Detecting and Tracking Hair Impurities in Mushroom Semi-product Images

A new area operator was proposed to extract axes feature of hair impurity in mushroom semi-product images. This operator was based on the local intensity comparison, and it had two advantages: 1) its outputs reached local maxima at the hair axes; 2) the accurate direction information of the hair axes could be derived by it. After the axes feature of the hairs was extracted using the proposed operator, an extended Kalman filter was applied to track the hairs. The searching path was established when a cost function was minimized. Starting points of the tracking path were found using dyadic wavelet analysis. The proposed algorithm had good adaptivity for the arbitrary orientation of the hairs. The experiment results showed that hairs could be tracked accurately. DOI: http://dx.doi.org/10.11591/telkomnika.v11i11.354

Expression of Ets-1, Ang-2 and maspin in ovarian cancer and their role in tumor angiogenesis

<p>Abstract</p> <p>Background</p> <p>Various angiogenic regulators are involved in angiogenesis cascade. Transcription factor Ets-1 plays important role in angiogenesis, remodeling of extracellular matrix, and tumor metastasis. Ets-1 target genes involved in various stages of new blood vessel formation include angiopoietin, matrix metalloproteinases (MMPs) and the protease inhibitor maspin.</p> <p>Methods</p> <p>We used immunohistochemistry (IHC) to detect the expression of Ets-1, angiopoietin-2 (Ang-2) and maspin in ovarian tumor and analyzed the relationship between the expression of these proteins and the clinical manifestation of ovarian cancer.</p> <p>Results</p> <p>Ets-1 expression was much stronger in ovarian cancer compared to benign tumors, but had no significant correlation with other pathological parameters of ovarian cancer. However, Ang-2 and maspin expression had no obvious correlation with pathological parameters of ovarian cancer. Ets-1 had a positive correlation with Ang-2 which showed their close relationship in angiogenesis. Although microvessel density (MVD) value had no significant correlation with the expression of Ets-1, Ang-2 or maspin, strong nuclear expression of maspin appeared to be correlated with high grade and MVD.</p> <p>Conclusions</p> <p>The expression of Ets-1, Ang2 and maspin showed close relationship with angiogenesis in ovarian cancer and expression of maspin appeared to be correlated with high grade and MVD. The mechanisms underlying the cross-talk of the three factors need further investigations.</p

Cucurbitacin B inhibits proliferation, induces G2/M cycle arrest and autophagy without affecting apoptosis but enhances MTT reduction in PC12 cells

In the present study, the effect of cucurbitacin B (a natural product with anti-cancer effect) was studied on PC12 cells. It significantly reduced the cell number, changed cell morphology and inhibited colony formation while MTT results showed increased cell viability. Cucurbitacin B treatment increased activity of succinode hydrogenase. No alteration in the integrity of mem-brane, the release of lactic dehydrogenase, the mitochondrial membrane potential, and the expression of apoptotic proteins suggested that cucurbitacin B did not induce apoptosis. The cell cycle was remarkably arrested at G2/M phase. Furthermore, cucurbitacin B induced autophagy as evidence by accumulation of autophagic vacuoles and the increase of LC3II. In addition, cucurbitacin B up-regulated the expression of p-beclin-1, p-ULK1, p-Wee1, p21 and down-regulated p-mTOR, p-p70S6K, CDC25C, CDK1, Cyclin B1. In conclusion, cucurbitacin B inhibited PC12 proliferation but caused MTT pitfall. Cucurbitacin B induced G2/M cell cycle arrest, autophagy, but not the apoptosis in PC12 cells

Neu-P11, a novel melatonin receptor agonist, could improve the features of type-2 diabetes mellitus in rats

