Evaluation of the performance of a 7-T 8 × 2 transceiver array.

The decoupled 8 × 2 transceiver array has been shown to achieve a mean B1 + of 11.7 uT with a coefficient of variation of ~11% over the intracranial brain volume for 7-T MR imaging. However, this array may be thought to give lower signal-to-noise ratio (SNR) and higher g-factors for parallel imaging compared with a radio frequency (RF) receive-only coil due to the latter's higher coil count and use of coil overlap to reduce the mutual impedance. Nonetheless, because the transceiver's highly decoupled design (pertinent for transmission) should also be constructive for reception, we measured the noise correlation, g-factors, and SNR for the decoupled transceiver in comparison with a commercial reference coil. We found that although the transceiver has half the number of receive elements in comparison with the reference coil (16 vs. 32), comparable g-factors and SNR over the head were obtained. From five subjects, the transceiver versus reference coil SNR was 65 ± 10 versus 67 ± 15. The mean noise correlation for all coil pairs was 10% ± 5% and 12% ± 9% (transceiver and reference coil, respectively). As changes in load impedance may alter the S parameters, we also examined the performance of the transceiver with tuned and matched (TM) versus untuned and unmatched (UTM) conditions on five subjects. We found that the noise correlation and SNR are robust to load variation; a noise correlation of 10% ± 5% and 10% ± 6% was determined with TM versus UTM conditions (SNRUTM/SNRTM = 0.97 ± 0.08). Finally, we demonstrate the performance of the array in human brain using T2-weighted turbo spin echo imaging, finding excellent SNR performance in both caudal and rostral brain regions.

Evaluation of the microbiological efficacy of cleaning agents for tracheostomy inner cannulas.

PURPOSE: Biofilms are a significant cause of morbidity in patients with indwelling medical devices. Biofilms pose a potential risk with reusable inner cannulas by increasing the risk of infections. Effective decontamination is thus vital in decreasing bioburden. The current guidelines for cleaning inner cannulas are varied, with multiple techniques being recommended, which are not supported by strong evidence. This randomized, controlled, cross-over study attempted to enumerate the bacterial count of inner cannulas used in tracheostomy patients (n = 60) pre-and post-decontamination with detergent (A) or sterile water (B). MATERIALS AND METHODS: The patients were randomly allocated to sequence A > B or B > A in 1:1 fashion. The saline flushing of the inner cannulas was plated on trypticase soy agar with 5 % sheep blood to enumerate the bacterial count. RESULTS: The mean ratio [Log (CFU)post/Log (CFU)pre]A/[Log (CFU)post/Log (CFU)pre]B based on 53 samples was 0.918 ± 0.470, two-sided 90 % confidence interval (CI) 0.812, 1.024. The equivalence criterion was met as the mean ratio after cleaning fell within the equivalence region of 0.8 and 1.25. CONCLUSION: This study demonstrated the microbiological efficacy of both detergent and sterile water in the decontamination of inner cannulas, and that sterile water was not less effective than detergent in reducing the bacterial load for safe re-use of inner cannulas. This has the potential to promote cost savings for patients with tracheostomy, both in the hospital and the community. The study findings may also be relevant in formulating tracheostomy care policies.

Map-based B0 shimming for single voxel brain spectroscopy at 7T.

