Spatiotemporal expression of regulatory kinases directs the transition from mitosis to cellular morphogenesis in Drosophila.

Embryogenesis depends on a tightly regulated balance between mitosis, differentiation, and morphogenesis. Understanding how the embryo uses a relatively small number of proteins to transition between growth and morphogenesis is a central question of developmental biology, but the mechanisms controlling mitosis and differentiation are considered to be fundamentally distinct. Here we show the mitotic kinase Polo, which regulates all steps of mitosis in Drosophila, also directs cellular morphogenesis after cell cycle exit. In mitotic cells, the Aurora kinases activate Polo to control a cytoskeletal regulatory module that directs cytokinesis. We show that in the post-mitotic mesoderm, the control of Polo activity transitions from the Aurora kinases to the uncharacterized kinase Back Seat Driver (Bsd), where Bsd and Polo cooperate to regulate muscle morphogenesis. Polo and its effectors therefore direct mitosis and cellular morphogenesis, but the transition from growth to morphogenesis is determined by the spatiotemporal expression of upstream activating kinases.

FGF signaling directs myotube guidance by regulating Rac activity.

Nascent myotubes undergo a dramatic morphological transformation during myogenesis, in which the myotubes elongate over several cell diameters and are directed to the correct muscle attachment sites. Although this process of myotube guidance is essential to pattern the musculoskeletal system, the mechanisms that control myotube guidance remain poorly understood. Using transcriptomics, we found that components of the Fibroblast Growth Factor (FGF) signaling pathway were enriched in nascent myotubes in Drosophila embryos. Null mutations in the FGF receptor heartless (htl), or its ligands, caused significant myotube guidance defects. The FGF ligand Pyramus is expressed broadly in the ectoderm, and ectopic Pyramus expression disrupted muscle patterning. Mechanistically, Htl regulates the activity of Rho/Rac GTPases in nascent myotubes and effects changes in the actin cytoskeleton. FGF signals are thus essential regulators of myotube guidance that act through cytoskeletal regulatory proteins to pattern the musculoskeletal system.

Mouse Embryonic Fibroblasts Protect ob/ob Mice From Obesity and Metabolic Complications.

The global obesity epidemic is fueling alarming rates of diabetes, associated with increased risk of cardiovascular disease and cancer. Leptin is a hormone secreted by adipose tissue that is a key regulator of body weight (BW) and energy expenditure. Leptin-deficient humans and mice are obese, diabetic, and infertile and have hepatic steatosis. Although leptin replacement therapy can alleviate the pathologies seen in leptin-deficient patients and mouse models, treatment is costly and requires daily injections. Because adipocytes are the source of leptin secretion, we investigated whether mouse embryonic fibroblasts (MEFs), capable of forming adipocytes, could be injected into ob/ob mice and prevent the metabolic phenotype seen in these leptin-deficient mice. We performed a single subcutaneous injection of MEFs into leptin-deficient ob/ob mice. The MEF injection formed a single fat pad that is histologically similar to white adipose tissue. The ob/ob mice receiving MEFs (obRs) had significantly lower BW compared with nontreated ob/ob mice, primarily because of decreased adipose tissue mass. Additionally, obR mice had significantly less liver steatosis and greater glucose tolerance and insulin sensitivity. obR mice also manifested lower food intake and greater energy expenditure than ob/ob mice, providing a mechanism underlying their metabolic improvement. Furthermore, obRs have sustained metabolic protection and restoration of fertility. Collectively, our studies show the importance of functional adipocytes in preventing metabolic abnormalities seen in leptin deficiency and point to the possibility of cell-based therapies for the treatment of leptin-deficient states.

Bioaccessible trace metals in lip cosmetics and their health risks to female consumers.

