Suppression of Ventilation-Induced Diaphragm Fibrosis through the Phosphoinositide 3-Kinase-γ in a Murine Bleomycin-Induced Acute Lung Injury Model.

Mechanical ventilation (MV), used in patients with acute lung injury (ALI), induces diaphragmatic myofiber atrophy and contractile inactivity, termed ventilator-induced diaphragm dysfunction. Phosphoinositide 3-kinase-γ (PI3K-γ) is crucial in modulating fibrogenesis during the reparative phase of ALI; however, the mechanisms regulating the interactions among MV, myofiber fibrosis, and PI3K-γ remain unclear. We hypothesized that MV with or without bleomycin treatment would increase diaphragm muscle fibrosis through the PI3K-γ pathway. Five days after receiving a single bolus of 0.075 units of bleomycin intratracheally, C57BL/6 mice were exposed to 6 or 10 mL/kg of MV for 8 h after receiving 5 mg/kg of AS605240 intraperitoneally. In wild-type mice, bleomycin exposure followed by MV 10 mL/kg prompted significant increases in disruptions of diaphragmatic myofibrillar organization, transforming growth factor-ß1, oxidative loads, Masson's trichrome staining, extracellular collagen levels, positive staining of α-smooth muscle actin, PI3K-γ expression, and myonuclear apoptosis (p < 0.05). Decreased diaphragm contractility and peroxisome proliferator-activated receptor-γ coactivator-1α levels were also observed (p < 0.05). MV-augmented bleomycin-induced diaphragm fibrosis and myonuclear apoptosis were attenuated in PI3K-γ-deficient mice and through AS605240-induced inhibition of PI3K-γ activity (p < 0.05). MV-augmented diaphragm fibrosis after bleomycin-induced ALI is partially mediated by PI3K-γ. Therapy targeting PI3K-γ may ameliorate MV-associated diaphragm fibrosis.

Attenuation of Ventilation-Enhanced Epithelial-Mesenchymal Transition through the Phosphoinositide 3-Kinase-γ in a Murine Bleomycin-Induced Acute Lung Injury Model.

Mechanical ventilation (MV) used in patients with acute lung injury (ALI) induces lung inflammation and causes fibroblast proliferation and excessive collagen deposition-a process termed epithelial-mesenchymal transition (EMT). Phosphoinositide 3-kinase-γ (PI3K-γ) is crucial in modulating EMT during the reparative phase of ALI; however, the mechanisms regulating the interactions among MV, EMT, and PI3K-γ remain unclear. We hypothesized that MV with or without bleomycin treatment would increase EMT through the PI3K-γ pathway. C57BL/6 mice, either wild-type or PI3K-γ-deficient, were exposed to 6 or 30 mL/kg MV for 5 h after receiving 5 mg/kg AS605240 intraperitoneally 5 days after bleomycin administration. We found that, after bleomycin exposure in wild-type mice, high-tidal-volume MV induced substantial increases in inflammatory cytokine production, oxidative loads, Masson's trichrome staining level, positive staining of α-smooth muscle actin, PI3K-γ expression, and bronchial epithelial apoptosis (p < 0.05). Decreased respiratory function, antioxidants, and staining of the epithelial marker Zonula occludens-1 were also observed (p < 0.05). MV-augmented bleomycin-induced pulmonary fibrogenesis and epithelial apoptosis were attenuated in PI3K-γ-deficient mice, and we found pharmacological inhibition of PI3K-γ activity through AS605240 (p < 0.05). Our data suggest that MV augmented EMT after bleomycin-induced ALI, partially through the PI3K-γ pathway. Therapy targeting PI3K-γ may ameliorate MV-associated EMT.

High-flow nasal cannula compared with continuous positive airway pressure in the treatment of obstructive sleep apnea.

