Effects of Seed Size and Frequency on Seed Dispersal and Predation by Small Mammals.

Frequency-dependent predation is common in predator-prey interactions. Size is an important characteristic of seeds and is crucial in the regeneration stage of plant seeds. However, the frequency dependence of animal predation on seed size has not been reported. In this study, we conducted a field experiment and used different sizes of Liaodong oak (Quercus wutaishanica) seeds to test the frequency dependence of intraspecific seed size selection in rodents. We used the number ratio of large to small seeds as the frequency. The results show that the rate of small seeds being eaten in situ was significantly higher than that of large seeds (p < 0.05). The rates of different-sized seeds being eaten after removal decreased with increasing frequencies, and there was no significant difference between frequencies except for 1:9 and 9:1. The rates of large seeds being scatter-hoarded were significantly higher than those of small seeds at different frequencies (p < 0.05). The eating distances after removal of large seeds were significantly longer than those of small seeds at the same frequencies (p < 0.05). Furthermore, the scatter-hoarding distances of large seeds were significantly longer than those of small seeds at three frequencies (1:9, 3:7, and 9:1) (p < 0.05). That is, rodents consumed more small seeds in situ, dispersed and scatter-hoarded more large seeds, and dispersed large seeds over longer distances. Rodents exhibited a negative frequency dependence for small seeds and a positive frequency dependence for large seeds on being eaten in situ. Moreover, rodents exhibited a negative frequency dependence for large seeds and a positive frequency dependence for small seeds on being eaten after removal and scatter-hoarding. These results reveal the frequency dependence of rodent selection on seed size and provide new insights into animal-mediated seed dispersal and the regeneration of plant populations.

Effects of Seed Size and Cache Density on the Seed Fate of Quercus wutaishanica Mediated by Rodents.

Animal-mediated seed dispersal is very important for plant population regeneration and the stability of forest ecosystems. Seed size and cache density are important factors for seed dispersal, but we still know little about seed size selection at different cache densities. Here, we conducted field experiments in a Larix principis-rupprechtii plantation in the Liupan Mountains in Ningxia province to investigate the effects of tag-marked Quercus wutaishanica seeds of different sizes and cache densities on predation and the scatter-hoarding behavior of rodents. The results showed lower proportions of intact in situ (IIS) and eaten in situ (EIS) large seeds than small seeds at all levels of cache density, with the exception of IIS seeds at a 6.25 seed·m-2 cache density. A higher proportion of small seeds were eaten after removal (EAR), but a higher proportion of large seeds were scatter-hoarded (SH) by rodents at most cache densities. Furthermore, rodents preferentially removed large seeds farther away for eating or scatter-hoarding. The IIS and EIS proportions of both large and small seeds declined, but the proportion of the two types of seeds that were EAR fluctuated, increasing with increasing cache density. Rodents preferred to increase the proportion of scatter-hoarding of large seeds with increasing cache density, whereas the proportion of scatter-hoarding of small seeds was maximized at a cache density of 6.25 seed·m-2. Both the eaten distance after removal (EDAR) and scatter-hoarded distance (SHD) increased with increasing cache density. These results suggest that large seeds are more likely to be scatter-hoarded and removed to longer distances than small ones. Rodents tended to reduce the seed proportion of EIS seeds and increased the proportion of seeds EAR and SH, and accordingly increased EDAR and SHD with increasing cache density. This study provides some scientific basis for animal-mediated seed dispersal.

Quercus wutaishanica shrub affects temperate forest community composition and soil properties under different restoration stage.

Quercus wutaishanica is the dominant tree species in the natural ecosystem restoration of temperate forests in China, and it plays an active role in maintaining ecological balance. However, little is known about how ecosystem versatility develops during the restoration of forest ecosystems dominated by Q. wutaishanica. In this study, we investigated the species composition of the Q. wutaishanica community, soil nutrients, and their functional traits at various restoration stages, and comprehensively analyzed the correlations among them. At the early stage of restoration (10 years of restoration), there were Spiraea pubescens and Syringa pubescens in Q. wutaishanica community (87% of the total species), while had a larger niche width. In the middle of restoration (30 years of restoration), shannon and evenness indices were the largest, while soil total carbon, ammonium nitrogen and chlorophyll content of Q. wutaishanica leaves were the highest; among them, soil total carbon was 15.7% higher than that in 10 years of restoration, 32.4% higher than that in 40 years of restoration, ammonium nitrogen was 71.7% higher than that in 40 years of restoration, and chlorophyll content was 217.9% higher than that in 10 years of restoration, and 51.8% higher than that in 40 years of restoration. At the later stage of restoration (40 years of restoration), Lonicera ferdinandii occupied the dominant ecological niche, and soil available nitrogen, available phosphorus content and leaf thickness were the largest; while AN was 10.9% higher than that of 10 years of restoration, 16.5% higher than that of 30 years of restoration, AP was 60.6% higher than that of 10 years of restoration, 21.6% higher than that of 30 years of restoration, leaf thickness was 22.3% higher than that of 10 years of restoration, 84.9% higher than that of 30 years of restoration. However, the restriction of various soil nutrients was reduced. Our study highlighted the effectiveness of soil resource availability in plant communities during restoration, reduced competition for light among plants, and altered species richness. Furthermore, changes in the interrelationship between plant community composition and leaf functional traits of the dominant species responded positively to community restoration. These results further deepen our understanding of forest management and restoration of forest communities. In the future, it is necessary to comprehensively consider the influence of various factors on forest community restoration.