While B0 shimming is an important requirement for in vivo brain spectroscopy, for single voxel spectroscopy (SVS), the role for advanced shim methods has been questioned. Specifically, with the small spatial dimensions of the voxel, the extent to which inhomogeneities higher than second order exist and the ability of higher order shims to correct them is controversial. To assess this, we acquired SVS from two loci of neurophysiological interest, the rostral prefrontal cortex (rPFC; 8 cc) and hippocampus (Hc; 9 cc). The rPFC voxel was placed using SUsceptibility Managed Optimization (SUMO) and an initial B0 map that covers the entire cerebrum to cerebellum. In each location, we compared map-based shimming (Bolero) with projection-based shimming (FAST(EST)MAP). We also compared vendor-provided spherical harmonic first- and second-order shims with additional third- and fourth-order shim hardware. The 7T SVS acquisition used stimulated echo acquisition mode (STEAM) TR/TM/TE of 6 s/20 ms/8 ms, a tissue water acquisition for concentration reference, and LCModel for spectral analysis. In the rPFC (n = 7 subjects), Bolero shimming with first- and second-order shims reduced the residual inhomogeneity σ B 0 from 9.8 ± 4.5 Hz with FAST(EST)MAP to 6.5 ± 2.0 Hz. The addition of third- and fourth-order shims further reduced σ B 0 to 4.0 ± 0.8 Hz. In the Hc (n = 7 subjects), FAST(EST)MAP, Bolero with first- and second-order shims, and Bolero with first- to fourth-order shims achieved σ B 0 values of 8.6 ± 1.9, 5.6 ± 1.0, and 4.6 ± 0.9 Hz, respectively. The spectral linewidth, Δ v σ B 0 , was estimated with a Voigt lineshape using σ B 0 and T2 = 130 ms. Δ v σ B 0 significantly correlated with the Cramer-Rao lower bounds and concentrations of several metabolites, including glutamate and glutamine in the rPFC. In both loci, if the B0 distribution is well described by a Gaussian model, the variance of the metabolite concentrations is reduced, consistent with the LCModel fit based on a unimodal lineshape. Overall, the use of the high order and map-based B0 shim methods improved the accuracy and consistency of spectroscopic data.

Sesquiterpenes from Streptomyces qinglanensis and Their Cytotoxic Activity.

Nine sesquiterpenes, including eight pentalenenes (1-8) and one bolinane derivative (9), were isolated from the culture broth of a marine-derived actinobacterium Streptomyces qinglanensis 213DD-006. Among them, 1, 4, 7, and 9 were new compounds. Their planar structures were determined by spectroscopic methods (HRMS, 1D, and 2D NMR), and the absolute configuration was established by biosynthesis consideration and electronic-circular-dichroism (ECD) calculations. All the isolated compounds were screened for their cytotoxicity against six solid and seven blood cancer cell lines. Compounds 4-6 and 8 showed a moderate activity against all of the tested solid cell lines, with GI50 values ranging from 1.97 to 3.46 µM.

Sediment pollutant exposures caused hepatotoxicity and disturbed glycogenesis.

Liver metabolic syndrome, which involves impaired hepatic glycogen synthesis, is persistently increased by exposure to environmental pollutants. Most studies have investigated the pathogenesis of liver damage caused by single metal species or pure organics. However, under normal circumstances, the pollutants that we are exposed to are usually chemical mixtures that accumulate over time. Sediments are long-term repositories for environmental pollutants due to their environmental cycles, which make them good samples for evaluating the effect of environmental pollutants on the liver via bioaccumulation. This study aimed to clarify the effects of sediment pollutants on liver damage. Our results indicate that industrial wastewater sediment (downstream) is more cytotoxic than sediments from other zones. Downstream sediment extract (DSE) causes hepatotoxicity, stimulates reactive oxygen species (ROS) generation, triggers mitochondrial dysfunction, induces cell apoptosis, and results in the release of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) proteins. Additionally, to elucidate the underlying mechanism by which sediment pollutants disturb hepatic glycogen synthesis, we investigated the effects of different sediment samples from different pollution situations on glycogen synthesis in liver cell lines. It was found that DSE induced multiple severe impairments in liver cells, and disturbed glycogen synthesis more than under other conditions. These impairments include decreased hepatic glycogen synthesis via inhibition and insulin receptor substrate 1 (IRS-1) /AKT /glycogen synthase kinase3ß (GSK3ß)-mediated glycogen synthase (GYS) inactivation. To our knowledge, this study provides the first detailed evidence of in vitro sediment-accumulated toxicity that interferes with liver glycogen synthesis, leading to hepatic cell damage through apoptosis.

Writing Multiple Choice Questions-Has the Student Become the Master?

