Training effects of set- and repetition-interval rest time on recumbent-boxing exercise: Could virtual reality improve further?

This study examined the influence of set-interval and repetition-interval rest time of virtual reality (VR) boxing game in supine-lying posture. Fifty healthy middle-aged adults were randomly assigned into VR and non-VR groups to perform six different exercise protocols with varying set-interval and repetition-interval rest times (S0R0, S0R1/3, S0R2/3, S40R0, S40R1/3, and S40R2/3). Analysis on the non-VR group showed significant differences between exercise protocols for average heart rate (p < 0.001), maximum ventilation volume (p < 0.001), respiratory quotient (p < 0.001), oxygen pulse (p < 0.001), and excess post-exercise oxygen consumption (EPOC) (p = 0.003). VR appeared to have no further improvement on physical training effects in middle-aged adults, while the participants reported negative experience that might be associated with the over-exertion. Future study might need to explore game design elements that can accommodate high-exertion exercises.

Whole-Brain Single-Cell Imaging and Analysis of Intact Neonatal Mouse Brains Using MRI, Tissue Clearing, and Light-Sheet Microscopy.

Tissue clearing followed by light-sheet microscopy (LSFM) enables cellular-resolution imaging of intact brain structure, allowing quantitative analysis of structural changes caused by genetic or environmental perturbations. Whole-brain imaging results in more accurate quantification of cells and the study of region-specific differences that may be missed with commonly used microscopy of physically sectioned tissue. Using light-sheet microscopy to image cleared brains greatly increases acquisition speed as compared to confocal microscopy. Although these images produce very large amounts of brain structural data, most computational tools that perform feature quantification in images of cleared tissue are limited to counting sparse cell populations, rather than all nuclei. Here, we demonstrate NuMorph (Nuclear-Based Morphometry), a group of analysis tools, to quantify all nuclei and nuclear markers within annotated regions of a postnatal day 4 (P4) mouse brain after clearing and imaging on a light-sheet microscope. We describe magnetic resonance imaging (MRI) to measure brain volume prior to shrinkage caused by tissue clearing dehydration steps, tissue clearing using the iDISCO+ method, including immunolabeling, followed by light-sheet microscopy using a commercially available platform to image mouse brains at cellular resolution. We then demonstrate this image analysis pipeline using NuMorph, which is used to correct intensity differences, stitch image tiles, align multiple channels, count nuclei, and annotate brain regions through registration to publicly available atlases. We designed this approach using publicly available protocols and software, allowing any researcher with the necessary microscope and computational resources to perform these techniques. These tissue clearing, imaging, and computational tools allow measurement and quantification of the three-dimensional (3D) organization of cell-types in the cortex and should be widely applicable to any wild-type/knockout mouse study design.

Achieving "Non-Foaming" Rhamnolipid Production and Productivity Rebounds of Pseudomonas aeruginosa under Weakly Acidic Fermentation.

The rhamnolipid production of Pseudomonas aeruginosa has been impeded by its severe foaming; overcoming the bottleneck of foaming has become the most urgent requirement for rhamnolipid production in recent decades. In this study, we performed rhamnolipid fermentation under weakly acidic conditions to address this bottleneck. The results showed that the foaming behavior of rhamnolipid fermentation broths was pH-dependent with the foaming ability decreasing from 162.8% to 28.6% from pH 8 to 4. The "non-foaming" rhamnolipid fermentation can be realized at pH 5.5, but the biosynthesis of rhamnolipids was significantly inhibited. Further, rhamnolipid yield rebounded from 8.1 g/L to 15.4 g/L after ultraviolet and ethyl methanesulfonate compound mutagenesis. The mechanism study showed that the species changes of rhamnolipid homologs did not affect the foaming behavior of the fermentation but had a slight effect on the bioactivity of rhamnolipids. At pH 8.0 to 5.0, increased surface tension, decreased viscosity and zeta potential, and aggregation of rhamnolipid molecules contributed to the "non-foaming" rhamnolipid fermentation. This study provides a promising avenue for the "non-foaming" rhamnolipid fermentation and elucidates the mechanisms involved, facilitating the understanding of pH-associated foaming behavior and developing a more efficient strategy for achieving rhamnolipid production.

