Rapid Low-Cost Microfluidic Detection in Point of Care Diagnostics.

Clinical diagnostics is a challenge in resource poor areas. Accessibility to diagnostic laboratories is severely curtailed in areas where resources and infrastructure are limited. There is a need to develop low cost and portable devices to promote healthcare services in such places. The development of low cost paper-based microfluidic devices (µPADS) and thread-based devices have the potential to revolutionize point of care diagnostics in poverty-ridden areas. In this report, we describe how frugal technologies can be used for the detection of biological fluids, and the need for developing low cost wireless and wearable technologies to be deployed in resource poor settings.

Nuclear Mitotic Apparatus (NuMA) Interacts with and Regulates Astrin at the Mitotic Spindle.

The large nuclear mitotic apparatus (NuMA) protein is an essential player in mitotic spindle assembly and maintenance. We report here the identification of Astrin, a spindle- and kinetochore-associated protein, as a novel interactor of NuMA. We show that the C-terminal tail of NuMA can directly bind to the C terminus of Astrin and that this interaction helps to recruit Astrin to microtubules. Knockdown of NuMA by RNA interference dramatically impaired Astrin recruitment to the mitotic spindle. Overexpression of the N terminus of mammalian homologue of Drosophila Pins (LGN), which blocks the microtubule binding of NuMA and competes with Astrin for NuMA binding, also led to similar results. Furthermore, we found that cytoplasmic dynein is required for the spindle pole accumulation of Astrin, and dynein-mediated transport is important for balanced distribution of Astrin between spindle poles and kinetochores. On the other hand, if Astrin levels are reduced, then NuMA could not efficiently concentrate at the spindle poles. Our findings reveal a direct physical link between two important regulators of mitotic progression and demonstrate the critical role of the NuMA-Astrin interaction for accurate cell division.

Trichostatin A protects against intestinal injury in rats with acute liver failure.

BACKGROUND: Histone deacetylase (HDAC) inhibitors have been widely applied in the clinic as anticancer drugs against multiple neoplasms and proved their anti-inflammation under different pathology recently. Trichostatin A (TSA) is an HDAC inhibitor specific in class I and II HDAC enzymes. The aim of the present study was to elucidate the protective effects of TSA on acute liver failure (ALF) in rats and its potential mechanism. METHODS: A total of 18 female Sprague-Dawley rats were separated into control, model, and TSA groups. We used Western blotting to determine the expression of HDACs, inflammatory cytokines, and acetylation of histone in liver and small intestine. The gene expression of inflammatory factors and Cox-2 was detected by a polymerase chain reaction. Colonic motility was assessed by spatiotemporal mapping. Histologic analysis and immunohistochemistry were performed. Intestinal permeability examination and levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin were also observed. RESULTS: ALF procedure caused harm to histology of liver and small intestine, increased the intestinal permeability and serum levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin. It also interrupted the normal organization of colonic motor patterns by hurting enteric nervous system and pacemaker cells. Along with the decrease of inflammatory factors in ALF rats by TSA administration, all the damage to the liver, the small intestine, and the colon was repaired. CONCLUSIONS: TSA alleviates the lesion in liver, as well as in small intestine and colon in ALF rats by directly inhibiting inflammatory response.

Aptamer and its potential applications for food safety.

Accompanied by industrial globalization, rapid urbanization, and population increment, mass production and staple trading for food consumption are upsoaring continuously, foodborne disease resulted from various food safety issues is currently a crucial public health concern worldwide, which has not only created a great burden on both economy and society, but also greatly threatened the sustainability of mankind's livelihood and human reproduction. In order to better ensure food safety and thus effectively curb the occurrence of foodborne diseases, the development and evolving of inspection strategies are indispensable measures for quality assurance and conformity assessment. Nowadays, as complementary measures to and with advantageous merits over classic analytical methods, highly specific and selective aptamer-based assays have found their increasingly important roles in various domains of food analysis. This critical review summarizes the advantages of aptamer as compared with antibody, introduces important evolving variants of systematic evolution of ligands by exponential enrichment (SELEX), and presents an overview of potential aptamer applications for food safety.

Amphiphilic polyethylenimine (PEI) as highly efficient non-viral gene carrier.