PURPOSE: Continuous positive airway pressure (CPAP) is a standard treatment for obstructive sleep apnea (OSA). However, CPAP has limitations. High-flow nasal cannula (HFNC) is already in use for various types of respiratory diseases. As HFNC generates positive airway pressure, it may be a potential candidate for OSA treatment. This prospective study compared the therapeutic effects of HFNC to CPAP in patients with OSA. METHODS: Patients whose apnea-hypopnea index (AHI) was > 5 events/h were enrolled in this study. All participants were randomly divided into two groups. The first group underwent CPAP the first night and HFNC the second night. Conversely, the second group received HFNC the first night and CPAP the second night. Their respiratory events and sleep quality were compared using baseline polysomnography, CPAP, and HFNC. RESULTS: In total, 28 participants completed this study. Median [interquartile range] AHI (35.0 [20.0-48.6] vs. 10.8 [5.5-20.6] events/h; p < 0.001) was significantly improved by the HFNC. However, sleep quality was not improved. When CPAP was compared directly with HFNC, CPAP demonstrated a more favorable effect for respiratory events (AHI 5.0 [2.0-7.0] vs. 10.8 [5.5-20.6] events/h; p < 0.001) and sleep efficiency (88.1 [79.9-92.5] vs. 77.9 [69.2-86.6] %; p = 0.001). CONCLUSION: The efficacy of CPAP was superior to HFNC for both respiratory events and sleep quality. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03843372; URL: www. CLINICALTRIALS: gov ; Date of registration: November 2, 2019.

Reduction in Ventilation-Induced Diaphragmatic Mitochondrial Injury through Hypoxia-Inducible Factor 1α in a Murine Endotoxemia Model.

Mechanical ventilation (MV) is essential for patients with sepsis-related respiratory failure but can cause ventilator-induced diaphragm dysfunction (VIDD), which involves diaphragmatic myofiber atrophy and contractile inactivity. Mitochondrial DNA, oxidative stress, mitochondrial dynamics, and biogenesis are associated with VIDD. Hypoxia-inducible factor 1α (HIF-1α) is crucial in the modulation of diaphragm immune responses. The mechanism through which HIF-1α and mitochondria affect sepsis-related diaphragm injury is unknown. We hypothesized that MV with or without endotoxin administration would aggravate diaphragmatic and mitochondrial injuries through HIF-1α. C57BL/6 mice, either wild-type or HIF-1α-deficient, were exposed to MV with or without endotoxemia for 8 h. MV with endotoxemia augmented VIDD and mitochondrial damage, which presented as increased oxidative loads, dynamin-related protein 1 level, mitochondrial DNA level, and the expressions of HIF-1α and light chain 3-II. Furthermore, disarrayed myofibrils; disorganized mitochondria; increased autophagosome numbers; and substantially decreased diaphragm contractility, electron transport chain activities, mitofusin 2, mitochondrial transcription factor A, peroxisome proliferator activated receptor-γ coactivator-1α, and prolyl hydroxylase domain 2 were observed (p < 0.05). Endotoxin-stimulated VIDD and mitochondrial injuries were alleviated in HIF-1α-deficient mice (p < 0.05). Our data revealed that endotoxin aggravated MV-induced diaphragmatic dysfunction and mitochondrial damages, partially through the HIF-1α signaling pathway.

Suppression of Hypoxia-Inducible Factor 1α by Low-Molecular-Weight Heparin Mitigates Ventilation-Induced Diaphragm Dysfunction in a Murine Endotoxemia Model.

Mechanical ventilation (MV) is required to maintain life for patients with sepsis-related acute lung injury but can cause diaphragmatic myotrauma with muscle damage and weakness, known as ventilator-induced diaphragm dysfunction (VIDD). Hypoxia-inducible factor 1α (HIF-1α) plays a crucial role in inducing inflammation and apoptosis. Low-molecular-weight heparin (LMWH) was proven to have anti-inflammatory properties. However, HIF-1α and LMWH affect sepsis-related diaphragm injury has not been investigated. We hypothesized that LMWH would reduce endotoxin-augmented VIDD through HIF-1α. C57BL/6 mice, either wild-type or HIF-1α-deficient, were exposed to MV with or without endotoxemia for 8 h. Enoxaparin (4 mg/kg) was administered subcutaneously 30 min before MV. MV with endotoxemia aggravated VIDD, as demonstrated by increased interleukin-6 and macrophage inflammatory protein-2 levels, oxidative loads, and the expression of HIF-1α, calpain, caspase-3, atrogin-1, muscle ring finger-1, and microtubule-associated protein light chain 3-II. Disorganized myofibrils, disrupted mitochondria, increased numbers of autophagic and apoptotic mediators, substantial apoptosis of diaphragm muscle fibers, and decreased diaphragm function were also observed (p < 0.05). Endotoxin-exacerbated VIDD and myonuclear apoptosis were attenuated by pharmacologic inhibition by LMWH and in HIF-1α-deficient mice (p < 0.05). Our data indicate that enoxaparin reduces endotoxin-augmented MV-induced diaphragmatic injury, partially through HIF-1α pathway inhibition.

