A novel Pik allele confers extended resistance to rice blast.

In the ongoing arms race between rice and Magnaporthe oryzae, the pathogen employs effectors to evade the immune response, while the host develops resistance genes to recognise these effectors and confer resistance. In this study, we identified a novel Pik allele, Pik-W25, from wild rice WR25 through bulked-segregant analysis, creating the Pik-W25 NIL (Near-isogenic Lines) named G9. Pik-W25 conferred resistance to isolates expressing AvrPik-C/D/E alleles. CRISPR-Cas9 editing was used to generate transgenic lines with a loss of function in Pik-W25-1 and Pik-W25-2, resulting in loss of resistance in G9 to isolates expressing the three alleles, confirming that Pik-W25-induced immunity required both Pik-W25-1 and Pik-W25-2. Yeast two-hybrid (Y2H) and split luciferase complementation assays showed interactions between Pik-W25-1 and the three alleles, while Pik-W25-2 could not interact with AvrPik-C, -D, and -E alleles with Y2H assay, indicating Pik-W25-1 acts as an adaptor and Pik-W25-2 transduces the signal to trigger resistance. The Pik-W25 NIL exhibited enhanced field resistance to leaf and panicle blast without significant changes in morphology or development compared to the parent variety CO39, suggesting its potential for resistance breeding. These findings advance our knowledge of rice blast resistance mechanisms and offer valuable resources for effective and sustainable control strategies.

First Report of 16SrII Group Related Phytoplasma Associated with Areca Palm Yellow Leaf Disease on Areca catechu in China.

The areca palm, Areca catechu L., family Arecaceae is an important herbal medicine which has potential for the treatment of parasitic diseases, digestive function disorders and depression (Peng et al. 2015). Yellow leaf disease (YLD), caused by phytoplasma, is a destructive disease of Areca catechu. In 1981, the YLD was first discovered in Tunchang, Hainan, China. According to the investigation in 2020, the occurrence area of YLD was 32 102.38 hm2 in Hainan, China, resulting in 50%-60% yield loss. Previous researchers based on 16S rDNA gene PCR amplification analysis showed that YLD in Hainan was caused by 16SrI group phytoplasma (Che et al. 2010). In August, 2022, yellow leaf symptoms were observed on middle and lower leaves of Areca catechu. Forty symptomatic plants and three asymptomatic samples were collected in Wenchang, Hainan, China (19°33'9″N, 110°48'5″E). Forty-three samples (0.1g each) were used to extract total DNA (TIANGEN plant genomic DNA extraction kit). Phytoplasma universal primers named P1/P7 (Schneider et al. 1995) and R16F2n/R16R2 (Gundersen and Lee 1996) for 16Sr DNA and primers named fTuf1/rTuf1 and fTufu/rTufu (Schneider et al. 1997) for tuf genes were used for amplifying phytoplasma sequences from isolated DNA samples by nested PCR. No fragment was amplified in asymptomatic plants and four out of forty symptomatic samples could amplify target fragment. R16F2n/R16R2 amplicons (1 248 bp) and fTufu/rTufu amplicons (845 bp) from four symptomatic Areca catechu samples were sequenced in BGI (https://genomics.cn/). The 16Sr DNA GenBank accession numbers of four positive strains (named HNWC5, HNDZ1, HNDZ3 and HNDZ6) were OQ586072, OQ586085, OQ586086, OQ586087, respectively and the tuf GenBank accession numbers were OQ595209, OQ595210, OQ595211, OQ595212, respectively. Sequence alignment showed that the 16S rDNA and tuf sequence of HNDZ1, HNDZ3 and HNDZ6 were 100% consistent. 16S rDNA of HNWC5 was 99.96% consistent with HNDZ1 and tuf of HNWC5 was 98.31% consistent with HNDZ1. Interestingly, blast search based on 16S rDNA gene of HNWC5 showed 100% sequence identity with that of 16SrII group phytoplasma such as 'Eclipta prostrata' phytoplasma strain Ep1(MH144204.1), 'Aeschynomene americana' phytoplasma isolate AA1(MH231157.1) and 'Acacia confusa' witches'-broom phytoplasma isolate HK6(ON408364.1). Blast search based on tuf gene of HNWC5 showed 98.7% sequence identity with that of bamboo witches'-broom phytoplasma (FJ853160.1) and 91.02% sequence identity with that of 'podocarpus nagi' fasciation phytoplasma (KR633146) and 90.78% sequence identity with that of 'Musa acuminata' elephantiasis disease phytoplasma (MF983708). The phylogenetic tree was constructed based on 16Sr DNA gene by MEGA 7.0 employing neighbor-joining (NJ) method with 1000 bootstrap value (Kumar et al. 2016). The result indicated that the HNWC5, HNDZ1, HNDZ3 and HNDZ6 phytoplasma strains clustered a subclade in 16SrII group. The virtual RFLP analysis based on the 16Sr DNA gene sequence was performed by the online phytoplasma classification tool iPhyClassifier (Zhao et al. 2009) using restriction endonucleases of AluI, BamHI, BfaI, BstUI, DraI, EcoRI, HaeIII, HhaI, HinfI, HpaI, HpaII, KpnI, Sau3AI, MseI, RsaI, SspI and TaqI. The result indicated that HNWC5 was most similar to the reference pattern of peanut witches'-broom phytoplasma (16SrII-A subgroup, GenBank accession: L33765) and the pattern similarity coefficient of HNWC5 is 1.00. However, the HpaII restriction endonuclease pattern of HNDZ1, HNDZ3 and HNDZ6 was different from L33765 and the similarity coefficient was 0.97, which indicated this strain may represent a new subgroup within the 16SrII group. To our knowledge, this is the first report of 16SrII group related phytoplasma associated with YLD on Areca catechu in China. Our study contributes to understanding the polymorphism of phytoplasma causing YLD and provides an important reference for pathogen specific detection.

