTSC = pyridine-2-carbaldehydethiosemicarbazone) are reported. The tin atom of this complex is coordinated to an N,N,S-tridentate PyTSC- anion, to a monodentate acetate ion, and to the two methyl groups in an approximately pentagonal bipyramidal environment with a vacant equatorial position. The complex partially evolves in DMSO and in DMSO/CHxCl4-x (X = 1, 2) mixtures, giving HPyTSC and SnMe2(OAc)2. [SnMe2 (PyTSC)(OAc)].HOAc, [SnMe2(DAPTSC)], and [SnPh2(DAPTSC)].2DMF (H2DAPTSC = 2,6-diacetylpyridine bis(thiosemicarbazone)) all suppress proliferation of Friend erythroleukaemia cells (FLC). DMSO-induced differentiation of FLC is slightly suppressed by [SnMe2(DAPTSC)] and is unaffected by [SnPh2(DAPTSC)].2DMF and [SnMe2(PyTSC)(OAc)].HOAc.
Subject(s)
Antineoplastic Agents/chemical synthesis , Organotin Compounds/chemical synthesis , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Cell Differentiation/drug effects , Cell Division/drug effects , Crystallography, X-Ray , Friend murine leukemia virus , Leukemia, Erythroblastic, Acute/drug therapy , Mice , Models, Molecular , Molecular Structure , Organotin Compounds/chemistry , Organotin Compounds/pharmacology , Solutions , Spectrophotometry, Infrared , Tumor Cells, CulturedABSTRACT
La combinación de métodos inmunológicos y ultraestructurales constituye un elemento fundamental para la identificación de las características del reconocimiento celular de los anticuerpos monoclonales. En el presente artículo se reporta la aplicación de la técnica de inmunoperoxidasa para microscopia electrónica, en la dilucidación del reconocimiento celular y la ubicación de las estructuras antigénicas identificadas por los anticuerpos monoclonales IOR-T1 e IOR-T2, utilizando como sustrato antigénico células mononucleares de sangre periférica de individuos normales y de pacientes con linfomas T cutáneos. Se determinó que el IOR-T1 reconoce un marcador de superficie celular característicos de los linfocito humanos T normales, que puede ser expresado también por células tumorales del mismo origen. El IOR-T2 identifica exclusivamente una parte de la población tumoral celular con características morfológicas compatibles con las células de Sézary, provenientes de la sangre periférica de dos pacientes con linfomas T cutáneos
