Improving the Comprehension of Pathogenicity and Phylogeny in 'Candidatus Phytoplasma meliae' through Genome Characterization.

'Candidatus Phytoplasma meliae' is a pathogen associated with chinaberry yellowing disease, which has become a major phytosanitary problem for chinaberry forestry production in Argentina. Despite its economic impact, no genome information of this phytoplasma has been published, which has hindered its characterization at the genomic level. In this study, we used a metagenomics approach to analyze the draft genome of the 'Ca. P. meliae' strain ChTYXIII. The draft assembly consisted of twenty-one contigs with a total length of 751.949 bp, and annotation revealed 669 CDSs, 34 tRNAs, and 1 set of rRNA operons. The metabolic pathways analysis showed that ChTYXIII contains the complete core genes for glycolysis and a functional Sec system for protein translocation. Our phylogenomic analysis based on 133 single-copy genes and genome-to-genome metrics supports the classification as unique 'Ca. P. species' within the MPV clade. We also identified 31 putative effectors, including a homolog to SAP11 and others that have only been described in this pathogen. Our ortholog analysis revealed 37 PMU core genes in the genome of 'Ca. P. meliae' ChTYXIII, leading to the identification of 2 intact PMUs. Our work provides important genomic information for 'Ca. P. meliae' and others phytoplasmas for the 16SrXIII (MPV) group.

Complete genome sequence of Agrobacterium pusense Bbcg2-2, a strain isolated from blueberry crown gall in Taiwan.

Agrobacterium pusense Bbcg2-2 is a strain isolated from a crown gall sample of blueberry (Vaccinium corymbosum) cultivar "Flicker" grown in Taiwan. The complete genome sequence of this bacterium consists of a 2,798,342-bp circular chromosome, a 2,140,031-bp linear chromid, and a 386,016-bp circular plasmid.

Complete genome sequence of Candidatus Phytoplasma australasiaticum WF_GM2021, a plant pathogen associated with soybean witches' broom disease in Taiwan.

The complete genome sequence of Candidatus Phytoplasma australasiaticum strain WF_GM2021, which consists of one 633,005-bp circular chromosome, is presented in this work. This uncultivated plant-pathogenic bacterium is associated with soybean (Glycine max) witches' broom disease in Wufeng District, Taichung City, Taiwan.

Complete genome sequence of "Candidatus Phytoplasma cynodontis" GY2015, a plant pathogen associated with Bermuda grass white leaf disease in Taiwan.

The complete genome sequence of "Candidatus Phytoplasma cynodontis" strain GY2015, which consists of one 498,922-bp circular chromosome, is presented in this work. This uncultivated plant-pathogenic bacterium is associated with Bermuda grass white leaf disease in Taoyuan, Taiwan.

Complete Genome Sequence of "Candidatus Phytoplasma aurantifolia" TB2022, a Plant Pathogen Associated with Sweet Potato Little Leaf Disease in China.

The complete genome sequence of "Candidatus Phytoplasma aurantifolia" TB2022, which consists of one 670,073-bp circular chromosome, is presented in this work. This bacterium is associated with sweet potato little leaf disease in Fujian Province, China.

Complete Genome Sequence of "Candidatus Phytoplasma asteris" QS2022, a Plant Pathogen Associated with Lettuce Chlorotic Leaf Rot Disease in China.

The complete genome sequence of "Candidatus Phytoplasma asteris" QS2022, which consists of one 834,303-bp circular chromosome, is presented in this work. This bacterium is associated with lettuce chlorotic leaf rot disease in Fujian Province, China.

Biological and Genetic Characterization of Physostegia Chlorotic Mottle Virus in Europe Based on Host Range, Location, and Time.

Application of high throughput sequencing (HTS) technologies enabled the first identification of Physostegia chlorotic mottle virus (PhCMoV) in 2018 in Austria. Subsequently, PhCMoV was detected in Germany and Serbia on tomatoes showing severe fruit mottling and ripening anomalies. We report here how prepublication data-sharing resulted in an international collaboration across eight laboratories in five countries, enabling an in-depth characterization of PhCMoV. The independent studies converged toward its recent identification in eight additional European countries and confirmed its presence in samples collected 20 years ago (2002). The natural plant host range was expanded from two to nine species across seven families, and we confirmed the association of PhCMoV presence with severe fruit symptoms on economically important crops such as tomato, eggplant, and cucumber. Mechanical inoculations of selected isolates in the greenhouse established the causality of the symptoms on a new indexing host range. In addition, phylogenetic analysis showed a low genomic variation across the 29 near-complete genome sequences available. Furthermore, a strong selection pressure within a specific ecosystem was suggested by nearly identical sequences recovered from different host plants through time. Overall, this study describes the European distribution of PhCMoV on multiple plant hosts, including economically important crops on which the virus can cause severe fruit symptoms. This work demonstrates how to efficiently improve knowledge on an emergent pathogen by sharing HTS data and provides a solid knowledge foundation for further studies on plant rhabdoviruses.[Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Baicalin protects neonatal rat brains against hypoxic-ischemic injury by upregulating glutamate transporter 1 via the phosphoinositide 3-kinase/protein kinase B signaling pathway.