CONSTRUCT: We compared the quality of clinician-authored and student-authored multiple choice questions (MCQs) using a formative, mock examination of clinical knowledge for medical students. BACKGROUND: Multiple choice questions are a popular format used in medical programs of assessment. A challenge for educators is creating high-quality items efficiently. For expediency's sake, a standard practice is for faculties to repeat items in examinations from year to year. This study aims to compare the quality of student-authored with clinician-authored items as a potential source of new items to include in faculty item banks. APPROACH: We invited Year IV and V medical students at the University of Adelaide to participate in a mock examination. The participants first completed an online instructional module on strategies for answering and writing MCQs, then submitted one original MCQ each for potential inclusion in the mock examination. Two 180-item mock examinations, one for each year level, were constructed. Each consisted of 90 student-authored items and 90 clinician-authored items. Participants were blinded to the author of each item. Each item was analyzed for item difficulty and discrimination, number of item-writing flaws (IWFs) and non-functioning distractors (NFDs), and cognitive skill level (using a modified version of Bloom's taxonomy). FINDINGS: Eighty-nine and 91 students completed the Year IV and V examinations, respectively. Student-authored items, compared with clinician-authored items, tended to be written at both a lower cognitive skill and difficulty level. They contained a significantly higher rate of IWFs (2-3.5 times) and NFDs (1.18 times). However, they were equally or better discriminating items than clinician-authored items. CONCLUSIONS: Students can author MCQ items with comparable discrimination to clinician-authored items, despite being inferior in other parameters. Student-authored items may be considered a potential source of material for faculty item banks; however, several barriers exist to their use in a summative setting. The overall quality of items remains suboptimal, regardless of author. This highlights the need for ongoing faculty training in item writing.

Components of a Behavior Change Model Drive Quality of Life in Community-Dwelling Older Persons.

This study aimed to inform a measurement approach for older persons who wish to engage in active living such as participating in a walking program. The Patient Generated Index, an individualized measurement approach, and directed and summative content analyses were carried out. A sample size of 204 participants (mean age 75 years; 62% women) was recruited; it generated 934 text threads mapped to 460 unique categories within 45 domains with similarities and differences for women and men. The Capability, Opportunity, Motivation, and Behaviors Model best linked the domains. The results suggest that older persons identify the need to overcome impaired capacity, low motivation, and barriers to engagement to live actively. These are all areas that active living programs could address. How to measure the outcomes of these programs remains elusive.

N-Acetylaspartate and Choline Metabolites in Cortical and Subcortical Regions in Clinical High Risk Relative to Healthy Control Subjects: An Exploratory 7T MRSI Study.

N-acetylaspartate (NAA) and choline (Cho) are two brain metabolites implicated in several key neuronal functions. Abnormalities in these metabolites have been reported in both early course and chronic patients with schizophrenia (SCZ). It is, however, unclear whether NAA and Cho's alterations occur even before the onset of the disorder. Clinical high risk (CHR) individuals are a population uniquely enriched for psychosis and SCZ. In this exploratory study, we utilized 7-Tesla magnetic resonance spectroscopic imaging (MRSI) to examine differences in total NAA (tNAA; NAA + N-acetylaspartylglutamate [NAAG]) and major choline-containing compounds, including glycerophosphorylcholine and phosphorylcholine [tCho], over the creatine (Cre) levels between 26 CHR and 32 healthy control (HC) subjects in the subcortical and cortical regions. While no tCho/Cre differences were found between groups in any of the regions of interest (ROIs), we found that CHR had significantly reduced tNAA/Cre in the right dorsal lateral prefrontal cortex (DLPFC) compared to HC, and that the right DLPFC tNAA/Cre reduction in CHR was negatively associated with their positive symptoms scores. No tNAA/Cre differences were found between CHR and HC in other ROIs. In conclusion, reduced tNAA/Cre in CHR vs. HC may represent a putative molecular biomarker for risk of psychosis and SCZ that is associated with symptom severity.

Association between Hyperuricemia and Hearing Impairment: Results from the Korean National Health and Nutrition Examination Survey.

