Novel Aggregation-Induced Emission Fluorescent Molecule for Platinum(IV) Ion-Selective Recognition and Imaging of Controlled Release in Cells.

The loading, delivery, and release of Pt(IV) precursors in living organisms are important aspects of exploring the development of platinum drugs. In recent years, the biological application of the fluorescent sensors to platinum drugs has been insufficient to meet the study of Pt(IV) precursors. It is urgent to design and develop a biocompatible, multifunctional fluorescent sensor for the study of loading, transport, and release of Pt(IV) ions. Herein, we report a fluorescent molecule (E)-6-(diethylamino)-N'-(4-(diphenylamino) benzylidene)-2-oxo-2H-chromene-3 carbohydrazide (CHTPA). CHTPA has good sensitivity and selectivity to Pt(IV) when the water content is 5%, and significant increase of the fluorescence emission intensity of CHTPA is observed with Pt(IV) concentration. The sensing mechanism is attributed to photo-induced electron transfer, which is verified by X-ray absorption near edge spectroscopy spectra, UV-vis absorption spectroscopy, 1H NMR spectra, and Fourier transform infrared spectra. Furthermore, the CHTPA-Pt(IV) complex is able to release Pt(IV) in aqueous solution, and the green fluorescence of CHTPA based on the aggregation-induced emission effect can be observed. Inspired by these, the amphiphilic block copolymer poly(ethyloxide)-block-polystyrene (PEO-b-PS) is used to prepare the nonconjugated polymer dots (Pdots). The experimental results show that Pdots can effectively slow down the release speed of Pt(IV) in aqueous solution and it has a great monodispersity in aqueous solution. Meanwhile, Pdots show low cytotoxicity, and this is favorable for intracellular applications. The investigation of cellular imaging indicates that these Pdots can act as a carrier to deliver Pt(IV) into MCF-7 cells for visualized delivery and sustained release of platinum(IV) ions. Therefore, this study provides a new avenue to design and develop a biocompatible multifunctional fluorescent sensor for studying the loading, delivery, and release of Pt(IV) in cells.

Data Mining Evidences Variabilities in Glucose and Lipid Metabolism among Fish Strains: A Case Study on Three Genotypes of Gibel Carp Fed by Different Carbohydrate Sources.

An 8-week feeding trial was conducted to evaluate the application of common carbohydrate sources, cornstarch (CS), wheat starch (WS), and wheat flour (WF), to different gibel carp genotypes, Dongting, CASIII, and CASV. The results of the growth and physical responses were analysed by data visualization and unsupervised machine learning. As revealed by a self-organizing map (SOM) and the cluster of growth and biochemical indicators, CASV had superior growth and feed utilization and better regulation of postprandial glucose, followed by CASIII, while Dongting showed a high level of plasma glucose with poor growth performance. CS, WS, and WF were differently utilized by the gibel carp, and WF was associated with greater zootechnical performance based on higher specific growth rate (SGR), feed efficiency (FE), protein retention efficiency (PRE), and lipid retention efficiency (LRE), induced hepatic lipogenesis, increased liver lipids, and enhanced muscle glycogen. Spearman's correlation analysis of the physiological responses indicated that plasma glucose had a significantly negative correlation with growth, feed utilization, glycogen storage, and plasma cholesterol level, and it was positively related to liver fat content in gibel carp. Transcriptional variabilities were observed: CASIII showed increased expression of pklr, which is involved in hepatic glycolysis, and pck and g6p, which are involved in gluconeogenesis. Interestingly, Dongting showed upregulation of genes involved in glycolysis and fatty acid oxidation in muscle. Furthermore, there were numerous interactions between carbohydrate sources and strains for growth, metabolites, and transcriptional control, confirming the existence of genetic polymorphisms in carbohydrate use in gibel carp. Globally, CASV showed relatively better growth and carbohydrate utilization, and wheat flour seemed to be more efficiently utilized by gibel carp.

Determining the phosphorus release curve for Sunphase HT phytase in nursery pig diets.

