Ultrasound treatment alleviates external pericarp browning and improves fruit quality of pomegranate during storage.

BACKGROUND: Ultrasound treatment has a beneficial role in horticultural production from harvest to consumption. The quality traits and microbiological load in pomegranate fruit were explored during 30 days' storage at 20 °C after 10 min and 30 min ultrasound treatments. RESULTS: Ultrasound treatment significantly reduced the microbiological load during storage, providing a relatively clean and suitable storage environment. This was especially true for the 30 min treatment, which also maintained relatively lower weight loss and kept the browning rate below 5% during storage. Meanwhile, the fruit treated with ultrasound had higher ascorbic acid and anthocyanin content, which provided better antibacterial properties and higher nutraceutical properties until the end of storage. The 30 min ultrasound treatment significantly delayed the decrease in catalase (CAT) enzyme activity and the increase in peroxidase (POD) enzyme activity. Combined with weighted gene co-expression network analysis (WGCNA), and correlation analysis, color indicators and antioxidant activity induced by ultrasound treatment were responsible for the relatively higher fruit quality of pomegranate. CONCLUSION: Ultrasound treatment can improve the sensory quality and nutritional characteristics of pomegranate fruits during storage, and reduce the microbiological load. Ultrasound for 30 min was better than 10 min for prolonging the storage life of pomegranate. Our results will provide valuable information for ultrasound application in other horticultural products. © 2023 Society of Chemical Industry.

A randomized control trial establishing the effectiveness of using interactive television-based art, music, and poetry therapies for treating the post-traumatic stress disorder of children exposed to traumatic events.

This study aimed to compare the effectiveness of interactive television-based music, art, and poetry therapies in reducing symptoms of posttraumatic stress disorder among school children who have experienced abduction. A randomized control trial was adopted using school children from three selected secondary schools in Northern Nigeria that have experienced the issue of kidnapping. The treatment is characterized by (1) group setting, (2) TV instruction (3) school collaboration, and (4) use of artistic mediation. The study sample comprised 470 students randomly divided into a control group of 235 and an experiment group of 235. The treatment group was further divided into art (N = 78), music (N = 78) and poetry (N = 79), respectively. Results suggest that interactive television art, music and poetry therapies are an effective intervention strategy for reducing the PTSD of school children who have experienced kidnapping in Nigeria. However, art therapy is more effective in reducing PTSD. This study has offered empirical evidence that could guide the planning and implementation of therapies aimed at assisting school children to recover from traumatic experiences.

The association between neuropeptide oxytocin and neuropsychiatric disorders after orthopedic surgery stress in older patients.

BACKGROUND: The health outcomes of geriatric patients exposed to surgery were found to be enhanced by social support and stress management. The aim of this study was to characterise the relationship between oxytocin and neuropsychiatric disorders after surgery. METHODS: A total of 132 geriatric patients aged ≥ 60 years received orthopedic surgery in the First Affiliated Hospital of Harbin Medical University (Harbin, China) were enrolled in the present study. The salivary levels of stress hormone cortisol and oxytocin were measured by enzyme-linked immunosorbent assay for the screening of the stress state and oxytocin function. Moreover, the Depression Anxiety and Stress Scale (DASS), the Geriatric Anxiety Inventory (GAI), the Geriatric Depression Scale (GDS) and the Montgomery-Åsberg Depression Rating Scale (MADRS) were conducted to identify the severity of anxiety and depression. The association between oxytocin and mental health was performed by linear regression analyses in older patients receiving orthopedic surgery. Finally, the Duke Social Support Index (DSSI) was selected to measure the social support and the potential link to mental outcomes. RESULTS: The scores from questionnaires showed that female patients with higher social support and higher levels of oxytocin demonstrated better stress-reducing responses as reflected by lower cortisol and decreased anxiety and depression symptoms. Regression analyses revealed that there was a significant association between oxytocin and scores in DASS, GAI, GDS, MADRS and DSSI, suggesting a potential link between peripheral oxytocin function and mood outcomes after orthopedic surgery. CONCLUSIONS: Our findings reveal that oxytocin enhances the stress-protective effects of social support and reduces anxiety and depression states under stressful circumstances, particularly in older women receiving orthopedic surgery.