Females can be exposed to toxic elements in lip cosmetics following ingestion. The bioaccessibility of Cr, Mn, Co, Ni, Cu, Cd, Sb and Pb in lip cosmetics (n = 32) were assessed via the dilute HCl extraction method, In Vitro Gastrointestinal protocol (IVG) and the United States Pharmacopeia Methodology (USPM), and then health risks were characterized. The total concentrations of trace metals (TMs) in lip cosmetics were in the range of 15.55-111.97 mg/kg (Mean: 60.99 mg/kg). Cu, Pb and Cr were the three major TMs and accounting for >75% of the total concentrations. Except Sb and Pb in 4/32 and 4/32 samples were higher than the US FDA (Food and Drug Administration of the United States) limits, the other TMs were lower than that limits. Only bioaccessible Pb in all samples significantly exceeded the FDA limit 0.1 mg/kg in candy. Using IVG or USPM might be preferable for evaluating the TMs exposure over HCl since they better represent gastrointestinal physiology. The estimated average daily intake (ADI) of bioaccessible ∑TMs through lip cosmetics ingestion of career women and female college students were under safety level. The long-term exposure of bioaccessible TMs by lip cosmetics using would inevitably cause non-carcinogenic health risk. This is the first report on the in vitro tests used for evaluating bioaccessible TMs in lip cosmetics.

Identifying the key factors that affect the formation of humic substance during different materials composting.

The aim of this work was to identify the factors which can affect humic substance (HS) formation. Composting periods, HS precursors, bacteria communities and environment factors were recognized as the key factors and few studies explored the potential relationships among them. During composting, HS precursors were mainly formed in the heating and thermophilic phases, but HS were polymerized in the cooling and mature phases. Moreover, bacterial species showed similar classification of community structure in the same composting period of different materials. Furthermore, structural equation model showed that NH4--N and NO3--N were the indirect environmental factors for regulating HS formation by the bacteria and precursors as the indirect and direct driver, respectively. Therefore, both environmental factors and HS precursors can be the regulating factors to promote HS formation. Given that, a new staging regulating method had been proposed to improve the amount of HS during different materials composting.

A new model of multi-visceral and bone metastatic prostate cancer with perivascular niche targeting by a novel endothelial specific adenoviral vector.

While modern therapies for metastatic prostate cancer (PCa) have improved survival they are associated with an increasingly prevalent entity, aggressive variant PCa (AVPCa), lacking androgen receptor (AR) expression, enriched for cancer stem cells (CSCs), and evidencing epithelial-mesenchymal plasticity with a varying extent of neuroendocrine transdifferentiation. Parallel work revealed that endothelial cells (ECs) create a perivascular CSC niche mediated by juxtacrine and membrane tethered signaling. There is increasing interest in pharmacological metastatic niche targeting, however, targeted access has been impossible. Here, we discovered that the Gleason 7 derived, androgen receptor negative, IGR-CaP1 cell line possessed some but not all of the molecular features of AVPCa. Intracardiac injection into NOD/SCID/IL2Rg -/- (NSG) mice produced a completely penetrant bone, liver, adrenal, and brain metastatic phenotype; noninvasively and histologically detectable at 2 weeks, and necessitating sacrifice 4-5 weeks post injection. Bone metastases were osteoblastic, and osteolytic. IGR-CaP1 cells expressed the neuroendocrine marker synaptophysin, near equivalent levels of vimentin and e-cadherin, all of the EMT transcription factors, and activation of NOTCH and WNT pathways. In parallel, we created a new triple-targeted adenoviral vector containing a fiber knob RGD peptide, a hexon mutation, and an EC specific ROBO4 promoter (Ad.RGD.H5/3.ROBO4). This vector was expressed in metastatic microvessels tightly juxtaposed to IGR-CaP1 cells in bone and visceral niches. Thus, the combination of IGR-CaP1 cells and NSG mice produces a completely penetrant metastatic PCa model emulating end-stage human disease. In addition, the metastatic niche access provided by our novel Ad vector could be therapeutically leveraged for future disease control or cure.

Exposure and health risk assessment of PM2.5-bound trace metals during winter in university campus in Northeast China.

In order to better understand the risk to students' health caused by pollution derived from fine particulate matter with an aerodynamic diameter <2.5mm (PM2.5), this study collected 189 samples in one outdoor and four different functional indoor environments of a research center in a university campus. Trace metals (TMs) bound to PM2.5 in outdoor and indoor environments were measured using X-ray fluorescence spectrometry. The TMs measured were: As, Co, Cd, Cr, Ni, Cu, Zn, Mn, Hg, and Pb. The measurements of PM2.5-bound TMs before and during the 2015 Spring Festival held in Northeast China were compared. Results showed that pollution due to PM2.5-bound TMs in outdoor and indoor environments was higher before than during the Spring Festival. Cu (in three indoor environments) and Zn (in an outdoor environment) showed the highest concentrations among the ten TMs that were measured. Hg showed the lowest concentrations in all the environments analyzed. The concentrations of PM2.5-bound TMs declined among four indoor environments in the following order: the atrium, the students' office (sampled just nine days before the Spring Festival), the laboratory, and an empty room. The potential carcinogenic and non-carcinogenic health risks derived from PM2.5-bound TMs were within safe limits for graduate and undergraduate students, according to the standards established by the United States Environmental Protection Agency (USEPA).

