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Animal studies have suggested that mushroom intake can alleviate non-alcoholic fatty liver disease (NAFLD) due to its anti-inflammatory and antioxidant properties. However, the association between mushroom intake and NAFLD is unknown in humans. We aimed to investigate the association of mushroom intake with NAFLD among Chinese adults. This is a cross-sectional study of 24 236 adults (mean (standard deviation) age: 40·7 (sd 11·9) years; 11 394 men (47·0 %)). Mushroom intake was assessed via a validated FFQ. Newly diagnosed NAFLD was identified based on the results of annual health examinations, including ultrasound findings and a self-reported history of the disease. Multiple logistic models were used to examine the association between mushroom intake and NAFLD. The prevalence of newly diagnosed NAFLD was 19·0 %. Compared with those consuming mushrooms less frequently (≤1 time/week), the fully adjusted OR of newly diagnosed NAFLD were 0·95 (95 % CI 0·86, 1·05) for those consuming 2–3 times/week and 0·76 (95 % CI 0·63, 0·92) for those consuming ≥4 times/week (Pfor trend = 0·01). The inverse association was consistent in subgroups defined by age, sex and BMI. In conclusion, higher mushroom intake was significantly associated with lower prevalence of NAFLD among Chinese adults. Future research is required to understand the causal association between mushroom intake and NAFLD.
Biochar conversion from corn stover was evaluated under various process conditions, and the absorption capacity of biochar was investigated for the removal of oxytetracycline in wastewater. Biochar was prepared at lower carbonization temperatures (200–500 °C) and was used in three different concentrations of chemical oxygen wastewater. The results showed that the biochar prepared at the temperature range of 200–500 °C had a faster sorption rate and shorter sorption equilibrium time compared to biochar produced at higher temperatures. The longest time to reach sorption equilibrium was 9 h for biochar obtained at 200 °C. However, the biochar prepared at 500 °C required only 0.5 h to reach the sorption equilibrium. The corn stover-biochar had the highest sorption capacity of 246.3 mg/g for oxytetracycline at 30 °C. The adsorption kinetics was consistent with pseudo–second-order kinetics. This study provides a theoretical basis for the conversion of corn stover into biochar as efficient sorbents.
The present study was undertaken to investigate the antiparasitic activity of extracellular products of Streptomyces albus. Bioactivity-guided isolation of chloroform extracts affording a compound showing potent activity. The structure of the compound was elucidated as salinomycin (SAL) by EI-MS, 1H NMR and 13C NMR. In vitro test showed that SAL has potent anti-parasitic efficacy against theronts of Ichthyophthirius multifiliis with 10 min, 1, 2, 3 and 4 h (effective concentration) EC50 (95% confidence intervals) of 2.12 (2.22–2.02), 1.93 (1.98–1.88), 1.42 (1.47–1.37), 1.35 (1.41–1.31) and 1.11 (1.21–1.01) mg L−1. In vitro antiparasitic assays revealed that SAL could be 100% effective against I. multifiliis encysted tomonts at a concentration of 8.0 mg L−1. In vivo test demonstrated that the number of I. multifiliis trophonts on Erythroculter ilishaeformis treated with SAL was markedly lower than that of control group at 10 days after exposed to theronts (P < 0.05). In the control group, 80% mortality was observed owing to heavy I. multifiliis infection at 10 days. On the other hand, only 30.0% mortality was recorded in the group treated with 8.0 mg L−1 SAL. The median lethal dose (LD50) of SAL for E. ilishaeformis was 32.9 mg L−1.
Many transgenic domestic animals have been developed to produce therapeutic proteins in the mammary gland, and this approach is one of the most important methods for agricultural and biomedical applications. However, expression and secretion of a protein varies because transgenes are integrated at random sites in the genome. In addition, distal enhancers are very important for transcriptional gene regulation and tissue-specific gene expression. Development of a vector system regulated accurately in the genome is needed to improve production of therapeutic proteins. The objective of this study was to develop a knock-in system for expression of human fibroblast growth factor 2 (FGF2) in the bovine β-casein gene locus. The F2A sequence was fused to the human FGF2 gene and inserted into exon 3 of the β-casein gene. We detected expression of human FGF2 mRNA in the HC11 mouse mammary epithelial cells by RT-PCR and human FGF2 protein in the culture media using western blot analysis when the knock-in vector was introduced. We transfected the knock-in vector into bovine ear fibroblasts and produced knock-in fibroblasts using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system. Moreover, the CRISPR/Cas9 system was more efficient than conventional methods. In addition, we produced knock-in blastocysts by somatic cell nuclear transfer using the knock-in fibroblasts. Our knock-in fibroblasts may help to create cloned embryos for development of transgenic dairy cattle expressing human FGF2 protein in the mammary gland via the expression system of the bovine β-casein gene.
