In situ Proteomic Profiling of Curcumin Targets in HCT116 Colon Cancer Cell Line

To date, the exact targets and mechanism of action of curcumin, a natural product with anti-inflammatory and anti-cancer properties, remain elusive. Here we synthesized a cell permeable curcumin probe (Cur-P) with an alkyne moiety, which can be tagged with biotin for affinity enrichment, or with a fluorescent dye for visualization of the direct-binding protein targets of curcumin in situ. iTRAQ™ quantitative proteomics approach was applied to distinguish the specific binding targets from the non-specific ones. In total, 197 proteins were confidently identified as curcumin binding targets from HCT116 colon cancer cell line. Gene Ontology analysis showed that the targets are broadly distributed and enriched in the nucleus, mitochondria and plasma membrane, and they are involved in various biological functions including metabolic process, regulation, response to stimulus and cellular process. Ingenuity Pathway Analysis™ (IPA) suggested that curcumin may exert its anticancer effects over multiple critical biological pathways including the EIF2, eIF4/p70S6K, mTOR signaling and mitochondrial dysfunction pathways. Functional validations confirmed that curcumin downregulates cellular protein synthesis, and induces autophagy, lysosomal activation and increased ROS production, thus leading to cell death

Deciphering chicken gut microbial dynamics based on high-throughput 16S rRNA metagenomics analyses

BACKGROUND: Chicken gut microbiota has paramount roles in host performance, health and immunity. Understanding the topological difference in gut microbial community composition is crucial to provide knowledge on the functions of each members of microbiota to the physiological maintenance of the host. The gut microbiota profiling of the chicken was commonly performed previously using culture-dependent and early culture-independent methods which had limited coverage and accuracy. Advances in technology based on next-generation sequencing (NGS), offers unparalleled coverage and depth in determining microbial gut dynamics. Thus, the aim of this study was to investigate the ileal and caecal microbiota development as chicken aged, which is important for future effective gut modulation. MATERIAL AND METHODS: Ileal and caecal contents of broiler chicken were extracted from 7, 14, 21 and 42-day old chicken. Genomic DNA was then extracted and amplified based on V3 hyper-variable region of 16S rRNA. Bioinformatics, ecological and statistical analyses such as Principal Coordinate Analysis (PCoA) was performed in mothur software and plotted using PRIMER 6. Additional analyses for predicted metagenomes were performed through PICRUSt and STAMP software package based on Greengenes databases. RESULTS: A distinctive difference in bacterial communities was observed between ilea and caeca as the chicken aged (P < 0.001). The microbial communities in the caeca were more diverse in comparison to the ilea communities. The potentially pathogenic bacteria such as Clostridium were elevated as the chicken aged and the population of beneficial microbe such as Lactobacillus was low at all intervals. On the other hand, based on predicted metagenomes analysed, clear distinction in functions and roles of gut microbiota such as gene pathways related to nutrient absorption (e.g. sugar and amino acid metabolism), and bacterial proliferation and colonization (e.g. bacterial motility proteins, two-component system and bacterial secretion system) were observed between ilea and caeca, respectively (P < 0.05). CONCLUSIONS: The caeca microbial communities were more diverse in comparison to ilea. The main functional differences between the two sites were found to be related to nutrient absorption and bacterial colonization. Based on the composition of the microbial community, future gut modulation with beneficial bacteria such as probiotics may benefit the host

Adaptation and validation of Richmond Compulsive Buying Scale in Chinese population

Background and aims Compulsive buying (CB) is a behavioral addiction that is conceptualized as an obsessive–compulsive and impulsive–control disorder. The Richmond Compulsive Buying Scale (RCBS), a six-item self-reporting instrument that has been validated worldwide, was developed based on this theoretical background. This study aimed to adapt RCBS to the Chinese population (RCBS-TC) to guide future national and international prevalence studies. Methods This methodological study was conducted in two phases. Phase 1 involved the forward and backward translation of RCBS, the content and face validation of the RCBS, and the evaluation of its translation adequacy. Phase 2 involved the psychometric testing of RCBS-TC for its internal consistency, stability, and construct validity using confirmatory factor analysis (CFA). Results In Phase 1, RCBS-TC obtained satisfactory item-level (I-CVI = 83.3%–100%) and scale-level content validity index (CVI/AVE = 97.2%), comprehensibility (100%), and translation adequacy [intraclass correlation coefficient (ICC) = 0.858]. In Phase 2, based on data collected from 821 adults, RCBS-TC demonstrated a satisfactory internal consistency (Cronbach’s α = .88; corrected item-total correlation coefficients = 0.61–0.78) 2-week test–retest reliability (ICC = 0.82 based on 61 university students). For construct validation, the CFA results indicated that the corrected first-order two-factor models were acceptable with the same goodness-of-fit indices (χ2/df = 8.56, CFI = 0.99, NFI = 0.98, IFI = 0.99, and RMSEA = 0.09). The 2-week test–retest reliability of RCBS-TC (n = 61) was also satisfactory (ICC = 0.82). Discussion and conclusions This methodological study adopted appropriate and stringent procedures to ensure that the translation and validation of RCBS-TC was of quality. The results indicate that this scale has a satisfactory reliability and validity for the Chinese population