Foaming of rhamnolipids fermentation: impact factors and fermentation strategies.

Rhamnolipids have recently attracted considerable attentions because of their excellent biosurfactant performance and potential applications in agriculture, environment, biomedicine, etc., but severe foaming causes the high cost of production, restraining their commercial production and applications. To reduce or eliminate the foaming, numerous explorations have been focused on foaming factors and fermentation strategies, but a systematic summary and discussion are still lacking. Additionally, although these studies have not broken through the bottleneck of foaming, they are conducive to understanding the foaming mechanism and developing more effective rhamnolipids production strategies. Therefore, this review focuses on the effects of fermentation components and control conditions on foaming behavior and fermentation strategies responded to the severe foaming in rhamnolipids fermentation and systematically summarizes 6 impact factors and 9 fermentation strategies. Furthermore, the potentialities of 9 fermentation strategies for large-scale production are discussed and some further strategies are suggested. We hope this review can further facilitate the understanding of foaming factors and fermentation strategies as well as conducive to developing the more effective large-scale production strategies to accelerate the commercial production process of rhamnolipids.

Optimization and scale-up of the production of rhamnolipid by Pseudomonas aeruginosa in solid-state fermentation using high-density polyurethane foam as an inert support.

To be competitive with common synthetic surfactants, the cost of production of rhamnolipid must be minimized by the fermentation process of non-foaming and low impurities. Herein, a novel solid-state fermentation process was developed for production of rhamnolipid by Pseudomonas aeruginosa SKY. The results were shown that high-density polyurethane foam is a satisfactory alternative to agro-industrial by-products for SSF of rhamnolipid. Palm oil and NaNO3 were promising carbon source and nitrogen source, respectively. Response surface methodology was employed to enhance the production of rhamnolipid. Palm oil, NaNO3 and liquid-to-solid ratios were significant factors. The optimal medium was developed as: 73.6 g/l palm oil; 3.0 g/l g NaNO3; 1.1 g NaCl; 1.1 g KCl; 3.4 g KH2PO4; 4.4 g K2HPO4; 0.5 g MgSO4·7H2O and 37.2 liquid-to-solid ratios. An overall 1.39-fold increase in rhamnolipid production was achieved in the optimized medium as compared with the unoptimized basal medium. Air pressure pulsation solid-state fermentation (APP-SSF) was applied to the experiment of scale-up for improving transfer efficiency of heat and mass. The yield of rhamnolipid reached 39.8 g/l in a 30 l APP-SSF fermenter. The crude extract of rhamnolipid lowered the surface tension of water to 28 mN/m and kept the critical micelle concentration at 50 mg/l. The work revealed the SSF with HPUF as an inert support was a promising fermentation system that could effectively produce rhamnolipid with low impurities, high productivity and low cost of production at a large scale.

Periploca forrestii Saponin Ameliorates Murine CFA-Induced Arthritis by Suppressing Cytokine Production.

Periploca forrestii Schltr. has been used as a Chinese folk medicine due to its versatile pharmacological effects such as promoting wounds and rheumatoid arthritis. However, the antiarthritic activity of Periploca forrestii saponin (PFS) and its active compound Periplocin has still not been demonstrated. Here, we evaluated the antiarthritic effects of PFS in adjuvant-induced arthritis (AIA) rats by intragastric administration at a dose of 50 mg/kg. The anti-inflammatory activities of Periplocin were also examined in LPS-induced AIA splenocytes and synoviocytes. PFS significantly ameliorated joint swelling; inhibited bone erosion in joints; lowered levels of IL-6 and TGF-ß1 in AIA rat splenocyte; and reduced joint protein expression levels of phospho-STAT3 and IKKα. Using LPS-induced AIA splenocytes, we demonstrate that Periplocin suppressed the key proinflammatory cytokines levels of IL-6, IFN-γ, TGF-ß1, and IL-13 and IL-22 and transcription factor levels of T-bet, GATA3, and C-Jun genes. Periplocin also suppressed LPS-induced cytokine secretion from synoviocytes. Our study highlights the antiarthritic activity of PFS and its derived Periplocin and the underlying mechanisms. These results provide a strong rationale for further testing and validation of the use of Periploca forrestii Schltr. as an alternative modality for the treatment of RA.