Background: Patients with the postural instability and gait difficulty (PIGD) subtype of Parkinson disease (PD) are at a higher risk of dysfunction and are less responsive to dopamine replacement therapy. The PIGD subtype was found to largely associate with white matter lesions, but details of the diffusion changes within these lesions have not been fully investigated. Voxel-based analysis for diffusion tensor imaging data is one of the preferred measures to compare diffusion changes in each voxel in any part of the brain. Methods: PD patients with the PIGD (n=12) and non-PIGD subtypes (n=12) were recruited to compare diffusion differences in fractional anisotropy, axial diffusivity, and radial diffusivity with voxel-based analysis. Results: Significantly reduced fractional anisotropy in bilateral superior longitudinal fasciculus, bilateral anterior corona radiata, and the left genu of the corpus callosum were shown in the PIGD subtype compared with the non-PIGD subtype. Increased radial diffusivity in the left superior longitudinal fasciculus was found in the PIGD subtype with no statistical differences in axial diffusivity found. Conclusions: Our study confirms previous findings that white matter abnormalities were greater in the PIGD subtype than in the non-PIGD subtype. Additionally, our findings suggested: (1) compared with the non-PIGD subtype, loss of white matter integrity was greater in the PIGD subtype; (2) bilateral superior longitudinal fasciculus may play a critical role in microstructural white matter abnormalities in the PIGD subtype; and (3) reduced white matter integrity in the PIGD subtype could be mainly attributed to demyelination rather than axonal loss.
The objective of this study was to explore the expression of urotensin II (UII), its receptor (GPR14), and vascular endothelial growth factor (VEGF), as well as their associations in the ischaemic brains of rats with focal cerebral ischaemia, under normal and diabetic conditions.
Diabetes mellitus (DM) was induced by injection of streptozotocin (STZ) into Sprague—Dawley rats. Focal cerebral ischaemia was induced by middle cerebral artery occlusion (MCAO) four weeks after DM onset by STZ. Rats (n=80) were divided into four groups: normal control, DM, MCAO, and DM/MCAO. Immunohistochemistry and reverse-transcriptase-polymerase chain reaction (RT-PCR) were used to detect the expression of UII, GPR14 and VEGF in the diabetic and ischaemic brain.
Expression of UII and GPR14 was increased at mRNA and protein levels in the DM and MCAO group compared with controls. In the DM/MCAO group, expression of UII and GPR14 was increased significantly in the ischaemic brain, and was accompanied by a significantly increased VEGF expression.
Diabetes mellitus was seen to aggravate brain lesions after ischaemia, and UII may have an important role.
The recent development of in-situ liquid stages for (scanning) transmission electron microscopes now makes it possible for us to study the details of electrochemical processes under operando conditions. As electrochemical processes are complex, care must be taken to calibrate the system before any in-situ/operando observations. In addition, as the electron beam can cause effects that look similar to electrochemical processes at the electrolyte/electrode interface, an understanding of the role of the electron beam in modifying the operando observations must also be understood. In this paper we describe the design, assembly, and operation of an in-situ electrochemical cell, paying particular attention to the method for controlling and quantifying the experimental parameters. The use of this system is then demonstrated for the lithiation/delithiation of silicon nanowires.
Altered lipid metabolism has been shown in fish fed plant protein sources. The present study aimed to gain further insights into how intestinal and hepatic lipid absorption and metabolism are modulated by plant meal (PM) and soya-saponin (SA) inclusion in salmon feed. Post-smolt Atlantic salmon were fed for 10 weeks one of four diets based on fishmeal or PM, with or without 10 g/kg SA. PM inclusion resulted in decreased growth performance, excessive lipid droplet accumulation in the pyloric caeca and liver, and reduced plasma cholesterol levels. Intestinal and hepatic gene expression profiling revealed an up-regulation of the expression of genes involved in lipid absorption and lipoprotein (LP) synthesis (apo, fatty acid transporters, microsomal TAG transfer protein, acyl-CoA cholesterol acyltransferase, choline kinase and choline-phosphate cytidylyltransferase A), cholesterol synthesis (3-hydroxy-3-methylglutaryl-CoA reductase) and associated transcription factors (sterol regulatory element-binding protein 2 and PPARγ). SA inclusion resulted in reduced body pools of cholesterol and bile salts. The hepatic gene expression of the rate-limiting enzyme in bile acid biosynthesis (cytochrome P450 7A1 (cyp7a1)) as well as the transcription factor liver X receptor and the bile acid transporter abcb11 (ATP-binding cassette B11) was down-regulated by SA inclusion. A significant interaction was observed between PM inclusion and SA inclusion for plasma cholesterol levels. In conclusion, gene expression profiling suggested that the capacity for LP assembly and cholesterol synthesis was up-regulated by PM exposure, probably as a compensatory mechanism for excessive lipid droplet accumulation and reduced plasma cholesterol levels. SA inclusion had hypocholesterolaemic effects on Atlantic salmon, accompanied by decreased bile salt metabolism.