The cancer control status and APACHE II score are prognostic factors for critically ill patients with cancer and sepsis.

BACKGROUND/PURPOSE: Patients with cancer are eligible for hospice care when their life expectancy is 180 days or shorter. This study investigated the prognostic factors of patients with cancer and sepsis who were admitted to an intensive care unit (ICU) to assist with clinical decisions of hospice care. METHODS: A series of 279 patients admitted to the medical ICU with cancer and sepsis were included. Another series of 109 patients with cancer and sepsis admitted to the other medical ICU in the different branch of our hospital was included to verify the results. RESULTS: Among 279 patients, the 30-, 90-, and 180-day mortality rates were 47.3%, 72.0%, and 81.0%, respectively. APACHE II score and the cancer control status (controlled or remission (CR), active newly diagnosed (AND) and active recurrent or progressive (ARP)) were significant predictors of 30- and 90-day mortality(30-day: AND(odds ratio: 5.66; 95% confidence interval: 2.12-15.15), ARP(6.24; 2.92-13.33), APACHE II( 1.07; 1.03-1.11); 90-day: AND(4.78; 1.91-11.99), ARP( 24.03; 11.11-51.99), APACHE II( 1.07; 1.02-1.19)) and were associated with a poor 180-day outcome. The 180-day mortality were significantly different among the patients with different cancer control status in the series of 279 patients (CR: 29.8%; AND: 69.4%; and ARP: 98.9 %) and that of 109 patients (46.4%; 96.8%; and 94.0%). CONCLUSION: APACHE II score and the cancer control status may be the prognostic factors for critically ill patients with cancer and sepsis, and they may be helpful for evaluating hospice care.

Thymidine Kinase-Negative Herpes Simplex Virus 1 Can Efficiently Establish Persistent Infection in Neural Tissues of Nude Mice.

Herpes simplex virus 1 (HSV-1) establishes latency in neural tissues of immunocompetent mice but persists in both peripheral and neural tissues of lymphocyte-deficient mice. Thymidine kinase (TK) is believed to be essential for HSV-1 to persist in neural tissues of immunocompromised mice, because infectious virus of a mutant with defects in both TK and UL24 is detected only in peripheral tissues, but not in neural tissues, of severe combined immunodeficiency mice (T. Valyi-Nagy, R. M. Gesser, B. Raengsakulrach, S. L. Deshmane, B. P. Randazzo, A. J. Dillner, and N. W. Fraser, Virology 199:484-490, 1994, https://doi.org/10.1006/viro.1994.1150). Here we find infiltration of CD4 and CD8 T cells in peripheral and neural tissues of mice infected with a TK-negative mutant. We therefore investigated the significance of viral TK and host T cells for HSV-1 to persist in neural tissues using three genetically engineered mutants with defects in only TK or in both TK and UL24 and two strains of nude mice. Surprisingly, all three mutants establish persistent infection in up to 100% of brain stems and 93% of trigeminal ganglia of adult nude mice at 28 days postinfection, as measured by the recovery of infectious virus. Thus, in mouse neural tissues, host T cells block persistent HSV-1 infection, and viral TK is dispensable for the virus to establish persistent infection. Furthermore, we found 30- to 200-fold more virus in neural tissues than in the eye and detected glycoprotein C, a true late viral antigen, in brainstem neurons of nude mice persistently infected with the TK-negative mutant, suggesting that adult mouse neurons can support the replication of TK-negative HSV-1. IMPORTANCE: Acyclovir is used to treat herpes simplex virus 1 (HSV-1)-infected immunocompromised patients, but treatment is hindered by the emergence of drug-resistant viruses, mostly those with mutations in viral thymidine kinase (TK), which activates acyclovir. TK mutants are detected in brains of immunocompromised patients with persistent infection. However, answers to the questions as to whether TK-negative (TK-) HSV-1 can establish persistent infection in brains of immunocompromised hosts and whether neurons in vivo are permissive for TK- HSV-1 remain elusive. Using three genetically engineered HSV-1 TK- mutants and two strains of nude mice deficient in T cells, we found that all three HSV-1 TK- mutants can efficiently establish persistent infection in the brain stem and trigeminal ganglion and detected glycoprotein C, a true late viral antigen, in brainstem neurons. Our study provides evidence that TK- HSV-1 can persist in neural tissues and replicate in brain neurons of immunocompromised hosts.