Background and Objectives: Hyperuricemia is associated with a variety of comorbidities. The objective of this study was to investigate the association between hyperuricemia and hearing impairment in Korean adults. Materials and Methods: Audiometric and laboratory test data from the 2019 to 2020 Korean National Health and Nutrition Examination Survey (KNHANES) were used for analysis. Hearing impairment was defined as a pure-tone average (0.5, 1, 2, 4 kHz) threshold level ≥ 41 decibels. The definition of hyperuricemia was different for males and females: >7 mg/dL for males vs. >6 mg/dL for females. Results: A total of 4857 (weight n = 17,990,725) subjects were analyzed. The mean age was 56.8 years old. The weighted prevalence was 12.1% for hyperuricemia and 2.5% for gout. The prevalence of hearing impairment was 13.4%. In the univariable analysis, hyperuricemia was significantly associated with hearing impairment. However, the diagnosis of gout was not associated with hearing impairment. In the multivariable analysis, hyperuricemia (odds ratios (OR): 1.41, 95% confidence interval [CI]: 1.03-1.92, p = 0.030) was associated with hearing impairment along with age (OR: 1.12, 95% CI: 1.10-1.14, p < 0.001), female sex (OR: 0.43, 95% CI: 0.34-0.64, p < 0.001), education (OR: 0.43, 95% CI: 0.30-0.63, p = 0.001), and occupational noise exposure (OR: 1.67, 95% CI: 1.25-2.22, p = 0.001). In the subgroup analysis, hyperuricemia was associated with hearing impairment in females (OR: 1.59, 95% CI: 1.02-2.48, p = 0.041) and the elderly aged 60 years or more (OR: 1.45, 95% CI: 1.05-1.99, p = 0.023). Conclusions: Hyperuricemia was independently associated with hearing impairment, especially in females and the elderly aged 60 years or more.

Roles of receptor-interacting protein kinase 1 in SH-SY5Y cells with beta amyloid-induced neurotoxicity.

Alzheimer's disease (AD), the major cause of dementia, affects the elderly population worldwide. Previous studies have shown that depletion of receptor-interacting protein kinase 1 (RIPK1) expression reverted the AD phenotype in murine AD models. Necroptosis, executed by mixed lineage kinase domain-like (MLKL) protein and activated by RIPK1 and RIPK3, has been shown to be involved in AD. However, the role of RIPK1 in beta-amyloid (Aß)-induced necroptosis is not yet fully understood. In this study, we explored the role of RIPK1 in the SH-SY5Y human neuroblastoma cells treated with Aß 1-40 or Aß 1-42. We showed that Aß-induced neuronal cell death was independent of apoptosis and autophagy pathways. Further analyses depicted that activation of RIPK1/MLKL-dependant necroptosis pathway was observed in vitro. We demonstrated that inhibition of RIPK1 expression rescued the cells from Aß-induced neuronal cell death and ectopic expression of RIPK1 was found to enhance the stability of the endogenous APP. In summary, our findings demonstrated that Aß can potentially drive necroptosis in an RIPK1-MLKL-dependent manner, proposing that RIPK1 plays an important role in the pathogenesis of AD.

Natural HSP90 inhibitors as a potential therapeutic intervention in treating cancers: A comprehensive review.

Heat shock protein 90 (Hsp90) has evolved as a cancerous cell growth regulator by stabilising various oncogenic kinases. Upon the Hsp90 inhibition, the expression of its client proteins is downregulated and thus leads to denaturation of cellular proteins and cancer cell death. Hsp90 inhibitors, particularly those naturally derived from plants, fungi and bacteria, have gained substantial interest as a feasible therapeutic approach for cancer treatment due to their diverse pharmacological properties. In order to gain insights into the potential development of more efficacious Hsp90 inhibitors for cancer treatment, this review is conducted to analyse both in vitro and in vivo data on the chemical and biological activities of natural Hsp90 inhibitors. The systematic search was conducted in databases (PubMed, Scopus and Web of Science) with terms "Hsp90 inhibitor" and "cancer", prompting a total of 61 articles after screening with inclusion criteria. This comprehensive review systematically summarised the efficacy of 14 different classes of naturally derived Hsp90 inhibitors in cancerous cell and animal tumour models by consolidating the primary outcomes in terms of IC50, reduction of tumour size and physicochemical properties. The detailed pharmacodynamic (the structure-activity relationship, mechanism of action) and pharmacokinetics (toxicity, oral bioavailability) of these Hsp90 inhibitors together with the study limitations were discussed. Collectively, these findings emphasise the necessity of comprehending the molecular mechanisms as well as the correlation of Hsp90 and its relative client proteins to drive the generation of viable Hsp90 inhibitors with improved pharmacodynamic and pharmacokinetic profiles.