Objective: Melatonin (Mel) and its receptors are promising for glycemic control in patients with type-2 diabetes mellitus (T2DM) and its complications, but there is significant heterogeneity among studies. This study aims to investigate the effects of Mel receptor agonist Neu-P11 on glucose metabolism, immunity and islet function in T2DM rats. Methods: In this study, SD rats were treated with high fat diet and streptozotocin (STZ) to establish T2DM model. Glucose oxidase method was used to measure blood glucose level. Glucose and insulin tolerance tests were used to assess glucose metabolism. HE staining was used to observe the pancreatic tissue injury. The apoptosis of islet β cells was analyzed by TUNEL and insulin staining. ROS levels and immune cell expression were analyzed by flow cytometry. IF was used to analyze the activation of microglia. The IgA, IgG, IgM, TNF-α, IL-10, IL-1β, IFN-γ, C-peptide and Insulin levels were determined by ELISA. The expression of CD11b, CD86, cleaved caspase3, p21, and P16 proteins were analyzed by western blot. Results: The results showed that the blood glucose level increased, insulin resistance occurred, spleen coefficient and ROS levels increased, humoral immunity in peripheral blood decreased, and inflammation increased in the model group compared to the control group. After Mel and Neu-P11 treatment, the blood glucose level decreased significantly, insulin sensitivity improved, spleen coefficient and ROS levels decreased, humoral immunity in peripheral blood enhanced, and inflammation improved in T2DM rats. Brain functional analysis of T2DM rats showed that microglia cells were activated, TNF-α and IL-β levels were increased, and IL-10 levels were decreased. Mel and Neu-P11 treatment reversed these indexes. Functional analysis of islet in T2DM rats showed that islet structure inflammation was impaired, islet β cells were apoptotic, p21 and p16 protein expressions were increased, and blood C-peptide and insulin were decreased. Mel and Neu-P11 treatment restored the function of pancreatic β cells and improved the damage of pancreatic tissue. Conclusion: Melatonin and its receptor Neu-P11 can reduce blood glucose level, enhance humoral and cellular immunity, inhibit microglia activation and inflammation, and repair islets β cell function, improve the characterization of T2DM related diseases

Determination of Phenolic Compounds from Lignin Decomposition Products in Marine Sediments by Ultra-High Performance Liquid Chromatography-High Resolution Mass Spectrometry

