Therapeutic potential of amniotic fluid stem cells to treat bilateral ovarian dystrophy in dairy cows in a subtropical region.

Amniotic fluid is a rich source of multipotent mesenchymal stem cells (MSCs). Amniotic fluid stem cells (AFSCs) have become a new source of stem cells; they have low immunogenicity and are easily harvested. For this reason, they may be useful in clinical tissue engineering. Moreover, AFSCs have anti-inflammatory properties and can repair tissues. This study evaluated the utility of AFSC injection to treat bilateral ovarian dystrophy in Holstein-Friesian cows. Bovine AFSCs (BAFSCs) were collected at slaughter from Holstein-Friesian cows during the third or fourth month of pregnancy and cultured in vitro. The BAFSCs began to show a fibroblast-like morphology. They were positive for ß-integrin, CD44, CD73, CD106 and Oct4 and negative for CD34 and CD45. After induction, the cells differentiated into mesodermal lineages. Bilateral ovarian dystrophy was confirmed by ultrasonography in 16 lactating cows. The subsequent experiment lasted 15 weeks. Serum was collected weekly to analyse progesterone concentrations, and weekly ultrasonography recorded ovarian changes. Each cow was equipped with an automatic heat detection system to facilitate oestrus observation and breeding records. The progesterone concentration of two cows in the treatment group (25%) significantly increased during weeks 10-15. On ultrasonography, the treatment group demonstrated mature follicles after BAFSCs injection, and foetuses were visualized approximately 40 days after artificial insemination (AI). Oestrus rates in the control and treatment groups were 0% (0/8) and 50% (4/8), respectively; pregnancy rates were 0% (0/8) and 25% (2/8), respectively. Calves were successfully delivered in both cases of pregnancy. These results show that BAFSCs can alleviate bovine ovarian dystrophy and restore fertility.

Acute toxicity and n-octanol/water partition coefficients of substituted thiophenols: determination and QSAR analysis.

The acute toxicity (-log EC(50)) to Photobacterium phosphoreum and the n-octanol/water partition coefficient (log K(ow)) of 31 kinds of substituted thiophenols were determined at 298.15K. The -log EC(50) values of studied chemicals are between 4.26 and 5.89. Their log K(ow) values are between 1.34 and 4.02. Comparative molecular field (CoMFA) and comparative molecular similarity index analysis (CoMSIA) models established were successful in predicting -log EC(50) and log K(ow) values of halogenated, methylic, amino and methoxy thiophenols. The size of molecule is the main factor influencing the properties. No correlation was found between the properties and their structural and thermodynamic descriptors from DFT calculation.

A saxitoxin-binding aptamer with higher affinity and inhibitory activity optimized by rational site-directed mutagenesis and truncation.

Saxitoxin (STX), a member of the family of paralytic shellfish poisoning toxins, poses toxicological and ecotoxicological risks. To develop an analytical recognition element for STX, a DNA aptamer (APT(STX1)) was previously discovered via an iterative process known as Systematic Evolution of Ligands by Exponential Enrichment (SELEX) by Handy et al. Our study focused on generating an improved aptamer based on APT(STX1) through rational site-directed mutation and truncation. In this study, we generated the aptamer, M-30f, with a 30-fold higher affinity for STX compared with APT(STX1). The Kd value for M-30f was 133 nM, which was calculated by Bio-Layer Interferometry. After optimization, we detected and compared the interaction of STX with aptamers (APT(STX1) or M-30f) through several techniques (ELISA, cell bioassay, and mouse bioassay). Both aptamers' STX-binding ability was demonstrated in all three methods. Moreover, M-30f performs better than its parent sequence with higher suppressive activity against STX. As a molecular recognition element, M-30f has good prospects for practical application.

Tissue expression and predicted protein structures of the bovine ANGPTL3 and association of novel SNPs with growth and meat quality traits.