Efficient and safe gene vectors are important for gene therapy. Here, a novel family of amphiphilic polyethylenimine (PEI) LD1-PEI bearing a polar group of branched PEI 25K and four dodecyl chains was developed. Agarose gel electrophoresis was used to confirm the formation of complexes. The transfection activity of the amphiphilic carrier was evaluated in different cell lines. The in vitro study showed that LD1-PEI showed a higher transfection efficiency with improved biocompatibility than PEI 25K. Serum showed almost no or only a slight effect on LD1-PEI/DNA transfection efficiency. In summary, LD1-PEI is a promising nonviral gene carrier.

Simultaneous determination of dyes in wines by HPLC coupled to quadrupole orbitrap mass spectrometry.

A new method combining the QuEChERS (quick, easy, cheap, effective, rugged, and safe) method with ultra high performance liquid chromatography and ESI quadrupole Orbitrap high-resolution MS was developed for the highly accurate and sensitive screening of 69 dyes in wines. Response surface methodology was employed to optimize the QuEChERS sample preparation method for the determination of 69 different analytes in wines for the first time. After optimization, the maximum predicted recovery was 99.48% rate for canacert indigo carmine under the optimized conditions of 10 mL acetonitrile, 1.45 g sodium acetate, 107 mg primary secondary amine, and 96 mg C18 . For the matrices studied, the recovery rates of the other 68 compounds ranged from 87.2-107.4%, with coefficient of variation < 6.4%. The mass accuracy typically obtained is routinely better than 1.6 ppm and only needed to be calibrated once a week. The LODs for the analytes are in the range 1-1000 µg/kg. This method has been successfully applied on screening of dyes in commercial wines, and it is very useful for the fast screening of different food additives.

Determination of 30 synthetic food additives in soft drinks by HPLC/electrospray ionization-tandem mass spectrometry.

A method combining SPE with HPLC/electrospray ionization-MS/MS was developed for simultaneous determination of 30 synthetic food additives, including synthetic colorants, preservatives, and sweeteners in soft drinks. All targets were efficiently separated using the optimized chromatographic and MS conditions and parameters in a single run within 18 min. The LOD of the analytes ranged from 0.01 to 20 microg/kg, and the method was validated with recoveries in the 80.8 to 106.4% range. This multisynthetic additive method was found to be accurate and reliable and will be useful to ensure the safety of food products, such as the labeling and proper use of synthetic food additives in soft drinks.

Regulation of NAD+ metabolism in aging and disease.

More than a century after discovering NAD+, information is still evolving on the role of this molecule in health and diseases. The biological functions of NAD+ and NAD+ precursors encompass pathways in cellular energetics, inflammation, metabolism, and cell survival. Several metabolic and neurological diseases exhibit reduced tissue NAD+ levels. Significantly reduced levels of NAD+ are also associated with aging, and enhancing NAD+ levels improved healthspan and lifespan in animal models. Recent studies suggest a causal link between senescence, age-associated reduction in tissue NAD+ and enzymatic degradation of NAD+. Furthermore, the discovery of transporters and receptors involved in NAD+ precursor (nicotinic acid, or niacin, nicotinamide, and nicotinamide riboside) metabolism allowed for a better understanding of their role in cellular homeostasis including signaling functions that are independent of their functions in redox reactions. We also review studies that demonstrate that the functional effect of niacin is partially due to the activation of its cell surface receptor, GPR109a. Based on the recent progress in understanding the mechanism and function of NAD+ and NAD+ precursors in cell metabolism, new strategies are evolving to exploit these molecules' pharmacological potential in the maintenance of metabolic balance.

Juvenile Plasma Factors Improve Organ Function and Survival following Injury by Promoting Antioxidant Response.

