Cell Exudation Research Articles

Electroacupuncture in promoting neural repair after spinal cord injury: Inhibiting the Notch signaling pathway and regulating downstream proteins expression.

Spinal cord injury (SCI) is one of the serious central nervous system injuries and the incidence of SCI continues to increase. Previous studies have indicated that electroacupuncture (EA) is beneficial for promoting recovery after SCI. In the present study, we attempted to evaluate how EA can promote the neural repair in SCI model rats by observing changes in the Notch signaling pathway. Experimental rats were randomly divided into four groups. Each group had its own intervention period: 1 day, 7 days, 14 days, and 28 days, and five randomized subgroups: blank control (B) group, blank electroacupuncture (BE) group, sham operation (S) group, model control (M) group and EA group. Animals in the EA group and the BE group were treated with EA at Dazhui (GV14) and Mingmen (GV4) acupoints for 20 min. After the intervention period, the Basso-Beattie-Bresnahan (BBB) score was used to evaluate the neurological function. We found that BBB score increased in EA-treated groups. Hematoxylin and eosin staining was used to observe pathological changes in the injured spinal cord and the results showed that EA therapy could promote the repair of injured spinal cord tissue. Immunohistochemistry and Western blot methods were used to detect the expression of proteins Delta1, Presenilin1, Hes1, and Hes5 in the injured spinal cord. The results showed that the expression levels of Delta1, Presenilin1, Hes1, and Hes5 increased significantly after SCI and decreased after EA treatment. Our study suggested that the possible mechanism by which EA could benefit the recovery after SCI in rats may include inhibiting the Notch signaling pathway and regulating the downstream proteins expression. In addition, our study can provide reference for selecting acupoints and treatment cycle in the treatment of SCI.

Comparison of virulence-related determinants between the ST59-t437 and ST239-t030 genotypes of methicillin-resistant\xa0Staphylococcus aureus

BackgroundMethicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen for human infection. Hospital-acquired (HA) and community-acquired (CA) MRSA infections are serious clinical problems worldwide. In this study, we selected typical HA-MRSA strain and CA-MRSA isolates from our previous research and compared their phenotypic and pathogenic abilities both in vitro and in vivo.ResultsST59-t437-SCCmecIVa (YNSA7) and ST59-t437-SCCmecVb (YNSA53) belonged to two prevalent subclones of CA-MRSA, while ST239-t030-SCCmecIII (YNSA163) was an HA-MRSA epidemic clone in Southwest China. ST59-t437 strains demonstrated faster growth ability, higher survival rate resistance to human blood, and more toxin secretion levels and cytotoxicity than ST239-t030. The virulence and regulatory genes of hld, psm-α, RNAIII, agrA, and crtN were highly expressed on CA-MRSA isolates, especially the ST59-t437-SCCmecIVa subclone. However, the ST239-t030 strain had the strongest adhesion and biofilm ability among these MRSA bacteria. Animal experiments revealed the most serious lethal effect on BALB/c mice caused by the YNSA7 strain infection. The survival rates of BALB/c mice infected with the three MRSA strains were 16.7, 50.0 and 100.0% for YNSA7, YNSA53 and YNSA163, respectively. Histopathological analyses of infected animals indicated that the lungs were the most seriously damaged organs, especially for ST59-t437 MRSA. Severe inflammatory reactions, tissue destruction, and massive exudation of inflammatory mediators and cells could be identified in ST59-t437 strain-infected animals.ConclusionsIn general, ST59-t437 strains showed higher pathogenic ability than the ST239-t030 isolate, while ST239-t030 MRSA revealed the features prevalent in hospital settings, specifically for adhesion and biofilm ability.

Heterogeneous ice nucleation ability of aerosol particles generated from Arctic sea surface microlayer and surface seawater samples at cirrus temperatures

Abstract. Sea spray aerosol particles are a recognised type of ice-nucleating particles under mixed-phase cloud conditions. Entities that are responsible for the heterogeneous ice nucleation ability include intact or fragmented cells of marine microorganisms as well as organic matter released by cell exudation. Only a small fraction of sea spray aerosol is transported to the upper troposphere, but there are indications from mass-spectrometric analyses of the residuals of sublimated cirrus particles that sea salt could also contribute to heterogeneous ice nucleation under cirrus conditions. Experimental studies on the heterogeneous ice nucleation ability of sea spray aerosol particles and their proxies at temperatures below 235 K are still scarce. In our article, we summarise previous measurements and present a new set of ice nucleation experiments at cirrus temperatures with particles generated from sea surface microlayer and surface seawater samples collected in three different regions of the Arctic and from a laboratory-grown diatom culture (Skeletonema marinoi). The particles were suspended in the Aerosol Interaction and Dynamics in the Atmosphere (AIDA) cloud chamber and ice formation was induced by expansion cooling. We confirmed that under cirrus conditions, apart from the ice-nucleating entities mentioned above, also crystalline inorganic salt constituents can contribute to heterogeneous ice formation. This takes place at temperatures below 220 K, where we observed in all experiments a strong immersion freezing mode due to the only partially deliquesced inorganic salts. The inferred ice nucleation active surface site densities for this nucleation mode reached a maximum of about 5×1010 m−2 at an ice saturation ratio of 1.3. Much smaller densities in the range of 108–109 m−2 were observed at temperatures between 220 and 235 K, where the inorganic salts fully deliquesced and only the organic matter and/or algal cells and cell debris could contribute to heterogeneous ice formation. These values are 2 orders of magnitude smaller than those previously reported for particles generated from microlayer suspensions collected in temperate and subtropical zones. While this difference might simply underline the strong variability of the number of ice-nucleating entities in the sea surface microlayer across different geographical regions, we also discuss how instrumental parameters like the aerosolisation method and the ice nucleation measurement technique might affect the comparability of the results amongst different studies.