The engineered CRISPR-Mb2Cas12a variant enables sensitive and fast nucleic acid-based pathogens diagnostics in the field.

Existing CRISPR/Cas12a-based diagnostic platforms offer accurate and vigorous monitoring of nucleic acid targets, but have the potential to be further optimized for more efficient detection. Here, we profiled 16 Cas12a orthologs, focusing on their trans-cleavage activity and their potential as diagnostic enzymes. We observed the Mb2Cas12a has more robust trans-cleavage activity than other orthologs, especially at lower temperatures. An engineered Mb2Cas12a-RRVRR variant presented robust trans-cleavage activity and looser PAM constraints. Moreover, we found the existing one-pot assay, which simultaneously performed Recombinase Polymerase Amplification (RPA) and Cas12a reaction in one system, resulted in the loss of single-base discrimination during diagnosis. Therefore, we designed a reaction vessel that physically separated the RPA and Cas12a steps while maintaining a closed system. This isolated but closed system made diagnostics more sensitive and specific and effectively prevented contamination. This shelved Mb2Cas12a-RRVRR variant-mediated assay detected various targets in less than 15 min and exhibited equal or greater sensitivity than qPCR when detecting bacterial pathogens, plant RNA viruses and genetically modified crops. Overall, our findings further improved the efficiency of the current CRISPR-based diagnostic system and undoubtedly have great potential for highly sensitive and specific detection of multiple sample types.

GW842166X Alleviates Osteoarthritis by Repressing LPS-mediated Chondrocyte Catabolism in Mice.

OBJECTIVE: To explore the role and underlying mechanism of GW842166X on osteoarthritis and osteoarthritis-associated abnormal catabolism. METHODS: The extracted mouse chondrocytes were treated with GW842166X followed by lipopolysaccharide (LPS). The chondrocytes were divided into the control group, LPS group, LPS+50 nmol/L GW842166X group, and LPS+100 nmol/L GW842166X group. The cytotoxicity of GW842166X was tested using the CCK-8 assay. Western blot, RT-qPCR, and ELISA were applied to evaluate the expression of the inflammatory biomarkers in mouse chondrocytes. The expression of extracellular matrix molecules was detected by the Western blot, RT-qPCR, and immunofluorescence. Additionally, the activity of NF-κB was checked by the Western blot and immunofluorescence. The mouse Hulth models were generated to examine the in vivo effects of GW842166X on osteoarthritis. Hematoxylin and eosin staining, safranin O/fast green staining, and immunohistochemistry were applied to detect the histological changes. RESULTS: GW842166X below 200 µmol/L had no cytotoxicity on the mouse chondrocytes. LPS-induced high expression of TGF-ß1, IL-10, TNF-α, and IL-6 was significantly reduced by GW842166X. In addition, GW842166X upregulated the expression of aggrecan and collagen type III, which was downregulated after the LPS stimulation. The upregulated expression of ADAMTS-5 and MMP-13 by LPS stimulation was dropped in response to the GW842166X treatment. Furthermore, LPS decreased the IκBα expression in the cytoplasm and increased the nuclear p65 expression. However, these changes were reversed by the GW842166X pretreatment. Moreover, the damages in the knees caused by the Hulth surgery in mice were restored by GW842166X. CONCLUSION: GW842166X impeded the LPS-mediated catabolism in mouse chondrocytes, thereby inhibiting the progression of osteoarthritis.

Overexpression of PgCBF3 and PgCBF7 Transcription Factors from Pomegranate Enhances Freezing Tolerance in Arabidopsis under the Promoter Activity Positively Regulated by PgICE1.