Production of phytase by Mitsuokella jalaludinii in semi-solid state fermentation of agricultural by-products

Phytase activity and growth of anaerobic rumen bacterium, Mitsuokella jalaludinii were investigated by semi-solid state fermentation. Carbon source (rice bran, yam and cassava), nitrogen sources (soya bean, offal meal, fish meal and feather meal) and growth factors (hemin, L-cysteine hydrochloride and minerals) were evaluated in a one-factor-at-atime approach. Rice bran and fish meal produced better growth and phytase enzyme activity. The removal of L-cysteine hydrochloride and minerals significantly decreased (p<0.05) phytase activity from 1178.72 U to 446.99 U and 902.54 U, respectively. The response surface methods (RSM) was conducted to optimize the phytase production and the results showed the combination of 7.7% of rice bran and 3.7% of fish meal in semi-solid state fermentation gave the highest phytase activity. Maximum phytase production and optimum growth of bacteria were detected at 12 h incubation in both MF medium (control) and agro-medium. In this agro-medium, M. jalaludinii produced 2.5 fold higher phytase activity compared to MF medium

Discrimination of Escherichia coli isolates recovered from mucosal contents of chicken intestines and different age by repetitive elements sequence-based PCR

Repetitive sequence-based PCR (rep-PCR) is a distinctive typing approach that is used to differentiate between bacterial strains. This method is also useful for studying bacterial diversity from different sources. In this study, four rep-PCR which are enterobacterial repetitive intergenic consensus PCR (ERIC-PCR), BOX-PCR, repetitive extragenic palindromic PCR (REP-PCR) and polytrinucleotide (GTG)5-PCR were evaluated for differentiation of eighteen Escherichia coli isolates to correct source based on part of intestine and age. These isolates were recovered earlier from ileal and caecal mucosal contents of chickens at a different age. The purpose of this study was to investigate the efficacy of four rep-PCR methods and composite of rep-PCR patterns to differentiate E. coli isolates to original sources of part of intestines and age based on the D index (discriminatory power determined based on Simpson’s index of diversity calculated at similarity coefficient of 90%). The (GTG)5-PCR had the highest D index (0.9804) for part of intestine and age factors. The similar D index was observed in the composite of rep-PCR patterns. The lowest D index was observed in ERIC- and BOX-PCR at 0.9020 and 0.8039 for part of intestine and age factors, respectively. (GTG)5-PCR was also the most discriminative rep-PCR observed due to its ability to cluster 14I 3E and 14I 2X isolates, and 14C 1E and 14C 3E isolates correctly in part of intestine and age factors. It was concluded that (GTG)5-PCR is a promising tool for discriminating E. coli isolates extracted from chicken intestines

Chronic adiponectin deficiency leads to Alzheimer’s disease-like cognitive impairments and pathologies through AMPK inactivation and cerebral insulin resistance in aged mice

(a) Immunoblotting analysis of IRβ in the hippocampus and frontal cortex of 18-month old wildtype and APN-KO mice. (b) Densitometric analysis of the ratio of IRβ. Mean ± S.E.M.; ***p < 0.001, n.s. statistically not significant; Scale bar: 100 μm. (JPG 30 kb

Effects of supplementing freeze-dried Mitsuokella jalaludinii phytase on the growth performance and gut microbial diversity of broiler chickens