Biochemical characterization of recombinant Enterovirus 71 3C protease with fluorogenic model peptide substrates and development of a biochemical assay.

Enterovirus 71 (EV71), a primary pathogen of hand, foot, and mouth disease (HFMD), affects primarily infants and children. Currently, there are no effective drugs against HFMD. EV71 3C protease performs multiple tasks in the viral replication, which makes it an ideal antiviral target. We synthesized a small set of fluorogenic model peptides derived from cleavage sites of EV71 polyprotein and examined their efficiencies of cleavage by EV71 3C protease. The novel peptide P08 [(2-(N-methylamino)benzoyl) (NMA)-IEALFQGPPK(DNP)FR] was determined to be the most efficiently cleaved by EV71 3C protease, with a kinetic constant kcat/Km of 11.8 ± 0.82 mM(-1) min(-1). Compared with literature reports, P08 gave significant improvement in the signal/background ratio, which makes it an attractive substrate for assay development. A Molecular dynamics simulation study elaborated the interactions between substrate P08 and EV71 3C protease. Arg39, which is located at the bottom of the S2 pocket of EV71 3C protease, may participate in the proteolysis process of substrates. With an aim to evaluate EV71 3C protease inhibitors, a reliable and robust biochemical assay with a Z' factor of 0.87 ± 0.05 was developed. A novel compound (compound 3) (50% inhibitory concentration [IC50] = 1.89 ± 0.25 µM) was discovered using this assay, which effectively suppressed the proliferation of EV 71 (strain Fuyang) in rhabdomyosarcoma (RD) cells with a highly selective index (50% effective concentration [EC50] = 4.54 ± 0.51 µM; 50% cytotoxic concentration [CC50] > 100 µM). This fast and efficient assay for lead discovery and optimization provides an ideal platform for anti-EV71 drug development targeting 3C protease.

The fast spiral-SelMQC technique for in vivo MR spectroscopic imaging of polyunsaturated fatty acids in human breast tissue.

The selective multiple-quantum coherence transfer method has been applied to image polyunsaturated fatty acids (PUFA) distributions in human breast tissues in vivo for cancer detection, with complete suppression of the unwanted lipid and water signals in a single scan. The Cartesian k-space mapping of PUFA in vivo using the selective multiple-quantum coherence transfer (Sel-MQC) chemical shift imaging (CSI) technique, however, requires excessive MR scan time. In this article, we report a fast Spiral-SelMQC sequence using a rapid spiral k-space sampling scheme. The Spiral-SelMQC images of PUFA distribution in human breast were acquired using two-interleaved spirals on a 3 T GE Signa magnetic resonance imaging scanner. Approximately 160-fold reduction of acquisition time was observed as compared with the corresponding selective multiple-quantum coherence transfer CSI method with an equivalent number of scans, permitting acquisition of high-resolution PUFA images in minutes. The reconstructed Spiral-SelMQC PUFA images of human breast tissues achieved a sub-millimeter resolution of 0.54 × 0.54 or 0.63 × 0.63 mm(2) /pixel for field of view = 14 or 16 cm, respectively. The Spiral-SelMQC parameters for PUFA detection were optimized in 2D selective multiple-quantum coherence transfer experiments to suppress monounsaturated fatty acids and other lipid signals. The fast in vivo Spiral-SelMQC imaging method will be applied to study human breast cancer and other human diseases in extracranial organs.

