Ralstonia solanacearum differentially modulates soil physicochemical properties and rhizospheric bacteriome of resistant and susceptible tobacco cultivars.

Ralstonia solanacearum is a devastating soilborne pathogen which poses significant yield and economic losses to tobacco production globally. The impact of R. solanacearum on rhizosphere bacteriome and soil physicochemical characteristics in resistant and susceptible tobacco cultivars is poorly understood. This study aims to determine the effect of R. solanacearum on soil physicochemical parameters and rhizosphere bacteriome of resistant (K326) and susceptible (Hongda) tobacco cultivars at various growth stages. Results demonstrated that the contents of available potassium and phosphorus, as well as soil pH were significantly increased in K326 soils (CK and T2) compared with Hongda (T1) after 21, 42, and 63 days post-inoculation (dpi) of R. solanacearum except for available nitrogen which showed an opposite trend. The qPCR results showed a significant decrease in R. solanacearum population in rhizosphere of K326 (T2) compared to the Hongda (T1) at 21 and 63 dpi than that after 42 dpi. The rhizosphere bacteriome analysis through 16S rRNA amplicon sequencing revealed that rhizosphere bacterial community composition was significantly different between two tobacco cultivars (Hongda and K326) and this effect was more prominent after 63 dpi (93 days after post-transplantation), suggesting that each cultivar recruits a unique set of bacterial communities. There was no obvious difference observed in the rhizosphere bacteriome of CK (K326) and T2 (K326), which might be attributed to the same genetic makeup and inherent resistance of K326 to bacterial wilt infection. Analysis of co-occurrence networks revealed that the microbial network in T1 (Hongda) was more complex than those in T2 (K326) and CK (K326), while the networks in CK and T2 were almost identical. The present research highlights the time-course relationship between environmental factors and rhizosphere bacteriome of tobacco cultivars showing different levels of resistance against R. solanacearum. Conclusively, studying the plant-soil-microbe interaction system in susceptible and resistant tobacco cultivars may enable us to develop effective integrated disease control plans for the healthy production of tobacco crops.

First Report of Twig Anthracnose of Acacia melanoxylon Caused by Colletotrichum siamense in China.

Acacia melanoxylon R. Br. native to Australia, is a high-quality timber tree with wide genetic and phenotypic diversity. In recent years, A. melanoxylon has been extensively cultivated in some provinces in southern China. In December 2019, anthracnose-like symptoms were observed on twigs of A. melanoxylon in China. In certain valleys in south China, the disease incidence on plants and shoots was 60-75% and 80-90%, respectively. The wither rate of disease branches was 30-40% in dry seasons from September to November. The appearance of symptoms occurred in a humid and warm valley. Symptoms were initially observed on the young branches as brown spindle shaped sunken spots. At later stages, the disease spots girdled the whole branch, which became wilted and its leaves turned reddish-brown. For pathogen isolation, diseased branches were sampled and 55 pieces (5× 5 mm) of these branches section were surface-sterilized in 75% ethanol for 30 seconds, followed by 0.5% NaClO for 5 min and then were rinsed three times in distilled water. After drying with sterilized filter paper, the surface-sterilized sections were transferred to potato dextrose agar medium (PDA) and incubated at 25 °C for 7 days in the dark. Three isolates were obtained as representatives for morphological characterization and were labeled as 1A912, 1B912, and 1C912. These specimens were deposited in the Guangdong Province Key Laboratory of Microbial Signals and Disease Control at the South China Agricultural University (China). Purified isolates were initially white, cottony and with dense aerial mycelium on PDA at 25 ℃, ten days later their colonies turned grayish white with orange conidial masses. Conidia were one-celled, hyaline, straight, cylindrical, with round obtuse ends, and measured 11.0 to16.3× 4.0 to 6.0 µm (n= 100), appressoria were 5.86 to 9.07 × 3.55 to 6.96 µm (n= 100). Morphological characteristics of selected isolates matched the Colletotrichum gloeosporioides species complex (Weir et al. 2012). For further identification, the internal transcribed spacer (ITS) region, and partial sequences of the actin (ACT), beta-tubulin (TUB2), and glycerol dehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified by PCR, and sequenced, using primer pairs ITS1/ ITS4 (White et al. 1990), Bt2a/ Bt2b (Donaldson and Glass 1995), ACT512F/ ACT783R, GDF1/ GDR1(Weir et al. 2012). The sequences were deposited in GenBank (ITS: MW228101-MW228103; TUB2: MW250346, MW320707, MW320708; ACT: MW250347, MW320703, MW320704; GAPDH: MW250348, MW320705, MW320706). The multilocus phylogenetic analysis distinguished the isolates 1A912, 1B912, and 1C912 as C. siamense. Pathogenicity of those three isolates of C. siamense was tested on healthy twigs of the one clone of A. melanoxylon. 27 young twigs of nine 1-year-old plants were inoculated with the mycelium of the 7 days-old isolates 1A912, 1B912, and 1C912(Each isolate infected three plants and each infected three young twigs) through an artificial wound. The same nine plants were inoculated with PDA medium alone (each infected three young twigs) as a negative control. Five days after inoculation, brown spindle spots similar to the field disease symptoms were observed on the twigs. No symptoms were observed on the control plants. The experiment was repeated twice. The fungus was successfully reisolated from the symptomatic plants, and had identical morphological and molecular characteristics to the initial isolates, fulfilling Koch´s postulates. To our knowledge, this is the first report of anthracnose caused by C. siamense on A. melanoxylon in China. Twig anthracnose can reduce the growth of A. melanoxylon. Further research on management options for this disease is required.