First Report of 16SrI-B Subgroup Related Phytoplasma Associated with Yellows Symptoms of Rubus cochinchinensis in China.

Rubus cochinchinensis, an important traditional Chinese medicine in China is used to treat rheumatic arthralgia, bruises and lumbocrural pain (He et al.2005). In January 2022, yellow leaves of R. cochinchinensis were found in Tunchang City, Hainan Province, a tropical island in China. Chlorosis spread along the direction of vascular tissue while the leaf veins remain green (Fig. 1). In addition, the leaves were slightly shrunken and the growth vigor is poor (Fig. 1). By survey, we found the incidence of this disease was about 30%. Three etiolated samples and three healthy samples (0.1g each) were used to extract total DNA (TIANGEN plant genomic DNA extraction kit). Using nested PCR method, phytoplasma universal primers P1 / P7 (Schneider et al., 1995) and R16F2n / R16R2 (Lee et al. 1993) were used to amplified phytoplasma 16S rDNA gene. Primers rp F1 / R1 (Lee et al. 1998) and rp F2 / R2 (Martini et al. 2007) were used to amplified rp gene. 16S rDNA gene and rp gene fragments were amplified from three leaf etiolated samples, but not from healthy samples. The amplified fragments were cloned and sequenced, and the sequences were assembled by DNASTAR11. By sequence alignment, we found the obtained 16S rDNA and rp gene sequences of three leaf etiolated samples were same. The length of 16S rDNA fragment was 1237 bp (accession number: ON944105) and the length of rp gene fragment was 1212 bp (accession number: ON960069). The phytoplasma strain was named as 'R. cochinchinensis' yellows leaf phytoplasma (RcT), RcT-HN1 strain. The 16S rDNA gene sequence of RcT-HN1is 99.8% consistent with 16SrI-B subgroup members such as the 'Brassica napus' dwarf phytoplasma strain WH3 (MG599470.1), Chinaberry yellows phytoplasma strain LJM-1(KX683297.1) and Arecanut yellow leaf disease phytoplasma strain B165 (FJ694685.1). The rp gene sequence of RcT-HN1 is 100% consistent with rpI-B subgroup members such as the 'Salix tetradenia' witches'-broom phytoplasma strain YM-1 (KC117314.1) and Chinaberry witches'-broom phytoplasma strain Hainan (EU348781.1). The phylogenetic tree analysis, based on concatenated 16S rDNA-rp gene sequence of same group phytoplasma by MEGA 7.0 employing neighbor-joining (NJ) method with 1000 bootstrap value, were performed (Kumar et al., 2016). The results showed that RcT-HN1 phytoplasma strain formed a subclade in aster yellows group B subgroup (Fig. 2). The virtual RFLP analysis based on the 16S rRNA gene fragment of RcT-HN1 phytoplasma strain was performed by the interactive online phytoplasma classification tool iPhyClassifier (Zhao et al., 2009). The results showed that the phytoplasma strain was same as the reference pattern of the onion yellows phytoplasma of 16SrI-B (GenBank accession: AP006628), and the similarity coefficient was 1.00. This is the first report that 16SrI-B subgroup related phytoplasma infected R. cochinchinensis and caused yellows symptoms in China. The discovery of the disease is helpful to the study of the spread of phytoplasma-related diseases and protect R. cochinchinensis resources.

