Cultured human umbilical cord mesenchymal stem cells (hUC-MSCs) are being tested in several clinical trials and encouraging outcomes have been observed. To determine whether in vitro expansion influences the genomic stability of hUC-MSCs, we maintained nine hUC-MSC clones in long-term culture and comparatively analyzed them at early and late passages. All of the clones senesced in culture, exhibiting decreased telomerase activity and shortened telomeres. Two clones showed no DNA copy number variations (CNVs) at passage 30 (P30). Seven clones had ≥1 CNVs at P30 compared with P3, and one of these clones appeared trisomic chromosome 10 at the late passage. No tumor developed in immunodeficient mice injected with hUC-MSCs, regardless of whether the cells had CNVs at the late passage. mRNA-Seq analysis indicated that pathways of cell cycle control and DNA damage response were downregulated during in vitro culture in hUC-MSC clones that showed genomic instability, but the same pathways were upregulated in the clones with good genomic stability. These results demonstrated that hUC-MSCs can be cultured for many passages and attain a large number of cells, but most of the cultured hUC-MSCs develop genomic alterations. Although hUC-MSCs with genomic alterations do not undergo malignant transformation, periodic genomic monitoring and donor management focusing on genomic stability are recommended before these cells are used for clinical applications.

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We compared meiotic pairing and recombination between tomato ( Lycopersicon esculentum) and homoeologous Solanum lycopersicoides chromosomes in monosomic additions (MAs) and substitution lines (SLs), each representing a single chromosome of the nightshade in a tomato background. Three configurations of each alien chromosome and its two tomato homoeologues were detected by genomic in situ hybridization in MA-7, -8, and -10 at diakinesis/metaphase-I: 1 trivalent (III), 1 bivalent + 1 univalent (II+I), and 3 univalents (3I). The II+I category was by far the most common, and the univalent was from S. lycopersicoides 91-99.5% of the time, indicating a high degree of preferential (homologous) pairing. In the corresponding substitution lines, association of homoeologous chromosomes was much higher (up to 90% of the cells), presumably due to the absence of homologous partners. However, SL-10 showed a surprisingly high frequency of univalents (about 73%). Genome-wide analysis of chromosome pairing revealed a decrease in the average chiasma frequency for both monosomic additions and substitution lines. Recombination between tomato and the nightshade was restricted in all cases, the reduction being more severe in each monosomic addition than in the corresponding substitution line. Recombination rates in the substitutions were less than those observed for the same chromosomes in the first backcross generation. Chromosomes 8 and 10 showed the highest and the lowest rates of homoeologous recombination, respectively. No recombination was detected between markers on the long arm of chromosome 10, presumably due to the presence of a paracentric inversion differentiating the two genomes in this region. The frequency of homoeologous pairing at diakinesis/metaphase-I was significantly higher than the rate of homoeologous recombination detected in the progeny, suggesting a strong selection against recombinant products in meiotic or post-meiotic stages.

ABSTRACT. We hypothesized that single nucleotide polymorphisms (SNPs) in certain microRNAs contribute to congenital heart disease (CHD) phenotypes. Five hundred and seventy-three subjects were enrolled in this study. DNA extracted from peripheral blood cells was used for SNP genotyping of miR-196a2 (rs11614913), miR27a (rs11671784, rs895819), and miR-499 (rs3746444). Allele and genotype association analyses were performed to evaluate the

ABSTRACT. The study aimed to investigate the effect of intrathecal injections of Tanshinone IIA on thermal hyperalgesia in a mouse model of bone cancer-pain. Spinal IL-1β, IL-6, TNF-α expression levels were analyzed. C3H/HeNCrlVr male mice were assigned to groups that either received dose-dependent injections of Tanshinone IIA, or the DMSO + Sham, Tanshinone IIA + Sham, DMSO + Tumor, and Control groups. Paw withdrawal thermal latency (PWTL) was measured with a radiant heat stimulus and mRNA expression levels were determined using real-time PCR. Fourteen days post-injection, PWTL in the DMSO + Tumor group was lower than that in the controls (P < 0.05). Twentyone days post-injection, compared with the Control group, there was no significant difference in PWTL and IL-1β, IL-6, and TNF-α expression levels between the Tanshinone IIA + Sham and DMSO + Sham groups (P > 0.05). PWTL in the DMSO + Tumor group was significantly lower than the Control group (P < 0.05), while the expression levels of IL-1β, IL-6, and TNF-α were significantly higher than controls. Compared with the DMSO + Tumor group, PWTLs were higher in the Tanshinone B.X. Ren et al. 2134©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (1): 2133-2138 (2015) IIA -20-μg and 40-μg groups, while expression levels of IL-1β, IL-6, and TNF-α were significantly lower (P < 0.05). These measures were not significantly different between the Tanshinone IIA 10 μg and the DMSO + Tumor groups (P > 0.05). In conclusion, Tanshinone IIA may inhibit the release of inflammatory cytokines, such as, IL-1 β, IL-6 α, TNF-α.