Cold stress limits plant growth, development and yields, and the C-repeat binding factors (CBFs) function in the cold resistance in plants. However, how pomegranate CBF transcription factors respond to cold signal remains unclear. Considering the significantly up-regulated expression of PgCBF3 and PgCBF7 in cold-tolerant Punica granatum 'Yudazi' in comparison with cold-sensitive 'Tunisia' under 4 °C, the present study focused on the two CBF genes. PgCBF3 was localized in the nucleus, while PgCBF7 was localized in the cell membrane, cytoplasm, and nucleus, both owning transcriptional activation activity in yeast. Yeast one-hybrid and dual-luciferase reporter assay further confirmed that PgICE1 could specifically bind to and significantly enhance the activation activity of the promoters of PgCBF3 and PgCBF7. Compared with the wild-type plants, the PgCBF3 and PgCBF7 transgenic Arabidopsis thaliana lines had the higher survival rate after cold treatment; exhibited increased the contents of soluble sugar and proline, while lower electrolyte leakage, malondialdehyde content, and reactive oxygen species production, accompanying with elevated enzyme activity of catalase, peroxidase, and superoxide dismutase; and upregulated the expression of AtCOR15A, AtCOR47, AtRD29A, and AtKIN1. Collectively, PgCBFs were positively regulated by the upstream PgICE1 and mediated the downstream COR genes expression, thereby enhancing freezing tolerance.

Self-Assembling Imageable Silk Hydrogels for the Focal Treatment of Osteosarcoma.

Background: The standard treatment for osteosarcoma comprises complete surgical resection and neoadjuvant chemotherapy, which may cause serious side effects and partial or total limb loss. Therefore, to avoid the disadvantages of traditional treatment, we developed self-assembling imageable silk hydrogels for osteosarcoma. Methods: We analysed whether iodine induced apoptosis in MG-63 and Saos-2 cells by using CCK-8 and flow cytometry assays and transmission electron microscopy. Western blotting was used to analyse the pathway of iodine-induced apoptosis in osteosarcoma cells. PEG400, silk fibroin solution, polyvinylpyrrolidone iodine (PVP-I), and meglumine diatrizoate (MD) were mixed to produce an imageable hydrogel. A nude mouse model of osteosarcoma was established, and the hydrogel was injected locally into the interior of the osteosarcoma with X-ray guidance. The therapeutic effect and biosafety of the hydrogel were evaluated. Results: Iodine treatment at 18 and 20 µM for 12 h resulted in cell survival rate reduced to 50 ± 2.1% and 50.5 ± 2.7% for MG-63 and Sao-2 cells, respectively (p < 0.01). The proportion of apoptotic cells was significantly higher in the iodine-treatment group than in the control group (p < 0.05), and apoptotic bodies were observed by transmission electron microscopy. Iodine could regulate the death receptor pathway and induce MG-63 and Saos-2 cell apoptosis. The hydrogels were simple to assemble, and gels could be formed within 38 min. A force of less than 50 N was required to inject the gels with a syringe. The hydrogels were readily loaded and led to sustained iodine release over 1 week. The osteosarcoma volume in the PEG-iodine-silk/MD hydrogel group was significantly smaller than that in the other three groups (p < 0.001). Caspase-3 and poly (ADP-ribose) polymerase (PARP) expression levels were significantly higher in the PEG-iodine-silk/MD hydrogel group than in the other three groups (p < 0.001). Haematoxylin and eosin (H&E) staining showed no abnormalities in the heart, liver, spleen, lung, kidney, pancreas or thyroid in any group. Conclusions: Self-assembling imageable silk hydrogels could be injected locally into osteosarcoma tissues with X-ray assistance. With the advantages of good biosafety, low systemic toxicity and minimal invasiveness, self-assembling imageable silk hydrogels provide a promising approach for improving the locoregional control of osteosarcoma.

Exogenous Arginine Treatment Maintains the Appearance and Nutraceutical Properties of Hard- and Soft-Seed Pomegranates in Cold Storage.