[Effects of exogenous dimethylarsinic acid on Brassica campestris growth and soil arsenic bioavailability].

A pot experiment was conducted to study the effects of exogenous dimethylarsinic acid (DMA) on the growth of Brassica campestris and the bioavailability of soil arsenic (As). With the increasing concentration of applied DMA, the emergence rate and biomass of B. campestris increased at low concentration DMA, but decreased at high concentration DMA. When the DMA concentration reached 90 mg x kg(-1), the emergence rate and biomass of B. campestris in the second cropping decreased by 9.5% and 57.0%, respectively, compared with those in the control, indicating that exogenous DMA had longer-term effects on the growth of B. campestris. The soil available As and the As uptake by B. campestris all increased with increasing concentration of exogenous DMA, and there existed significant correlations among them. After applied into soil, the exogenous DMA demethylated, with As(V) as the main product and lesser amount of As (III), and the concentrations of soil As(V) and As(III) increased with increasing application rate of exogenous DMA.

[Influence of flooding on form transformation of soil arsenic].

An incubation test was conducted to study the dynamics of exogenously supplied dimethylarsenic acid (DMA), monomethylarsonic acid (MMA), and As(V) in soil under flooding. With the increasing time of incubation, the exogenously supplied DMA and MMA were mainly transformed into As(V), and the As(V) concentration increased, being significantly higher after incubated for 150 days, compared with that after incubated for 1 day. The exogenously supplied DMA was demethylated into As(V) within 30 days, accompanied by a little As(III), while the transformation rate of exogenously supplied MMA was rather slow within 60 days, accompanied by a little As(III) and DMA. The exogenously supplied As(V) decreased with increasing time of incubation, and its form had less change except that a little As(III) occurred.

[Transformation of exogenous dimethyl arsenic in soil].

A pot experiment was conducted to study the speciation transformation of exogenous dimethyl arsenic (DMA) in soil. The added DMA was mainly transformed into arsenate [As(V)], accompanied with a small amount of monomethyl arsenic (MMA). When the concentration of added DMA was 30 mg x kg(-1), the transformation rate was the highest, being 6.71%, 8.11%, 11.33%, and 19.32% when the cultivation time was 10, 15, 30, and 40 days, respectively. With increasing concentration of added DMA, soil soluble arsenic (AE-As) had an increasing trend, but decreased as the cultivation time increased. Comparing with CK, the addition of DMA increased the concentrations of soil arsenic bounded to aluminum (Al-As), iron (Fe-As), calcium (Ca-As), which was possibly due to the adsorption or fixation of added DMA and its transformation products by the oxides or hydroxides of soil aluminum, iron, and calcium.

[Transformation of different exogenous arsenic forms in soil under aerobic condition].

An incubation test was conducted to study the transformation of exogenous dimethylarsenic acid (DMA), monomethylarsenic acid (MMA), and arsenate [As(V)] in soil under the condition of 35% of water-holding capacity. After added into soil, the concentrations of test arsenic forms all showed a decreasing trend with time. The DMA and MMA were mainly demethylated, and after 120 days incubation at constant temperature and humidity, transformed into As(V). A small amount of MMA was detected in the treatment with added DMA on the 120th day of incubation, and a small amount of DMA was detected in the treatment with added MMA during the period of 7-60 days incubation. By the end of the incubation test, the concentrations of added DMA and MMA in soil decreased significantly (P < 0.01), with the decrement being 99.5% and 94.3% and the concentration of transformed As(V) increased by 4.61 and 5.15 times, respectively. Comparing with DMA and MMA, the As(V) after added into soil had less form transformation.

[Form transformation of arsenic in soil and corresponding analyzing methods].