Early predictors of lung necrosis severity in children with community-acquired necrotizing pneumonia.

OBJECTIVE: To analyze baseline clinical and laboratory characteristics and explore the possible predictors of lung necrosis severity in children with community-acquired necrotizing pneumonia (NP). METHODOLOGY: This retrospective observational study was performed in a tertiary referral center. A total of 104 patients aged <15 years with community-acquired pneumonia and radiologically confirmed NP by computed tomography (CT) were included. Patients were classified into the mild, moderate, or massive necrosis groups. RESULTS: Among them, 29, 41, and 34 patients had mild, moderate, and massive necrosis, respectively. Moreover, 34.6% of the patients were admitted to pediatric intensive care unit. Massive necrosis was more likely to occur during winter (p < 0.05) and was associated with more severe clinical outcomes, such as longer duration of fever, longer hospitalization, increased mortality, and a higher risk of subsequent surgical intervention (p < 0.05). Multivariate analysis demonstrated that the following were independent risk factors for massive necrosis in this study: C-reactive protein (CRP) (p = 0.036), serum albumin (p = 0.009), and immunoglobulin M (IgM) (p = 0.022). Receiver operating characteristic analysis showed that when the cut-off value for CRP, serum albumin, and IgM were set at 122 mg/L, 30.8 g/L, and 95.7 mg/dl, respectively, they showed good diagnostic performance for differentiating patients with massive necrosis from all patients with NP. CONCLUSION: NP is a potentially severe complication of pediatric community-acquired pneumonia. Different severities of lung necrosis can lead to different clinical outcomes. CRP, serum albumin, and IgM levels are independent predictors of the degree of lung necrosis.

Correlations Between Endocrine Hormones and Cognitive Function in Patients with Obesity: a Cross-sectional Study.

BACKGROUND: Obesity not only affects physical and mental conditions, but also influences cognitive function. Endocrine hormones may influence the risk of obesity and severe obesity. Our study investigated the influences of changes in levels of endocrine hormones on cognitive function in patients with obesity and severe obesity. METHODS: We used the MATRICS Consensus Cognitive Battery (MCCB) to evaluate cognitive function in the groups of healthy control (HC), obesity (OB), and severe obesity (SOB). We detected the levels of endocrine hormones and cortisol at 8:00 am in the three groups. We statistically analyzed differences in cognitive function and levels of endocrine hormones among the three groups, and performed linear correlation analysis of cortisol level and cognitive function. Using mediation analysis, we assessed the influences of body mass index (BMI) on endocrine levels and cognitive function. RESULTS: Our findings revealed that OB and SOB groups exhibited a lower level of cortisol than HC group. Among the three groups, we found significant differences in verbal learning (F = 4.433, P = 0.014), social cognition (F = 4.778, P = 0.010), and total cognition (F = 2.989, P = 0.008). After post hoc Bonferroni correction, we noted that SOB group had worse scores in the above-mentioned three areas than OB and HC groups. Moreover, except for working memory and visual cognition, we identified that the degree of cognitive impairment in SOB group was more severe than that in OB group. In our study, mediation analysis showed a mediating effect of BMI on cortisol level and verbal learning. We also found correlations between cortisol level and attention/alertness (r = 0.277, P = 0.003), between cortisol level and verbal learning (r = 0.205, P = 0.030), and between BMI and verbal learning (r = - 0.192, P = 0.043). CONCLUSION: In our study, patients in both OB and SOB groups experienced a widespread cognitive impairment. We also found that patients with severe obesity had more severe cognitive impairment. Cognitive impairment could be mediated by abnormal cortisol metabolism, and BMI could be a mediating factor in regulation of cortisol level.