Arginine is a natural preservative; however, its effects on the storage of different cultivars of pomegranates have not been investigated extensively. Therefore, the fruit quality of soft-seed Tunisia and hard-seed Yudazi pomegranates was investigated after treatment with arginine at four concentrations during cold storage for 80 days. Pomegranates treated with 1.0 mM arginine exhibited a relatively lower loss of vitamin C, soluble solid, total phenol, and anthocyanin contents in arils, together with a better fruit appearance. Combined with principal component analysis (PCA), the storage life of fruits treated with 1.0 mM arginine showed a higher correlation with antioxidant enzyme activity (e.g., superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT)) during the first 40 days of cold storage, whereas after 40 days of cold storage, storage life was more dependent on the integrity of the cell membrane affected by malondialdehyde (MDA) content, electrolyte leakage (EL), and hydrogen peroxide (H2O2) accumulation. Arginine treatment contributed significantly to the appearance and inner quality of the hard-seed pomegranate cv. Yudazi fruit during cold storage compared to those of soft-seed Tunisia. Taken together, arginine application combined with cold storage enhanced the nutraceutical properties and marketability of pomegranate fruits.

Genome-Wide Identification and Comprehensive Analysis of the AP2/ERF Gene Family in Pomegranate Fruit Development and Postharvest Preservation.

Pomegranate (Punica granatum L.) is a kind of fruit with significant economic, ecological and health values. AP2/ERF transcription factors belong to a large group of factors mainly found in plants and play key roles in plant growth and development. However, AP2/ERF genes in pomegranate and their implication in development and postharvest preservation have been little described. In this study, 116 PgAP2/ERF genes in pomegranate were identified and renamed based on their chromosomal distributions. Phylogenetic relationship with genes from other species, structures, duplications, annotations, cis-elements in promoter sequences, and protein-protein interaction networks among PgAP2/ERF proteins were comprehensively explored. Expression analysis revealed several PgAP2/ERFs associated with the phenotypes of pomegranate seed hardness, including PgAP2/ERF5, PgAP2/ERF36, PgAP2/ERF58, and PgAP2/ERF86. Subsequent analysis indicated that many differentially expressed PgAP2/ERF genes are potentially important regulators of pomegranate fruit development. Furthermore, expression of more than one-half of PgAP2/ERFs was repressed in 'Tunisian soft seed' pomegranate fruit under low-temperature cold storage. The results showed that 1-MCP implicated in promoting postharvest preservation of 'Tunisian soft seed' pomegranate upregulated the PgAP2/ERF4, PgAP2/ERF15, PgAP2/ERF26, PgAP2/ERF30, PgAP2/ERF35 and PgAP2/ERF45 genes compared to those under low-temperature cold storage. This indicates that these genes are important candidate genes involved in pomegranate postharvest preservation. In summary, the findings of the present study provide an important basis for characterizing the PgAP2/ERF family genes and provide information on the candidate genes involved in pomegranate fruit development and postharvest preservation.

Large, star-shaped retinal tear associated with orbital cosmetic filler.

PURPOSE: Presentation of a unique case of large, star-shaped retinal tear associated with orbital cosmetic filler injection. OBSERVATION: A 55-year-old healthy female presented to emergency department with sudden onset of blurred vision on her left eye occurred after an orbital cosmetic filler injection containing hyaluronic acid (HA) performed by an aesthetic nurse. On fundus examination, the left eye showed a mild - moderate, unusual appearance vitreous haze and a large, star-shaped retinal tear temporal to macula. Optical coherence tomography (OCT) examination demonstrated a normal foveal contour without any structural damage on the retinal layers. Vitrectomy, endolaser and silicon oil tamponade were performed. Visual acuity was maintained at 6/7.5 following silicon oil removal at 6 weeks after the initial surgery. CONCLUSIONS AND IMPORTANCE: In this particular case, immediate vitrectomy is key to avoid further complications such as retinal toxicity and detachment and to maintain optimal visual acuity. Importantly, facial cosmetic filler procedure should be performed by an adequately trained individual to avoid such complications.

Ultrastructural Observations of Botrytis cinerea and Physical Changes in Resistant and Susceptible Grapevines.