Based on the analysis of the sources and existing forms of soil arsenic, this paper approached the inter-transformationr of different arsenic forms in soil. In the meanwhile, the extraction and determination methods of different soil arsenic forms were also compared. It was considered that HPLC-HG-AFS had the advantages of high sensitivity, low detection limit, better selectivity, low operation cost, and less inter-transformation of different arsenic forms, being able to be used as the prior method for the detection of different arsenic forms. Combining with previous research results, the form transformation of arsenic in soil and the promising research aspects were also discussed.

In vivo MR spectroscopic imaging of polyunsaturated fatty acids (PUFA) in healthy and cancerous breast tissues by selective multiple-quantum coherence transfer (Sel-MQC): a preliminary study.

The spatial distribution of polyunsaturated fatty acids (PUFA) in healthy and cancerous human breast tissues was measured in vivo with a selective multiple-quantum coherence transfer (Sel-MQC) technique. This method selectively detected the olefinic methylene protons (-CH = CH-) of PUFA at 5.3 ppm that were coupled with allylic methylene protons (-CH(2)-CH(2)-CH=) of unsaturated acyl chain at 2.8 ppm. Unwanted lipid coherences and tissue water signal were dephased in a single scan. Breast PUFA were mapped at 1 cm(3) voxel resolution in sagittal slices of nine breasts in six healthy female volunteers that were compared to invasive ductal carcinoma (IDC) in one breast cancer patient. The healthy breast tissue displayed continuous PUFA distribution. In some individuals, PUFA appeared throughout the breast tissue; in others they were only located in the central breast area. Decreased PUFA levels were detected in the IDC of the breast cancer patient. The magnetic resonance spectroscopic imaging (MRSI) measurement was consistent with the histological findings, ultrasound, and mammography images. PUFA patterns are sensitive to abnormal breast tissue changes including malignant transformations, and thus may serve as a biomarker for early diagnosis and therapeutic monitoring of breast disease.

High flow-resolution for mobility estimation in 2D-ENMR of proteins using maximum entropy method (MEM-ENMR).

Multidimensional electrophoretic NMR (nD-ENMR) is a potentially powerful tool for structural characterization of co-existing proteins and protein conformations. By applying a DC electric field pulse, the electrophoretic migration rates of different proteins were detected experimentally in a new dimension of electrophoretic flow. The electrophoretic mobilities were employed to differentiate protein signals. In U-shaped ENMR sample chambers, individual protein components in a solution mixture followed a cosinusoidal electrophoretic interferogram as a function of its unique electrophoretic migration rate. After Fourier transformation in the electrophoretic flow dimension, the protein signals were resolved at different resonant frequencies proportional to their electrophoretic mobilities. Currently, the mobility resolution of the proteins in the electrophoretic flow dimension is limited by severe truncations of the electrophoretic interferograms due to the finite electric field strength available before the onset of heat-induced convection. In this article, we present a successful signal processing method, the Burg's maximum entropy method (MEM), to analyze the truncated ENMR signals (MEM-ENMR). Significant enhancement in flow resolution was demonstrated using two-dimensional ENMR of two protein samples: a lysozyme solution and a solution mixture of bovine serum albumin (BSA) and ubiquitin. The electrophoretic mobilities of lysozyme, BSA and ubiquitin were measured from the MEM analysis as 7.5x10(-5), 1.9x10(-4) and 8.7x10(-5) cm2 V-1 s-1, respectively. Results from computer simulations confirmed a complete removal of truncation artifacts in the MEM-ENMR spectra with 3- to 6-fold resolution enhancement.

In vivo tumor lactate relaxation measurements by selective multiple-quantum-coherence (Sel-MQC) transfer.