EFFECT OF ORAL CARBONIC ANHYDRASE INHIBITOR ON CYSTOID MACULAR EDEMA ASSOCIATED WITH RETINITIS PIGMENTOSA: An OCT and OCT Angiography Study.

PURPOSE: To investigate the factors associated with visual improvement in response to oral carbonic anhydrase inhibitors (CAIs) and the occurrence of microvascular changes in patients with retinitis pigmentosa-associated cystoid macular edema (RP-CME). METHODS: This retrospective cohort study included 59 eyes from 39 patients with RP-CME who underwent at least 3 months of oral CAI treatment. The eyes were divided into responding and nonresponding groups based on optical coherence tomography (OCT) criteria (resolution of cyst and reduction of foveal or parafoveal volume). All eyes were assessed before and after treatment using OCT and OCT angiography. RESULTS: Thirty-three eyes (55.9%) demonstrated a positive response to treatment, and 26 eyes (44.1%) did not. Compared with nonresponding eyes, responding eyes had a significantly higher frequency of multilayer CME than CME limited to the inner nuclear layer ( P = 0.016). Subgroup analysis within the responding group revealed that improvements in visual acuity were more likely in eyes with fovea-involving CME and a higher baseline external limiting membrane and ellipsoid zone width. Microvascular parameters showed no significant changes after treatment. CONCLUSION: Eyes with CME extending to the outer nuclear layer or central fovea, and higher initial photoreceptor integrity may be prognostic factors associated with structural and functional improvements after carbonic anhydrase inhibitors treatment. Early treatment of multilayer CME with foveal involvement seems to be crucial in preventing irreversible photoreceptor damage.

MACULAR MICROVASCULATURE IN X-LINKED RETINOSCHISIS: Optical Coherence Tomography and Optical Coherence Tomography Angiography Study.

PURPOSE: The study aimed to evaluate the macular microvasculature of X-linked retinoschisis (XLRS) and identify correlations between vascular changes, structural changes, and functional outcome. METHODS: Genetically confirmed XLRS patients and heathy control subjects underwent complete ophthalmic examination, dilated funduscopic examination, optical coherence tomography, and optical coherence tomography angiography. Schisis distribution, outer plexiform layer discontinuation, photoreceptor layer thickness, and photoreceptor outer segment length were reviewed using optical coherence tomography. Vascular flow density and foveal thickness at foveal and parafoveal area were measured using optical coherence tomography angiography. RESULTS: A total of 17 eyes of 9 XLRS patients and 22 eyes of 11 control subjects were examined from July 2018 to August 2020. Flow density in the deep capillary plexus at foveal and parafoveal area decreased in XLRS patients compared with control subjects (P = 0.014 and 0.001, respectively), whereas foveal avascular zone area and perimeter remarkably increased (P = 0.015 and 0.001, respectively). Although outer and total retinal layers were significantly thicker in XLRS, inner retinal layer was thinner with reduced photoreceptor layer thickness and shortened photoreceptor outer segment length (P < 0.001 and P < 0.001, respectively). Foveal flow loss in deep capillary plexus, foveal avascular zone enlargement, thinner inner retina and photoreceptor layer thickness, and shortened photoreceptor outer segment length correlated with best-corrected visual acuity. CONCLUSION: X-linked retinoschisis eyes exhibit decreased flow density in the deep capillary plexus and variable foveal avascular zone with enlarged perimeter. Structural deterioration of the photoreceptor best reflects the degenerative changes, whereas microvascular alteration shows considerable correlation with functional outcome in XLRS.