The putative Gγ subunit gene MGG1 is required for conidiation, appressorium formation, mating and pathogenicity in Magnaporthe oryzae.

Heterotrimeric G-proteins play key roles in the transduction of extracellular signals to various downstream effectors in eukaryotes. In our previous study, a T-DNA insertional mutant A1-412, in which the promoter of a putative Gγ subunit gene MGG1 was disrupted, was impaired in asexual/sexual sporulation, appressorium formation, and pathogenicity in Magnaporthe oryzae. Here the roles of MGG1 in regulating fungal development and plant infection were further investigated and verified using a gene deletion strategy. Targeted gene deletion mutants of MGG1 exhibited similar phenotypes to those of A1-412. The Δmgg1 mutants were unable to differentiate appressorium on hydrophobic surfaces and nonpathogenic to susceptible hosts. The defects of the Δmgg1 mutants in appressorium formation were partially restored by adding exogenous cAMP or IBMX (a phosphodiesterase inhibitor), although the induced appressoria were still nonfunctional. Expressing Mgg1-GFP fusion protein in an Δmgg1 mutant could complement all phenotypes of the mutant, and bright GFP fluorescence was observed at the periphery of fungal cells, indicating that Mgg1 mainly localizes to plasma membrane. Quantitative RT-PCR analysis revealed that deletion of MGG1 resulted in a significant reduction in mRNA levels of the genes encoding Gα (MagA, MagB, and MagC), Gß (Mgb1), and adenylate cyclase (Mac1). Moreover, intracellular cAMP accumulation was significantly reduced in Δmgg1 mutants compared to that in the wild-type strain. Taken together, our results suggested that Gγ subunit Mgg1 might act upstream of cAMP signaling pathway and play critical roles in regulation of conidiation, appressorium formation, mating, and plant infection in M. oryzae.

[Influence of different sevoflurane concentration on somatosensory evoked potentials monitoring in spinal cord surgery].

