The clinical role of serum cell division control 42 in coronary heart disease.

Cell division control 42 (CDC42) regulates blood lipids, atherosclerosis, T cell differentiation and inflammation, which is involved in the process of coronary heart disease (CHD). This study aimed to evaluate the CDC42 level and its correlation with clinical features, the T-helper 17 (Th17)/regulatory-T (Treg) cell ratio and prognosis in CHD patients. In total, 210 CHD patients, 20 healthy controls and 20 disease controls were enrolled. Serum CDC42 levels of all participants were measured by enzyme-linked immunosorbent assay. In CHD patients, Th17 and Treg cells were discovered by flow cytometry; CHD patients were followed-up for a median of 16.9 months (range of 2.5-38.2 months). CDC42 level was lowest in CHD patients (median (interquartile range (IQR)): 402.5 (287.3-599.0) pg/mL), moderate in disease controls (median (IQR): 543.5 (413.0-676.3) pg/mL) and highest in healthy controls (median (IQR): 668.0 (506.5-841.3) pg/mL) (p < .001). Moreover, in CHD patients, lower CDC42 level was related to more prevalent diabetes mellitus (p = .021), and higher levels of C-reactive protein (p = .001), Gensini score (p = .006), Th17 cells (p = .001) and Th17/Treg ratio (p < .001) but was associated with lower Treg cells (p = .018). Furthermore, CDC42 low level [below the median level (402.5 pg/mL) of CDC42 in CHD patients] was correlated with higher accumulating major adverse cardiovascular event (MACE) risk (p = .029), while no correlation was found between the quartile of CDC42 level and accumulating MACE risk in CHD patients (p = .102). The serum CDC42 level is decreased and its low level is related to higher Th17/Treg ratio and increased accumulating MACE risk in CHD patients.

Genome-Wide Analysis of AGC Kinases Reveals that MoFpk1 Is Required for Development, Lipid Metabolism, and Autophagy in Hyperosmotic Stress of the Rice Blast Fungus Magnaporthe oryzae.

During eukaryotic evolution, the TOR-AGC kinase signaling module is involved in the coordinated regulation of cell growth and survival. However, the AGC kinases in plant-pathogenic fungi remain poorly understood. In this study, we have identified 20 members of the AGC family of protein kinases. Evolutionary and biological studies have revealed that AGC kinases are highly conserved and involved in the growth (8 genes), conidiation (13 genes), conidial germination (9 genes), appressorium formation (9 genes), and pathogenicity (5 genes) of Magnaporthe oryzae, in which a subfamily protein of the AGC kinases, MoFpk1, the activator of flippase, specifically exhibited diverse roles. Two kinase sites were screened and found to be critical for MoFpk1: 230K and 326D. Moreover, MoFpk1 is involved in cell wall integrity through the negative regulation of MoMps1 phosphorylation. The deletion of MoFpk1 resulted in defective phosphatidylacetamide (PE) and phosphatidylserine (PS) turnover and a series of lipid metabolism disorders. Under hyperosmotic stress, since the ΔMofpk1 mutant is unable to maintain membrane asymmetry, MoYpk1 phosphorylation and MoTor activity were downregulated, thus enhancing autophagy. Our results provide insights into the evolutionary and biological relationships of AGC kinases and new insight into plasma membrane (PM) homeostasis, i.e., responses to membrane stress and autophagy through lipid asymmetry maintenance. IMPORTANCE Our identification and analysis of evolutionary and biological relationships provide us with an unprecedented high-resolution view of the flexible and conserved roles of the AGC family in the topmost fungal pathogens that infect rice, wheat, barley, and millet. Guided by these insights, an AGC member, MoFpk1, was found to be indispensable for M. oryzae development. Our study defined a novel mechanism of plasma membrane homeostasis, i.e., adaptation to stress through the asymmetric distribution of phospholipids. Furthermore, defects in the asymmetric distribution of phospholipids in the membrane enhanced autophagy under hyperosmotic stress. This study provides a new mechanism for the internal linkage between lipid metabolism and autophagy, which may help new fungicide target development for controlling this devastating disease.