Urban sediment pollutants alternate human cell essential behaviour through promoting oxidative damage.

The main objective of this study was to establish a human cell-based platform to assess the effects of sediment toxicity on oxidative damage and cell essential behaviour. Since sediment pollution has increased as a consequence of including but not limited to industrialisation, the contaminants accumulated in sediments have already led to human health concerns. The Hsinchu Science Park is one of the most prominent semiconductor manufacturing centres in the world, and the Ke-Ya River flows through Hsinchu Science Park and the Hsinchu urban district. Because semiconductor wastes potentially contribute to higher-than-normal rates of cancers, birth defects, and serious diseases, the quality assessment of the Ke-Ya River has prompted widespread concerns. While previous studies have shown an association between the degradation of fish populations and sediment pollutants, very little is known about the issues on human health. Herein, the effects of sediment from three sediment sampling sites of the Ke-Ya River on 11 different human cell lines were directly evaluated. The upstream represents the undeveloped zone, the middle-stream represents the household/industrial wastewater zone, and the downstream represents the accumulation zone. Our results indicated that the sediment pollution of the downstream Ke-Ya River was more cytotoxic than that of the middle stream and upstream. Downstream sediment extract (DSE) significantly increased reactive oxygen species (ROS) levels across all cell types. Accordingly, oxidative stress can trigger redox-sensitive pathways and alter essential biological processes such as cell viability, cell adhesion, and cell motility. Importantly, the MTT assay indicated that DSE significantly decreased the viability of brain, oral, lung, breast, liver, pancreatic, cervical, prostate, and colorectal cells. Furthermore, the adhesive ability and wound healing ability of most cells were greatly reduced in the presence of DSE compared to other conditions. Thus, this study shows the results of the first analyses completed on the sediment cytotoxicity in human cells, and stimulated ROS levels are crucial for cellular life. In future research, the detailed cause and effect mechanisms of the abundant ROS generated in DSE will be further investigated. We sincerely hope that our study provides a scientific basis for further investigations with a global perspective on public health challenges.

Sonogenetic-Based Neuromodulation for the Amelioration of Parkinson's Disease.

Sonogenetics is a promising strategy allowing the noninvasive and selective activation of targeted neurons in deep brain regions; nevertheless, its therapeutic outcome for neurodegeneration diseases that need long-term treatment remains to be verified. We previously enhanced the ultrasound (US) sensitivity of targeted cells by genetic modification with an engineered auditory-sensing protein, mPrestin (N7T, N308S). In this study, we expressed mPrestin in the dopaminergic neurons of the substantia nigra in Parkinson's disease (PD) mice and used 0.5 MHz US for repeated and localized brain stimulation. The mPrestin expression in dopaminergic neurons persisted for at least 56 days after a single shot of adeno-associated virus, suggesting that the period of expression was long enough for US treatment in mice. Compared to untreated mice, US stimulation ameliorated the dopaminergic neurodegeneration 10-fold and mitigated the PD symptoms of the mice 4-fold, suggesting that this sonogenetic strategy has the clinical potential to treat neurodegenerative diseases.

Dynamic B0 shimming for multiband imaging using high order spherical harmonic shims.