OBJECTIVE: To evaluate the effects of different end-tidal concentrations of sevoflurane on somatosensory evoked potentials, to explore the feasibility of sevoflurane applied in such kind of surgery, so as to provide useful information in making anesthesia plan for spinal cord surgery. METHODS: Thirty-two patients scheduled for spinal cord surgery (ASA I-II, 18-65 years old) were enrolled. After induction of anesthesia, they were assigned to receive sevoflurane anesthesia of increment end-tidal concentration in the sequence of 0.0%, 0.5%, 1.0% and 1.5% respectively, under a background intravenous infusion of propofol and remifentanil. Remifentanil infusion rate was 0.2 µg·kg⁻¹ ·min⁻¹, while the propofol infusion rate was adjusted to maintain BIS values within the range of 30-50. The amplitude and latency of each SSEPs were recorded and compared. RESULTS: Sevoflurane inhibited SSEPs in a dose-dependent manner, SSEPs amplitude significantly decreased following increased end-tidal sevoflurane concentration. The amplitudes of the left side were 2.36 (0.42-9.87), 2.14 (0.52-9.44), 1.94 (0.47-9.44), 1.64 (0.36-7.46) µV respectively (F = 21.66, P < 0.01). The amplitudes of the right side were 2.71 (0.43-7.1), 2.73 (0.43-7.1), 2.34 (0.44-6.6), 1.64 (0.39-6.15) µV respectively (F = 33.94, P < 0.01). SSEPs latencies were significantly prolonged, on the left side, the latencies were (41.48 ± 3.45), (42.45 ± 3.60), (43.20 ± 3.42), (44.38 ± 3.78) ms, respectively (F = 68.07, P < 0.01). On the right side, the latencies were (40.65 ± 4.91), (41.53 ± 4.76), (42.31 ± 4.93), (43.39 ± 4.79)ms, respectively (F = 56.52, P < 0.01). Yet, as a monitoring modality for dynamic observation, SSEPs still could be monitored successfully under sevoflurane anesthesia in all these 32 patients. CONCLUSIONS: Sevoflurane has depression effects on SSEPs in a dose-dependent manner. It can decrease the amplitudes and prolong the latencies. Considering significant individual difference, the feasibility of sevoflurane in such kind of surgery can be determined by measuring base amplitude of SSEPs under total intravenous anesthesia.

Widely targeted metabolomics combined with E-tongue and E-nose reveal dynamic changes of tender coconut water in responses to the infection of Ceratocystis paradoxa.

Ceratocystis paradoxa is a major cause of postharvest disease in tender coconuts worldwide. We conducted a comprehensive study using widely targeted metabolomics, electronic tongue (E-tongue), and electronic nose (E-nose) analyses to investigate the impacts of C. paradoxa invasion on the quality of tender coconut water (TCW) from fresh control (FC), uninoculated (UN), skin-inoculated (SI), and deep-inoculated (DI) nuts. DI exhibited significantly higher taste indicators associated with bitterness, saltiness, astringency aftertaste, and bitter aftertaste, as well as odor sensor values related to various compounds such as long-chain alkanes, hydrides, methane, organic sulfides, etc. Invasion of C. paradoxa into the endosperm altered the flavor characteristics of TCW mainly through the modulation of carbohydrate and secondary metabolite pathways. Furthermore, significant correlations were observed between the differentially expressed flavorful metabolites and the sensor indicators of the E-nose and E-tongue. These findings offer valuable insights into understanding the impact of C. paradoxa infection on coconuts.

Acupuncture vs Sham Acupuncture for Chronic Sciatica From Herniated Disk: A Randomized Clinical Trial.