De Novo Purine Nucleotide Biosynthesis Pathway Is Required for Development and Pathogenicity in Magnaporthe oryzae.

Purine nucleotides are indispensable compounds for many organisms and participate in basic vital activities such as heredity, development, and growth. Blocking of purine nucleotide biosynthesis may inhibit proliferation and development and is commonly used in cancer therapy. However, the function of the purine nucleotide biosynthesis pathway in the pathogenic fungus Magnaporthe oryzae is not clear. In this study, we focused on the de novo purine biosynthesis (DNPB) pathway and characterized MoAde8, a phosphoribosylglycinamide formyltransferase, catalyzing the third step of the DNPB pathway in M. oryzae. MoAde8 was knocked out, and the mutant (∆Moade8) exhibited purine auxotroph, defects in aerial hyphal growth, conidiation, and pathogenicity, and was more sensitive to hyperosmotic stress and oxidative stress. Moreover, ∆Moade8 caused decreased activity of MoTor kinase due to blocked purine nucleotide synthesis. The autophagy level was also impaired in ∆Moade8. Additionally, MoAde5, 7, 6, and 12, which are involved in de novo purine nucleotide biosynthesis, were also analyzed, and the mutants showed defects similar to the defects of ∆Moade8. In summary, de novo purine nucleotide biosynthesis is essential for conidiation, development, and pathogenicity in M. oryzae.

A Subunit of ESCRT-III, MoIst1, Is Involved in Fungal Development, Pathogenicity, and Autophagy in Magnaporthe oryzae.

The culprit of rice blast, Magnaporthe oryzae, is a filamentous fungus that seriously affects the yield and quality of rice worldwide. MoIst1, a subunit of ESCRT-III, is involved in identified ubiquitinated proteins and transports them into the intraluminal vesicles of multivesicular bodies (MVBs) for degradation in lysosomes. Here, we identify and characterize MoIst1 in M. oryzae. Disruption of MoIst1 leads to a significant decrease in sporulation and formation of appressoria, defects in response to oxidative stress, cell wall stress, hyperosmotic stress, and reduced pathogenicity. Deletion of MoIst1 also caused the decreased Pmk1 phosphorylation levels, appressorium formation, the delayed translocation and degradation of lipid droplets and glycogen, resulting in a decreased appressorium turgor. In addition, deletion of MoIst1 leads to an abnormal autophagy. In summary, our results indicate that MoIst1 is involved in sporulation, appressorium development, plant penetration, pathogenicity, and autophagy in M. oryzae.

A Putative D-Arabinono-1,4-lactone Oxidase, MoAlo1, Is Required for Fungal Growth, Conidiogenesis, and Pathogenicity in Magnaporthe oryzae.

Magnaporthe oryzae is the causal agent of rice blast outbreaks. L-ascorbic acid (ASC) is a famous antioxidant found in nature. However, while ASC is rare or absent in fungi, a five-carbon analog, D-erythroascorbic acid (EASC), seems to appear to be a substitute for ASC. Although the antioxidant function of ASC has been widely described, the specific properties and physiological functions of EASC remain poorly understood. In this study, we identified a D-arabinono-1,4-lactone oxidase (ALO) domain-containing protein, MoAlo1, and found that MoAlo1 was localized to mitochondria. Disruption of MoALO1 (ΔMoalo1) exhibited defects in vegetative growth as well as conidiogenesis. The ΔMoalo1 mutant was found to be more sensitive to exogenous H2O2. Additionally, the pathogenicity of conidia in the ΔMoalo1 null mutant was reduced deeply in rice, and defective penetration of appressorium-like structures (ALS) formed by the hyphal tips was also observed in the ΔMoalo1 null mutant. When exogenous EASC was added to the conidial suspension, the defective pathogenicity of the ΔMoalo1 mutant was restored. Collectively, MoAlo1 is essential for growth, conidiogenesis, and pathogenicity in M. oryzae.