Density-Dependent Seed Predation of Quercus wutaishanica by Rodents in Response to Different Seed States.

The predation and/or dispersal of Quercus seeds by rodents play an important role in the creation of the tree species. The present study examined the effects of community habitats on the predation and dispersal of Quercus wutaishanica seeds by rodents. We released seeds with densities set at 2, 4, 8, 16, and 32 seed square meter with litter cover, soil burial, and bare ground in the Liupan Mountains National Nature Reserve in the Ningxia Hui Autonomous Region, northwest China. The results showed that (1) the litter cover and soil burial significantly increased the seed survival probability compared with bare ground treatments, especially the predation in situ (PIS) (p < 0.05). Both the scatter hoarding (SH) and larder hoarding (LH) for litter cover and soil burial were significantly increased compared with bare ground (p < 0.05). (2) The large seeds are preferentially predated after dispersal and their long-distance dispersal (>5 m) was significantly greater than that of small seeds (p < 0.05), while small seeds are more likely to be preyed on in situ or during short-distance dispersal (<3 m). (3) The Q. wutaishanica seed predation by rodents increased at a high density rather than at a low density, indicating a negative density-dependent predation. These findings provide insights into the ecological characteristics of Quercus tree regeneration and shed light on the coexistence between rodents and different-sized seeds.

Characterization of immortalized human mammary epithelial cell line HMEC 2.6.

Primary human mammary epithelial cells have a limited life span which makes it difficult to study them in vitro for most purposes. To overcome this problem, we have developed a cell line that was immortalized using defined genetic elements, and we have characterized this immortalized non-tumorigenic human mammary epithelial cell line to establish it as a potential model system. human mammary epithelial cells were obtained from a healthy individual undergoing reduction mammoplasty at SIU School of Medicine. The cells were transduced with CDK4R24C followed by transduction with human telomerase reverse transcriptase. Post all manipulation, the cells displayed a normal cell cycle phase distribution and were near diploid in nature, which was confirmed by flow cytometry and karyotyping. In vitro studies showed that the cells were anchorage dependent and were non-invasive in nature. The cell line expressed basal epithelial markers such as cytokeratin 7, CD10, and p63 and was negative for the expression of estrogen receptor and progesterone receptor. Upon G-band karyotyping, the cell line displayed the presence of a few cytogenic abnormalities, including trisomy 20 and trisomy 7, which are also commonly present in other immortalized mammary cell lines. Furthermore, the benign nature of these cells was confirmed by multiple in vitro and in vivo experiments. Therefore, we think that this cell line could serve as a good model to understand the molecular mechanisms involved in the development and progression of breast cancer and to also assess the effect of novel therapeutics on human mammary epithelial cells.

Static pressure drives proliferation of vascular smooth muscle cells via caveolin-1/ERK1/2 pathway.

Intimal hyperplasia plays an important role in various types of vascular remodeling. Mechanical forces derived from blood flow are associated with the proliferation of vascular smooth muscle cells (VSMC). This contributes to many vascular disorders such as hypertension, atherosclerosis and restenosis after percutaneous transluminal angioplasty (PTA). In this study, we show that static pressure induces the proliferation of VSMC and activates its related signal pathway. VSMC from a rat aorta were treated with different pressures (0, 60, 90, 120, 150 and 180 mm Hg) in a custom-made pressure incubator for 24h. The most active proliferation of VSMC was detected at a pressure of 120 mm Hg. VSMC was also incubated under a static pressure of 120 mm Hg for different time intervals (0, 2, 4, 8, 12 and 24h). We found that static pressure significantly stimulates VSMC proliferation. Extracellular signal-regulated kinases 1/2 (ERK1/2) activation showed a peak at the pressure of 120 mm Hg at 4-h time point. Moreover, caveolin-1 expression was significantly inhibited by rising static pressure. Downregulation of VSMC proliferation could be found after PD98059 (ERK1/2 phosphorylation inhibitor) treatment. Our data also showed that a siRNA-mediated caveolin-1 knock down increased ERK1/2 phosphorylation and VSMC proliferation. These results demonstrate that static pressure promotes VSMC proliferation via the Caveolin-1/ERK1/2 pathway.

