Priming of Immune System in Tomato by Treatment with Low Concentration of L-Methionine.

Various metabolites, including phytohormones, phytoalexins, and amino acids, take part in the plant immune system. Herein, we analyzed the effects of L-methionine (Met), a sulfur-containing amino acid, on the plant immune system in tomato. Treatment with low concentrations of Met enhanced the resistance of tomato to a broad range of diseases caused by the hemi-biotrophic bacterial pathogen Pseudomonas syringae pv. tomato (Pst) and the necrotrophic fungal pathogen Botrytis cinerea (Bc), although it did not induce the production of any antimicrobial substances against these pathogens in tomato leaf tissues. Analyses of gene expression and phytohormone accumulation indicated that Met treatment alone did not activate the defense signals mediated by salicylic acid, jasmonic acid, and ethylene. However, the salicylic acid-responsive defense gene and the jasmonic acid-responsive gene were induced more rapidly in Met-treated plants after infection with Pst and Bc, respectively. These findings suggest that low concentrations of Met have a priming effect on the phytohormone-mediated immune system in tomato.

Response of tomatoes primed by mycorrhizal colonization to virulent and avirulent bacterial pathogens.

Most plants interact with arbuscular mycorrhizal fungi, which enhance disease resistance in the host plant. Because the effects of resistance against bacterial pathogens are poorly understood, we investigated the effects of mycorrhizal colonization on virulent and avirulent pathogens using phytopathological and molecular biology techniques. Tomato plants colonized by Gigaspora margarita acquired resistance not only against the fungal pathogen, Botrytis cinerea, but also against a virulent bacterial pathogen, Pseudomonas syringae pv. tomato DC3000 (Pst). In G. margarita-colonized tomato, salicylic acid (SA)- and jasmonic acid (JA)-related defense genes were expressed more rapidly and strongly compared to those in the control plants when challenged by Pst, indicating that the plant immunity system was primed by mycorrhizal colonization. Gene expression analysis indicated that primed tomato plants responded to the avirulent pathogen, Pseudomonas syringae pv. oryzae, more rapidly and strongly compared to the control plant, where the effect on the JA-mediated signals was stronger than in the case with Pst. We found that the resistance induced by mycorrhizal colonization was effective against both fungal and bacterial pathogens including virulent and avirulent pathogens. Moreover, the activation of both SA- and JA-mediated signaling pathways can be enhanced in the primed plant by mycorrhizal colonization.

Effects of colonization of a bacterial endophyte, Azospirillum sp. B510, on disease resistance in tomato.

A plant growth-promoting bacteria, Azospirillum sp. B510, isolated from rice, can enhance growth and yield and induce disease resistance against various types of diseases in rice. Because little is known about the interaction between other plant species and this strain, we have investigated the effect of its colonization on disease resistance in tomato plants. Treatment with this strain by soil-drenching method established endophytic colonization in root tissues in tomato plant. The endophytic colonization with this strain-induced disease resistance in tomato plant against bacterial leaf spot caused by Pseudomonas syringae pv. tomato and gray mold caused by Botrytis cinerea. In Azospirillum-treated plants, neither the accumulation of SA nor the expression of defense-related genes was observed. These indicate that endophytic colonization with Azospirillum sp. B510 is able to activate the innate immune system also in tomato, which does not seem to be systemic acquired resistance.

Diabetic phenotype of transgenic pigs introduced by dominant-negative mutant hepatocyte nuclear factor 1α.

AIM: The present study aimed to identify the characteristics of genetically modified pigs carrying a mutant human gene as a research model for diabetes and its complications. METHODS: We developed a transgenic cloned pig (founder, male) carrying a mutant gene, i.e., human HNF-1α (P291fsinsC), which is responsible for maturity-onset diabetes of the young type 3. Transgenic progeny obtained via the artificial insemination of wild type (WT) sows with the cryopreserved sperm derived from the founder pig was pathologically examined. RESULTS: The transgenic progeny maintained a high blood glucose level (>200mg/dL). Additionally, the oral glucose tolerance test results showed that the recovery of blood glucose levels in the transgenic progeny was significantly delayed compared with that in the WT semi-siblings. Hypoplasia of the islets of Langerhans was confirmed by the histopathological image of the pancreas, based on the hyperglycemia noted in the progeny being ascribed to decreased insulin secretion. Retinal hemorrhage and cotton-wool spots, i.e., findings consistent with non-proliferative diabetic retinopathy, were detected, and these progressed over time. The histopathological image of the renal glomeruli showed a nodular lesion that is characteristic of diabetic nephropathy in humans. CONCLUSIONS: These data demonstrated that the genetically modified pig that we developed is a promising model for research on diabetes and its complications.

[Postural and Conditional Variations of Nasal Patency in Patients under General Anesthesia Approximating Sleep as Assessed with Acoustic Rhinometry].