The necrotrophic fungus Botrytis cinerea is a major threat to grapevine cultivation worldwide. Here, a highly resistant Chinese wild grapevine, Vitis amurensis 'Shuangyou' (SY), and the susceptible V. vinifera 'Red Globe' (RG) were selected for study, and their pathogenic infection and biochemical responses to B. cinerea were evaluated. The results revealed more trichomes on and a thicker cuticle for leaves of SY than RG under scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Both SEM and TEM also showed that conidial germination, appressorium formation, and hyphal development of B. cinerea were delayed on the leaves of resistant SY. Fewer infected hyphae were also observed in leaves of resistant SY when compared with susceptible RG. The infected leaves of resistant SY harbored higher levels of cellulase and pectinase activity during the early infection stages of B. cinerea at 4 h postinoculation (hpi), and higher glucanase and chitinase activity were maintained in the inoculated leaves of SY from 4 through 18 hpi. Lignin was deposited in the infected leaves of susceptible RG but not in resistant SY. Taken together, these results provide insights into the ultrastructural characterizations and physical changes in resistant and susceptible grapevines.

Comparative transcriptomic analysis highlights contrasting levels of resistance of Vitis vinifera and Vitis amurensis to Botrytis cinerea.

Botrytis cinerea is a major grapevine (Vitis spp.) pathogen, but some genotypes differ in their degree of resistance. For example, the Vitis vinifera cultivar Red Globe (RG) is highly susceptible, but V. amurensis Rupr Shuangyou (SY) is highly resistant. Here, we used RNA sequencing analysis to characterize the transcriptome responses of these two genotypes to B. cinerea inoculation at an early infection stage. Approximately a quarter of the genes in RG presented significant changes in transcript levels during infection, the number of which was greater than that in the SY leaves. The genes differentially expressed between infected leaves of SY and RG included those associated with cell surface structure, oxidation, cell death and C/N metabolism. We found evidence that an imbalance in the levels of reactive oxygen species (ROS) and redox homeostasis probably contributed to the susceptibility of RG to B. cinerea. SY leaves had strong antioxidant capacities and improved ROS homeostasis following infection. Regulatory network prediction suggested that WRKY and MYB transcription factors are associated with the abscisic acid pathway. Weighted gene correlation network analysis highlighted preinfection features of SY that might contribute to its increased resistance. Moreover, overexpression of VaWRKY10 in Arabidopsis thaliana and V. vinifera Thompson Seedless enhanced resistance to B. cinerea. Collectively, our study provides a high-resolution view of the transcriptional changes of grapevine in response to B. cinerea infection and novel insights into the underlying resistance mechanisms.

Pathomechanism of Lower-level Discogenic Groin Pain and Clinical Outcomes of Percutaneous Endoscopic Discectomy for the Treatment of Discogenic Groin Pain.