Bacillus cereus from refulgent corneal ulcer: A case report

A Gram-positive spore forming bacilli, aerobe or facultative anaerobe is . It is ubiquitous which disperse in the environment. is a potential pathogen accountable for fulminant human infectious disease and hardly ever contribute to eye infections. A 35-year-old farmer came to ophthalmology OPD on February 2019, and the patient complained of acute onset left eye pain with diminution of vision. There was history of eye injury by the vegetation (thorn). There was presence of corneal ulcer. The patient was not immunocompromised.Corneal scrapping was collected from left eye. On Gram staining from corneal scrapping only two Gram positive bacilli, with occasional pus cell and fibrinous exudate was seen. Culture on blood agar medium it showed beta haemolytic, about 2-5 mm in diameter, big flat, irregular edges and greyish white colonies which were oval, slightly granular but not dry. On Mac -Conkey agar medium it showed non lactose fermenting, big flat, irregular colonies.From colonies Gram staining showed Gram positive bacilli with spore. They were motile, catalase positive, oxidase positive, reduced nitrate to nitrite and were resistant to penicillin. The identification was also confirmed by Vitek. This case highlights the importance of cultivation, detection and antibiotic susceptibility test rather than treating patients with ocular infection empirically.

Examining the Effects of Microalgal Metabolites on Ciliary Activity of the Eastern Oyster Crassostrea virginica

Suspension-feeding bivalve molluscs have evolved a highly effective mechanism for particle discrimination, which allows them to process efficiently the material to which they are exposed. The mechanisms controlling this preingestive sorting process have been described as either passive or active. Evidence of a passive selection mechanism has been demonstrated in several species of suspension-feeding bivalves. In contrast, to date there has been no evidence that active selection mechanisms underlay particle selection in bivalves, although the possibility of such mechanisms has been hypothesized numerous times. The present study was designed to examine active selection on the gill of the eastern oyster Crassostrea virginica. The gill was chosen for study because in oysters this organ is involved in particle selection. Two in vivo assays were designed in which the gill was exposed to dissolved cell exudate or extracts from the microalga Tetraselmis chuii, and the transport of polystyrene microspheres (25 µm) by the frontal cilia of the ordinary filament quantified by means of video endoscopy. Results demonstrated that the addition of exudates or extracts of T. chuii cells had no significant effect on the percentage of particles being transported dorsally (likely ingested) or ventrally (more likely rejected), and no differences in the number of particles in the ventral groove between control and experimental treatments. Results of follow-up experiments using covalently bound neoglycoproteins commonly found on cell surfaces of microalgal species demonstrated differences in transport of captured microspheres (10 µm, carboxylated) depending on the sugar type. Although chemoreception cannot be completely ruled out, these findings further indicate that physicochemical properties of particles, and not an active behavioral or physiological response (i.e., chemoreception of dissolved metabolites) by the animal, mediate particle selection in oysters.

A Win-Loss Interaction on Fe0 Between Methanogens and Acetogens From a Climate Lake.

Diverse physiological groups congregate into environmental corrosive biofilms, yet the interspecies interactions between these corrosive physiological groups are seldom examined. We, therefore, explored Fe0-dependent cross-group interactions between acetogens and methanogens from lake sediments. On Fe0, acetogens were more corrosive and metabolically active when decoupled from methanogens, whereas methanogens were more metabolically active when coupled with acetogens. This suggests an opportunistic (win–loss) interaction on Fe0 between acetogens (loss) and methanogens (win). Clostridia and Methanobacterium were the major candidates doing acetogenesis and methanogenesis after four transfers (metagenome sequencing) and the only groups detected after 11 transfers (amplicon sequencing) on Fe0. Since abiotic H2 failed to explain the high metabolic rates on Fe0, we examined whether cell exudates (spent media filtrate) promoted the H2-evolving reaction on Fe0 above abiotic controls. Undeniably, spent media filtrate generated three- to four-fold more H2 than abiotic controls, which could be partly explained by thermolabile enzymes and partly by non-thermolabile constituents released by cells. Next, we examined the metagenome for candidate enzymes/shuttles that could catalyze H2 evolution from Fe0 and found candidate H2-evolving hydrogenases and an almost complete pathway for flavin biosynthesis in Clostridium. Clostridial ferredoxin-dependent [FeFe]-hydrogenases may be catalyzing the H2-evolving reaction on Fe0, explaining the significant H2 evolved by spent media exposed to Fe0. It is typical of Clostridia to secrete enzymes and other small molecules for lytic purposes. Here, they may secrete such molecules to enhance their own electron uptake from extracellular electron donors but indirectly make their H2-consuming neighbors—Methanobacterium—fare five times better in their presence. The particular enzymes and constituents promoting H2 evolution from Fe0 remain to be determined. However, we postulate that in a static environment like corrosive crust biofilms in lake sediments, less corrosive methanogens like Methanobacterium could extend corrosion long after acetogenesis ceased, by exploiting the constituents secreted by acetogens.