Baicalin is a flavonoid compound extracted from Scutellaria baicalensis root. Recent evidence indicates that baicalin is neuroprotective in models of ischemic stroke. Here, we investigate the neuroprotective effect of baicalin in a neonatal rat model of hypoxic-ischemic encephalopathy. Seven-day-old pups underwent left common carotid artery ligation followed by hypoxia (8% oxygen at 37°C) for 2 hours, before being injected with baicalin (120 mg/kg intraperitoneally) and examined 24 hours later. Baicalin effectively reduced cerebral infarct volume and neuronal loss, inhibited apoptosis, and upregulated the expression of p-Akt and glutamate transporter 1. Intracerebroventricular injection of the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) inhibitor LY294002 30 minutes before injury blocked the effect of baicalin on p-Akt and glutamate transporter 1, and weakened the associated neuroprotective effect. Our findings provide the first evidence, to our knowledge that baicalin can protect neonatal rat brains against hypoxic-ischemic injury by upregulating glutamate transporter 1 via the PI3K/Akt signaling pathway.

Ultrastructural Changes of Brain Tissues Surrounding Hematomas after Intracerebral Hemorrhage.

Our knowledge about pathophysiology of intracerebral hemorrhage (ICH) mainly originates from preclinical models of ICH. In this study, cerebral ultrastructure surrounding hematoma and its correlation with clinical severity were investigated in ICH patients. Thirty patients with basal ganglia hemorrhage and 6 control subjects were enrolled. Surgical evacuation was performed for patients with a blood loss >30 ml. Stroke severity was assessed using the Glasgow Coma Scale (GCS) and the National Institute of Health Stroke Scale (NIHSS). Transmission electron microscopy (TEM) was used to evaluate the ultrastructural characteristics of tissue specimens. Neural cells surrounding the hematomas showed evidence of cell swelling and necrosis. Decreased numbers of organelles and mitochondrial cristae were accompanied by cytoplasmic vacuolization, nuclear membrane invagination and breakdown, and intranuclear chromatic agglutination. These changes resulted in disintegration together with malacia, disappearance of the nucleus and nucleolus, and karyopyknosis. More serious ultrastructural damage was seen in patients with greater NIHSS scores, lower GCS scores, and greater bleeding volumes (p < 0.001). These findings suggest that neural cells undergo unfavorable ultrastructural changes that are responsible for dysfunction after ICH.

Activation of large-conductance Ca(2+)-activated K(+) channels inhibits glutamate-induced oxidative stress through attenuating ER stress and mitochondrial dysfunction.

Large-conductance Ca(2+)-activated K(+) channels (BK channels) are widely expressed throughout the vertebrate nervous system, and are involved in the regulation of neurotransmitter release and neuronal excitability. Here, the neuroprotective effects of NS11021, a selective and chemically unrelated BK channel activator, and potential molecular mechanism involved have been studied in rat cortical neurons exposed to glutamate in vitro. Pretreatment with NS11021 significantly inhibited the loss of neuronal viability, LDH release and neuronal apoptosis in a dose-dependent manner. All these protective effects were fully antagonized by the BK-channel inhibitor paxilline. NS11021-induced neuroprotection was associated with reduced oxidative stress, as evidenced by decreased reactive oxygen species (ROS) generation, lipid peroxidation and preserved activity of antioxidant enzymes. Moreover, NS11021 significantly attenuated the glutamate-induced endoplasmic reticulum (ER) calcium release and activation of ER stress markers, including glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP) and caspase-12. Pretreatment with NS11021 also mitigated the mitochondrial membrane potential (MMP) collapse, cytochrome c release, and preserved mitochondrial Ca(2+) buffering capacity and ATP synthesis after glutamate exposure. Taken together, these results suggest that activation of BK channels via NS11021 protects cortical neurons against glutamate-induced excitatory damage, which may be dependent on the inhibition of ER stress and preservation of mitochondrial dysfunction.

[Expression of matrix metalloproteinase MMP-9 in the plasma and hematoma fluid of intracerebral hemorrhage patients].