BACKGROUNDLignin is an important component of marine organic carbon. It is also an important biomarker for extracting information on the evolution of the land and marine environment and tracking the source of organic marine matter. However, the existing analytical techniques are difficult to determine lignin directly. So, the content of phenolic compounds in the decomposition products of lignin in marine sediments were generally determined to indicate the content of lignin and the source of organic matter. The content of phenolic compounds in the decomposition products of lignin in marine sediments is often used to reflect the content of lignin. In addition, by calculating the diagnostic ratio of individual phenolic compounds, it also provides important information about the classification, source, and diagenesis of terrestrial organic matter in marine sediments. However, phenolic compounds in the decomposition products of lignin have the characteristics of strong polarity and low volatility, so they cannot be directly detected by gas chromatography and need to be derivatized first, which makes the sample processing complicated and often results in incomplete derivatization. Therefore, it is of great significance to develop a simple and reliable method for determination of phenolic compounds of the lignin decomposition products in marine sediments to explore the source of organic matter and understand the environmental evolution process.OBJECTIVESTo establish a simple and reliable method for the determination of phenolic compounds of lignin decomposition products in marine sediments using solid phase extraction (SPE) combined with ultra-high performance liquid chromatography-high resolution mass spectrometry, and to trace the content level and source of lignin in the sediments of Laizhou Bay in China.METHODSMarine sediment samples were first decomposed with oxidative-alkaline CuO and extracted by solid phase extraction. Briefly, the oxidation was carried out in a polytetrafluoroethylene digestion tank. 1.00g of sediment sample, 500mg of copper oxide, and 100mg of ammonium ferrous sulfate were accurately weighed and placed in the tank. The components were thoroughly mixed with the sample and then the digestion tank was transferred to a glove box filled with nitrogen. 8.0mL of aqueous sodium hydroxide solution with a concentration of 8.0% (bubbled with N2 to remove dissolved oxygen) was added to the tank. The digestion tank was covered tightly and transferred to an oven heating to 150℃ for reaction, which was terminated after 3h. After the digestion tank cooled to room temperature, it was carefully unscrewed, and an internal standard (ethyl vanillin) solution was added. Subsequently, the hydrolysate was transferred to a centrifuge tube, spun at 8000r/min for 10min, and the supernatant and reaction residue was separated. 2.0mL of 1.0% sodium hydroxide solution was added to rinse the residue, and centrifuged at 8000r/min for 10min. Combining the centrifuged supernatant obtained twice, the solution was acidified to pH=1 with hydrochloric acid. After the solution was left to stand for 30 minutes, solid phase extraction was performed.　　The SPE procedure was as follows: A hydrophilic-lipophilic balance (HLB) SPE cartridge (200mg, 6mL) was conditioned with 5mL of methanol and 5mL of ultrapure water. Sample solution was passed through the cartridge in a flow rate 1.0mL/min, and then the cartridges were rinsed with 10mL water, and dried under vacuum for about 3min. Phenolic compounds were eluted with 10mL ethyl acetate, and were evaporated by a rotary evaporator, reconstituted with sample solvent. Then, ultra-high performance liquid chromatography using ZORBAX Eclipse XDB-C18 column with packing particle size of 1.8μm was used to directly separate all target compounds at 28℃, with gradient elution. The mobile phase was composed of ultrapure water with 0.1% formic acid (V/V) and acetonitrile/methanol (9:1, V/V) , and the flow rate was set to 0.25mL/min. Electrospray ionization (in positive) time of flight mass spectrometry was applied to detect target compounds in full scan mode, and quantification was performed using an internal standard determination.RESULTSFirstly, chromatographic conditions and solid phase extraction conditions were systematically optimized. Ultra-high performance liquid chromatography was used for the chromatographic separation of phenolic compounds from lignin decomposition products in marine sediments. The separation effects of three mobile phase systems, namely, water-acetonitrile, water- methanol, and water-methanol-acetonitrile, were compared. When using a water-methanol -acetonitrile ternary mobile phase system, the resolution of various phenolic compounds was superior to the commonly used water-acetonitrile or water-methanol binary mobile phase systems in the literature. In addition, the effects of mobile phase acidity (trifluoroacetic acid, formic acid, and acetic acid were added into the mobile phase) on the separation of various phenolic compounds were investigated. The results showed that adding a certain concentration of all three acids to the mobile phase provided better separation results. Considering the compatibility with mass spectrometry, it was finally determined that adding 0.1% formic acid into the mobile phase achieved good peak patterns and resolution.　　In order to determine the ionization mode suitable for the analysis of phenolic compounds from lignin decomposition products in marine sediment, electrospray ionization (ESI) mass spectrometry was performed on each target phenolic compound in ESI+ and ESI− mode, respectively. Under ESI+ mode, various target phenolic compounds were less affected by interfering substances in the sample matrix, and the MS response value for most of the phenolic compounds was higher than that found in ESI− mode. Hence, ESI-TOF/MS in positive mode was selected to determine phenolic compounds of lignin decomposition products in marine sediment. Subsequently, the fragmentation voltage was optimized to obtain the highest sensitivity for all target phenolic compounds, which was the main mass spectrometric condition that affected the quantification accuracy and sensitivity. The effect of fragmentation voltage on the MS response signal of each target phenolic compound was investigated in the range of 80V to 200V. Overall, considering the detection sensitivity of the [M+H]+ ion peak of each target compound, 130V was selected as the optimal fragmentation voltage to determine phenolic compounds of lignin decomposition products in marine sediment.　　The effect of pH (1.0-2.5) of the loading solution for solid phase extraction on the extraction efficiency of various target phenolic compounds was systematically investigated, to ensure that the phenolic compounds of lignin decomposition products in marine sediments have a good recovery rate during the SPE process. When the pH of the loading solution was 1.0 and 1.5, the recovery rate of various phenolic compounds by using HLB solid phase extraction column was significantly higher than that of the loading solution adjusted pH to 2.0 and 2.5. When the pH of the sample solution was 1.0 and 1.5, although the recoveries of syringaldehyde and acetovanillone were relatively similar, the recoveries of other phenolic compounds were the highest at a pH of 1.0. Considering the recovery rate of all the target phenolic compounds and applicability of the method, the pH of the sample solution was confirmed to adjust to 1.0. In this study, HLB SPE column with 200mg of packing material was used to enrich phenolic compounds in sample extraction solution. Generally, 5-10mL of eluting solvent can ensure the full elution of all target phenolic compounds adsorbed on the SPE column. Therefore, based on the results of literature research, ethyl acetate was finally selected as the eluting solvent, with a dosage of 10mL.　　Under the optimum experimental conditions, the 11 main decomposition phenol compounds of lignin in marine sediments were well separated within 20 minutes. The proposed method had good precision (RSD was less than 9.0%), the correlation coefficient (R2) was not less than 0.9989 in the linear range, and the recovery rate of all spiked phenol compounds in blank marine sediment was in the range of 86.8%-93.2%, thereby indicating that the developed method would be suitable to determine the target decomposition phenol compounds of lignin in marine sediment. Subsequently, the method was used to determine the phenolic compounds of lignin decomposition products in the surface sediments of Laizhou Bay. The detection rate of 11 target phenolic compounds in 12 surface sediment samples was 100%, and the concentration of Σ8 in 12 surface sediment samples ranged from 0.001mg/10gds to 0.019mg/10gds. The value of C/V was between 0.18 and 0.81, with an average of 0.38; the value of S/V was between 0.18 and 0.45, with an average of 0.26; PON/P value was between 0.01 and 0.07, with an average of 0.03; P/(V+S) value was between 0.55 and 3.77, with an average of 1.44; (Ad/Al)v value was between 0.12 and 1.07, with an average of 0.48; the value of (Ad/Al)s was between 0.15 and 1.26, with an average of 1.02.CONCLUSIONSThe above diagnostic ratios indicate that the lignin in the surface sediments of Laizhou Bay originate mainly from the herbaceous tissue of angiosperms, while the proportion of organic matter in vascular plants is relatively low. The degradation degree of terrestrial organic matter in most sampling stations is medium or high, but there is still a small amount of fresh plant debris. The proposed method has the advantages of high efficiency, simple for sample pretreatment, and is a powerful technique for the determination of main decomposition product phenolic compounds of lignin in marine sediments