Multiplexing surface anchored functionalized iron carbide nanoparticle: A low molecular weight proteome responsive nano-tracer

Harvesting the low molecular weight (LMW) proteins from the cellular exudates is a big challenge for early disease detection. Here, we introduce a unique probe composed of surface-functionalized Fe2C NPs with different functional groups to harvest, identify and profile differentially expressed biomarker proteins. Three different functionalization of Fe2C NPs with Fe2C@NH2, Fe2C@COOH and Fe2C@PEG enabled to harvest 119 differentially expressed proteins from HeLa cell exudates. Among these proteins, 57 were LMW which 82.46 % were up-regulated and 17.54 % were down-regulated. The Fe2C@NH2 were able to separate 60S ribosomal proteins L7a, and L11, and leucine-rich repeat-containing protein 59. These proteins play a vital role in the maturation of large subunit ribosomal ribonucleic acid, mRNA splicing via spliceosome and cancer cell inhibitor, respectively. While, Fe2C@COOH identifies the 60S ribosomal protein types L7, 40S ribosomal protein S11, and 60S ribosomal protein L24. These proteins were important for large ribosomal subunit biogenesis, translational initiation, and assembly of large subunit precursor of pre-ribosome. Finally, the Fe2C@PEG extracted 40S ribosomal protein S2, splicing factor, arginine/serine-rich and 40S ribosomal protein S4, X isoform which were responsible for nonsense-mediated decay, oligodendrocyte differentiation and multicellular organism development. Thus, these results help us in defining oncogenic biomarkers for early disease detection.

Inflammation-Related Carcinogenesis: Lessons from Animal Models to Clinical Aspects.

Simple SummaryIn multicellular organisms, inflammation is the body’s most primitive and essential protective response against any external agent. Inflammation, however, not only causes various modern diseases such as cardiovascular disorders, neurological disorders, autoimmune diseases, metabolic syndrome, infectious diseases, and cancer but also shortens the healthy life expectancy. This review focuses on the onset of carcinogenesis due to chronic inflammation caused by pathogen infections and inhalation/ingestion of foreign substances. This study summarizes animal models associated with inflammation-related carcinogenesis by organ. By determining factors common to inflammatory carcinogenesis models, we examined strategies for the prevention and treatment of inflammatory carcinogenesis in humans.Inflammation-related carcinogenesis has long been known as one of the carcinogenesis patterns in humans. Common carcinogenic factors are inflammation caused by infection with pathogens or the uptake of foreign substances from the environment into the body. Inflammation-related carcinogenesis as a cause for cancer-related death worldwide accounts for approximately 20%, and the incidence varies widely by continent, country, and even region of the country and can be affected by economic status or development. Many novel approaches are currently available concerning the development of animal models to elucidate inflammation-related carcinogenesis. By learning from the oldest to the latest animal models for each organ, we sought to uncover the essential common causes of inflammation-related carcinogenesis. This review confirmed that a common etiology of organ-specific animal models that mimic human inflammation-related carcinogenesis is prolonged exudation of inflammatory cells. Genotoxicity or epigenetic modifications by inflammatory cells resulted in gene mutations or altered gene expression, respectively. Inflammatory cytokines/growth factors released from inflammatory cells promote cell proliferation and repair tissue injury, and inflammation serves as a “carcinogenic niche”, because these fundamental biological events are common to all types of carcinogenesis, not just inflammation-related carcinogenesis. Since clinical strategies are needed to prevent carcinogenesis, we propose the therapeutic apheresis of inflammatory cells as a means of eliminating fundamental cause of inflammation-related carcinogenesis.

The protective role of Zingerone in a murine asthma model via activation of the AMPK/Nrf2/HO-1 pathway.