OBJECTIVE: To investigate the matrix metalloproteinase (MMP)-9 played in secondary brain injury following intracerebral hemorrhage (ICH). METHODS: Hematoma fluid and peripheral blood samples were collected from 60 ICH patients, 34 males and 26 females, aged 60 +/- 13 (37 - 81) n the days 1, 4, and 7 after evacuation of hematoma. Peripheral blood samples were collected form. 30 sex, and age-matched healthy adults as normal controls. Cerebrospinal fluid (SCF) samples were collected from 10 sex, and age-matched patients to undergo operation during lumbar anesthesia. ELISA was used to detect the content of MMP-9. Tada formula was used to calculate the perihematomal edema volume. The National Institutes of Health Stroke Scale (NIHHS) and Glasgow Coma Score (GCS) were used to assess the condition of patients. RESULTS: (1) The MMP-9 levels in the plasma and hematoma fluid of the ICH patients at all time points were all significantly higher than those of the normal controls (all P < 0.01). MMP-9 was not found in the normal CSF. (2) The plasma and hematoma fluid MMP-9 levels were increased already in the day 1, peaked in the day 4, and then kept at a high level until the day 7. (3) The MMP-9 levels in hematoma fluid t all time points were all significantly higher than those in the plasma (all P < 0.01). (4) The MMP-9 level was positively correlated with the hematoma volume and NIHSS score, and negatively correlated with the GCS score (both P < 0.01). CONCLUSIONS: MMP-9 may takes part in the secondary injury after ICH, and its change is correlated with the hydrocephalus of patients. The dynamical change of the plasma MMP-9 level is consistent with the hematoma fluid MMP-9 level after ICH. There is a positive correlation among the MMP-9 level, perihematomal edema volume, and severity of ICH.

[Cloning and characterization of gerM gene involved in germination in Bacillus thuringiensis].

In Bacilli, gerM is a very conservative gene. Primers were designed according to the gerM gene sequence of Bacillus cereus, and a 640bp DNA fragment was obtained from Bacillus thuringiensis subsp. kustaki 1. 175 by PCR. Using this fragment as a probe, a 4.5kb DNA fragment was cloned from the partial DNA library of Bacillus thuringiensis subsp. kustaki 1.175. Sequence analysis showed that the fragment contains one complete open reading frame (ORF) that encodes a 349-amino acid (aa) protein, which has high homology with GerM protein from Bacillus subtilis. This gene was designated gerM (GenBank Accession No. DQ537381 ) . RT-PCR analysis showed that gerM gene was only expressed in the process of sporulation, suggesting gerM is not required for the vegetative growth. The function of the gerM gene was studied by a strategy of gene disruption, and the resulting gerM disruption mutant did show normal growth and sporulation. However, gerM disruption mutant spores germinate slower than wild-type spores when triggered by L-alanine or inosine, indicating that gerM is required for the spore normal germination initiated by L-alanine or inosine in Bacillus thuringiensis.

[Differentiation of human umbilical cord blood mesenchymal stem cells toward neurons induced by baicalin in vitro].

OBJECTIVE: To study the possibility and mechanism by which the human umbilical blood mesenchymal stem cells (MSCs) are induced to differentiate into neuron in vitro by baicalin, a kind of flavonoid isolated from an important medicinal plant Scutellariae Radix. METHODS: Human cord blood was obtained via sterile procedure with 20 U/ml of preservative-free heparin. MSCs were isolated by the two-step assay of gelatin sedimentation plus Ficoll centrifugation separation, purified and amplified in the liquid culture medium. According to the kind of antioxidants, the experiment was conducted in five groups: induction group, control group I, control group II, control group III and control group IV. Five subgroups of MSCs amplificated ex vivo for 2 weeks in each group were induced by the media containing baicalin (BC, 200 - 400 micromol/L) or baicalin-free for seven days. The media consisted of induction medium (DMEM plus 200 - 400 micromol/L of baicalin) and post-induction medium (DMEM plus 200 - 400 micromol/L of baicalin, B27). The expression of neuronal or glia specific markers was evaluated by using indirect immunofluorescence cytochemistry staining. The percentage of differentiated cells and living cells was measured by Hoechest 33,258 staining assay. RESULTS: After induction for 7 days, MSCs displayed neuronal morphologies, such as pyramidal cell bodies and processes formed extensive network. The undifferentiated cells did not exhibit a neuronal or glial phenotype, while the differentiating cells expressed NSE and MAP-2, the specific markers of neuron, but did not express GFAP, the specific marker of glia on the seventh day after induced by baicalin. The percentage of NSE and MAP-2 expressed on the seventh day after induced by baicalin was (76.3 +/- 9.2)% and (78.5 +/- 5.5)%, respectively, which was significantly higher compared with control group I, control group II and control group III (P < 0.01), respectively. In addition, the ratio of living cells after induced for seven days in the BC group was (85.3 +/- 4.8)%, which increased significantly compared with control group I, control group II and control group III (P < 0.01), respectively. CONCLUSIONS: Baicalin may induce the umbilical blood MSCs to neuron or neuron-alike cells in vitro in a moderate and stable manner. The mechanism of such an induction may be related with its controlling the activity of NF-kappaB which regulates the production of many kinds of cytokines, such as brain-derived neurotrophic factor (BDNF), etc.