Endosomal sorting complexes required for transport-0 (ESCRT-0) are essential for fungal development, pathogenicity, autophagy and ER-phagy in Magnaporthe oryzae.

Magnaporthe oryzae is an important plant pathogen that causes rice blast. Hse1 and Vps27 are components of ESCRT-0 involved in the multivesicular body (MVB) sorting pathway and biogenesis. To date, the biological functions of ESCRT-0 in M. oryzae have not been determined. In this study, we identified and characterized Hse1 and Vps27 in M. oryzae. Disruption of MoHse1 and MoVps27 caused pleiotropic defects in growth, conidiation, sexual development and pathogenicity, thereby resulting in loss of virulence in rice and barley leaves. Disruption of MoHse1 and MoVps27 triggered increased lipidation of MoAtg8 and degradation of GFP-MoAtg8, indicating that ESCRT-0 is involved in the regulation of autophagy. ESCRT-0 was determined to interact with coat protein complex II (COPII), a regulator functioning in homeostasis of the endoplasmic reticulum (ER homeostasis), and disruption of MoHse1 and MoVps27 also blocked activation of the unfolded protein response (UPR) and autophagy of the endoplasmic reticulum (ER-phagy). Overall, our results indicate that ESCRT-0 plays critical roles in regulating fungal development, virulence, autophagy and ER-phagy in M. oryzae.

The COPII subunit MoSec24B is involved in development, pathogenicity and autophagy in the rice blast fungus.

The endoplasmic reticulum (ER) acts as the starting point of the secretory pathway, where approximately one-third of the proteins are correctly folded and modified, loaded into vesicles, and transported to the Golgi for further processing and modification. In this process, COPII vesicles are responsible for transporting cargo proteins from the ER to the Golgi. Here, we identified the inner shell subunit of COPII vesicles (MoSec24B) and explored the importance of MoSec24B in the rice blast fungus. The targeted disruption of MoSec24B led to decreased growth, reduced conidiation, restricted glycogen and lipids utilization, sensitivity to the cell wall and hypertonic stress, the failure of septin-mediated repolarization of appressorium, impaired appressorium turgor pressure, and decreased ability to infect, which resulted in reduced pathogenicity to the host plant. Furthermore, MoSec24B functions in the three mitogen-activated protein kinase (MAPK) signaling pathways by acting with MoMst50. Deletion of MoSec24B caused reduced lipidation of MoAtg8, accelerated degradation of exogenously introduced GFP-MoAtg8, and increased lipidation of MoAtg8 upon treatment with a late inhibitor of autophagy (BafA1), suggesting that MoSec24B regulates the fusion of late autophagosomes with vacuoles. Together, these results suggest that MoSec24B exerts a significant role in fungal development, the pathogenesis of filamentous fungi and autophagy.

Afterload-related cardiac performance predicts prognosis in critical ill patients with sepsis: A prospective observational pilot study.

ABSTRACT: To investigate the usefulness of afterload-related cardiac performance (ACP) for assessing cardiac impairment and predicting prognosis in septic patients.Adult patients with sepsis in the intensive care unit were included. Cardiac output, cardiac index, cardiac power index, and ACP were calculated at the time of admission (D0) and 48-72 h after admission (D3). They were correlated with Acute Physiology and Chronic Health Evaluation II and sequential organ failure assessment scores, then the prognostic values were analyzed.A total of 41 patients with sepsis were selected. ACP showed a stronger negative correlation with Acute Physiology and Chronic Health Evaluation II and sequential organ failure assessment scores than cardiac output, cardiac index, and cardiac power index. ACP predicted 28-day mortality with an area under the curve of 0.775 and 0.976 on D0 and D3, respectively. In addition, most non-survivors had emergent cardiac impairment (ACP ≤ 80%) on D0, and cardiac function was deteriorated on D3. Survival analysis showed that the patients with a decreased ACP from D0 to D3 had the highest mortality. The decrease of ACP on D3 was an independent risk factor for mortality (hazard ratio, 11.89; P = .0028).ACP can be used to assess the severity of cardiac impairment in sepsis. Continued decline of ACP during the first 3 days strongly suggests a poor prognosis.