Asthma is one of the most common illnesses associated with chronic airway inflammation; however, there are currently no effective therapies apart from glucocorticoids. Zingerone (ZIN), an active compound isolated from ginger, has been reported to have a broad spectrum of pharmacological properties. In this study, Zingerone was administrated to H2O2-stimulated mouse airway epithelial cell line MLE12 cells and asthmatic mice. The concentration of cytokines was evaluated using enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE), Periodic Acid-Schiff (PAS) and Masson staining were used for histological analyses. Protein levels in cells or lung tissues were determined using western blot, immunohistochemistry staining. The results showed that treatment with Zingerone dramatically inhibited oxidative stress and the inflammatory response in MLE12 cells stimulated with H2O2 and asthmatic mice. Furthermore, Zingerone treatment could decrease the expression of phosphorylated (p)-IκBα and p65 (nuclear) and increase the expression of phosphorylation of AMP-activated protein kinase (p-AMPK), nuclear factor erythroid-2-related factor 2 (Nrf2), and hemeoxygenase-1 (HO-1) to alleviate oxidative damage and inflammation both in vivo and in vitro. In addition, Zingerone treatment reduced the exudation and infiltration of inflammatory cells and suppressed goblet cell hyperplasia in a murine asthma model. Treatment with Zingerone also decreased the level of interleukin (IL)-4, IL-5, IL-13, and increased the level of interferon gamma (IFN-γ) in the BALF and attenuated airway hyperresponsiveness (AHR). However, inhibition of AMPK or Nrf2 suppressed the cellular protective, antioxidative, and anti-inflammatory properties of Zingerone. Taken together, these results demonstrate that Zingerone possesses the potential to relieve asthma via upregulating the AMPK/Nrf2/HO-1 signaling pathway.

Effects of Dexmedetomidine on Inflammatory Factors and Mitogen Activated Protein Kinase-Related Signaling Pathways of Alveolar Lavage Fluid in Mice with Lipopolysaccharide-Induced Acute Lung Injury

To investigate the effects and mechanisms of Dexmedetomidine on inflammatory factors and mitogenactivated protein kinase related signaling pathways in alveolar lavage fluid of mice with acute lung injury induced by lipopolysaccharide. 36 clean-grade male Kunming mice were randomly divided into blank control group (normal control group), lipopolysaccharide-induced acute lung injury model group, and lipopolysaccharide+dexmedetomidine group, with 12 mice in each group. The lipopolysaccharide group mice were intraperitoneally injected with 0.1 ml 5 mg kg-1 Lipopolysaccharide, lipopolysaccharide+Dexmedetomidine mice were injected with 25 μg kg-1. Dexmedetomidine intraperitoneally with 0.1 ml and 1 h later with 5 mg kg-1. Lipopolysaccharide injected with 0.1 ml intraperitoneally and the blank control group mice were intraperitoneally injected with normal saline of the same volume as the lipopolysaccharide group. All mice in each group were sacrificed after intraperitoneal injection for 6 h, and their lung tissues were separated by thoracotomy and their bronchoalveolar lavage fluid was recovered for subsequent detection. Analysis groups of mice lung tissue pathology, compare groups of mice lung tissue pathology score, lung wet weight/dry weight (W)/(D), mitogen-activated protein kinase pathways (p38 lightning, extracellular signal regulating kinase, c-jun amino terminal kinase) and bronchoalveolar lavage fluid protein expression level of tumor necrosis factor alpha beta, interleukin-1β, myeloperoxidase, protein concentration. In the blank control group, lung tissue structure was basically normal and alveolar structure was intact. lipopolysaccharide group mice had obvious lung tissue damage, and inflammatory cells appeared in the alveoli, with significantly increased exudation of red blood cells and proteins. Lipopolysaccharide+Dexmedetomidine group mice lung tissue damage significantly improved, inflammatory cell infiltration decreased, and red blood cell, protein exudation significantly decreased. Lung histopathological scores and bronchoalveolar lavage fluid levels of tumor necrosis factor alpha, interleukin- 1β and myeloperoxidase in lipopolysaccharide group were significantly higher than those in blank control group (p<0.05). The lung histopathological scores and the levels of tumor necrosis factor alpha, interleukin- 1β and myeloperoxidase in bronchoalveolar lavage fluid of lipopolysaccharide+Dexmedetomidine group were significantly lower than those of lipopolysaccharide group (p<0.05). The protein concentration in W/D and bronchoalveolar lavage fluid in lipopolysaccharide group was significantly higher than that in blank control group (p<0.05). Protein concentration in W/D and bronchoalveolar lavage fluid of lipopolysaccharide+Dexmedetomidine group was significantly lower than that of lipopolysaccharide group (p<0.05). The protein expression levels of extracellular signal regulating kinase, c-jun amino terminal kinase and P-P38 in THE lipopolysaccharide group were significantly higher than those in the blank control group (p<0.05). The protein expression levels of extracellular signal regulating kinase, c-jun amino terminal kinase and P-P38 in the lipopolysaccharide+Dexmedetomidine group were significantly lower than those in the lipopolysaccharide group (p<0.05). There was no significant difference in extracellular signal regulating kinase, c-jun amino terminal kinase and p38 protein expression levels in each group (p>0.05). Dexmedetomidine can significantly alleviate acute lung injury induced by lipopolysaccharide in mice, and its mechanism may be realized by blocking the activation of mitogen-activated protein kinaserelated signaling pathway and thereby reducing inflammatory response, providing a new target for the treatment of sepsis induced acute lung injury in the futurey.

Canine Olfactory Detection of a Non-Systemic Phytobacterial Citrus Pathogen of International Quarantine Significance.