Case Report: A case of ALS type 6 associated with a FUS gene variant and right limb muscle weakness and atrophy as the initial symptom

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive degeneration of upper and lower motor neurons. This degeneration results in increasing muscle weakness, ultimately culminating in respiratory failure and death. Mutations in the fused in sarcoma (FUS) gene have been identified as a significant cause of ALS. Here, we present the case of a 40-year-old woman who exhibited right limb muscle weakness and atrophy as her initial symptom. Whole genome sequencing revealed a mutation in the FUS gene, specifically c.1450_1456delinsCCC (p.Tyr484Profs*44), leading to a diagnosis of ALS type 6 (ALS6). The c.1450_1456delinsCCC (p.Tyr484Profs*44) mutation is a frameshift mutation resulting from a non-triplet base deletion in the coding region of the FUS gene. This mutation is novel and has not been previously reported in China or internationally. Furthermore, the onset of muscle weakness and atrophy exclusively in the ipsilateral limb is very rare among ALS patients, and we have found no related reports. This case report aims to enhance medical professionals’ understanding of the complexities associated with ALS caused by FUS gene mutations and the onset of ALS symptoms, thereby facilitating more accurate clinical diagnosis and treatment

The distributive and structural characteristics of bronchus-associated lymphoid tissue (BALT) in Bactrian camels (Camelus bactrianus)

Background Bronchus-associated lymphoid tissue (BALT), distributed in the bronchial mucosa, plays a critical role in maintaining the mucosal immune homeostasis of the lower respiratory tract. The bronchial tree is a functional structure for gas exchange with the outside environment and maintains basic lung morphology. Methods To explore the structural and distributive characteristics of BALT in Bactrian camels, twelve healthy adult Bactrian camels were divided into two groups (six in each group). The lungs, bronchial tree and BALT were observed and analysed systematically through anatomical and histological methods. Results The results showed that Bactrian camel lungs were constituted by the left cranial lobe, left caudal lobe, right cranial lobe, right caudal lobe and accessory lobe, but lacked the middle lobe. The cranial lobe was narrow and small, the caudal lobe was extremely developed (almost four times the cranial lobe in size), and the accessory lobe was smaller than the cranial lobe; the bronchial tree, an unequal dichotomy with a tracheobronchial branch, was composed of dorsal, ventral, lateral and medial bronchiole systems. Isolated lymphoid follicles (the chief type) and aggregates of lymphoid follicles revealed two types of BALT, and germinal centres, follicle-associated epithelium and high endothelial venules could be observed in some well-developed BALT. Additionally, BALT was scattered along the bronchial tree in the entire lung, and the density increased from the trachea to the lower graded branches (densest in the bronchioles) and then decreased, with the occasional location around respiratory bronchioles or among the pulmonary mesenchyme. In the conducting portion, BALT was primarily located in the mucosa lamina propria but was also found in the submucosa, under the muscular layer, and around the submucosal glands and cartilage. Conclusion The results demonstrated that the lung morphology of Bactrian camels was similar to that of horses, but the bronchial branches were more closely related to those of ruminants. These characteristics were in accordance with the morphological and structural variation regularity of lungs with species evolution. BALT was mainly scattered in the conducting portion, and bronchioles, as the final “checkpoint” in the surveillance, capture and recognition of antigens before pulmonary exchange, were the pivotal locational position of BALT. However, BALT at different depths of the bronchial wall of the conducting portion might be at different developmental stages. Our study provided evidence for further insight into the mucosal immunomodulatory mechanism of BALT in the respiratory system of Bactrian camels