BACKGROUND: Groin pain can be induced by high-level (L1-L2 or L2-L3) lumbar disc herniation. However, 4.1% of patients with lower-level (L4-L5 or L5-S1) lumbar disc herniation also complained of groin pain. The pathomechanism of groin pain occurring due to lumbar disc herniation at and below the L4-5 levels is still unclear. OBJECTIVE: To investigate the afferent pathways of lower-level lumbar disc herniation induced groin pain. And evaluate the clinical results of transforaminal endoscopic discectomy treatment for discogenic groin pain. STUDY DESIGN: This retrospective observational study used an experimental design (institutional review board: HROH 201-C2-100). SETTING: The research took place in the Laboratory Research Center and spine center at The First Affiliated Hospital of Harbin Medical University. METHODS: Firstly, 14 adult Wistar rats were randomly divided into 2 groups: control group (the paravertebral sympathetic trunks were preserved) and experimental group (the paravertebral sympathetic trunks were resected). All Wistar rats were intraperitoneally anesthetized, and then 1 (mu)L of fast blue was injected into the dorsal rami of L2 spinal nerves on the right side. Forty hours later, 2 (mu)L of nuclear yellow was injected into the right posterior portion of the L5-L6 intervertebral disc. The L1 and L2 spinal ganglia were sectioned 8 hours later to observe fluorescently double-labeled cells and the effect of paravertebral sympathetic trunk resection. Secondly, 14 adult Wistar rats were anesthetized, and the right posterior portion of the L5-L6 intervertebral disc was electrostimulated to observe potential changes in the genitocrural nerve in the ipsilateral inguinal region. To evaluate the clinical outcomes of transforaminal endoscopic discectomy for the treatment of discogenic groin pain, between September 2015 and May 2017, transforaminal endoscopic discectomy was performed on 30 patients with lower-level discogenic groin pain. Outcomes were analyzed utilizing the visual analog scale, Oswestry disability index, and MacNab Criteria. RESULTS: The total proportion of cells in the right L1 and L2 spinal ganglia with fast blue/nuclear yellow double labeling was 3.33% and 3.41% (48 and 56), respectively. The number of fluorescently double-labeled cells in the resected paravertebral sympathetic trunk group was significantly less (P < 0.01). Electrical stimulation of the right posterior portion of the L5-L6 intervertebral disc could elicit action potentials in the ipsilateral genitofemoral nerve. All patients were followed for 12 months, and the visual analog scale score at 1 week, 1 month, 3 months, 6 months, and 12 months after the operation was 0.79 ± 0.55, 0.54 ± 0.55, 0.47 ± 0.65, 0.51 ± 0.65, and 0.69 ± 0.55, respectively, showing a significant decrease compared with the preoperative visual analog scale score (P < 0.01). Based on the MacNab scoring system, the effective rate was 100%, and the rate of good and excellent results was 93.3%. LIMITATIONS: A relatively small number of patients and a short follow-up period. CONCLUSIONS: Discogenic groin pain is transmitted by sympathetic nerves and appears in the area segmentally innervated by the anterior rami of the L1 and L2 spinal nerves. Posterolateral percutaneous transforaminal endoscopic discectomy and radiofrequency thermal annuloplasty are effective minimally invasive alternative treatments for discogenic groin pain.

Optic disc pit maculopathy: a review of diagnosis and treatment.

Optic disc pit is a rare congenital defect which appears as a circumscribed greyish depression in the lamina cribrosa. Serous macular detachment is one of the most visually debilitating complications of optic disc pit, affecting 25-75 per cent of patients. Although there is a wide variety of treatment modalities available with varying degrees of success, there is yet no consensus in the optimal management of optic disc pit-associated maculopathy. This review discusses the literature on the pathogenesis, clinical presentation, diagnosis and treatment options.

Percutaneous Endoscopic Interlaminar Lumbar Discectomy with Local Anesthesia for L5-S1 Disc Herniation: A Feasibility Study.

BACKGROUND: General anesthesia (GA), which is routinely applied in patients who undergo percutaneous endoscopic interlaminar lumbar discectomy (PEILD) of L5-S1 disc herniation, is closely associated with postoperative cognitive dysfunction (POCD) in the elderly. Local anesthesia (LA) is an alternative pain control protocol that has not yet been fully evaluated. OBJECTIVES: To evaluate the feasibility of LA in PEILD compared with GA. STUDY DESIGN: A retrospective study. SETTING: This study took place at the First Affiliated Hospital of Harbin Medical University. METHODS: A total of 120 patients (aged 60-85 years) diagnosed with L5-S1 disc herniation and with American Society of Anesthesiologists fitness grade I or II between March 2016 and August 2017 were enrolled in the current study. Patients were randomly divided into LA group and GA group. For LA, 0.25% lidocaine was injected layer-by-layer into skin, subcutaneous tissue, fasciae, lumbar facet joint, muscle, and ligamentum flavum followed by injection of 1.33% lidocaine into epidural space; for GA, propofol, sufentanil, and cisatracurium were infused intravenously at 1 to 2 mg/kg, 0.3 µg/kg, and 0.15 mg/kg, respectively. Visual Analog Scale (VAS), Oswestry Disability Index (ODI), and MacNab Criteria (MNC) evaluated the feasibility of LA as pain control protocol in comparison to GA before and after operation. The development of POCD was assessed by the Mini-Mental State Examination 1 and 7 days postsurgery. Feasibility of LA as a pain control protocol was also evaluated by patient's willingness to receive the same surgical procedure immediately and 24 hours after the surgery, and intraoperative fluoroscopy use, blood loss, surgery duration, postoperative bed confinement, and duration and cost of hospital stay were also evaluated. RESULTS: Patients in both LA and GA groups had comparable VAS grade, ODI, and MNC pre- and post-PEILD, with significant pain reduction after operation. However, POCD developed only in GA group but not in LA group. In addition, compared with GA, LA group did not require postoperative bed confinement, had significantly shorter hospital stay, and lower hospital cost. Low intraoperative VAS grade and willingness to receive the same procedure reflected the acceptance of LA by patients. LIMITATIONS: The development of POCD was examined only 7 days after operation. The follow-up should be extended to 3 months and 2 years postoperation. CONCLUSIONS: LA has satisfactory pain control and low-risk of POCD in PEILD and is well accepted by patients. The benefits of LA are no postoperative bed confinement, faster recovery, shorter hospital stay, and lower hospital cost. KEY WORDS: L5-S1 disc herniation, older patients, percutaneous endoscopic interlaminar lumbar discectomy, local anesthesia, general anesthesia, postoperative cognitive dysfunction, American Society of Anesthesiologists grade, Oswestry Disability Index, MacNab Criteria, Mini-Mental State Examination.