Importance: Sciatica is commonly caused by herniated lumbar disc and contributes to severe pain and prolonged disability. Although acupuncture is widely used by patients with chronic sciatica, the evidence of its efficacy is scarce. Objective: To investigate the efficacy and safety of acupuncture compared with sham acupuncture in patients with chronic sciatica from herniated disk. Design, Settings, and Participants: This was a multicenter 2-arm randomized clinical trial conducted in 6 tertiary-level hospitals in China of patients with chronic sciatica from herniated disk. Participants were recruited from March 25, 2021, to September 23, 2021, with a final follow-up through September 22, 2022. Data analyses were performed from December 2022 to March 2023. Interventions: Participants were randomly assigned to receive 10 sessions of acupuncture (n = 110) or sham acupuncture (n = 110) over 4 weeks. Participants, outcome assessors, and statisticians were blinded. Main Outcomes and Measures: The 2 coprimary outcomes were changes in visual analog scale (VAS) for leg pain and Oswestry Disability Index (ODI) from baseline to week 4. Secondary outcomes were adverse events. Results: A total of 216 patients (mean [SD] age, 51.3 [15.2] years; 147 females [68.1%] and 69 males [31.9%]) were included in the analyses. The VAS for leg pain decreased 30.8 mm in the acupuncture group and 14.9 mm in the sham acupuncture group at week 4 (mean difference, -16.0; 95% CI, -21.3 to -10.6; P < .001). The ODI decreased 13.0 points in the acupuncture group and 4.9 points in the sham acupuncture group at week 4 (mean difference, -8.1; 95% CI, -11.1 to -5.1; P < .001). For both VAS and ODI, the between-group difference became apparent starting in week 2 (mean difference, -7.8; 95% CI, -13.0 to -2.5; P = .004 and -5.3; 95% CI, -8.4 to -2.3; P = .001, respectively) and persisted through week 52 (mean difference, -10.8; [95% CI, -16.3 to -5.2; P < .001; and -4.8; 95% CI, -7.8 to -1.7; P = .003, respectively). No serious adverse events occurred. Conclusions and Relevance: This randomized clinical trial found that in patients with chronic sciatica from herniated disk, acupuncture resulted in less pain and better function compared with sham acupuncture at week 4, and these benefits persisted through week 52. Acupuncture should be considered as a potential treatment option for patients with chronic sciatica from a herniated disk. Trial Registration: Chictr.org Identifier: ChiCTR2100044585.

[Comparative study on two total intravenous anesthesia techniques in complex spine surgery].

OBJECTIVE: To compare anesthesia profiles of target-controlled infusion (TCI) and manual-controlled infusion (MCI) of propofol and remifentanil in kyphosis correction or scoliosis correction surgery, in which intraoperative spinal cord monitoring was employed. METHODS: In the study, 160 patients scheduled for kyphosis correction surgery or scoliosis correction were enrolled and randomly allocated into 2 groups, group TCI and group MCI. In group TCI, induction and maintenance of general anesthesia were carried out by target-controlled infusion of propofol and remifentanil. Marsh and Minto, three-compartment pharmacokinetic models for propofol and remifentanil were used respectively. In group MCI, the patients received propofol and remifentanil by conventional dose-weight infusion method. Muscle relaxants were only applied for ease of induction. In both the groups, anesthesia depth was monitored by bispectral index (BIS), and the change of hemodynamic parameters was kept in the range of 20% of the baseline. The control convenience of anesthesia depth and change amplitude of BIS in the main steps of the procedure were compared. The total amounts of propofol and remifentanil, influence on the hemodynamics, influence on the successful rates of somatosensory evoked potentials (SSEPs) and motor evoked potentials (MEPs) and electromyography (EMG) were compared too. RESULTS: The time of pump regulation in group TCI was significantly less than in group MCI (P<0.05). The BIS fluctuation in the main steps of the procedure was significantly smaller in Group TCI than in group MCI (P<0.05). There was no significant difference in the amounts of propofol and remifentanil(P=0.158 and P=0.168). The time to awake (P=0.972) and time to extubation (P=0.944) had no significant difference. The successful rates of SSEPs and MEPs had no significant difference between groups TCI amd MCI (P>0.05). CONCLUSION: Both TCI and MCI can offer practical anesthesia for spinal deformity correction surgery. TCI has the advantage in keeping smooth and steady depth of anesthesia.

Acupuncture for chronic sciatica: protocol for a multicenter randomised controlled trial.

BACKGROUND: Chronic Sciatica is a disabling condition causing considerable medical, social and financial implications. Currently, there is no recognised long-term effective treatment to alleviate sciatica. Acupuncture has been widely used for treating chronic pains with persistent analgesic effects. We aim to evaluate the efficacy and safety of acupuncture for chronic sciatica with follow-up in 52 weeks. METHODS AND ANALYSIS: This is a multicenter randomised sham-controlled trial. A total of 216 patients with chronic sciatica will be enrolled and randomly assigned to the acupuncture or sham acupuncture group. There will be 10 treatment sessions applied in 4 weeks with frequency decreased over time. Patients will complete follow-ups during 52 weeks. The primary outcomes are changes in leg pain intensity and disability from baseline to week 4. Secondary outcomes include back pain intensity, frequency and bothersomeness, quality of life, and global perceived effect. Adverse events will be recorded in detail. ETHICS AND DISSEMINATION: Ethical approval of this trial was granted from the ethics committee of Beijing University of Chinese Medicine and all study centres (No. 2020BZYLL0803). Written informed consent will be obtained from enrolled patients. Trial results will be disseminated in peer-reviewed publications. TRIAL REGISTRATION NUMBER: ChiCTR2100044585 (Chinese Clinical Trial Registry, http://www.chictr.org.cn, registered on 24 March 2021); preresults.