Modeling of surface topography on diamond-turned spherical and freeform surfaces.

Single-point diamond turning machines are capable of generating high-quality spherical and freeform surfaces. However, there are inevitable tool marks on the diamond-turned surfaces. Although several models have been proposed to describe the surface topography of a flat surface, there is a lack of a more general model to describe spherical and freeform surfaces. In this paper, we propose a model to estimate the surface topography of the diamond-turned spherical and freeform surfaces. The model takes into consideration the basic cutting parameters as well as the dominant relative vibration components between the diamond tool and the workpiece in both infeed and feeding directions. We first discuss the principles and create a model for spherical surfaces. The model is then extended to describe more general freeform surfaces. We also show how the micro-waviness of the diamond tool impacts the surface topography. Finally, we conduct a series of face cutting experiments and conclude that there is good correlation between the model and the experimental results.

Computer-aided high-accuracy testing of reflective surface with reverse Hartmann test.

The deflectometry provides a feasible way for surface testing with a high dynamic range, and the calibration is a key issue in the testing. A computer-aided testing method based on reverse Hartmann test, a fringe-illumination deflectometry, is proposed for high-accuracy testing of reflective surfaces. The virtual "null" testing of surface error is achieved based on ray tracing of the modeled test system. Due to the off-axis configuration in the test system, it places ultra-high requirement on the calibration of system geometry. The system modeling error can introduce significant residual systematic error in the testing results, especially in the cases of convex surface and small working distance. A calibration method based on the computer-aided reverse optimization with iterative ray tracing is proposed for the high-accuracy testing of reflective surface. Both the computer simulation and experiments have been carried out to demonstrate the feasibility of the proposed measurement method, and good measurement accuracy has been achieved. The proposed method can achieve the measurement accuracy comparable to the interferometric method, even with the large system geometry calibration error, providing a feasible way to address the uncertainty on the calibration of system geometry.

Modeling of surface topography in single-point diamond turning machine.

Surface roughness is an important factor in characterizing the performance of high-precision optical surfaces. In this paper, we propose a model to estimate the surface roughness generated by a single-point diamond turning machine. In this model, we take into consideration the basic tool-cutting parameters as well as the relative vibration between the tool and the workpiece in both the infeed and feeding directions. Current models focus on the relative tool-workpiece vibration in the infeed direction. However, based on our experimental measurements, the contribution of relative tool-workpiece vibration in the feeding direction is significant and cannot be ignored in the model. The proposed model is able to describe the surface topography for flat as well as cylindrical surfaces of the workpiece. It has the potential to describe more complex spherical surfaces or freeform surfaces. Our experimental study with metal materials shows good correlation between the model and the diamond-turned surfaces.

Creating unconventional geometric beams with large depth of field using double freeform-surface optics.

We consider here creation of an unconventional flattop beam with a large depth of field by employing double freeform optical surfaces. The output beam is designed with continuous variations from the flattop to almost zero near the edges to resist the influence of diffraction on its propagation. We solve this challenging problem by naturally incorporating an optimal transport map computation scheme for unconventional boundary conditions with a simultaneous point-by-point double surface construction procedure. We demonstrate experimentally the generation of a long-range propagated triangular beam through a plano-freeform lens pair fabricated by a diamond-tuning machine.

Effect of interleukin-17 on in vitro cytokine production in healthy controls and patients with severe sepsis.