For millennia humans have benefitted from application of the acute canine sense of smell to hunt, track and find targets of importance. In this report, canines were evaluated for their ability to detect the severe exotic phytobacterial arboreal pathogen Xanthomonas citri pv. citri (Xcc), which is the causal agent of Asiatic citrus canker (Acc). Since Xcc causes only local lesions, infections are non-systemic, limiting the use of serological and molecular diagnostic tools for field-level detection. This necessitates reliance on human visual surveys for Acc symptoms, which is highly inefficient at low disease incidence, and thus for early detection. In simulated orchards the overall combined performance metrics for a pair of canines were 0.9856, 0.9974, 0.9257 and 0.9970, for sensitivity, specificity, precision, and accuracy, respectively, with 1–2 s/tree detection time. Detection of trace Xcc infections on commercial packinghouse fruit resulted in 0.7313, 0.9947, 0.8750, and 0.9821 for the same performance metrics across a range of cartons with 0–10% Xcc-infected fruit despite the noisy, hot and potentially distracting environment. In orchards, the sensitivity of canines increased with lesion incidence, whereas the specificity and overall accuracy was >0.99 across all incidence levels; i.e., false positive rates were uniformly low. Canines also alerted to a range of 1–12-week-old infections with equal accuracy. When trained to either Xcc-infected trees or Xcc axenic cultures, canines inherently detected the homologous and heterologous targets, suggesting they can detect Xcc directly rather than only volatiles produced by the host following infection. Canines were able to detect the Xcc scent signature at very low concentrations (10,000× less than 1 bacterial cell per sample), which implies that the scent signature is composed of bacterial cell volatile organic compound constituents or exudates that occur at concentrations many fold that of the bacterial cells. The results imply that canines can be trained as viable early detectors of Xcc and deployed across citrus orchards, packinghouses, and nurseries.

The progression of pulmonary arterial hypertension induced by monocrotaline is characterized by lung nitrosative and oxidative stress, and impaired pulmonary artery reactivity

The time-course of pulmonary arterial hypertension in the monocrotaline (MCT) model was investigated. Male rats were divided into two groups: MCT (received a 60 mg/kg i.p. injection) and control (received saline). The MCT and control groups were further divided into three cohorts, based on the follow-up interval: 1, 2, and 3 weeks. Right ventricle (RV) catheterization was performed and RV hypertrophy (RVH) was estimated. The lungs were used for biochemical, histological, molecular, and immunohistochemical analysis, while pulmonary artery rings were used for vascular reactivity. MCT promoted lung perivascular edema, inflammatory cells exudation, greater neutrophils and lymphocytes profile, and arteriolar wall thickness, compared to CTR group. Increases in pulmonary artery pressure and in RVH were observed in the MCT 2- and 3-week groups. The first week was marked by the presence of nitrosative stress (50% moderate and 33% accentuated staining by nitrotyrosine). These alterations lead to an adaptation of NO production by NO synthase activity after 2 weeks. Oxidative stress was evident in the third week, probably by an imbalance between endothelin-1 receptors, resulting in extracellular matrix remodeling, endothelial dysfunction, and RVH. Also, it was found a reduced pulmonary arterial vasodilatory response to acetylcholine after 2 (55%) and 3 (45%) weeks in MCT groups. The relevance of this study is precisely to show that nitrosative and oxidative stress predominate in distinct time windows of the disease progression.

Impaired spermatogenesis in COVID-19 patients.

BackgroundThe current study aimed to determine the impact of SARS-CoV-2 infection on male fertility.MethodsThis is a single-center, hospital-based observational study that included autopsied testicular and epididymal specimens of deceased COVID-19 male patients (n=6) and recruited recovering COVID-19 inpatients (n=23) with an equal number of age-matched controls, respectively. We performed histopathological examinations on testicular and epididymal specimens, and also performed TUNEL assay and immunohistochemistry. Whereas, we investigated the semen specimen for sperm parameters and immune factors.FindingsAutopsied testicular and epididymal specimens of COVID-19 showed the presence of interstitial edema, congestion, red blood cell exudation in testes, and epididymides. Thinning of seminiferous tubules was observed. The number of apoptotic cells within seminiferous tubules was significantly higher in COVID-19 compared to control cases. It also showed an increased concentration of CD3+ and CD68+ in the interstitial cells of testicular tissue and the presence of IgG within seminiferous tubules. Semen from COVID-19 inpatients showed that 39.1% (n=9) of them have oligozoospermia, and 60.9% (n=14) showed a significant increase in leucocytes in semen. Decreased sperm concentration, and increased seminal levels of IL-6, TNF-α, and MCP-1 compared to control males were observed.InterpretationImpairment of spermatogenesis was observed in COVID-19 patients, which could be partially explained as a result of an elevated immune response in testis. Additionally, autoimmune orchitis occurred in some COVID-19 patients. Further research on the reversibility of impairment and developing treatment are warranted.FundingThis study was supported by Ministry of Science and Technology of China Plan, Hubei Science and Technology Plan, National Key Research and Development Program of China, HUST COVID-19 Rapid Response Call, China and National Natural Science Foundation of China; these funding bodies are public institutions, and they had no role in study conception, design, interpretation of results, and manuscript preparation.