A total of 280 pigs (DNA 241 × 600, initially 10.4 ±â€…0.24 kg) were used in a 21-d study to determine the available P (aP) release curve for Sunphase HT phytase (Wuhan Sunhy Biology Co., Ltd., Wuhan, P.R. China) when fed diets with a high phytate concentration. On day 21 post-weaning, considered day 0 of the study, pigs were blocked by average pen body weight (BW) and randomly allotted to 1 of 7 dietary treatments with 5 pigs per pen and 8 pens per treatment. Dietary treatments were derived from a single basal diet, and ingredients including phytase, monocalcium P, limestone, and sand were added to create the treatment diets. Treatments included three diets with increasing (0.11%, 0.19%, and 0.27%) aP from monocalcium P, or four diets with increasing phytase (250, 500, 1,000, or 2,000 phytase unit (FTU)/kg) added to the diet formulated to 0.11% aP. All diets were corn-soybean meal-canola meal-based and were formulated to contain 1.24% SID Lys, a 1.10:1 total calcium-to-phosphorus ratio, and a calculated 0.32% phytate P. Prior to the beginning of the study, all pigs were fed a diet containing 0.11% aP from days 18 to 21 post-weaning. At the conclusion of the study, 1 pig, closest to the mean weight of each pen, was euthanized, and the right fibula, 10th rib, and metacarpal were collected to determine bone ash and density. After cleaning, bones were submerged in ultra-purified water under a vacuum for 4 h and then weighed to calculate the density (Archimedes principle). For bone ash, bones were processed using the non-defatted method. From days 0 to 21, increasing aP from monocalcium P increased (linear, P ≤ 0.014) average daily gain (ADG), gain-to-feed (G:F), and final BW. Pigs fed increasing phytase had increased (linear, P ≤ 0.045) ADG, final BW, and plasma inositol concentration as well as improved (quadratic, P = 0.023) G:F. For bone characteristics, pigs fed increasing aP from inorganic P had a linear improvement (P ≤ 0.019) in fibula bone ash weight and percentage bone ash, rib bone ash weight and bone density, and all metacarpal bone properties, with a quadratic response (P ≤ 0.030) for fibula bone density and rib percentage ash. Additionally, pigs fed increasing phytase had increased (P < 0.05) bone ash weight, percentage bone ash, and bone density in either a linear or quadratic fashion depending on the bone analyzed. The available P release curve generated for Sunphase HT phytase for percentage bone ash combining values from the right fibula, 10th rib, and metacarpal is aP release, % = (0.360 × FTU) ÷ (2,330.250 + FTU).

Effects of reduced-protein diets with protease supplementation on growth, carcass yield, intestinal morphology, organ development, nutrient digestibility, and blood biochemical of broiler chickens.

This study was conducted to evaluate growth performance, carcass yield, intestinal morphology, organ development, nutrient digestibility, and blood biochemical parameters of broiler fed 1% reduced-protein diets with/without protease supplementation. A total of 1,120 one-day-old male broiler chickens with average initial body weight (BW), 46.45 ±â€…0.49 g, were divided into five groups with seven replications and 32 birds per replication. The treatment varied according to the protein and protease enzyme levels: positive control (PC), negative control (NC, PC with reduction of 1% protein), PC supplemented with 50 g/t protease (PC + 50), NC supplemented with 50 g/t protease (NC + 50), and NC supplemented with 100 g/t protease (NC + 100). The results showed that there was no significant effect of 1% reduced-protein diets, with or without protease on feed intake, final BW, average daily gain, feed conversion ratio, and nutrient digestibility. The changes in dietary protein level and supplementation of protease did not affect carcass yield, but significantly affected abdominal fat content, PC + 50 group had significantly lower abdominal fat content than NC-based diet including NC, NC + 50, NC + 100. Reduced-protein with protease supplementation strongly affected organ weight, especially on day 21: the pancreas was heavier in PC and NC + 50 group than other groups, spleen was heaver in NC group than in NC + 100 group, thymus was heavier in NC + 50 group than in PC, NC and NC + 100 group, small intestine was heavier in NC + 50 and NC + 100 group than in PC group, and large intestine was also heavier in NC + 50 group than in NC group. Villus height sampled at 35-d was significantly increased with protease supplement, and which was significantly higher in NC + 100 group than NC group. Regarding on blood metabolites, only urea and uric acid were affected by the reduction of dietary protein, broiler fed PC diet had higher urea and uric acid content than fed NC diet. In conclusion, supplementation of 50 g/t protease in 1% reduced-protein diets does not negatively affect on growth, nutrient digestibility, carcass yield, organ development, and blood metabolites. Moreover, supplementation of protease in low-protein diet could effectively promote organ development and benefit intestine morphology.

Efficacy of enhanced Escherichia coli phytase on growth performance, bone quality, nutrient digestibility, and metabolism in nursery pigs fed corn-soybean meal diet low in calcium and digestible phosphorous.