Efficacy of acupuncture for sciatica: study protocol for a randomized controlled pilot trial.

BACKGROUND: Acupuncture is widely used for pain diseases while evidence of its efficacy for sciatica is insufficient. We aim to explore the feasibility and efficacy of acupuncture with different acupoint selecting strategies for sciatica induced by lumbar disc herniation. METHODS: This is a multicenter, three-arm, patient-assessor-blinded randomized controlled pilot trial. Ninety patients will be assigned randomly into 3 groups including disease-affected meridians (DAM) group, non-affected meridians (NAM) group, and sham acupuncture (SA) group in a 1:1:1 ratio. The trial involves a 4-week treatment along with follow-up for 22 weeks. The primary outcome is the change of leg pain intensity measured by the visual analogue scale (VAS) from baseline to week 4 after randomization. Secondary outcomes include functional status, back pain intensity, and quality of life. Adverse events will also be recorded. DISCUSSION: The results will inspire the optimal acupuncture strategy for sciatica and help establish a better design as well as power calculation for a full-scale study. TRIAL REGISTRATION: ChiCTR2000030680 (Chinese Clinical Trial Registry, http://www.chictr.org.cn , registered on 9 March 2020).

[Anesthesia for cesarean section in pregnancy complicated with severe scoliosis: a case report].

Scoliosis refers to a complex three-dimensional spine deformity. Onset of scoliosis can occur in an infantile, juvenile, adolescent, or adult period. Severe scoliosis not only affects the appearance of the patient, but also impairs the functions of circulation and respiratory systems. Significant changes of the cardiopulmonary function can occur during pregnancy. In this report, general anesthesia was needed for emergent cesarean section of the puerperant complicated with severe scoliosis. The puerperant had already suffered type II respiratory failure preoperatively. There might be managements of difficult airway, depression with circulation and respiration of the neonate, and the risk for inhibition of the uterine contraction and awareness. Facing the challenges, the anesthesiologists must work out a detail anesthesia plan quickly, and make sure the safety of the puerperant and the neonate.

The broad-spectrum rice blast resistance (R) gene Pita2 encodes a novel R protein unique from Pita.