Role of Human 15-Lipoxygenase-2 in the Biosynthesis of the Lipoxin Intermediate, 5S,15S-diHpETE, Implicated with the Altered Positional Specificity of Human 15-Lipoxygenase-1.

The oxylipins, 5S,12S-dihydroxy-6E,8Z,10E,14Z-eicosatetraenoic acid (5S,12S-diHETE) and 5S,15S-dihydroxy-6E,8Z,11Z,13E-eicosatetraenoic acid (5S,15S-diHETE), have been identified in cell exudates and have chemotactic activity toward eosinophils and neutrophils. Their biosynthesis has been proposed to occur by sequential oxidations of arachidonic acid (AA) by lipoxygenase enzymes, specifically through oxidation of AA by h5-LOX followed by h12-LOX, h15-LOX-1, or h15-LOX-2. In this work, h15-LOX-1 demonstrates altered positional specificity when reacting with 5S-HETE, producing 90% 5S,12S-diHETE, instead of 5S,15S-diHETE, with kinetics 5-fold greater than that of h12-LOX. This is consistent with previous work in which h15-LOX-1 reacts with 7S-HDHA, producing the noncanonical, DHA-derived, specialized pro-resolving mediator, 7S,14S-diHDHA. It is also determined that oxygenation of 5S-HETE by h15-LOX-2 produces 5S,15S-diHETE and its biosynthetic kcat/KM flux is 2-fold greater than that of h15-LOX-1, suggesting that h15-LOX-2 may have a greater role in lipoxin biosynthesis than previously thought. In addition, it is shown that oxygenation of 12S-HETE and 15S-HETE by h5-LOX is kinetically slow, suggesting that the first step in the in vitro biosynthesis of both 5S,12S-diHETE and 5S,15S-diHETE is the production of 5S-HETE.

Protective effect of Shenfu injection on the endothelium of severe sepsis by inhibiting CD11b+ cell paralysis induced by high mobility group protein B1

To investigate the relationship between the biochemical parameters, the pulmonary pathologic injury and the immune mechanism of severe sepsis in infant porcine, and the intervention effect of Shenfu injection. Panamanian infant porcine (2-3 months old) were divided into sham operation group (Sham group; intravenous injection of normal saline), lipopolysaccharide (LPS) induced severe sepsis model group (LPS group; intravenous injection of LPS 1 mg/kg, and continuing at 0.5 mg×kg-1×h-1 for 12 hours), and Shenfu injection intervention group (SF group; intravenous injection of Shenfu injection 10 mL/kg at the same time of modeling, twice a day) according to the random number table method, with 5 in each group. Forty-eight hours after the challenge, the changes of C-reactive protein (CRP), procalcitonin (PCT), oxygenation index (PaO2/FiO2), base excess (BE), blood lactate (Lac) and other biochemical indexes were detected with blood sampling; the number of neutrophils [myeloperoxidase positive (MPO+)] and their activated subsets CD11b+CD64+, M1 macrophages (CD80+CD64+), CD4+ and CD8+ T cells were analyzed in peripheral blood by flow cytometry. The levels of plasma cytokines were detected by enzyme linked immunosorbent assay (ELISA). The pathological damage of lung tissue was observed by hematoxylin-eosin (HE) staining. The mRNA expression of lung injury related molecules and their receptors, chemokines, cytokines, vascular endothelial related molecules and tight junction protein were detected by reverse transcription-polymerase chain reaction (RT-PCR). Compared with Sham group, the levels of CRP, PCT and Lac in LPS group significantly increased, and PaO2/FiO2 and BE significantly decreased. In the peripheral blood, CD80+CD64+ macrophages, CD11b+CD64+ and MPO+ neutrophils, CD4+, CD8+ T cells and tumor necrosis factor-α (TNF-α) significantly decreased, but interleukin-10 (IL-10) and vascular endothelial cell growth factor (VEGF) significantly increased; the mRNA expressions of high mobility group protein B1 (HMGB1), receptor for advanced glycation end products (RAGE), angiopoietin 2/angiopoietin 1 (Ang2/Ang1) significantly increased, Toll like receptor 9 (TLR9), chemokines (CXCL9 and CXCL10), TNF-α, IL-27, Tek tyrosine kinase 2 (TIE2), vascular endothelial cadherin (VE-CAD) and Occludin significantly decreased in lung tissue. HE staining showed inflammatory cell infiltration and exudation in the alveolar cavity, alveoli consolidation, thickening of the alveolar interstitial layer and emphysema. Compared with LPS group, Lac in SF group significantly decreased (mmol/L: 4.2±1.0 vs. 6.3±1.1, P < 0.05), while BE and PaO2/FiO2 significantly increased [BE (mmol/L): -6.4±2.6 vs. -11.6±2.5, PaO2/FiO2 (mmHg, 1 mmHg = 0.133 kPa): 180±36 vs. 105±35, both P < 0.05]; the percentage of CD80+CD64+ macrophages, CD11b+CD64+ and MPO+ neutrophils significantly increased [CD80+CD64+: (7.13±2.01)% vs. (3.80±0.46)%, CD11b+CD64+: (8.33±2.55)% vs. (2.15±0.47)%, MPO+: (21.22±2.33)% vs. (8.31±0.46)%, all P < 0.05]; the mRNA expressions of HMGB1 and RAGE decreased to some extent [HMGB1 mRNA (2-ΔΔCT): 1.81±0.45 vs. 2.23±0.85, RAGE mRNA (2-ΔΔCT): 6.69±3.48 vs. 11.60±6.91, both P < 0.05], the mRNA expressions of CXCL9 and TIE2 increased to a certain extent [CXCL9 mRNA (2-ΔΔCT): 1.06±0.63 vs. 0.50±0.12, TIE2 mRNA (2-ΔΔCT): 1.42±0.68 vs. 0.27±0.16, both P < 0.05]; the pathological damage of lung tissue were significantly alleviated. The increased abnormality of BE, Lac, and PaO2/FiO2, the immune exhausting and paralysis may be important factors leading to the fatal lung injury in infant porcine with severe sepsis. The possible mechanism is that the excessive activation of HMGB1 and its receptor RAGE, the suppression of TLR9 and corresponding chemokine and inflammatory factors lead to increased endothelial damage and decreased tight connection, which in turn induces capillary leakage. The intervention of immune disorders by Shenfu injection in the severe pneumonia accompanied by sepsis may be related to elevated M1 macrophages and activated neutrophils in the peripheral blood, inhibition of HMGB1 and its receptor RAGE mRNA expression, and elevated CXCL9 and TIE2 expression.