This study aimed to investigate whether aquaporin 3 (Aqp3) mRNAs are expressed in immature oocytes and altered during in vitro maturation process. Five- to 6-week-old female ICR mice were primed by gonadotropin for 24 and 48 h. Immature oocytes obtained 48 h after priming were also matured in vitro for 17 to 18 h. In vivo matured oocytes were obtained after 48 h priming followed by hCG injection. Total RNAs were extracted from 80 to 150 oocytes in each experimental group, and the levels of Aqp3 mRNA were quantified by real-time reverse transcriptase polymerase chain reaction. The experiments were repeated twice using different oocytes. The Aqp3 mRNA was expressed in immature oocytes, as well as in in vitro and in vivo matured oocytes. The expression level was higher in immature oocytes obtained 48 h after priming (17.2 ± 8.6, mean ± SD) than those with no priming (5.7 ± 0.8) or obtained 24 h after priming (2.5 ± 0.8). The expression of Aqp3 mRNA decreased after in vitro maturation (1.2 ± 0.5), which was similar to in vivo matured oocytes (1.0 ± 0.0). Our work demonstrated that Aqp3 mRNA expression increased during the development of immature oocyte but decreased after completion of in vitro maturation. The results indicate that AQP3 is certainly needed for the acquisition of immature oocytes’ full growing potential within antral follicles.
In 1995, the first author of this book joined Victoria University. Immediately after that, he established a new research group called the Optoelectronic Imaging Group (OIG), with a focus on the introduction of femtosecond lasers into optical microscopy. While the first two-photon fluorescence microscope was reported in 1990, it was not until 1996 that the first two-photon fluorescence microscope in Australia was constructed by a group of OIG Ph.D. students with a femtosecond laser supported by the major equipment fund of Victoria University. It was this new instrument that gave the OIG research students and staff a powerful tool to conduct biophotonic research. At the beginning of 2000, most of the OIG members moved to Swinburne University of Technology to form a new research centre called the Centre for Micro-Photonics (CMP). Since 1995, research students of the OIG and the CMP, including four of the authors of the book, Damian Bird, Daniel Day, Ling Fu and Dru Morrish, have made many significant contributions to femtosecond biophotonic methods. The aim of this book is to provide a systematic introduction into these methods. Chapters 1–3, 6 and 8 were completed by Min Gu and Chapters 4, 5, 7 and 9 were written by Damian Bird, Ling Fu, Dru Morrish and Daniel Day, respectively. All the authors participated in the final editing of the book.
In this chapter, we introduce a new trapping and excitation technique, which utilises a single femtosecond pulse infrared illumination source to simultaneously trap and excite a microsphere probe. The induction of morphology dependent resonance (MDR) in the trapped probe is achieved under two-photon excitation. Monitoring of the MDR in the trapped probe provides a contrast mechanism for imaging and sensing. The experimental measurement of MDR within a laser trapped microsphere excited under two-photon absorption is confirmed in Section 7.2. The effect of the laser power as well as the pulse width on the transverse trapping force is investigated in Section 7.3. The dependence of two-photon induced MDR on the scanning velocity of a trapped particle is then experimentally determined. These parameters are fundamental to the acquisition of images and sensing with femtosecond laser tweezers as described in Section 7.4.
Laser trapping is an ideal method for the remote, non-invasive manipulation of a morphology dependent resonance microcavity. Controlled scanning and manipulation of the microcavity is possible via laser trapping. The microcavity has an enhanced evanescent field at its surface due to the resonant circumferential propagation of radiation at glancing angles greater than the critical angle. Freely suspended in a medium, the cavity becomes increasingly sensitive to its surrounding environment. The interaction of the cavity with its local environment during scanning dynamically alters the coupling to and leakage from the cavity. Monitoring the change in coupling to and leakage from the cavity over time enables imaging and sensing.