BACKGROUND/PURPOSE: Interleukin (IL)-17 family members (IL-17A to IL-17F) are appearing to play key roles in host defense and inflammatory disease. Recently, several cytokines, such as IL-6, IL-10, IL-12, and transforming growth factor (TGF)-ß1, were shown to have vital roles in severe sepsis. However, the influence of IL-17 on these cytokine responses from peripheral blood mononuclear cells (PBMCs) is unclear. METHODS: Fifty-two patients who were admitted to our intensive care unit (ICU) because of severe sepsis were enrolled into this study. To validate experimental findings, 25 healthy controls were enrolled. Lipopolysaccharide-stimulated PBMCs with IL-17 or anti-IL-17 treatments were cultured for 24 hours. IL-6, IL-10, IL-12, and TGF-ß1 levels in supernatants were measured. RESULTS: The IL-12 production from stimulated PBMCs was increased after IL-17 treatment in both control and patient groups. Additional treatment of anti-IL-17 enhanced IL-10 production but decreased IL-12 production from stimulated PBMCs of healthy controls and patients with severe sepsis. CONCLUSION: IL-17 was helpful for inflammation in severe sepsis. Lack of IL-17 decreased IL-12 and enhanced IL-10 production from PBMCs, which resulted in immune imbalance.

A peroxiredoxin cDNA from Taiwanofungus camphorata: role of Cys31 in dimerization.

Peroxiredoxins (Prxs) play important roles in antioxidant defense and redox signaling pathways. A Prx isozyme cDNA (TcPrx2, 745 bp, EF552425) was cloned from Taiwanofungus camphorata and its recombinant protein was overexpressed. The purified protein was shown to exist predominantly as a dimer by sodium dodecyl sulfate-polyacrylamide gel electrolysis in the absence of a reducing agent. The protein in its dimeric form showed no detectable Prx activity. However, the protein showed increased Prx activity with increasing dithiothreitol concentration which correlates with dissociation of the dimer into monomer. The TcPrx2 contains two Cys residues. The Cys(60) located in the conserved active site is the putative active peroxidatic Cys. The role of Cys(31) was investigated by site-directed mutagenesis. The C31S mutant (C(31) → S(31)) exists predominantly as a monomer with noticeable Prx activity. The Prx activity of the mutant was higher than that of the corresponding wild-type protein by nearly twofold at 12 µg/mL. The substrate preference of the mutant was H2O2 > cumene peroxide > t-butyl peroxide. The Michaelis constant (K M) value for H2O2 of the mutant was 0.11 mM. The mutant enzyme was active under a broad pH range from 6 to 10. The results suggest a role of Cys(31) in dimerization of the TcPrx2, a role which, at least in part, may be involved in determining the activity of Prx. The C(31) residue does not function as a resolving Cys and therefore the TcPrx2 must follow the reaction mechanism of 1-Cys Prx. This TcPrx2 represents a new isoform of Prx family.

Modulation of nitrosative stress via glutathione-dependent formaldehyde dehydrogenase and S-nitrosoglutathione reductase.

Glutathione-dependent formaldehyde dehydrogenase (GFD) from Taiwanofungus camphorata plays important roles in formaldehyde detoxification and antioxidation. The enzyme is bifunctional. In addition to the GFD activity, it also functions as an effective S-nitrosoglutathione reductase (GSNOR) against nitrosative stress. We investigated the modulation of HEK (human embryonic kidney) 293T cells under nitrosative stress by transfecting a codon optimized GFD cDNA from Taiwanofungus camphorata (Tc-GFD-O) to these cells. The parental and transfected HEK 293T cells were then subjected to S-nitrosoglutathione treatment to induce nitrosative stress. The results showed that in Tc-GFD-O-transfected 293T cells, the expression and activity of GFD increased. Additionally, these cells under the nitrosative stress induced by S-nitrosoglutathione showed both higher viability and less apoptosis than the parental 293T cells. This finding suggests that the Tc-GFD-O in HEK 293T cells may provide a protective function under nitrosative stress.

Cell death of alveolar lymphocytes and monocytes is negatively correlated with driving pressure and mechanical power in patients with acute respiratory distress syndrome.