Studies have shown that factors in the blood of young organisms can rejuvenate the old ones. Studies using heterochronic parabiosis models further reinforced the hypothesis that juvenile factors can rejuvenate aged systems. We sought to determine the effect of juvenile plasma-derived factors on the outcome following hemorrhagic shock injury in aged mice. We discovered that pre-pubertal (young) mice subjected to hemorrhagic shock survived for a prolonged period, in the absence of fluid resuscitation, compared to mature or aged mice. To further understand the mechanism of maturational dependence of injury resolution, extracellular vesicles isolated from the plasma of young mice were administered to aged mice subjected to hemorrhagic shock. The extracellular vesicle treatment prolonged life in the aged mice. The treatment resulted in reduced oxidative stress in the liver and in the circulation, along with an enhanced expression of the nuclear factor erythroid factor 2-related factor 2 (Nrf2) and its target genes, and a reduction in the expression of the transcription factor BTB and CNC homology 1 (Bach1). We propose that plasma factors in the juvenile mice have a reparative effect in the aged mice in injury resolution by modulating the Nrf2/Bach1 axis in the antioxidant response pathway.

Unraveling proteome changes of irradiated goat meat and its relationship to off-flavor analyzed by high-throughput proteomics analysis.

Irradiation has been regarded as a perspective preservation technology to prolong the shelf life of goat meat. In order to evaluate the effect of irradiation on goat meat, the high-throughput proteomics was applied to observe the proteome profiles changes and off-flavor formation of irradiated goat meat. A total of 207 proteins with significant changes (fold change > 2 or < 0.5, P < 0.05, VIP > 1) was determined in five irradiation doses (0, 1, 2, 4 and 6 kGy). Among them, 26 proteins involving 7 functional enrichments terms were found to be related to off-flavor. Particularly, proteins involved in protein oxidation, cysteine and methionine metabolism lead to irradiation off-flavor formation. However, the irradiation off-flavor will gradually dissipate due to metabolic process during storage. This study was designed to provide a proteomic insight into irradiation off-flavor of goat meat and suggest an underlying connection between off-flavor formation and biological processes.

[Expression and subcellular localization of APOBEC3G in peripheral blood mononuclear cells and liver tissues of chronic HBV patients].

OBJECTIVE: To study the expression level and intracellular localization of APOBEC3G in peripheral blood mononuclear cells (PBMCs) and liver tissues of chronic HBV patients. METHODS: The expression level and intracellular localization of APOBEC3G in PBMCs and liver tissues were detected using the western blot and confocal laser scanning microscope (CLSM). RESULTS: Western-blot showed that the expression level of APOBEC3G in PBMCs of healthy controls was very low. The relative expression levels of APOBEC3G in PBMC of patients with chronic hepatitis B, chronic severe hepatitis, liver cirrhosis, or liver cancer were 4.12+/-0.21, 4.07+/-0.28, 4.16+/-0.36 or 4.21+/-0.39 respectively, which were higher than that in the healthy controls. However, there was no significant difference in APOBEC3G expression among different chronic HBV patients (q = 0.931, 0.744, 1.675, 1.675, 2.606 or 0.931, respectively, all P values more than 0.05). In addition, there was no significant difference on APOBEC3G in liver tissues between chronic hepatitis B patients and hepatocellular carcinoma patients (4.40+/-0.34 vs 4.34+/-0.43, q = 0.588, P more than 0.05). CLSM indicated that the localization of APOBEC3G protein was in cytoplasm of PBMCs and hepatocytes. CONCLUSION: APOBEC3G is upregulated in the PBMCs of chronic hepatitis B patients.

Rapid senescence-like response after acute injury.

Cellular senescence is a state of irreversible growth arrest. Short-term programmed senescence such as in embryonic development and slowly progressing senescence as in aging are both well described. However, acute senescence in living organisms is not well understood. We hypothesized that hemorrhagic shock injury (HI) caused by whole body hypoxia and nutrient deprivation, resulting in organ dysfunction due to severe blood loss, could lead to acute senescence in vivo. Our experiments, for the first time, demonstrate a rapidly emerged, senolytics-responsive, senescence-like response in the rat liver in less than five hr following hemorrhagic shock. We conclude that the senescence, or pseudosenescence, observed is necessary to maintain tissue homeostasis following the injury.

Discrimination of Milk from Different Animal Species by a Foodomics Approach Based on High-Resolution Mass Spectrometry.