PURPOSE: To describe and implement a strategy for dynamic slice-by-slice and multiband B0 shimming using spherical harmonic shims in the human brain at 7T. THEORY: For thin axial slices, spherical harmonic shims can be divided into pairs of shims (z-degenerate and non-z-degenerate) that are spatially degenerate, such that only ½ of the shims (non-z-degenerate) are required for single slice optimizations. However, when combined, the pairs of shims can be used to simultaneously generate the same in-plane symmetries but with different amplitudes as a function of their z location. This enables multiband shimming equivalent to that achievable by single slice-by-slice optimization. METHODS: All data were acquired at 7T using a spherical harmonic shim insert enabling shimming up through 4th order with two additional 5th order shims (1st-4th+). Dynamic shim updating was achieved using a 10A shim power supply with 2 ms ramps and constrained optimizations to minimize eddy currents. RESULTS: In groups of eight subjects, we demonstrated that: 1) dynamic updating using 1st-4th+ order shims reduced the SD of the B0 field over the whole brain from 32.4 ± 2.6 and 24.9 ± 2 Hz with 1st-2nd and 1st-4th+ static global shimming to 15.1 ± 1.7 Hz; 2) near equivalent performance was achieved when dynamically updating only the non-z-degenerate shims (14.3 ± 1.5 Hz), or when a using multiband shim factor of 2, MBs = 2, and all shims (14.4 ± 2.0 Hz). CONCLUSION: High order spherical harmonics provide substantial improvements over static global shimming and enable dynamic multiband shimming with near equivalent performance to that of dynamic slice-by-slice shimming. This reduces distortion in echo planar imaging.

Proteomic and microbial assessments on the effect of Antrodia cinnamomea in C57BL/6 mice.

Antrodia cinnamomea (AC) is a nutraceutical fungus and studies have suggested that AC has the potential to prevent or alleviate diseases. However, little is known about the AC-induced phenotypes on the intestine-liver axis and gut microbial alterations. Here, we performed two-dimensional difference gel electrophoresis (2D-DIGE) and MALDI-Biotyper to elaborate the AC-induced phenotypes on the intestine-liver axis and gut microbial distribution of C57BL/6 mice. The experimental outcomes showed that the hepatic density may increase by elevating hepatic redox regulation, lipid degradation and glycolysis-related proteins and alleviating cholesterol biosynthesis and transport-related proteins in C57BL/6 mice with AC treatment. Moreover, AC facilitates intestinal glycolysis, TCA cycle, redox and cytoskeleton regulation-related proteins, but also reduces intestinal vesicle transport-related proteins in C57BL/6 mice. However, the body weight, GTT, daily food/water intake, and fecal/urine weight were unaffected by AC supplementation in C57BL/6 mice. Notably, the C57BL/6-AC mice had a higher gut microbial abundance of Alistipes shahii (AS) than C57BL/6-Ctrl mice. In summary, the AC treatment affects intestinal permeability by regulating redox and cytoskeleton-related proteins and elevates the gut microbial abundance of AS in C57BL/6 mice that might be associated with increasing hepatic density and metabolism-related proteins of the liver in C57BL/6 mice. Our study provides an insight into the mechanisms of AC-induced phenotypes and a comprehensive assessment of AC's nutraceutical effect in C57BL/6 mice.

Biomarker discovery in highly invasive lung cancer cell through proteomics approaches.

Lung cancer is one of the leading causes of cancer-related death worldwide. The most common type of lung cancer is non-small cell lung cancer (NSCLC). When NSCLC is detected, patients are typically already in a metastatic stage. Metastasized cancer is a major obstacle of effective treatment and understanding the mechanisms underlying metastasis is critical to treat cancer. Herein, we selected an invasive subpopulation from the human lung cancer cell line A549 using the transwell system and named it as A549-I5. Invasive and migratory activities of this cell line were analysed using wound healing, invasion, and migration assays. In addition, epithelial-mesenchymal transition (EMT) markers, such as Snail 1, Twist, Vimentin, N-cadherin and E-cadherin, were assessed through immunoblotting. In comparison to A549 cells, the invasive A549-I5 lung cancer cells had enhanced invasiveness, motility and EMT marker expression. Proteomic analysis identified 83 significantly differentially expressed proteins in A549-I5 cells. These identified proteins were classified according to their cellular functions and most were involved in cytoskeleton, redox regulation, protein degradation and protein folding. In summary, our results provide potential diagnostic markers and therapeutic candidates for the treatment of NSCLC metastasis. SIGNIFICANCE OF THE STUDY: When NSCLC is detected, most patients are already in a metastatic stage. Herein, we selected an invasive subpopulation from a human lung cancer cell line which had increased EMT markers as well as high wound healing, invasion and migration abilities. Proteomic analysis identified numerous proteins associated with functions in cytoskeleton, redox regulation, protein degradation and protein folding that were differentially expressed in these cells. These results may provide potential diagnostic markers and therapeutic candidates for the treatment of NSCLC metastasis.