BACKGROUND: Pathogenesis of acute respiratory distress syndrome (ARDS) involves immune cell death and removal from the injured lungs. ARDS severity is related to lung compliance. However, the correlation between the respiratory mechanics and alveolar immune cell death in patients with ARDS remains unclear. METHODS: Twenty-four patients with respiratory failure and ARDS were enrolled in the intensive care unit between November 2019 and November 2021. Neutrophil extracellular traps (NETs) and cell death of lymphocytes and monocytes in bronchoalveolar lavage fluid were detected on days 1 and 8. RESULTS: Lung compliance was positively correlated with the cell death percentage of alveolar CD4/CD8 lymphocytes and monocytes on day 8 (Pearson's correlation coefficient (r) = 0.554, p = 0.005; r = 0.422, p = 0.040; r = 0.569, p = 0.004, respectively). There was no association between lung compliance and the percentage of alveolar NETs on days 1 and 8. The cell death percentages of alveolar CD4/CD8 lymphocytes and monocytes were negatively correlated with driving pressure (DP) on days 1 (r = - 0.440, p = 0.032; r = - 0.613, p = 0.001; r = -0.557, p = 0.005, respectively) and 8 (r = - 0.459, p = 0.024; r = - 0.407, p = 0.048; r = - 0.607, p = 0.002, respectively). The cell death percentages of alveolar CD4/CD8 lymphocytes and monocytes were also negatively correlated with mechanical power (MP) on days 1 (r = - 0.558, p = 0.005; r = - 0.593, p = 0.002; r = - 0.571, p = 0.004, respectively) and 8 (r = - 0.539, p = 0.007; r = - 0.338, p = 0.107; r = - 0.649, p < 0.001, respectively). The percentage of alveolar NETs on days 1 and 8 was not associated with DP or MP. CONCLUSION: Patients with higher cell death rates of alveolar CD4/CD8 lymphocytes and monocytes exhibited lower DP and MP. Patients with less cell death of alveolar CD4/CD8 lymphocytes and monocytes required more DP or MP to maintain adequate ventilation.

Embedded corrugated long-period fiber gratings for sensing applications.

We demonstrate a method to make possible the mass production of corrugated long-period fiber gratings (C-LPFGs) by utilizing imprint lithography on polycarbonate (PC) substrates. For such C-LPFGs whose working principle is based on photoelastic effect, pretensile tension is required to be applied to inducing periodical refractive index variation. We then present an attempt to use PC as embedding material for providing internal compressive stress for C-LPFGs to have a photoelastic effect. This type of LPFG, termed embedded corrugated long-period fiber gratings (EC-LPFGs), is obtained after reimprinting the C-LPFGs into other PC substrates. Since compressive stress is retained due to the materials of different coefficients of thermal expansion (CTE), unlike C-LPFGs, EC-LPFGs can serve as strain, bending, and temperature sensors without the need of pretensile strain. The two most troublesome problems, the fragility of an etched fiber grating and the requirement of pretensile strain, can be simultaneously alleviated or solved by EC-LPFGs.

Prolonged Mechanical Ventilation: Outcomes and Management.

The number of patients requiring prolonged mechanical ventilation (PMV) is increasing worldwide, placing a burden on healthcare systems. Therefore, investigating the pathophysiology, risk factors, and treatment for PMV is crucial. Various underlying comorbidities have been associated with PMV. The pathophysiology of PMV includes the presence of an abnormal respiratory drive or ventilator-induced diaphragm dysfunction. Numerous studies have demonstrated that ventilator-induced diaphragm dysfunction is related to increases in in-hospital deaths, nosocomial pneumonia, oxidative stress, lung tissue hypoxia, ventilator dependence, and costs. Thus far, the pathophysiologic evidence for PMV has been derived from clinical human studies and experimental studies in animals. Moreover, recent studies have demonstrated the outcome benefits of pharmacological agents and rehabilitative programs for patients requiring PMV. However, methodological limitations affected these studies. Controlled prospective studies with an adequate number of participants are necessary to provide evidence of the mechanism, prognosis, and treatment of PMV. The great epidemiologic impact of PMV and the potential development of treatment make this a key research field.

Serial Increases in Human Leukocyte Antigen-DR Expression and Decreases in Interleukin-10 Expression in Alveolar Monocytes of Survivors of Pneumonia-Related Acute Respiratory Distress Syndrome.