The myeloid-binding peptide adenoviral vector enables multi-organ vascular endothelial gene targeting.

Vascular endothelial cells (ECs) are ideal gene therapy targets as they provide widespread tissue access and are the first contact surfaces following intravenous vector administration. Human recombinant adenovirus serotype 5 (Ad5) is the most frequently used gene transfer system because of its appreciable transgene payload capacity and lack of somatic mutation risk. However, standard Ad5 vectors predominantly transduce liver but not the vasculature following intravenous administration. We recently developed an Ad5 vector with a myeloid cell-binding peptide (MBP) incorporated into the knob-deleted, T4 fibritin chimeric fiber (Ad.MBP). This vector was shown to transduce pulmonary ECs presumably via a vector handoff mechanism. Here we tested the body-wide tropism of the Ad.MBP vector, its myeloid cell necessity, and vector-EC expression dose response. Using comprehensive multi-organ co-immunofluorescence analysis, we discovered that Ad.MBP produced widespread EC transduction in the lung, heart, kidney, skeletal muscle, pancreas, small bowel, and brain. Surprisingly, Ad.MBP retained hepatocyte tropism albeit at a reduced frequency compared with the standard Ad5. While binding specifically to myeloid cells ex vivo, multi-organ Ad.MBP expression was not dependent on circulating monocytes or macrophages. Ad.MBP dose de-escalation maintained full lung-targeting capacity but drastically reduced transgene expression in other organs. Swapping the EC-specific ROBO4 for the CMV promoter/enhancer abrogated hepatocyte expression but also reduced gene expression in other organs. Collectively, our multilevel targeting strategy could enable therapeutic biological production in previously inaccessible organs that pertain to the most debilitating or lethal human diseases.

[Study on the method of nanometer-size zirconium dioxide enrichment separation and inductively coupled plasma mass spectrometry determination of trace barium(II) in water].

Monoclinic fusiform zirconium dioxide (ZrO2) nanoparticles were synthesized by hydrothermal method, a method was established based on monoclinic fusiform nanometer-sized ZrO2 enrichment separation, and trace barium in water was determined by inductively coupled plasma mass spectrometry (ICP-MS). The detection limit for Ba(II) was 0.007 ng x mL(-1), and the relative standard deviation was 0.13% (n = 11). The optimal enrichment separation conditions of nanometer-sized ZrO2 for Ba(II) were studied in detail, it was found that the percentage of adsorbed Ba(II) was more than 99% under pH 10.0 and 2 mL 0.5 mol x L(-1) HCl was sufficient for elution of Ba(II) by more than 98%. The static adsorption capacity of ZrO2 to Ba(II) was 196.6 microg x g(-1) and enrichment factor was 250. Properties of nanometer-sized ZrO2 were discussed through regeneration experiment and effects of co-existing ions and contrast experiment to ordinary ZrO2, adsorption properties of nanometer-sized ZrO2 were applied to real samples in the analysis of Ba(II) and the determination was carried out by ICP-MS with satisfactory results.

[Method study on calcium and phosphorus simultaneous determination by microwave-digestion and ICP-MS].

As a pre-digestion method of breast milk microwave-digestion was adopted in this method, and the contents of calcium and phosphorus were determined by ICP-MS at the same time. By optimizing conditions of digestion and ICP-MS, appropriate internal elements and the isotope mass number and EPA200.8 interference calibration equation were chosen. The accuracy and reliability of method were verified through the recovery and milk powder analysis of national standard substances (GBW10017). Determination results are in the permissible range. The relative deviation (RSD) is less than 2.40%, while the recovery is between 102.8% and 104.0%. ICP-MS method has wide dynamic range, and doesn't need to dilute samples. It is suitable for lots of samples and multi-element analyzed at the same time, and making up default of different method and different element determined respectively in national standard. It is a determination method for calcium and phosphorus supply of breast milk.