Localization of Pasteurella multocida antigens in the brains of pigs naturally infected with Pasteurellosis revealing a newer aspect of pathogenesis

Pasteurella multocida is an economically important respiratory pathogen of pigs confronting swine industry worldwide. Despite extensive research over the decades, its pathogenesis is still poorly understood. Recent reports have demonstrated the nervous system affection as a newer aspect of pathogenesis by Pasteurella multocida type B:2 in Haemorrhagic Septicemia, but there are no reports of the involvement of nervous system by P. multocida in pigs. Therefore, the study was aimed to explore the neurovirulence of Pasteurella multocida in naturally infected pigs. A total of 15 brains were collected from the natural cases of pig mortality suggestive of Pasteurellosis. Grossly, the leptomeninges were markedly congested and brains were oedematously swollen. Histologically, there was moderate to severe fibrinohaemorrhagic and mononuclear cells exudates present in the leptomeningeal tissue and cerebrospinal spaces. Similar vascular inflammatory lesions (perivascular and perineuronal) along with gliosis, neuronal degeneration and necrosis were noted in various subanatomical sites of the brain (cerebrum, cerebellum, brainstem and spinal cord). The culture and biochemical tests showed the presence of P. multocida within the brain tissue. P. multocida type specific antibody staining in the brain tissues revealed intense distribution of antigens in the inflammatory exudates of meningeal vessels, neurons, glial cells and endothelial cells of the blood vessels contributing its association with neuropathological lesions. Pasteurella multocida specific PCR amplification of capsular polysaccharide gene yielded 460 bp and multiplex PCR showed the involvement of capsular serogroups A &D. All the isolates showed the presence of 10 genes for virulence factors. The disease confirmation of both serotypes was proven by Koch's postulates using Swiss albino mice. Further, histopathological brain lesions along with the immunohistochemical detection of bacterial antigens were corroborated with natural cases of P. multocida as described above. To the best of our knowledge, we first time report the neuroinvasion of P. multocida in naturally infected pigs.

Role of miR-128, Sp1 and CDKN1A mRNA in Repairing Muscle Injury in a Rat Skeletal Muscle Injury Model

In order to detect the changes in expression of miR-128 during skeletal muscle repair in a rat model of muscle injury and to provide a basis for treating skeletal muscle injury, a rat skeletal muscle injury model was established. Rats were randomly divided into the quiet control group, 0 h after exercise group, 6 h after exercise group, 12 h after exercise group, 24 h after exercise group, 48 h after exercise group, 72 h after exercise group, 1 w after exercise group and 2 w after exercise groups to explore the expression pattern of miR-128, Sp1, CDKN1A during skeletal muscle contusion repair. Results showed that muscle fibers in the control group were polygonal, closely arranged and regular in shape. The myocyte membrane was intact and the nucleus was located under the myometrium. At 0 h after exercise, cells were swollen and round, cell volume increased and the cell gap increased. At 6, 12, 24 and 48 h after exercise, the cells continued to swell and the intercellular space became smaller than that in group C. In the 72 h after exercise group, cell swelling was very high and there were a large number of infiltrated inflammatory cells and exudation of blood cells. In 1 w after exercise group, inflammatory cell infiltration decreased since exercise. When it reaches to 2w, the morphology and structure of the cells return to normal, and the repair process was completed. Successful establishment of the rat skeletal muscle repair model, it is found that the expression of muscle-specific miR-128 did not change significantly during the process of muscle repair. miR-128 might be involved in the process of regeneration after skeletal muscle injury.