BACKGROUND: Rice blast is generally considered the most devastating rice disease worldwide. The development of resistant varieties has been proven to be the most economical strategy to control the disease. A cluster of resistant (R) genes on rice chromosome 12 including Pita, Pita2 and Ptr has been studies for decades. However, the relationship between these R genes has not been well established. RESULTS: In this study, we compared the resistance spectra controlled by Pita2 and Pita by testing their monogenic lines (MLs) in four hotspots found in the Philippines and Burundi from 2014 to 2018. The reaction patterns were distinct in two countries and that Pita2-mediated field resistance was relatively prevalent. Pathogenicity tests using 328 single-spore isolates in greenhouse further verified that IRBLta2-Re for Pita2 conferred a relatively broader spectrum resistance than those of Pita. Rough and fine mapping of Pita2 were conducted using F2 and F3 populations derived from IRBLta2-Re [CO] and CO 39 consisting of 4344 progeny to delimit Pita2 in a genomic interval flanked by two markers 12 g18530 and 12 g18920 proximal to the centromere of chromosome 12. Alignment of the markers to the genomic sequence of IR64, which harbors Pita2 verified by genetic analysis, approximately delimited the candidate gene(s) within 313-kb genomic fragment. The two Pita2 suppressive mutants that contain mutations within Pita2 were verified and identified. Comparative sequence analysis in these two mutants further identified that each individual allele contains a single nucleotide substitution at a different position resulting in nonsense and missense mutations in the protein product of LOC_Os12g18729. On the contrary, no sequence mutation was detected in other candidate genes, indicating that mutations in LOC_Os12g18729 were responsible for the loss of function of Pita2. Pita2 encodes a novel R protein unique from Pita, which is exactly identical to the previously cloned Ptr. Moreover, based on the resistance gene analysis of rice varieties and mutants containing Pita, it was found that Pita2 rather than Pita was responsible for the specificity to some differential isolates with AvrPita. The diagnosis and survey of Pita2 in IRRI released varieties showed relatively low frequency, implying a high value of its application for breeding resistant varieties against rice blast via marker assisted selection. CONCLUSION: Our study clarified the relationship between Pita, Pita2 and Ptr. Pita2 is identical to Ptr and distinct from Pita in both sequence and chromosomal location although Pita2 and Pita are genetically linked to each other. The loss of function of Pita2 but not Pita eliminate the specificity to some AvrPita containing isolates, however, the mechanism underlying the recognition between Pita2/Pita and AvrPita remains elusive.

Functional characterization of electron-transferring flavoprotein and its dehydrogenase required for fungal development and plant infection by the rice blast fungus.

Electron-transferring flavoprotein (ETF) and its dehydrogenase (ETFDH) are highly conserved electron carriers which mainly function in mitochondrial fatty acid ß oxidation. Here, we report the identification and characterization of ETF α and ß subunit encoding genes (ETFA and ETFB) and ETFDH encoding gene (ETFDH) in the rice blast fungus Magnaporthe oryzae. It was demonstrated that, by impacting fatty acid metabolism, ETF and ETFDH mutations led to severe growth and conidiation defects, which could be largely rescued by exogenous acetate or carbonate. Furthermore, although conidium germination and appressorium formation appeared to be normal in ETF and ETFDH mutants, most appressoria failed to penetrate the host epidermis due to low turgor pressure. The few appressoria that succeeded in penetration were severely restricted in invasive growth and consequently failed to cause disease. Moreover, ETF mutant etfb(-) induced ROS accumulation in infected host cells and exogenous antioxidant GSH accelerated mutant invading growth without increasing the penetration rate. In addition, mutant etfb(-) displayed elevated lipid body accumulation and reduced ATP synthesis. Taken together, ETF and ETFDH play an important role in fungal development and plant infection in M. oryzae by regulation of fatty acid metabolism, turgor establishment and induction of host ROS accumulation.

The antiallodynic action of pregabalin may depend on the suppression of spinal neuronal hyperexcitability in rats with spared nerve injury.

BACKGROUND: Pregabalin (PGB) is a novel antiepileptic drug and is also used as a first-line medication for the treatment of neuropathic pain. However, the mechanisms of its analgesic effects remain largely unknown. OBJECTIVES: To elucidate the mechanisms underlying the antiallodynic action of PGB in rats with neuropathic pain. METHODS: In a rat model of neuropathic pain induced by spared nerve injury, mechanical allodynia, as a behavioural sign of neuropathic pain, was assessed by measuring 50% paw withdrawal threshold with von Frey filaments. Activities of dorsal horn wide dynamic range (WDR) neurons were examined by extracellular electrophysiological recording in vivo. RESULTS: Spinal administration of PGB exerted a significant antiallodynic effect and a prominent inhibitory effect on the hypersensitivity of dorsal horn WDR neurons in rats with spared nerve injury. CONCLUSION: The antiallodynic action of PGB is likely dependent on the suppression of WDR neuron hyperexcitability in rats with neuropathic pain.

Performance of biotrickling filters for hydrogen sulfide removal under starvation and shock loads conditions.

