Simultaneous Quantitation of Free Amino Acids, Nucleosides and Nucleobases in Sipunculus nudus by Ultra-High Performance Liquid Chromatography with Triple Quadrupole Mass Spectrometry

To evaluate the nutritional and functional value of Sipunculus nudus, a rapid, simple and sensitive analytical method was developed using ultra-high performance liquid chromatography coupled with a triple quadrupole mass detection in multiple-reaction monitoring mode for the simultaneous quantitative determination of 25 free amino acids and 16 nucleosides and nucleobases in S. nudus within 20 min, which was confirmed to be reproducible and accurate. The limits of detection (LODs) and quantification (LOQs) were between 0.003–0.229 μg/mL and 0.008–0.763 μg/mL for the 41 analytes, respectively. The established method was applied to analyze 19 batches of S. nudus samples from four habitats with two different processing methods. The results showed that S. nudus contained a variety of free amino acids, nucleosides and nucleobases in sufficient quantity and reasonable proportion. They also demonstrated that the contents of these compounds in different parts of S. nudus were significantly discriminating, which were in the order: (highest) coelomic fluid > body wall > intestine (lowest). The method is simple and accurate, and could serve as a technical support for establishing quality control of S. nudus and other functional seafoods. Moreover, the research results also laid foundation for further exploitation and development of S. nudus

Identification of Novel Genomic Aberrations in AML-M5 in a Level of Array CGH

<div><p>To assess the possible existence of unbalanced chromosomal abnormalities and delineate the characterization of copy number alterations (CNAs) of acute myeloid leukemia-M5 (AML-M5), R-banding karyotype, oligonucelotide array CGH and FISH were performed in 24 patients with AML-M5. A total of 117 CNAs with size ranging from 0.004 to 146.263 Mb was recognized in 12 of 24 cases, involving all chromosomes other than chromosome 1, 4, X and Y. Cryptic CNAs with size less than 5 Mb accounted for 59.8% of all the CNAs. 12 recurrent chromosomal alterations were mapped. Seven out of them were described in the previous AML studies and five were new candidate AML-M5 associated CNAs, including gains of 3q26.2-qter and 13q31.3 as well as losses of 2q24.2, 8p12 and 14q32. Amplication of 3q26.2-qter was the sole large recurrent chromosomal anomaly and the pathogenic mechanism in AML-M5 was possibly different from the classical recurrent 3q21q26 abnormality in AML. As a tumor suppressor gene, <i>FOXN3</i>, was singled out from the small recurrent CNA of 14q32, however, it is proved that deletion of <i>FOXN3</i> is a common marker of myeloid leukemia rather than a specific marker for AML-M5 subtype. Moreover, the concurrent amplication of <i>MLL</i> and deletion of <i>CDKN2A</i> were noted and it might be associated with AML-M5. The number of CNA did not show a significant association with clinico-biological parameters and CR number of the 22 patients received chemotherapy. This study provided the evidence that array CGH served as a complementary platform for routine cytogenetic analysis to identify those cryptic alterations in the patients with AML-M5. As a subtype of AML, AML-M5 carries both common recurrent CNAs and unique CNAs, which may harbor novel oncogenes or tumor suppressor genes. Clarifying the role of these genes will contribute to the understanding of leukemogenic network of AML-M5.</p></div

Comparative clinico-biological parameters and number of CR by number of CNAs.

<p>Comparative clinico-biological parameters and number of CR by number of CNAs.</p

<i>FOXN3</i> gene (14q31.3-q32.11) deletions in two patients with AML-M5.

<p>(A) Array CGH showed a lower signal ratio spanning 14q22.3-q32.33 (55,109,046–106,342,076 bp). <i>FOXN3</i> gene was one of the genes located at the deleted region. (B) The microdeletion of 14q32.11 (89,027,534–89,236,808 bp) involving <i>FOXN3</i> gene was inferred by array CGH.</p

The recurrent chromosomal abnormalities of 24 cases of AML-M5.

<p>The recurrent chromosomal abnormalities of 24 cases of AML-M5.</p

Candidate AML-M5 associated genes inferred by array CGH.

<p>Candidate AML-M5 associated genes inferred by array CGH.</p

Distribution of CNAs recognized by array CGH in 24 patients with AML-M5.

<p>A total of 117 CNAs with 51 genomic gains (43.1%) and 66 genomic losses (56.9%) was observed. 70 CNAs (59.8%) were cryptic with size less than 5 Mb and invisible to karyotype. The arrows indicated recurrent CNAs and harbored interesting genes.</p

Comparative clinico-biological parameters and number of CR by number of CNAs.

<p>Comparative clinico-biological parameters and number of CR by number of CNAs.</p

Recurrent CNAs reported by previous studies of MDS/AML and this study.

<p>Recurrent CNAs reported by previous studies of MDS/AML and this study.</p