word summaryFibroblast Growth Factor 21 (FGF21) is a hormone that induces weight loss in model organisms. These findings have led to trials in humans of FGF21 analogues with some showing weight loss and lipid lowering effects. Recent genetic studies have shown that a common allele in the FGF21 gene alters the balance of macronutrients consumed but there was little evidence of an effect on metabolic traits. We studied a common FGF21 allele (A:rs838133) in 451,099 people from the UK Biobank study. We replicated the association between the A allele and higher percentage carbohydrate intake. We then showed that this allele is more strongly associated with body fat distribution, with less fat in the lower body, and higher blood pressure, than it is with BMI, where there is only nominal evidence of an effect. These human phenotypes of naturally occurring variation in the FGF21 gene will inform decisions about FGF21's therapeutic potential. IntroductionFGF21 is a hormone secreted primarily by the liver whose multiple functions include signalling to the paraventricular nucleus of the hypothalamus to suppress sugar and alcohol intake [1,2], stimulating insulin-independent glucose uptake by adipocytes [3] and acting as an insulin sensitizer [4]. These features and several other lines of evidence have prompted the development of FGF21 based therapies as potential treatments for obesity and type 2 diabetes, with consistent effects on triglyceride lowering, some effects on weight loss but little effect on glucose tolerance [5,6]. An early trial showed lipid lowering effects in people with type 2 diabetes and obesity but there was only suggestive evidence for effects on weight and glucose tolerance [7]. A recent study suggested that FGF21 analogues may alter not peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission.The copyright holder for this preprint (which was . http://dx.doi.org/10.1101/214700 doi: bioRxiv preprint first posted online Nov. 6, 2017; blood pressure in humans [8], although changes in blood pressure were not observed in a previous trial [9]. Pre-clinical evidence of FGF21's potential role in metabolism includes resistance to diet induced obesity in mice overexpressing FGF21 [3] and improved glucose tolerance in obese mice through administration of recombinant FGF21 [3]. Subsequent studies have confirmed these findings in mice [10] and shown similar effects in non human primates, including improvement of glucose tolerance and slight weight loss in diabetic rhesus monkeys [11], but other studies are less conclusive [5].Recent studies have shown that FGF21 affects the balance of macronutrients consumed. Studies in mice and non human primates show that genetically and pharmacologically raising FGF21 levels suppresses sugar and alcohol intake [1,2]. Three human genetic studies have shown that the minor A allele at rs838133 (A/G, Minor Allele Frequency=44.7%), which results in a synonymous change to the first exon of FGF21, is associated with higher carbohydrate and lower...

Chiasmata constitute one of the cornerstones of sexual reproduction in most eukaryotes. They mediate the reciprocal genetic exchange between homologues and are essential to the proper orientation of the homologous centromeres in meiosis I. As markers of recombination, they offer a cytological means of mapping. Rather than trying to accurately count individual chiasmata, we have examined properties of the mathematical relationship between frequencies of nonadorned disomic configurations in meiosis (ring, rods, and univalents) and the probabilities at which arms of the respective chromosomes are chiasmate (one or more chiasma per arm). Numerical analyses indicated that conventionally analyzed bivalents with nonidentified arms yield statistically biased estimates of chiasma probabilities under a broad range of circumstances. We subsequently analyzed estimators derived from adorned configurations with ISH-marked arms, which were found to be statistically far superior, and with no assumptions concerning interference across the centromere. We applied this methodology in the study of chromosomes 16 and 23 of cotton (Gossypium hirsutum), and estimated their arm lengths in centimorgans. The results for chromosome 23, the only one of the two chromosomes with a documented RFLP map, were consistent with the literature. Similar molecular-meiotic configuration analyses can be used for a wide variety of eukaryotic organisms and purposes: for example, providing far more powerful meiotic comparisons of genomes of chromosomes, and a rapid means of evaluating effects on recombination. Key words : meiotic configurations, chiasma frequencies, in situ hybridization, cotton.

Tumor cell invasion surrounding intracerebrally implanted tumors in rats was studied by comparing the results of cerebral microangiography, fluorescence imaging of blood-brain barrier (BBB) disruption and histopathology. Each comparison was on subsequent sections taken from an initial 1 mm coronal slice of brain taken through the cell injection site containing tumor using the RT-2 glioma model. Tumor extension was assessed at 3, 5, 7 and 9 days after tumor implantation. Analysis of the brain adjacent to tumor shows that the actual tumor cell invasion area is greater than the area of BBB disruption at later stages of tumor growth and the extent of tumor vascularization lies well within the area defined by the extent of tumor cell invasion. Furthermore, this study found that the size of the area of tumor cell invasion remains relatively stable in proportion to the solid tumor mass at various stages of growth such that the area of tumor invaded brain was approximately 2.5 times greater than the area outlined by solid tumor mass. We conclude that measurement of the solid tumor mass, tumor vascular area and region of blood-brain barrier disruption due to tumor, grossly underestimate the total tumor volume. Therapies aimed at controlling glial tumor growth must, therefore, include normal appearing regions of brain peripheral to the abnormal region defined as tumor and tumor invaded brain as defined by radiographic and imaging studies.