The hepatocyte growth factor-expressing character is required for mesenchymal stem cells to protect the lung injured by lipopolysaccharide in vivo

Abstract Background Acute respiratory distress syndrome (ARDS) is a life-threatening condition in critically ill patients. Recently, we have found that mesenchymal stem cells (MSC) improved the permeability of human lung microvascular endothelial cells by secreting hepatocyte growth factor (HGF) in vitro. However, the properties and functions of MSC may change under complex circumstances in vivo. Here, we sought to determine the role of the HGF-expressing character of MSC in the therapeutic effects of MSC on ARDS in vivo. Methods MSC with HGF gene knockdown (MSC-ShHGF) were constructed using lentiviral transduction. The HGF mRNA and protein levels in MSC-ShHGF were detected using quantitative real-time polymerase chain reaction and Western blotting analysis, respectively. HGF levels in the MSC culture medium were measured by enzyme-linked immunosorbent assay (ELISA). Rats with ARDS induced by lipopolysaccharide received MSC infusion via the tail vein. After 1, 6, and 24 h, rats were sacrificed. MSC retention in the lung was assessed by immunohistochemical assay. The lung wet weight to body weight ratio (LWW/BW) and Evans blue dye extravasation were obtained to reflect lung permeability. The VE-cadherin was detected with inmmunofluorescence, and the lung endothelial cell apoptosis was assessed by TUNEL assay. The severity of lung injury was evaluated using histopathology. The cytokines and HGF levels in the lung were measured by ELISA. Results MSC-ShHGF with markedly lower HGF expression were successfully constructed. Treatment with MSC or MSC carrying green fluorescent protein (MSC-GFP) maintained HGF expression at relatively high levels in the lung at 24 h. MSC or MSC-GFP decreased the LWW/BW and the Evans Blue Dye extravasation, protected adherens junction VE-cadherin, and reduced the lung endothelial cell apoptosis. Furthermore, MSC or MSC-GFP reduced the inflammation and alleviated lung injury based on histopathology. However, HGF gene knockdown significantly decreased the HGF levels without any changes in the MSC retention in the lung, and diminished the protective effects of MSC on the injured lung, indicating the therapeutic effects of MSC on ARDS were partly associated with the HGF-expressing character of MSC. Conclusions MSC restores lung permeability and lung injury in part by maintaining HGF levels in the lung and the HGF-expressing character is required for MSC to protect the injured lung. </jats:sec

Overexpressing CsSABP2 enhances tolerance to Huanglongbing and citrus canker in C. sinensis

Huanglongbing (HLB) and citrus canker, arising from Candidatus Liberibacter asiaticus (CaLas) and Xanthomonas citri pv. Citri (Xcc), respectively, have been imposing tremendous losses to the global citrus industry. Systemic acquired resistance (SAR) has been shown to be crucial for priming defense against pathogen in citrus. Salicylic acid (SA) binding protein 2 (SABP2), which is responsible for converting methyl salicylate (MeSA) to SA, is essential for full SAR establishment. Here, we characterized the functions of four citrus SABP2 genes (CsSABP2-1, CsSABP2-1V18A, CsSABP2-2 and CsSABP2-3) against HLB and citrus canker. In vitro enzymatic assay revealed that all four proteins had MeSA esterase activities, and CsSABP2-1 and CsSABP2-1V18A has the strongest activity. Their activities were inhibited by SA except for CsSABP2-1V18A. Four genes controlled by a strong promoter 35S were induced into Wanjincheng orange (Citrus sinensis Osbeck) to generate transgenic plants overexpressing CsSABP2. Overexpressing CsSABP2 increased SA and MeSA content and CsSABP2-1V18A had the strongest action on SA. Resistance evaluation demonstrated that only CsSABP2-1V18A had significantly enhanced tolerance to HLB, although all four CsSABP2s had increased tolerance to citrus canker. The data suggested the amino acid Val-18 in the active site of CsSABP2 plays a key role in protein function. Our study emphasized that balancing the levels of SA and MeSA is crucial for regulating SAR and conferring broad-spectrum resistance to HLB and citrus canker. This finding offers valuable insights for enhancing resistance through SAR engineering