Angiopoietin-like protein 3 (ANGPTL3) is a secreted protein that regulates lipid, glucose and energy metabolism. This study was conducted to better understand the effect of ANGPTL3 on important economic traits in cattle. First, transcript profiles for ANGPTL3 were measured in nine different Jiaxian cattle tissues. Second, polymorphisms were identified in the complete coding region and promoter region of the bovine ANGPTL3 gene in 707 cattle samples. Finally, an association study was carried out utilizing these single nucleotide polymorphisms (SNPs) to determine the effect of these SNPs on the growth and meat quality traits. Quantitative real-time PCR analysis showed that ANGPTL3 was mainly expressed in the liver. The promoter of the bovine ANGPTL3 contained several putative transcription factor binding sites (SF1, HNF-1, LXRα, NFκß, HNF-3 and C/EBP). In total, four SNPs of the bovine ANGPTL3 gene were identified by direct sequencing. SNP1 (rs469906272: g.-38T>C) was identified in the promoter, SNP2 (rs451104723:g.104A>T) and SNP3 (rs482516226: g.509A>G) were identified in exon 1, and SNP4 (rs477165942: g.8661T>C) was identified in exon 6. Changes in predicted protein structures due to non-synonymous SNPs were analyzed. Haplotype frequencies and linkage disequilibrium were also investigated. Analysis of four SNPs in cattle from different native Chinese breeds (Nanyang (NY) and Jiaxian (JX)) and commercial breeds (Angus (AG), Hereford (HF), Limousin (LM), Luxi (LX), Simmental (ST) and Jinnan (JN)) revealed a significant association with growth traits (including: BW and hipbone width) and meat quality traits (including: Warner-Bratzler shear force and ribeye area). Therefore, implementation of these four mutations in selection indices in the beef industry may be beneficial in selecting individuals with superior growth and meat quality traits.

Radon measurements in underground dwellings from two prefectures in China.

Radon, an established lung carcinogen, remains the single most important environmental radiation exposure. Yet, an excess of lung cancer from breathing radon in homes has not been consistently demonstrated in studies conducted to date. To address several major problems that have hindered previous studies of lung cancer and radon in homes, we have embarked upon a lung cancer case-control study in Gansu Province, China, where a substantial proportion of the population live in underground dwellings. In this paper, we report on results of a pilot study in which radon measurements were made for 3 days in the summer in 40 homes under normal occupancy conditions using short-term E-PERM detectors and for 6 months from February through August in 49 homes using long-term alpha-track detectors. Useable E-PERM data were obtained from 38 homes and useable alpha-track data from 47 homes. For both types of detectors, measurements were approximately log-normally distributed. Arithmetic and geometric means were 233 and 185 Bq m-3 (range 74-1,590 Bq m-3) for E-PERM measurements and 165 and 158 Bq m-3 (range 74-592 Bq m-3) for alpha-track measurements, respectively; 68% of E-PERM measured homes and 55% of alpha-track measured homes exceeded 148 Bq m-3. Alpha-track measurements made at the entry, middle, and rear areas of the underground dwellings did not differ significantly (arithmetic means of 168, 162, and 165 Bq m-3 with standard deviations 63, 73, and 48, respectively), which suggests that air circulation may be minimal. The underground dwellings measured in the pilot study had high radon levels and the underground dwellers may provide an excellent population for studying indoor radon and risk of lung cancer.

Biochemical and enzymological study of lactate dehydrogenase isoenzymes from commercial quality control sera and several animal tissue sources.

We assayed the isoenzymes of lactate dehydrogenase (EC 1.1.1.27) in commercial quality control sea and several animal tissue extracts, using electrophoresis. We compared the Km values and activation energies of the isoenzymes, in order to find suitable animal tissue sources with a similar isoenzyme profile to that of human serum lactate dehydrogenase. Some of the control sera contained only one isoenzyme fraction corresponding to porcine heart isoenzyme-1 or chicken heart isoenzyme-1, which showed essentially no changes of enzyme activity as a function of pyruvate substrate concentration. Other control sera, which contained isoenzymes from human red cells haemolysates, or from animal tissue extracts with a human serum matrix, showed significant changes of enzyme activity was a function of substrate concentration, and showed different Km values and activation energies from those of human serum. Of the serum and tissue samples from several animal sources, rat heart and kidney extracts showed the greatest similarity to human serum, with respect to the electrophoretic pattern and the Km, pH optimum and activation energy of lactate dehydrogenase isoenzymes.