The effects of Mn²⁺-, Zn²⁺- or Cu²⁺-nanosuccinate added to freezing extender on select post-thaw semen characteristics were determined in six Texel rams (aged 2-4 years) during seasonal anestrus (April-May). Ejaculates (n = 6 per ram) collected into an artificial vagina were divided into ten isovolumetric fractions each. Semen was diluted in lactose-yolk-tris-citrate-glycerin medium and nanosuccinates (Mn²⁺- and Zn²⁺-nanosuccinate: 0.0 (control), 2.5, 5.0 and 7.5 μg/l; Cu²⁺-nanosuccinate: 0.0 (control), 1.25, 2.5 and 3.75 μg/l) were added to semen extender. Extended semen was loaded into 0.25-ml straws and frozen in liquid nitrogen. After thawing, sperm motility parameters were determined with computer assisted semen analysis (CASA), and the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) was measured with a spectrophotometric technique. The addition of 5.0 μg/l of Mn²⁺- and Zn²⁺-nanosuccinate significantly increased the sperm progressive motility and both 2.5 and 5.0 μg/l improved sperm motion kinetics. Further, both nanosuccinates at a dose of 5.0 μg/l significantly decreased SOD activity and stimulated an increase in GPx and CAT activity in semen samples. Alternatively, the addition of Cu²⁺-nanosuccinate (highest dose) significantly reduced the progressive motility and velocity of ram spermatozoa, increased the percentage of sperm with acrosomal/head defects and seminal SOD activity, and depressed CAT (highest dose) and GPx (all doses) activity. In summary, the addition of Mn²⁺- and Zn²⁺-nanosuccinate to semen extender had beneficial effects on sperm motility/motion kinetics and structural integrity, whereas Cu²⁺-nanosuccinate generally had debilitating effects on the post-thaw semen characteristics in rams.

2023-11-02·Neuro-oncology

Ex vivo to in vivo model of malignant peripheral nerve sheath tumors for precision oncology

Article

作者: He, Kevin ; Conniff, Eric ; Hirbe, Angela C ; Jubenville, Tyler ; Largaespada, David A ; Larsson, Alex T ; Osum, Sara H ; Gosline, Sara J C ; Williams, Kyle B ; Rono, Elizabeth ; Yang, Kuangying ; Tibbitts, Justin F ; Pratilas, Christine A ; Bade, Jessica ; Lyu, Yang ; Pollard, Kai ; Pino, James C ; Chrisinger, John S A ; Wood, David K ; Cummins, Katherine ; Dehner, Carina A ; Calizo, Ana ; Bhatia, Himanshi ; Lisok, Alla ; Crampton, Alexandra L ; Zhang, Xiaochun ; Schefer, Daniel ; Gross, John M

Abstract:

Background:

Malignant peripheral nerve sheath tumors (MPNST) are aggressive soft tissue sarcomas that often develop in patients with neurofibromatosis type 1 (NF1). To address the critical need for novel therapeutics in MPNST, we aimed to establish an ex vivo 3D platform that accurately captured the genomic diversity of MPNST and could be utilized in a medium-throughput manner for drug screening studies to be validated in vivo using patient-derived xenografts (PDX).

Methods:

Genomic analysis was performed on all PDX-tumor pairs. Selected PDX were harvested for assembly into 3D microtissues. Based on prior work in our labs, we evaluated drugs (trabectedin, olaparib, and mirdametinib) ex vivo and in vivo. For 3D microtissue studies, cell viability was the endpoint as assessed by Zeiss Axio Observer. For PDX drug studies, tumor volume was measured twice weekly. Bulk RNA sequencing was performed to identify pathways enriched in cells.

Results:

We developed 13 NF1-associated MPNST-PDX and identified mutations or structural abnormalities in NF1 (100%), SUZ12 (85%), EED (15%), TP53 (15%), CDKN2A (85%), and chromosome 8 gain (77%). We successfully assembled PDX into 3D microtissues, categorized as robust (>90% viability at 48 h), good (>50%), or unusable (<50%). We evaluated drug response to “robust” or “good” microtissues, namely MN-2, JH-2-002, JH-2-079-c, and WU-225. Drug response ex vivo predicted drug response in vivo, and enhanced drug effects were observed in select models.

Conclusions:

These data support the successful establishment of a novel 3D platform for drug discovery and MPNST biology exploration in a system representative of the human condition.

2019-05-01·Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry

Structural analysis and biological functionalities of iron(III)– and manganese(III)–thiosemicarbazone complexes: in vitro anti-proliferative activity on human cancer cells, DNA binding and cleavage studies

Article

作者: Şahin, Onur ; Aslim, Belma ; Yılmaz, Zehra Kübra ; Tükenmez, Ümmügülsüm ; Ülküseven, Bahri ; Kaya, Büşra

One iron(III) and two manganese(III) complexes based on thiosemicarbazone were synthesized and characterized using analytical and spectroscopic data. The crystallographic analysis showed the square pyramid structures of the complexes. Electronic spectra analysis was performed to determine the nature of the interaction between the complexes and calf thymus DNA (CT-DNA). DNA cleavage activities of the complexes were examined by gel electrophoresis (pBR322 DNA). The cytotoxicity of the complexes was determined against human cervical carcinoma (HeLa) and human colorectal adenocarcinoma (HT-29) cell lines by MTT assay. The results indicated that complex Fe1 is bound to CT-DNA via the intercalation mode, while complexes Mn1 and Mn2 are bound to CT-DNA via groove binding and/or electrostatic interactions rather than the intercalation mode. In addition, they showed good binding activity, which followed the order of Fe1 > Mn2 > Mn1. Complexes were found to promote the cleavage of DNA from supercoiled form (SC, Form I) to nicked circular form (NC, Form II) without concurrent formation of Form III, revealing the single-strand DNA cleavage. No significant cleavage was found in the presence of Mn1 and Mn2; however, it was observed at 2000 and 3000 µM concentrations of Fe1. The ability of Fe1 to cleave DNA was greater than that of other complexes and these results are in conformity with their DNA-binding affinities. Cytotoxicity determination tests revealed that the complex Fe1 on HeLa and HT-29 cells exhibited a higher anti-proliferative effect than Mn1 and Mn2 (Fe1 > Mn2 > Mn1). These studies suggested that the complex Fe1 could be a good candidate as a chemotherapeutic drug targeting DNA.

100 项与 MN-2 (MISSION) 相关的药物交易

登录后查看更多信息