An untargeted foodomics strategy based on ultra-high-performance liquid chromatography coupled with quadrupole orbitrap and chemometrics was used to observe subtle differences in the molecule profiles of raw milk from different animal species (cow milk, goat milk, and water buffalo milk), which could prevent the fraud activities in the dairy industry. In data-dependent acquisition (DIA), spectra for all precursor ions facilitated the comprehensive identification of unknown compounds in untargeted foodomics. Chemometrics techniques were used to analyze large amounts of complex data to observe the separation of different sample groups and find the potential markers of sample groups. Finally, five markers were putatively identified by the potential marker identification workflow. The quantification results showed that ß-carotene was found only in cow milk; ergocalciferol was found only in water buffalo milk; and the contents of nonanoic acid, decanoic acid, and octanoic acid were higher in goat milk than those in cow milk and water buffalo milk. The quantification of ß-carotene enabled the detection of cow milk with a sensitivity threshold of 5% (w/w). This work provided an efficient approach for the discrimination of cow milk, goat milk, and water buffalo milk. Compared with proteomics and genomics, the simpler analytical procedures, lower costs, and higher speed of this work make it of great benefit for routine operations.

Investigation of Differentially Expressed Proteins Induced by Alteration of Natural Se Uptake with Ultrahigh-Performance Liquid Chromatography Quadrupole Orbitrap Uncovers the Potential Nutritional Value in Se-Enriched Green Tea.

Se-enriched green tea, with an increasing consumption, is the shoot of tea plants grown naturally in a seleniferous region. A label-free proteomic strategy based on ultrahigh-performance liquid chromatography quadrupole Orbitrap was applied to characterize and distinguish the difference between the Se-enriched and normal green tea with a total of 283 proteins identified and 264 proteins quantified, in which 96 proteins were observed different. The expressions of 10 proteins were upregulated and 40 proteins were downregulated (p < 0.05) in Se-enriched samples. Gene ontology, Kyoto Encyclopedia of Genes and Genomes pathway, and protein-protein interaction (PPI) network analysis results indicated that these differentially expressed proteins significantly interacted and were involved in secondary metabolites and inflammatory response biological processes. Furthermore, the expression of methyl-jasmonate- and ethylene-related genes changed significantly in Se-enriched green tea, and catalase proteins were employed as the center of the pathway that changed significantly in the PPI network. These results associating with the current knowledge of selenium in soil-plant cycling revealed that organic selenium was synthesized in green tea, which provided novel information on Se assimilation in Camellia sinensis and improved the understanding of Se-enriched green tea as a possible ideal selenium supplement in daily life.

A strategy for the determination of flavor substances in goat milk by liquid chromatography-high resolution mass spectrometry.

A new analytical method for untargeted screening of flavor compounds in goat milk and milk flavor using mass spectrometry was developed and validated. All compounds of low molecular weight were considered as candidate chemicals, instead of focusing only on compounds that already have a known affect on the flavor quality of goat milk. High performance liquid chromatography coupled with high resolution quadrupole orbitrap mass spectrometry based on external standard was used to analyse flavor compounds in complex samples. The untargeted screening strategy is data pre-processing, automated componentization, suspect library spectra searching and the retained analytes confirming based on the reference standard solutions. Forty-two flavor compounds were confirmed and analyzed. The established method was validated by considering the guidelines specified in Commission Decision 2002/657/EC and European SANCO/12571/2013 Guideline 2013. This validated method was successfully applied to samples detection, and some unique flavor compounds were found between goat milk and milk flavor samples, which can provide some useful references for the illicit addition of flavors and fragrances in dairy products.

Effects of spices on the formation of biogenic amines during the fermentation of dry fermented mutton sausage.

A precise and unambiguous quantitative strategy for six biogenic amines in dry fermented mutton sausage with liquid chromatography tandem quadrupole Orbitrap mass spectrometry was developed to investigate the inhibitory effects of star anise, amomum tsao-ko, clove, cassia, fennel, bay leaf, and nutmeg on the accumulation of biogenic amines. Compared the data-dependent MS2 fragmentation (dd-MS2), variable data-independent acquisition (vDIA), and multiplex data-independent acquisition (mDIA), and the mDIA acquisition mode was selected for further analysis. The LODs and LOQs were 0.9-1.5 µg kg-1 and 2.9-5.0 µg kg-1. The maximum inhibition of spice on tryptamine, putrescine, spermidine, 2-phenylethylamine, tyrosamine, and histamine were 21.8%, 19.3%, 27.5%, 24.6%, 18.7% and 24.4%, which revealed that the accumulation of BAs was significantly inhibited by spice extracts, while cassia and fennel extracts showed the best effect. Thus, the established method can provide a better reference for dry fermented mutton sausage quality, safety and trade analysis.