Isolation and Functional Analysis of Effector Proteins of Magnaporthe oryzae.

In nature, plants have evolved a myriad of preformed and induced defenses to protect themselves from microbes. Upon microbial infection, the recognition of the microbe-associated molecular patterns (MAMPs) by the pattern recognition receptors (PRRs) triggers the first stage of defense response (Dodds and Rathjen, Nat Rev Genet 11:539-548, 2010). However, in order to develop microbial delivery, effectors target PRRs for deregulating immune responses and facilitating host colonization (Thomma et al., Plant Cell 23:4-15, 2011). Here, we contribute a protocol for the screening system of Magnaporthe oryzae effectors and construct a fluorescent system to trace secretory proteins in the sheath infection samples. Using the tobacco rattle virus (TRV) system, the proteins including LysM, Chitin, Cutinase, and CFEM domains were selected and divided into two kinds according to the results of cell death induced or inhibited test in Nicotiana benthamiana. Then, candidate effectors can be deleted or overexpressed in M. oryzae. The barley or rice infection with M. oryzae, rice leaf sheath inoculation, and subcellular localization during infection can be performed to explore the functions of these effectors.

Pulmonary edema following diuretic therapy: A case report.

INTRODUCTION: Diuretics are a commonly used for the treatment of acute pulmonary edema. However, inappropriate administration of diuretic drugs can result in clinical treatment failure and cause acute pulmonary edema. This is due to rapid decreases in intravascular volume as a result of diuretic treatment. To date, the clinical phenomenon of inappropriate use of diuretics leading to acute pulmonary edema remains unexplored and unrecognized. Here, we report the first case of this problem-pulmonary edema following diuretic therapy. PATIENT CONCERNS: A 71-year-old male patient who was intubated and transferred to the intensive care unit (ICU) due to respiratory failure was initially diagnosed with pneumonia as a complication of acute respiratory distress syndrome (ARDS). After treatments including antibiotics, lung protective ventilation strategies, and restrictive fluid management, his respiratory symptoms improved. However, the patient's dyspnea became more severe after experimental diuretic therapy. DIAGNOSIS: A point-of-care ultrasound (POCUS) examination showed increased extravascular lung water retention during a hypovolemic state. After full examinations and analysis, the diagnosis of acute pulmonary edema was determined. INTERVENTIONS: The most likely cause of acute pulmonary edema was left ventricular (LV) hyperdynamic status due to a hypovolemic status caused by excessive diuretic therapy. Consequently, we administrated intravenous fluids and a ß-receptor blocker to the patient. OUTCOMES: Following these treatment, the patient's respiratory distress improved remarkably. CONCLUSION: We report the first case of pulmonary edema following diuretic therapy to stress the need of physicians to follow guidelines of clinical practice. Maintaining an appropriate volume status and treatment of ß-receptor blockers is the key to reversing the progress of this adverse effect. In this process, POCUS is a reliable diagnostic tool to identify the cause of acute pulmonary edema and can increase the accuracy of clinical evaluations. It is likely that a wider use of POCUS will help physicians to obtain a faster, and more accurate, diagnosis of the etiology of acute pulmonary edema, thus allowing a more appropriate therapy.

[The prognostic value of extravascular lung water index in patients with acute respiratory distress syndrome].