The frequency-selective multiple-quantum-coherence (Sel-MQC) lactate (Lac) filter offers complete lipid and water suppression in a single scan for robust in vivo detection of tumor Lac, even in the presence of abundant lipids. Conversion of the detected signal into accurate tissue concentrations of Lac requires knowledge of in vivo Lac T1 and T2 relaxation times. This work reports modifications to the Sel-MQC pulse sequence, T1- and T2-Sel-MQC, that facilitate relaxation measurements of Lac. The T1-Sel-MQC sequence combines an inversion prepulse with the Sel-MQC filter. The T2-Sel-MQC sequence incorporates a CH3-selective 180 degrees pulse during the MQ preparation period to overcome the J-modulation effects and allow the insertion of variable echo delays. The performance of these sequences was evaluated with the use of phantoms and subcutaneous murine tumor models in vivo. The present approach will allow investigators to correct for the relaxation-induced Lac signal loss in Sel-MQC experiments for the quantitative mapping of in vivo tumor Lac distribution.

Magnetic resonance spectroscopic imaging of tumor metabolic markers for cancer diagnosis, metabolic phenotyping, and characterization of tumor microenvironment.

Cancer cells display heterogeneous genetic characteristics, depending on the tumor dynamic microenvironment. Abnormal tumor vasculature and poor tissue oxygenation generate a fraction of hypoxic tumor cells that have selective advantages in metastasis and invasion and often resist chemo- and radiation therapies. The genetic alterations acquired by tumors modify their biochemical pathways, which results in abnormal tumor metabolism. An elevation in glycolysis known as the "Warburg effect" and changes in lipid synthesis and oxidation occur. Magnetic resonance spectroscopy (MRS) has been used to study tumor metabolism in preclinical animal models and in clinical research on human breast, brain, and prostate cancers. This technique can identify specific genetic and metabolic changes that occur in malignant tumors. Therefore, the metabolic markers, detectable by MRS, not only provide information on biochemical changes but also define different metabolic tumor phenotypes. When combined with the contrast-enhanced Magnetic Resonance Imaging (MRI), which has a high sensitivity for cancer diagnosis, in vivo magnetic resonance spectroscopic imaging (MRSI) improves the diagnostic specificity of malignant human cancers and is becoming an important clinical tool for cancer management and care. This article reviews the MRSI techniques as molecular imaging methods to detect and quantify metabolic changes in various tumor tissue types, especially in extracranial tumor tissues that contain high concentrations of fat. MRI/MRSI methods have been used to characterize tumor microenvironments in terms of blood volume and vessel permeability. Measurements of tissue oxygenation and glycolytic rates by MRS also are described to illustrate the capability of the MR technology in probing molecular information non-invasively in tumor tissues and its important potential for studying molecular mechanisms of human cancers in physiological conditions.

Constant-time multidimensional electrophoretic NMR.

Multidimensional electrophoretic NMR (ENMR) has been introduced to determine structures of coexisting proteins and protein conformations in solution. Signals of different proteins are separated in a new dimension of electrophoretic flow according to their characteristic electrophoretic mobilities. The electrophoretic interferograms have been generated in the flow dimension in two approaches by incrementing either the amplitude or the duration of the electric field. The ENMR method of incrementing the duration of the electric field, however, introduces severe signal decays due to molecular diffusion and spin relaxation, limiting the effectiveness of the method. In this study, an improved method of constant-time multidimensional ENMR (CT-ENMR) has been proposed and successfully tested. The time delays between the magnetic field gradients and the RF pulses are kept constant in this new method so that the molecular diffusion and spin relaxation processes contribute to only a constant factor of signal amplitude. As an alternative approach of incrementing the amplitude of the electric field, this novel method significantly enhances our capability and potential in characterizing structural changes of interacting proteins during biological signaling processes. The CT-ENMR method is particularly useful in studies where the amplitude-incrementing of the electric field is not optimal. For example, the CT-ENMR method is superior when the electric field is applied in the direction not parallel to the static magnetic field B(0) to the xy-magnetization. The new method was successfully demonstrated with a sample solution containing 100 mM 4,9-dioxa-1,12-dodecanediamine and 100 mM L-aspartic acid in D(2)O.