A strategy of untargeted foodomics profiling for dynamic changes during Fu brick tea fermentation using ultrahigh-performance liquid chromatography-high resolution mass spectrometry.

During a microbial fermentation process, the chemical composition of Fu brick tea changed substantially. To better profile the unique changes of metabolites and lipids in Fu brick tea at different fermentation stages, a multifaceted strategy combining untargeted foodomics with UHPLC-Q-Orbitrap analysis was developed. The discriminative tea metabolites and lipids were determined by statistical analysis and online database matching. A total of 12 characteristic components (3 metabolic and 9 lipid variables) were found to differ significantly among samples. Quantitative analysis showed that, except for trimyristin and incensole acetate, the content of these compounds gradually increased and finally stabilized with increasing fermentation time. The simultaneous determination of metabolites and lipids in this study provided detailed information for the analysis of multi-component changes in a complex matrix.

Recent advances in Baijiu analysis by chromatography based technology-A review.

Baijiu, a conventional fermented beverage with thousands history, plays a crucial part in physical health of people and social culture in China. The application of high-throughput screening and confirmation of flavor components in the field of authenticity identification and potential function has attracted more and more attention and interest of researchers. With more attention to health, pesticides residues, phthalates, biogenic amines and other hazardous substances remained in Baijiu have also become the quality parameters concerned by consumers. This review aims to present updated and critical overview on the substances analysis of Baijiu by means of hyphenated chromatographic techniques based on various pretreatment approaches in recent years. Subsequently, the advance and main direction of Baijiu composition analysis were evaluated and prospected.

Foodomics analysis of natural aging and gamma irradiation maturation in Chinese distilled Baijiu by UPLC-Orbitrap-MS/MS.

Classification of Feng-flavor Baijiu considering aging category was furnished with a comprehensive fingerprinting strategy which used UPLC-Orbitrap and foodomics, and the most discriminant 29 compounds related to aging, 15 organic acids, 8 esters as well as some carbonyl compounds were discovered. Increase of aromatic organic acids, decline of carcinogenic dibutyl phthalate and generation of numerous aromatic substances in Baijiu were caused by irradiation until the system reached a relatively stable state which needed 28 days. It is similarity of physical and chemical reaction process of natural aging and irradiation maturation that both can facilitate flavor of base Baijiu, while the aging time can be plainly shorten utilizing irradiation for Baijiu, whose outcome has been demonstrated by LC-MS and sensory evaluation. The combination of UPLC-Orbitrap and foodomics was applied as a valid tactic to analyze a complex system and gamma irradiation can be a powerful tool to promote Baijiu aging.

NLRX1 Regulation Following Acute Mitochondrial Injury.

Several metabolic, cardiovascular, and neurological disorders are characterized by mitochondrial dysfunction followed by dysregulation of cellular energetics. Mitochondria play an important role in ATP production and cell death regulation. NLRX1, a mitochondria-targeted protein, is known to negatively regulate innate immunity, and cell death responses. However, the role of this protein in cellular homeostasis following mitochondrial injury is not well-understood. To understand the mechanisms underlying the effect of acute injury in regulating NLRX1 signaling pathways, we used an in vitro model of mitochondrial injury wherein, rat pulmonary microvascular endothelial cells were subjected to sodium azide treatment or glucose starvation. Both sodium azide and glucose starvation activated NF-κB and TBK1 associated innate immune response. Moreover, increased TBK1, IKK, IκB, and TRAF6 were recruited to mitochondria and interacted with NLRX1. Depletion of endogenous NLRX1 resulted in exacerbated NF-κB and TBK1 associated innate immune response and apoptosis. Our results suggest that NLRX1 participates in the regulation of innate immune response in mitochondria, and plays an important role in the maintenance of cellular homeostasis following acute mitochondrial injury. We propose that the mitochondrial recruitment of inflammatory mediators and their interaction with NLRX1 are protective responses to maintain cellular homeostasis following injury.