Screening Vitis Genotypes for Responses to Botrytis cinerea and Evaluation of Antioxidant Enzymes, Reactive Oxygen Species and Jasmonic Acid in Resistant and Susceptible Hosts.

Botrytis cinerea is a necrotrophic fungal phytopathogen with devastating effects on many Vitis genotypes. Here, a screening of 81 Vitis genotypes for leaf resistance to B. cinerea revealed two highly resistant (HR), twelve resistant (R), twenty-five susceptible (S) and forty-two highly susceptible (HS) genotypes. We focused on the HR genotype, 'Zi Qiu' (Vitis davidii), and the HS genotype 'Riesling' (V. vinifera), to elucidate mechanisms of host resistance and susceptibility against B. cinerea, using detached leaf assays. These involved a comparison of fungal growth, reactive oxygen species (ROS) responses, jasmonic acid (JA) levels, and changes in the anti-oxidative system between the two genotypes after inoculation with B. cinerea. Our results indicated that the high-level resistance of 'Zi Qiu' can be attributed to insignificant fungal development, low ROS production, timely elevation of anti-oxidative functions, and high JA levels. Moreover, severe fungal infection of 'Riesling' and sustained ROS production coincided with relatively unchanged anti-oxidative activity, as well as low JA levels. This study provides insights into B. cinerea infection in grape, which can be valuable for breeders by providing information for selecting suitable germplasm with enhanced disease resistance.

The Grape VlWRKY3 Gene Promotes Abiotic and Biotic Stress Tolerance in Transgenic Arabidopsis thaliana.

WRKY transcription factors are known to play important roles in plant responses to various abiotic and biotic stresses. The grape WRKY gene, WRKY3 was previously reported to respond to salt and drought stress, as well as methyl jasmonate and ethylene treatments in Vitis labrusca × V. vinifera cv. 'Kyoho.' In the current study, WRKY3 from the 'Kyoho' grape cultivar was constitutively expressed in Arabidopsis thaliana under control of the cauliflower mosaic virus 35S promoter. The 35S::VlWRKY3 transgenic A. thaliana plants showed improved salt and drought stress tolerance during the germination, seedling and the mature plant stages. Various physiological traits related to abiotic stress responses were evaluated to gain further insight into the role of VlWRKY3, and it was found that abiotic stress caused less damage to the transgenic seedlings than to the wild-type (WT) plants. VlWRKY3 over-expression also resulted in altered expression levels of abiotic stress-responsive genes. Moreover, the 35S::VlWRKY3 transgenic A. thaliana lines showed improved resistance to Golovinomyces cichoracearum, but increased susceptibility to Botrytis cinerea, compared with the WT plants. Collectively, these results indicate that VlWRKY3 plays important roles in responses to both abiotic and biotic stress, and modification of its expression may represent a strategy to enhance stress tolerance in crops.