Inactivation of the ß (1, 2)-xylosyltransferase and the α (1, 3)-fucosyltransferase gene in rice (Oryza sativa) by multiplex CRISPR/Cas9 strategy.

KEY MESSAGE: CRISPR/Cas9-mediated OsXylT and OsFucT mutation caused the elimination of plant-specific ß1,2-xylose and α1,3-fucose residues on glycoproteins in rice, which is the first report of OsXylT/OsFucT double KO mutation in rice. N-glycosylation pathway is the one of post-translational mechanism and is known as highly conserved in eukaryotes. However, the process for complex-N-glycan modification is different between mammals and plants. In plant-specific manner, ß1,2-xylose and α1,3-fucose residues are transferred to N-glycan core structure on glycoproteins by ß1,2-xylosyltransferase (ß1,2-XylT) and α1,3-fucosyltransferase (α1,3-FucT), respectively. As an effort to use plants as a platform to produce biopharmaceuticals, the plant-specific N-glycan genes of rice (Oryza sativa), ß1,2-xylT (OsXylT) and α1,3-FucT (OsFucT), were knocked out using multiplex CRISPR/Cas9 technology. The double knock-out lines were found to have frameshift mutations by INDELs. Both ß1,2-xylose and α1,3-fucose residues in the lines were not detected in Western blot analysis. Consistently, there was no peak corresponding to the N-glycans in MALDI-TOF/MS analysis. Although α1,3-fucose and ß1,2-xylose residues were not detected in the line, other plant-specific residues of ß1,3-galactose and α1,4-fucose were detected. Thus, we suggest that each enzymes working on the process for complex N-glycan biosynthesis might independently act in rice, hence the double knock-out of both OsXylT and OsFucT might be not enough to humanize N-glycan structure in rice.

Doxorubicin-Resistant TNBC Cells Exhibit Rapid Growth with Cancer Stem Cell-like Properties and EMT Phenotype, Which Can Be Transferred to Parental Cells through Autocrine Signaling.

Emerging evidence suggests that breast cancer stem cells (BCSCs), and epithelial-mesenchymal transition (EMT) may be involved in resistance to doxorubicin. However, it is unlear whether the doxorubicin-induced EMT and expansion of BCSCs is related to cancer dormancy, or outgrowing cancer cells with maintaining resistance to doxorubicin, or whether the phenotypes can be transferred to other doxorubicin-sensitive cells. Here, we characterized the phenotype of doxorubicin-resistant TNBC cells while monitoring the EMT process and expansion of CSCs during the establishment of doxorubicin-resistant MDA-MB-231 human breast cancer cells (DRM cells). In addition, we assessed the potential signaling associated with the EMT process and expansion of CSCs in doxorubicin-resistance of DRM cells. DRM cells exhibited morphological changes from spindle-shaped MDA-MB-231 cells into round-shaped giant cells. They exhibited highly proliferative, EMT, adhesive, and invasive phenotypes. Molecularly, they showed up-regulation of Cyclin D1, mesenchymal markers (ß-catenin, and N-cadherin), MMP-2, MMP-9, ICAM-1 and down-regulation of E-cadherin. As the molecular mechanisms responsible for the resistance to doxorubicin, up-regulation of EGFR and its downstream signaling, were suggested. AKT and ERK1/2 expression were also increased in DRM cells with the advancement of resistance to doxorubicin. Furthermore, doxorubicin resistance of DRM cells can be transferred by autocrine signaling. In conclusion, DRM cells harbored EMT features with CSC properties possessing increased proliferation, invasion, migration, and adhesion ability. The doxorubicin resistance, and doxorubicin-induced EMT and CSC properties of DRM cells, can be transferred to parental cells through autocrine signaling. Lastly, this feature of DRM cells might be associated with the up-regulation of EGFR.