As discussed in Chapters 1 and 2, biological tissue is a highly scattering medium which will affect image resolution, contrast and signal level. This chapter discusses the effect of multiple scattering in a tissue-like turbid medium on two-photon fluorescence microscopy. Section 3.1 discusses a model based on imaging of microspheres embedded in a turbid medium. A quantitative study of the limiting factors on image quality is given in Section 3.2. In particular, the limitation on the penetration depth in turbid media, revealed from Monte-Carlo simulation and experimental measurements, is presented in Section 3.3.
Two-photon fluorescence microscopy of microspheres embedded in turbid media
Two-photon fluorescence microscopy has been extensively used due to its significant advantages over single-photon fluorescence microscopy. This technology has been used for in vivo imaging of thick biological samples. Since the required image information is taken at a large depth within a biological specimen, optical multiple scattering within tissue may result in a severe distortion on images obtained in this situation. Thus, the effect of optical multiple scattering on fluorescence image quality should be understood if high quality images are to be obtained at significant depths into a biological specimen. In this section, we present measured images of small fluorescent microspheres embedded in a turbidmedium which has different scattering characteristics under singlephoton and two-photon excitation. Imaging of small spheres embedded in a turbid medium has practical importance since it can be considered to be an approximate model of imaging small tumours embedded in biological tissue.
Ever since researchers realised that microscopy based on nonlinear optical effects can provide information that is blind to conventional linear techniques, applying nonlinear optical imaging to in vivo medical diagnosis in humans has been the ultimate goal. The development of nonlinear optical endoscopy that permits imaging under conditions in which a conventional nonlinear optical microscope cannot be used is the primary method to extend applications of nonlinear optical microscopy toward this goal. Fibreoptic approaches that allow for remote delivery and collection in a minimally invasive manner are normally used in nonlinear optical endoscopy. In Chapter 4, a compact nonlinear optical microscope based on a single-mode fibre (SMF) coupler to replace complicated bulk optics was described.
There are several key challenges involved in the pursuit of in vivo nonlinear optical endoscopy. First, an excitation laser beam with an ultrashort pulse width should be delivered efficiently to a remote place where efficient collection of faint nonlinear optical signals from biological samples is required. Second, laser-scanning mechanisms adopted in such a miniaturised instrumentation should permit size reduction to a millimetre scale and enable fast scanning rates for monitoring biological processes. Finally, the design of a nonlinear optical endoscope based on micro-optics must maintain great flexibility and compact size to be incorporated into endoscopes to image internal organs.
The techniques introduced in Chapters 1 and 2 are emerging technologies that offer significant promise as tools for diagnostic imaging at the cellular level. Using devices founded on well established techniques such as confocal microscopy and confocal fluorescence microscopy, instruments capable of providing point-of-care pathological analysis of malignant and cancer causing tissues are becoming practical realities. Through examination of the physical properties of inherent autofluorescence or fluorescent dyes that are used as markers in conjugation with biological samples, very good detection of cellular processes can be achieved. Tagging of target biological cells makes it possible to examine cells in vivo and achieve real time three-dimensional (3D) visualisation for diagnosis of the pathological state.
However, the inherent nature of these devices is such that the conditions under which these techniques can be applied is fundamentally limited. In most cases (for definitive analysis) a surgical biopsy is performed on the patient and the sample is extensively prepared for observation by the pathologist on bulk, bench-top imaging apparatus. Ideally, examination of whole, intact specimens within internal cavities of the body would be the preferred method that may decrease patient trauma and eliminate diagnosis lag time.
One of the recent developments in confocal fluorescence microscopy is the introduction of optical fibres and fibre-optical components into the microscope geometry. Optical fibre couplers in particular offer the most compact and cost effective solution.
As discussed in Chapter 6, the trapping volume of a far-field laser trapping geometry is approximately three times larger in the axial direction than that in the transverse direction. Such trapping volume elongation leads to a significant background and poses difficulties in the observations of nano-particle dynamics. In this chapter, we deal with near-field optics using focused evanescent illumination. The recent development of near-field optical tweezers is reviewed in Section 8.1. Section 8.2 introduces the new concept of near-field laser tweezing with a focused evanescent field. This technology is characterised both experimentally and theoretically in Section 8.3. Section 8.4 presents the utilisation of a femtosecond laser beam in a near-field optical trap. Finally, some discussions on this new method are given in Section 8.5.
Near-field optical tweezers
Near-field laser trapping or tweezers means that radiation force that is used for trapping and manipulating a micro-object results from the interaction with an evanescent wave. Recently, a new trapping modality based on the evanescent wave illumination, also called near-field illumination, has been proposed and demonstrated. This trapping technique results in a significantly reduced trapping volume due to the fact that the strength of an evanescent wave decays rapidly with the distance from the surface at which the field is generated. In this section, the near-field trapping mechanism based on the different ways to generate a localised near-field is reviewed.