[Neural apoptosis and apoptosis-related genes in intracerebral hemorrhage patients].

OBJECTIVE: To observe the neural apoptosis and expression of apoptosis-related genes in brain in order to elucidate the regulation mechanism in the perihematomal region of intracerebral hemorrhage (ICH) patients. METHODS: Specimens of perihematomal region in human brain were obtained from 29 patients undergoing surgical evacuation of an intracerebral hematoma. Specimens of brain tissue were collected from the corpses of 6 persons within 3 hours after the accidental death. Neural apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated deoxyuridine 5' triphosphate nick end labeling (TUNEL) method and immunohistochemistry was used to detect the expression of Bcl-2, Bax, P53, and caspase-3 genes. RESULTS: The apoptosis rates of the ICH group was 4.10 +/- 0.28, significantly higher than that of the control group (0.57 +/- 0.43, P < 0.01). The expression rate of Bcl-2 the ICH group was 2.68 +/- 0.52, significantly higher than that of the control group (1.54 +/- 0.56, P < 0.01). The expression rate of Bax of the ICH group was 3.49 +/- 0.18, significantly higher than that of the control group (0.96 +/- 0.27, P < 0.01). The expression of P53 was 4.12 +/- 0.63, significantly higher than that of the control group (0.96 +/- 0.71, P < 0.01). The expression of caspase-3 of the ICH group was 3.50 +/- 0.25, significantly higher than that of the control group (0.74 +/- 0.73, P < 0.01). The expression levels of Bcl-2 and P53 were negatively correlated with the apoptosis rate (both P < 0.01), while the expression levels of Bax and caspase-3, and the Bax/Bcl-2 ratio were positively correlated with the apoptosis rate in perihematomal region of ICH patients (all P < 0.01). CONCLUSION: Apoptosis is involved in the delayed brain injury after ICH in human and is the main factor of delayed neural death. Some of the genes take part in the regulation of neural apoptosis: Bax and caspase-3 hasten the apoptosis while Bcl-2 and P53 restrain it.

[Prophylactic effect of artesunate against experimental infection of Schistosoma mansoni].

OBJECTIVE: To study the prophylactic effect of artesunate against the infection of Schistosoma mansoni in mice and its optimal scheme for preventing schistosomiasis mansoni. METHODS: BALB/c mice were infected by tail dipping method with S. mansoni cercariae. Mice were administered orally with artesunate at different developmental stage of the parasite, with different regimens. The reduction rates of total and female worms, the number of eggs in the liver and intestine, and the fecundity were calculated and treated statistically. RESULTS: The optimal dosage of artesunate to prevent murine schistosomiasis was 300 mg/kg. The parasite was found to be especially susceptible to artesunate in its schistosomula stage of 14 and 21 d after infection, resulting in worm reduction rate of 84% and 93% respectively compared with control. High protection was reached with worm reduction rate of 99% by the regimens of 300 mg/kg once a week for 4 consecutive weeks beginning 14 d after infection. The fecundity was significantly suppressed, suggesting that the drug inhibited sexual maturation of female worms. The effective protection could also be gained with prolonged interval time of two weeks with worm reduction rate of 97% and 96% beginning 14 or 21 d after infection. CONCLUSION: Artesunate kills schistosomula and reduces the fecundity of females effectively, the infected mice do not develop schistosomiasis mansoni when treated with artesunate. It's proposed that an optimal scheme for field use be the first administration 14 or 21 days after infection with 1 or 2 weeks interval.

[Association study of five pedigree with high incidence of schizophrenia].

This study is to explore the relationship between polymorphism of Catechol O-methyltransferase gene and schizophrenia. Search for a gene predisposing to schizophrenia in the Han nationality in China. Five pedigrees with high incidence of schizophrenia were studied by polymerize chain reaction(PCR) and restriction fragment length polymorphism(RFLP) technique. Statistics analysis of the transmission/disequilibrium test (TDT) showed that the COMT gene is associated with schizophrenia in the five pedigree with high incidence schizophrenia(P=0.0455). The results suggest that there might have a schizophrenia liability gene on 22 chromosome (22q11.2) in our studied pedigrees.