ARDS is a potentially lethal syndrome. HLA-DR expression in monocytes reflects their activation and antigen-presenting capacity. However, the correlation between clinical outcomes and HLA-DR expression in alveolar monocytes/macrophages in patients with pneumonia-related ARDS remains unclear. Thus, we determined the trends of HLA-DR and cytokine expressions in alveolar monocytes using repeated measurements to answer this question. Thirty-one pneumonia patients with respiratory failure and ARDS without coronavirus disease 2019 between November 2019 and November 2021 were enrolled in our intensive care unit and three without complete data were excluded. Interleukin (IL)-10, IL-12, and HLA-DR expression in bronchoalveolar lavage (BAL) monocytes were determined on days one and eight. Monocyte HLA-DR expression (mHLA-DR) and CD4 T lymphocytes percentages in BAL cells of survivors increased remarkably after seven days. Monocyte IL-10 expression and monocytes percentages in BAL cells of survivors decreased substantially after seven days. The mHLA-DR was negatively correlated with disease severity scores on day one and eight. In conclusion, serial increases in HLA-DR expression and decreases in IL-10 expression were observed in BAL monocytes of survivors of pneumonia-related ARDS. More studies are needed to confirm this point of view, and then development of a therapeutic agent restoring mHLA-DR and preventing IL-10 production can be considered.

Effect of molecular mass and sulfate content of fucoidan from Sargassum siliquosum on antioxidant, anti-lipogenesis, and anti-inflammatory activity.

Macroalgae (seaweeds) are abundant in functional polysaccharides known for their unique biochemical activities. In this study, the antioxidant, anti-lipogenic, and anti-inflammatory activities of the fucoidan extracted from brown seaweed Sargassum siliquosum were investigated by 1,1-diphenyl-2-picrylhydrazyl (DPPH)-scavenging ability, lipid synthesis inhibition, and suppression of pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-α) production, respectively. To examine the effect of molecular mass on fucoidan's bioactivities above, the extracted fucoidan was subject to hydrogen peroxide-mediated partial hydrolysis to obtain lower molecular mass compounds within the range of 107.3-3.2 kDa. Results indicated that fucoidan's antioxidant activity increased with a corresponding decrease in molecular mass; the dosage for the half-maximal response (EC50) dropped from 2.58 to 1.82 mg/mL when the molecular mass decreased from 107.3 to 3.2 kDa. In addition, both the anti-lipogenesis and anti-inflammatory activities of fucoidan were significantly enhanced by 71.1% and 36.7%, respectively, when the molecular mass decreased to about 3 kDa. To further test the effect of sulfation on fucoidan's bioactivities, low molecular mass fucoidan was treated with SO3-DMF to increase the sulfate content. The results indicated that when sulfate content increased from 18.7% to 32.1%, EC50 of DPPH decreased from 1.82 mg/mL to 0.86 mg/mL and the anti-inflammatory activity also increased by 35.2%; however, the anti-lipogenesis activity decreased.

Whole-cell biocatalyst for cadaverine production using stable, constitutive and high expression of lysine decarboxylase in recombinant Escherichia coli W3110.

Microbial production of industrial chemicals is a sustainable approach to reduce the dependence on petroleum-based chemicals such as acids, alcohols, and amines, in which the cadaverine is a natural diamide and serves as one of the key monomers for biopolymer production. In this study, the constitutive promoter J23100 driven lysine decarboxylase (CadA) for cadaverine production was established and compared in different Escherichia coli strains. The best chassis designed as JW, expressed the highest amount of CadA by using J23100 promoter, showing stable and high copy numbers (i.e., PCN > 100) when culture in the antibiotic-free medium. JW attained a CadA activity of 167 g-DAP/g-DCW-h and had the maximum biocatalyst of 45.6 g-DCW/L in fed-batch fermentation. In addition, JW was able to convert 2.5 M L-lysine to 221 g/L cadaverine, with 86 % yield and 55.3 g/L-h productivity. The whole-cell biocatalyst could be reused over four times at an average of 97 % conversion when supplied half of fresh cells in the reaction. This work developed a stable, constitutive expression, long-term preservation, high-level expression of CadA for DAP production, and paved an alternative opportunity of bio-nylon for industry in the future.