Efficacy of Escherichia coli phytase (ASP) was evaluated in nursery pigs fed low Ca and digestible P corn and soybean meal diet. Piglets were weaned on day 21, fed a common commercial starter diet for 7 d, and assigned to pens (4 pigs/pen: 2 ♀ and 2 ♂) based on day 7 BW. Positive control (PC) and negative (NC) diets were formulated with similar energy and nutrients with exception of total Ca, total P, and digestible P concentrations being 79%, 67%, and 55% that of PC diet, respectively. Two other diets were formulated by adding ASP in NC at 500 and 1,000 FTU/kg. All diets had 0.2% TiO2 indigestible marker. The diets were allocated to pens to give 6 replicates per diet and fed for 42 d. Feed intake and body weight were monitored at 14-d intervals. On day 42, 1 pig/pen was bled and euthanized to access blood and tissue samples. Analyzed total Ca and P in NC diet was 71% and 69% of concentration in PC diet. Recovery of phytase in pelleted diets was 66.2% and 73.5% for NC+500 FTU/kg and NC+1,000 FTU/kg diets, respectively. Between days 15 and 42, pigs fed NC diet grew slower and ate less feed than pigs fed the other diets. Overall (days 0-42), phytase in NC increased (P ≤ 0.05) ADG linearly and quadratically. On day 42, pigs fed PC, NC+500 FTU/kg, and NC+1,000 FTU/kg were +6.1, +5.9, and +7.1 kg heavier (P < 0.05) than pigs fed NC, respectively. Pigs fed PC and NC plus phytase exhibited higher (P = 0.003) G:F relative to NC pigs between days 15 and 28. Pigs fed NC diet had lower (P < 0.001) plasma P concentration, apparent total tract digestibility (ATTD) of Ca and P, and metacarpal and metatarsal bone attributes than pigs fed any other diets. Supplementation of phytase in NC linearly increased (P < 0.05) plasma P concentration, ATTD of Ca and P, and bone attributes. Specifically, phytase increased (P ≤ 0.025) dry weight, length, and ash weight in metacarpals and metatarsals. In conclusion, low total Ca and digestible P diet depressed growth and P utilization in piglets. Supplemental phytase improved performance in pigs fed NC linked to enhanced nutrients uptake and metabolism commensurate to pigs fed adequate total Ca and digestible P from inorganic source.

Hepatic glucose metabolic responses to digestible dietary carbohydrates in two isogenic lines of rainbow trout.

Rainbow trout (Oncorhynchus mykiss) was recognized as a typical 'glucose-intolerant' fish and poor dietary carbohydrate user. Our first objective was to test the effect of dietary carbohydrates themselves (without modification of dietary protein intake) on hepatic glucose gene expression (taking into account the paralogs). The second aim was to research if two isogenic trout lines had different responses to carbohydrate intake, showing one with a better use dietary carbohydrates. Thus, we used two isogenic lines of rainbow trout (named A32h and AB1h) fed with either a high carbohydrate diet or a low carbohydrate diet for 12 weeks. We analysed the zootechnical parameters, the plasma metabolites, the hepatic glucose metabolism at the molecular level and the hormonal-nutrient sensing pathway. Globally, dietary carbohydrate intake was associated with hyperglycaemia and down regulation of the energy sensor Ampk, but also with atypical regulation of glycolysis and gluconeogenesis in the liver. Indeed, the first steps of glycolysis and gluconeogenesis catalysed by the glucokinase and the phospenolpyruvate carboxykinase are regulated at the molecular level by dietary carbohydrates as expected (i.e. induction of the glycolytic gck and repression of the gluconeogenic pck); by contrast, and surprisingly, for two other key glycolytic enzymes (phosphofructokinase enzyme - pfkl and pyruvate kinase - pk) some of the paralogs (pfklb and pklr) are inhibited by carbohydrates whereas some of the genes coding gluconeogenic enzymes (the glucose-6-phosphatase enzyme g6pcb1b and g6pcb2a gene and the fructose1-6 biphosphatase paralog fbp1a) are induced. On the other hand, some differences for the zootechnical parameters and metabolic genes were also found between the two isogenic lines, confirming the existence of genetic polymorphisms for nutritional regulation of intermediary metabolism in rainbow trout. In conclusion, our study determines some new and unexpected molecular regulations of the glucose metabolism in rainbow trout which may partly lead to the poor utilization of dietary carbohydrates and it underlines the existence of differences in molecular regulation of glucose metabolism between two isogenic lines which provides arguments for future selection of rainbow trout.

Regulation by Dietary Carbohydrates of Intermediary Metabolism in Liver and Muscle of Two Isogenic Lines of Rainbow Trout.

Rainbow trout (Oncorhynchus mykiss) is recognized as a typical "glucose-intolerant" fish, and the limits of dietary carbohydrate utilization have been investigated for many years. In this study, the objective was to test the molecular effects of dietary carbohydrates on intermediary metabolism in two major metabolic tissues, liver and muscle. Another objective was also to study if the response to carbohydrate intake depended on the genetic background. We fed two isogenic lines of rainbow trout (named A22h and N38h) with high carbohydrate diet (carbohydrate, 22.9%) or low carbohydrate diet (carbohydrate, 3.6%) for 12 weeks. Carbohydrates were associated with higher feed utilization owned by the well-known protein-sparing effect, with better fish growth performance. However, atypical regulation of glycolysis and gluconeogenesis in liver and absence of hk and glut4 induction in muscle, were also observed. Regarding the effects of carbohydrates on other metabolism, we observed an increased, at a molecular level, of hepatic cholesterol biosynthesis, fatty acid oxidation and mitochondrial energy metabolism. Genetic variability (revealed by the differences between the two isogenic lines) was observed for some metabolic genes especially for those involved in the EPA and DHA biosynthetic capacity. Finally, our study demonstrates that dietary carbohydrate not only affect glucose metabolism but also strongly impact the lipid and energy metabolism in liver and muscle of trout.