In the industrial operation of biotrickling filters for hydrogen sulfide (H(2)S) removal, shock loads or starvation was common due to process variations or equipment malfunctions. In this study, effects of starvation and shock loads on the performance of biotrickling filters for H(2)S removal were investigated. Four experiments were conducted to evaluate the changes of biomass and viable bacteria numbers in the biotrickling filters during a 24-d starvation. Compared to biomass, viable bacteria numbers decreased significantly during the starvation, especially when airflow was maintained in the absence of spray liquid. During the subsequent re-acclimation, all the bioreactors could resume high removal efficiencies within 4 d regardless of the previous starvation conditions. The results show that the re-acclimation time, in the case of biotrickling filters for H(2)S removal, is mainly controlled by viable H(2)S oxidizing bacteria numbers. On the other hand, the biotrickling filters can protect against shock loads in inlet fluctuating H(2)S concentration after resuming normal operation. When the biotrickling filters were supplied with H(2)S at an input of lower than 1700 mg/m(3), their removal efficiencies were nearly 98% regardless of previous H(2)S input.

Cellular basis of itch sensation.

Itch and pain are two distinct sensations. Although our previous study suggested that gastrin-releasing peptide receptor (GRPR) is an itch-specific gene in the spinal cord, a long-standing question of whether there are separate neuronal pathways for itch and pain remains unsettled. We selectively ablated lamina I neurons expressing GRPR in the spinal cord of mice. These mice showed profound scratching deficits in response to all of the itching (pruritogenic) stimuli tested, irrespective of their histamine dependence. In contrast, pain behaviors were unaffected. Our data also suggest that GRPR+ neurons are different from the spinothalamic tract neurons that have been the focus of the debate. Together, the present study suggests that GRPR+ neurons constitute a long-sought labeled line for itch sensation in the spinal cord.

Two-stage analysis of pharmacokinetics of sufentanil administered by target-controlled infusion in Chinese patients.

BACKGROUND: Sufentanil is a suitable choice for target-controlled infusion (TCI) because of its shorter context-sensitive half-time. The current study was to estimate the pharmacokinetics of sufentanil TCI in Chinese patients using the two-stage analysis. METHODS: Twelve adult patients with American Society of Anesthesiologists (ASA) physical status I or II undergoing elective surgery under general anesthesia were included. Anesthesia was induced with propofol, rocuronium and sufentanil administered by TCI lasting for 30 minutes, with target effect-site concentration of sufentanil 4 or 6 ng/ml. Frequent arterial blood samples (1.5 ml) were taken during and up to 24 hours after sufentanil TCI. Before the end of surgery, another arterial blood sample (1.0 ml) was drawn for the blood-gas analysis. Plasma sufentanil concentrations were determined by liquid chromatography-tandem mass spectrometry (limit of quantitation was 5 pg/ml). The data were analyzed with the two-stage approach, linear regression and correlation analysis. RESULTS: The pharmacokinetics of sufentanil TCI were adequately described by a three-compartment model. The variables were derived as follows: the volume of central compartment (V(1)) was 5.4 L, volume of distribution at steady-state (Vdss) was 222.6 L, metabolic clearance (Cl(1)) was 0.84 L/min and elimination half-life (t(1/2Y)) was 389 minutes. Patients' age, gender and PaCO2 correlated significantly with the pharmacokinetic parameters. The Vdss, volume of slowly equilibrating compartment (V(3)) and t(1/2Y) increased, and rapid distribution clearance (Cl(2)) decreased with increasing patient age. Male patients had larger values of Vdss, volume of rapidly equilibrating compartment (V(2)) and V(3) than female patients. The Vdss and V(3) increased with higher PaCO2 values. There were no significant correlations between the pharmacokinetic variables and body weight, height, lean body mass, plasma albumin, sufentanil dose, duration of surgery, pH or base excess of blood (BE-B). CONCLUSIONS: The pharmacokinetics of sufentanil TCI in Chinese patients can be optimally described by a three-compartment model. The pharmacokinetic analysis technique may affect the pharmacokinetic parameters and correlations.