Tumor suppressor function of Rab25 in triple-negative breast cancer.

Rab proteins are a group of ubiquitously expressed proteins that are responsible for intracellular transport of vesicles. Recent evidence has shown that certain Rab proteins are involved in the pathogenesis of cancer. We have recently shown that Rab25 is lost in a large fraction of breast cancer samples, particularly those derived from hormonally insensitive tumors. We have further investigated the role of Rab25 by re-expressing Rab25 in tumorigenic cell lines and measuring the impact on tumor formation as well as on various molecular pathways through PCR array analysis. In vivo tumor growth of cell lines with re-expressed Rab25 was markedly suppressed. Our data suggest that Rab25 acts through multiple pathways to enhance apoptosis and to suppress angiogenesis and invasion by modulating VEGF-A and VEGFR-1 expression. These findings suggest that Rab25 represents a novel class of cellular modulators that can influence both tumor initiation and the progression of the established tumors, thus ultimately affecting the biology of the malignant disease.

Aldo-keto reductase family 1 B10 affects fatty acid synthesis by regulating the stability of acetyl-CoA carboxylase-alpha in breast cancer cells.

Recent studies have demonstrated that aldo-keto reductase family 1 B10 (AKR1B10), a novel protein overexpressed in human hepatocellular carcinoma and non-small cell lung carcinoma, may facilitate cancer cell growth by detoxifying intracellular reactive carbonyls. This study presents a novel function of AKR1B10 in tumorigenic mammary epithelial cells (RAO-3), regulating fatty acid synthesis. In RAO-3 cells, Sephacryl-S 300 gel filtration and DEAE-Sepharose ion exchange chromatography demonstrated that AKR1B10 exists in two distinct forms, monomers (approximately 40 kDa) bound to DEAE-Sepharose column and protein complexes (approximately 300 kDa) remaining in flow-through. Co-immunoprecipitation with AKR1B10 antibody and protein mass spectrometry analysis identified that AKR1B10 associates with acetyl-CoA carboxylase-alpha (ACCA), a rate-limiting enzyme of de novo fatty acid synthesis. This association between AKR1B10 and ACCA proteins was further confirmed by co-immunoprecipitation with ACCA antibody and pulldown assays with recombinant AKR1B10 protein. Intracellular fluorescent studies showed that AKR1B10 and ACCA proteins co-localize in the cytoplasm of RAO-3 cells. More interestingly, small interfering RNA-mediated AKR1B10 knock down increased ACCA degradation through ubiquitination-proteasome pathway and resulted in >50% decrease of fatty acid synthesis in RAO-3 cells. These data suggest that AKR1B10 is a novel regulator of the biosynthesis of fatty acid, an essential component of the cell membrane, in breast cancer cells.

A mutation in Tpst2 encoding tyrosylprotein sulfotransferase causes dwarfism associated with hypothyroidism.

The growth-retarded (grt) mouse has an autosomal recessive, fetal-onset, severe thyroid hypoplasia related to TSH hyporesponsiveness. Through genetic mapping and complementation experiments, we show that grt is a missense mutation of a highly conserved region of the tyrosylprotein sulfotransferase 2 (Tpst2) gene, encoding one of the two Tpst genes implicated in posttranslational tyrosine O-sulfation. We present evidence that the grt mutation leads to a loss of TPST2 activity, and TPST2 isoform has a high degree of substrate preference for TSH receptor (TSHR). The expression of TPST2 can restore TSH-TSHR-mediated cAMP production in fibroblasts derived from grt mice. Therefore, we propose that the tyrosine sulfation of TSHR by TPST2 is crucial for TSH signaling and resultant thyroid gland function.

Loss of RAB25 expression in breast cancer.

A novel breast cancer cell line (RAO-3) was established by transduction of the Q61L mutant RAS into human mammary epithelial cells that were immortalized with catalytic subunit of telomerase (hTERT). The cells displayed anchorage-independent growth and proliferation, and formed human mammary spindle cell carcinoma when injected into nude mice. Chromosome locus 1q22-23 was partially duplicated and inverted on one of the 3 chromosomes present in the cell line. We report here that mutations of chromosome 1q22-23 locus have resulted in the loss of RAB25 expression in the breast cancer cell line. Transduction of RAB25 into the breast cancer cell line arrests anchorage-independent growth. We have also demonstrated loss of RAB25 in human breast tumor tissue. These data suggest that loss of RAB25 might contribute to tumorigenesis of breast cancer, and RAB25 is likely to be an important factor in the development of breast cancer. RAB25 could be used as biological marker of breast cancer and provides a target for gene replacement therapy.