Efficacy of Lenvatinib, a multitargeted tyrosine kinase inhibitor, on laser-induced CNV mouse model of neovascular AMD.

Neovascular age-related macular degeneration (AMD) is a leading cause of vision loss worldwide. Although intravitreal injection of anti-VEGF antibodies and VEGF Trap have significant clinical benefits, the complications of intravitreal injection, drug resistance and patient compliance still need to be concerned. In this study, the effects of an orally administered multi-targeted tyrosine kinase inhibitor (Lenvatinib, E7080) were evaluated in vitro and in vivo on neovascular AMD mouse model. The results showed that E7080 effectively inhibited the proliferation, migration and tubule formation of human choroidal microvascular endothelial cells (HCMECs), and suppressed the angiogenesis of zebrafish subintestinal vessels without causing malformation. The anti-angiogenic effect of E7080 on the laser-induced choroidal neovascularization (CNV) mouse model by oral administration of 10 mg/kg/day was observed. The fluorescein angiography showed CNV leakage area in treatment group vs control group was 3.407 ±â€¯0.2939 vs 5.202 ±â€¯0.9001 (P = .0133) at day 7th post laser-induced CNV, 1.138 ±â€¯0.4334 vs 3.122 ±â€¯0.3466 (P = .0064) at day 14th, 1.401 ±â€¯0.6577 vs 2.781 ±â€¯0.9815 (P = .00262) at day 21th respectively. Moreover, pharmacokinetics analysis in rat retina showed that E7080 rapidly penetrated the blood-retina barrier to retina through oral administration. The T1/2 in retina was 3.81 ±â€¯0.77 h, the Tmax was 4.60 ±â€¯0.73 h, the AUC0-∞ was 110448.51 ± 18532.51 h*ng/g after a single dose administration analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS). In conclusion, our study suggested that orally administered E7080 can be a novel therapeutic strategy for neovascular AMD.

Pharmacotherapy of Vitreomacular Traction.

Vitreomacular traction occurs due to incomplete or anomalous posterior vitreous detachment. Over time, the vitreous pulls anteriorly and causes retinal distortion and eventually reduced vision. Traditionally, vitreomacular traction was treated with vitrectomy surgery. In the past few years, there is a paradigm shift towards pharmacologic vitreolysis, which involves the intravitreal injection of enzymatic and non-enzymatic agents that facilitate posterior vitreous detachment. Many agents have been investigated and trialled including plasmin, microplasmin (Ocriplasmin), hyaluronidase, nattokinase, chondroitinase and dispase. This review will focus on the progress and current status in this research.

Ectopic Expression of the Wild Grape WRKY Transcription Factor VqWRKY52 in Arabidopsis thaliana Enhances Resistance to the Biotrophic Pathogen Powdery Mildew But Not to the Necrotrophic Pathogen Botrytis cinerea.

WRKY transcription factors are known to play important roles in plant responses to biotic stresses. We previously showed that the expression of the WRKY gene, VqWRKY52, from Chinese wild Vitis quinquangularis was strongly induced 24 h post inoculation with powdery mildew. In this study, we analyzed the expression levels of VqWRKY52 following treatment with the defense related hormones salicylic acid (SA) and methyl jasmonate, revealing that VqWRKY52 was strongly induced by SA but not JA. We characterized the VqWRKY52 gene, which encodes a WRKY III gene family member, and found that ectopic expression in Arabidopsis thaliana enhanced resistance to powdery mildew and Pseudomonas syringae pv. tomato DC3000, but increased susceptibility to Botrytis cinerea, compared with wild type (WT) plants. The transgenic A. thaliana lines displayed strong cell death induced by the biotrophic powdery mildew pathogen, the hemibiotrophic P. syringe pathogen and the necrotrophic pathogen B. cinerea. In addition, the relative expression levels of various defense-related genes were compared between the transgenic A. thaliana lines and WT plants following the infection by different pathogens. Collectively, the results indicated that VqWRKY52 plays essential roles in the SA dependent signal transduction pathway and that it can enhance the hypersensitive response cell death triggered by microbial pathogens.