OBJECTIVE: To evaluate the relationship between extravascular lung water index (EVLWI), pulmonary vascular permeability index (PVPI) and the severity categories defined by the Berlin definition, and investigate the prognostic value of EVLWI in patients with acute respiratory distress syndrome (ARDS). METHODS: Fifty-four patients with ARDS who received pulse index continuous cardiac output (PiCCO) monitoring were analyzed retrospectively. The patients were divided into mild, moderate and severe groups according to oxygenation index (OI) at admission. Characteristics and acute physiology and chronic health evaluation II (APACHEII) score in mild, moderate and severe groups were recorded, and the differences in EVLWI and PVPI among the severity categories of ARDS patients were analyzed. The correlation between EVLWI and intrathoracic blood volume index (ITBVI), PVPI and OI was analyzed. The receiver operating characteristic curve (ROC curve) was used to evaluate the prognosis accuracy of these indicators. RESULTS: Of 54 patients, there were 18 ARDS patients in mild group, 22 in moderate group and 14 in severe group. The 28-day mortality rate was gradually increased with severity categories of ARDS, and mortality in moderate and severe groups were significantly higher than that in mild group [54.5% (12/22), 57.1% (8/14) vs. 22.2% (4/18), both P<0.05]. 216 independent evaluation were measured in 54 patients within 72 hours, EVLWI was increased with severity categories of ARDS [EVLWI in mild, moderate and severe groups were 11 (10, 13) mL/kg, 14 (11, 16) mL/kg, 17 (12, 23) mL/kg, U=15.556, P=0.000], as well as the change tendency in PVPI [PVPI in mild, moderate and severe groups were 2.0 (1.8, 2.5), 2.7 (2.0, 3.9), 3.7 (2.1, 4.8), U=16.552, P=0.000] . The negative correlation between EVLWI and OI was found (r=-0.599, P=0.000), as well as between PVPI and OI (r=-0.541, P=0.000). The positive correlation between EVLWI and PVPI was found (r=0.811, P=0.000), as well as between EVLWI and ITBVI (r=0.168, P=0.041). The area under ROC curve (AUC) for EVLWI evaluating the prognosis at 0, 24, 48, 72 hours was 0.639 ± 0.082, 0.642 ± 0.082, 0.702 ± 0.078, 0.719 ± 0.076 respectively, 95% confidence interval (95%CI) was 0.479-0.799, 0.483-0.802, 0.551-0.852, 0.570-0.869 respectively, AUC for APACHEII was 0.783 ± 0.068, and 95%CI was 0.648-0.918. As EVLWI>13 mL/kg at 72 hours, the sensitivity and the specificity of accurate judgment were 81.8% and 61.5%. CONCLUSIONS: Combining the Berlin definition with EVLWI and PVPI, the severity of ARDS can be assessed accurately. Dynamic observation of EVLWI can be one of prognostic factors in the patients with ARDS.

[Effects of continuous veno-venous hemofiltration on the hemodynamics of sheep with endotoxic shock].

OBJECTIVE: To study the influence of continuous veno-venous hemofiltration (CVVH) on the hemodynamics and the prognosis of sheep with endotoxic shock. METHODS: Endotoxic shock was induced by intravenous infusion of lipopolysaccharide, L-2880, to sheep, and the animals were randomly divided to control group (NHF, endotoxin only) or treatment group (HF, endotoxin+CVVH). In the latter groups a zero-balanced CVVH was instituted with an ultrafiltration rate of 40 ml x kg(-1) x h(-1) and a blood flow rate of 80 ml/min. The hemodynamic parameters were monitored. RESULTS: After the administration of endotoxin, the mean arterial pressure (MAP) was decreased significantly, while mean pulmonary artery pressure (MPAP) and cardiac index (CI) increased significantly in both groups (P<0.05 or P<0.01). With CVVH, MAP and stroke volume (SV) were increased significantly, and were significantly higher than that of NHF group (P<0.05 or P<0.01). CI declined progressively in NHF group and significantly lower than that of HF group, in which there was no significant change(P<0.05). No significant difference in MPAP was found between two groups (P>0.05). Moreover, plasma lactate levels were lower in NHF group than in HF group (P<0.05). After 5 hours, the sheep in HF group all survived, and 2 sheep of NHF group died. CONCLUSION: Intravenous infusion of endotoxin could produce endotoxic shock in sheep. CVVH was shown to have a beneficial effect on the hemodynamics, and it gave a better prognosis in endotoxic, indicating that it might be important method to treat endotoxic shock.