Inclusion of phytase in animal feedstuff is a common practice to enhance nutrients availability. However, little is known about the effects of phytase supplementation on the microbial ecology of the gastrointestinal tract. In this study, freeze-dried Mitsuokella jalaludinii phytase (MJ) was evaluated in a feeding trial with broilers fed a low available phosphorus (aP) diet. A total of 180 male broiler chicks (day-old Cobb) were assigned into three dietary treatments: Control fed with 0.4% (w/w) of available phosphorus (aP); Group T1 fed low aP [0.2% (w/w)] supplemented with MJ; and T2 fed low aP and deactivated MJ. The source of readily available P, dicalcium phosphate (DCP), was removed from low aP diet, whereby additional limestone was provided to replace the amount of Ca normally found in DCP. For each treatment, 4 replicate pens were used, where each pen consisted of 15 animals. The animals' energy intake and caecal bacterial community were monitored weekly for up to 3 weeks. The apparent metabolizable energy (AME) and apparent digestibility of dry matter (ADDM) of broilers fed with different diets were determined. In addition, the caecal microbial diversities of broilers were assessed using high-throughput next-generation sequencing targeting the V3-V4 region of bacterial 16S rRNA. The results showed that broilers fed with T1 diet have better feed conversion ratio (FCR) when compared to the Control (p < .05) and T2 diets (p < .05), demonstrating the efficiency of MJ as a supplement to low aP diet. Nevertheless, MJ did not significantly affect the microbial population and diversity in broilers' caeca, which mainly consists of members from Bacteroidetes, Firmicutes, and Proteobacteria. Regardless, significant variations in the caecal bacterial composition were observed over time, probably due to succession as the broilers aged. This is the first reported study on the effect of MJ on the microbial diversity of broiler's caeca

Alterations in Brain Network Topology and Structural-Functional Connectome Coupling Relate to Cognitive Impairment

According to the network-based neurodegeneration hypothesis, neurodegenerative diseases target specific large-scale neural networks, such as the default mode network, and may propagate along the structural and functional connections within and between these brain networks. Cognitive impairment no dementia (CIND) represents an early prodromal stage but few studies have examined brain topological changes within and between brain structural and functional networks. To this end, we studied the structural networks [diffusion magnetic resonance imaging (MRI)] and functional networks (task-free functional MRI) in CIND (61 mild, 56 moderate) and healthy older adults (97 controls). Structurally, compared with controls, moderate CIND had lower global efficiency, and lower nodal centrality and nodal efficiency in the thalamus, somatomotor network, and higher-order cognitive networks. Mild CIND only had higher nodal degree centrality in dorsal parietal regions. Functional differences were more subtle, with both CIND groups showing lower nodal centrality and efficiency in temporal and somatomotor regions. Importantly, CIND generally had higher structural-functional connectome correlation than controls. The higher structural-functional topological similarity was undesirable as higher correlation was associated with poorer verbal memory, executive function, and visuoconstruction. Our findings highlighted the distinct and progressive changes in brain structural-functional networks at the prodromal stage of neurodegenerative diseases

Toward controllable and predictable synthesis of high-entropy alloy nanocrystals.

High-entropy alloy (HEA) nanocrystals have attracted extensive attention in catalysis. However, there are no effective strategies for synthesizing them in a controllable and predictable manner. With quinary HEA nanocrystals made of platinum-group metals as an example, we demonstrate that their structures with spatial compositions can be predicted by quantitatively knowing the reduction kinetics of metal precursors and entropy of mixing in the nanocrystals under dropwise addition of the mixing five-metal precursor solution. The time to reach a steady state for each precursor plays a pivotal role in determining the structures of HEA nanocrystals with homogeneous alloy and core-shell features. Compared to the commercial platinum/carbon and phase-separated counterparts, the dendritic HEA nanocrystals with a defect-rich surface show substantial enhancement in catalytic activity and durability toward both hydrogen evolution and oxidation. This quantitative study will lead to a paradigm shift in the design of HEA nanocrystals, pushing away from the trial-and-error approach

Meta-analysis Followed by Replication Identifies Loci in or near CDKN1B, TET3, CD80, DRAM1, and ARID5B as Associated with Systemic Lupus Erythematosus in Asians

Systemic lupus erythematosus (SLE) is a prototype autoimmune disease with a strong genetic involvement and ethnic differences. Susceptibility genes identified so far only explain a small portion of the genetic heritability of SLE, suggesting that many more loci are yet to be uncovered for this disease. In this study, we performed a meta-analysis of genome-wide association studies on SLE in Chinese Han populations and followed up the findings by replication in four additional Asian cohorts with a total of 5,365 cases and 10,054 corresponding controls. We identified genetic variants in or near CDKN1B, TET3, CD80, DRAM1, and ARID5B as associated with the disease. These findings point to potential roles of cell-cycle regulation, autophagy, and DNA demethylation in SLE pathogenesis. For the region involving TET3 and that involving CDKN1B, multiple independent SNPs were identified, highlighting a phenomenon that might partially explain the missing heritability of complex diseases