Evaluation of lipopolysaccharide-induced acute lung injury attenuation in mice by Glycyrrhiza glabra

Objective: The medicinal importance of Glycyrrhiza glabra is well reported in Asian folk core medicine. In the current study, we have investigated the treatment effects of ethanolic extract of G. glabra in acute lung injury (ALI) mice models. Materials and Methods: ALI murine models were established by intratracheal instillation of bacterial lipopolysaccharide (LPS). Bronchoalveolar lavage fluid (BALF) was prepared for measuring cell migration and cell count, protein content, and superoxidase dismutase (SOD) activity. Lung tissues were collected for evaluating the wet-to-dry weight ratio, messenger ribonucleic acid (mRNA) expression of pro-inflammatory cytokine, and histological change. Results: It was observed that the treatment of ALI mice significantly decreased the total cell count and exudation of protein into BALF. Further, SOD activity in BALF of LPS-induced ALI mice was greatly improved when treated with G. glabra extract at 200 or 400 mg/kg of body weight, and the SOD activity increased in a dose-response manner. Evaluation of lung wet-to-dry weight ratio, pro-inflammatory mRNA expression levels, and histologic examination of lung tissues indicates that plant extract has significantly attenuated the tissue injury. Conclusions: The results showed that G. glabra had a protective effect on LPS-induced ALI in mice which might be contributed due to diverse phytoconstituents of the plant extract. To fulfill the unmet needs of treating ALI, the active principles in G. glabra could be promising lead-alternative therapy to develop high-potent anti-inflammation agents for ALI.

Mitochondrial Coenzyme Q Protects Sepsis-Induced Acute Lung Injury by Activating PI3K/Akt/GSK-3β/mTOR Pathway in Rats.

The aim of our study was to assess the effects of mitochondrial coenzyme Q (MitoQ) on sepsis-induced acute lung injury (ALI) and investigate its possible mechanisms. The cecal ligation and puncture (CLP) method was used to establish a septic ALI model. Rats were randomly divided into Con group, CLP group, MitoQ group, and MitoQ + LY294002 group. The survival rate of the rats was recorded, and the survival rate curve was plotted. Moreover, the ratio of wet/dry weight (W/D) in lung tissue was measured. The activity of myeloperoxidase (MPO) was measured by using the MPO colorimetric activity assay kit. The levels of high-mobility group box 1 (HMGB1) and interleukin-6 (IL-6), macrophage inflammatory protein 2 (MIP2), and keratinocyte chemoattractant (KC) were analyzed by ELISA. The histopathological changes were measured by HE staining, and the lung injury was scored. TUNEL assay was applied to detect the apoptotic cells in lung tissue. The protein expressions were detected by western blot. MitoQ increased the survival rate and alleviated pulmonary edema in septic ALI rats. In addition, MitoQ inhibited the MPO activity and decreased the levels of HMGB1 and IL-6. After treatment with MitoQ, alveolar wall edema, inflammatory cell infiltration, and red blood cell exudation were relieved. MitoQ inhibited cell apoptosis in lung tissue of septic ALI rats. Meanwhile, MitoQ treatment remarkedly increased the expression of p-Akt, p-GSK-3β, and p-mTOR but decreased Bax, caspase-3, caspase-9, Beclin-1, and LC-3II/LC-3I. The effects of MitoQ were significantly reversed by the PI3K inhibitor (LY294002). Our study demonstrated that MitoQ could protect sepsis-induced acute lung injury by activating the PI3K/Akt/GSK-3β/mTOR pathway in rats.

POTENSI RIPASUDIL SEBAGAI TERAPI TERBARUKAN UNTUK MENGATASI UVEITIS GLAUCOMA MELALUI INHIBISI RHO KINASE PATHWAY (ROCK)

Uveitis is an inflammatory disease that accounts for 300 thousand new cases of blindness each year in America, and is responsible for 5-10% of vision changes worldwide. First-line therapy for uveitis is topical corticosteroids. Unfortunately, corticosteroid use is a risk factor for glaucoma. About 20-40% of patients experience an increase in pathological intra-ocular pressure that requires secondary glaucoma therapy. To reduce this pressure, the first therapy is beta-blockers and adrenergic agonists. But in inflammatory conditions, therapeutic side effects decrease. Over the years, a new glaucoma therapy agent, Ripasudil, has emerged. Research shows that Ripasudil can not only reduce intraocular pressure in glaucoma, but also has the anti-inflammatory effect needed to treat uveitis. The study of the potential of Ripasudil using literature obtained from the database publications of the journal Pubmed, Clinical Trials and Science Direct. Supporting articles are sought based on specific keywords by the inclusion criteria of articles won in the UK, publications of the past 10 years (except basic science), EBM articles level 1-3 and support the discussion of literature review. Ripasudil can release inflammatory cell infiltration and exudation proteins in aqueous humor. In addition, MCPI-1 protein levels and IL-1, IL-6, TNF-dan, and MCP-1 mRNA levels in the iris-ciliary body decreased after Ripasudil administration. Clinical trials also show that Ripasudil can be used as a glaucoma therapy agent. Ripasudil can be an updated therapy to treat glaucoma uveitis through inhibition of the ROCK pathway and treatment of proinflammatory mediators.