3-Bromopyruvate reverses hypoxia-induced pulmonary arterial hypertension through inhibiting glycolysis: In vitro and in vivo studies.

BACKGROUND: Pulmonary arterial smooth muscle cell (PASMC) proliferation is vital to pulmonary vascular remodeling in pulmonary arterial hypertension (PAH) pathogenesis, and inhibiting PASMC metabolism could serve as a new possible therapy to reverse the process. 3-Bromopyruvate (3-BrPA) is an effective glycolysis inhibitor with its effect in PAH remains unclear. Our study aims to assess the therapeutic effect of 3-BrPA in PAH rats and investigate the possible mechanism of 3-BrPA in PASMC proliferation and apoptosis. METHODS: 27 healthy SD rats were grouped and treated with hypoxia/normoxia and administration of 3-BrPA/physiological saline. Mean pulmonary artery pressure (mPAP) and cardiac output (CO) were measured and pulmonary vascular resistance (PVR) was calculated. Right ventricular hypertrophy index (RVHI) was calculated to evaluate the right ventricular hypertrophy degree. The percentage of medial wall area (WA%) and medial wall thickness (WT%) were measured by image analysis. PASMCs groups received hypoxia/normoxia treatments and 3-BrPA/physiological saline. PASMC proliferation and migration were respectively detected by CCK-8 and cell wound scratch assay. Hexokinase II (HK-2) expression and lactate level were respectively measured by Western Blotting and lactate test kit to detect glycolysis. RESULTS: mPAP, PVR, PVHI, WA% and WT% in rats increased after the hypoxia treatment, but were lower compared to rats received 3-BrPA in hypoxia environment. HK-2 expression, lactate concentration, OD value and scratch areas in PASMCs increased after the hypoxia treatment, but were decreased after the administration of 3-BrPA. CONCLUSION: 3-BrPA can inhibit PASMC proliferation and migration by inhibiting glycolysis, and is effective in reversing the vascular remodeling in hypoxia-induced PAH rats.

Grape seed proanthocyanidin reverses pulmonary vascular remodeling in monocrotaline-induced pulmonary arterial hypertension by down-regulating HSP70.

Heat shock protein 70 (HSP70) is a molecular chaperone which has a low content in cytoplasm under normal physiological conditions. A higher intracytoplasmic HSP70 level can be observed in pulmonary arterial smooth muscle cell (PASMC) in pulmonary arterial hypertension (PAH), and this up-regulation can promote pho-IκBα expression, which is an NF-κB signaling pathway inhibitor. NF-κB signaling pathway up-regulation can promote PASMC proliferation and pulmonary vascular remodeling in PAH, resulting in elevation of pulmonary pressure and the subsequent right heart failure caused by right ventricular hypertrophy. Grape seed proanthocyanidin (GSP) is effective in vascular protection and several tumor treatments, and its effect on PAH treatment remains to be elucidated. In this study, we made observations and contrasts in monocrotaline(MCT) -induced PAH rats, and found decrease in mPAP, PVR and RVHI after GSP administration. Our study also proved GSP's effect on down-regulating the intracytoplasmic HSP70 content both in cellular and animal levels. The results indicate a possible mechanism of GSP reversing pulmonary vascular remodeling by down-regulating HSP70, and this change may influence pho-IκBα expression. Therefore, inhibition of NF-κB signaling pathway caused by GSP can lead to inhibition of PASMC proliferation in PAH.