Nasal obstruction is a known risk factor associated with obstructive sleep apnea (OSA). Multiple methods of measuring nasal abnormalities including anterior and posterior rhinomanometry and acoustic rhinometry (AR) have been used to better quantify the relationship. These measurements are usually used in the sitting and awakening situation, which differ from the situations in the supine position and while asleep. The goal of this study was to evaluate the nasal patency in patients under general anesthesia approximating sleep as assessed with AR. The nasal minimal cross-sectional area (MCSA) and nasal volume in 50 patients who underwent general anesthesia were compared with subjects in the sitting, supine, and supine position under general anesthesia using AR to measure the nasal dimensions and patency. MCSA and nasal volume decreased more significantly in the supine than in the sitting subjects and these decrements were shown more in the supine position under general anesthesia than in the supine position without anesthesia. Furthermore, regarding nasal volume, the difference between in the sitting and in the supine was larger than the difference between in the supine position with and without general anesthesia. These results suggested that the decrements regarding nasal volume under positional change were greater than those under conditional change. The positional variation can affect the airway patency hydrodynamically and these alterations indicate the relationship of the pathophysiology in patients with OSA.

[Two cases of allergic fungal rhinosinusitis with bone destruction].

We report herein two cases of allergic fungal rhinosinusitis accompanied by bone destruction of the adjacent nasal sinuses. The first case involved a 21-year-old man who presented with left exophthalmos. Computed tomography (CT) showed soft tissue lesions in the left paranasal sinuses and destruction of the left lamina papyraceae, as well as infiltration of the lesion into the orbit. In the second case, a 39-year-old man, CT showed bone destruction of the skull base and medial wall of the left orbit. In both cases, total serum immunoglobulin (Ig)E level was >1000 IU/mL and fungus-specific IgEs were increased. Fungal hyphae were identified within the mucus on histopathological examination in both cases; however, no fungal invasion of the mucosa was apparent. Final diagnosis was allergic fungal rhinosinusitis (AFRS) in both cases. AFRS is a relatively new disease concept that was proposed in the early 1980s, with disease characteristics very similar to eosinophilic rhinosinusitis. Occasionally, AFRS must be differentiated from malignant disease or invasive fungal rhinosinusitis, so an understanding of the clinical features is important.

[Effect of posterior nasal neurectomy combined with the inferior turbinate surgery for Japanese cedar pollinosis].

INTRODUCTION: Up to 30% of the Japanese population suffers from Japanese cedar pollinosis, and some of these patients also have severe perennial allergies or other pollen allergies. Posterior nasal neurectomy has recently been reported as effective treatment for severe perennial allergic and intrinsic rhinitis. However, the efficacy of this surgery for seasonal allergic rhinitis has not been shown. In this study, the effectiveness of posterior nasal neurectomy combined with inferior turbinate surgery for patients with Japanese cedar pollinosis with concomitant intractable perennial allergies was evaluated with a questionnaire. SUBJECTS AND METHODS: Nine patients with Japanese cedar pollinosis, who also had perennial allergies, and undergone posterior nasal neurectomy combined with inferior turbinate surgery from April in 2005 to July in 2008, were enrolled. A numeric rating scale (NMR) was used to evaluate clinical symptoms in the pre- and post-surgery periods. The frequency of medication (oral administration and nasal spray) during the cedar pollinosis season was also evaluated. RESULTS: The NMR scores for all nasal and eye symptoms during cedar pollinosis season were significantly reduced after the surgery. The frequency of medication during the season was dramatically decreased after surgery. CONCLUSION: The present study demonstrates that posterior nasal neurectomy combined with inferior turbinate surgery is a highly effective treatment for Japanese cedar pollinosis. Since the Japanese cedar pollinosis season is limited to 2 to 3 months, such surgery is indicated for patients who suffer from severe perennial allergies and are also sensitized to some allergens such as Japanese cedar and ragweed pollen.

A case of 25 gauge vitrectomy for malignant glaucoma with microphthalmos.

UNLABELLED: We report the first case report of pars plana lensectomy (PPL) and pars plana vitrectomy using a 25 gauge vitrectomy system for microphthalmos. RESULTS: A 45-year-old woman reported repeated pain in her left eye, despite the use of medications. Ultrasound biomicroscopy revealed a narrow angle and thickened sclera. The lens and the iris showed anterior displacement with a flattening of the ciliary body. The axial length was 15.16 mm in the right eye and 15.04 mm in the left eye. She was diagnosed with a malignant glaucoma with microphthalmos. The patient underwent PPL and pars plana vitrectomy, using a 25 gauge vitrectomy system. The intraocular pressure was well controlled after surgery without medication. CONCLUSION: We considered the PPL and 25 gauge vitrectomy effective in microphthalmos when performed safely.

Strong resistance against Rice grassy stunt virus is induced in transgenic rice plants expressing double-stranded RNA of the viral genes for nucleocapsid or movement proteins as targets for RNA interference.

Rice grassy stunt virus (RGSV), a member of the genus Tenuivirus, causes significant economic losses in rice production in South, Southeast, and East Asian countries. Growing resistant varieties is the most efficient method to control RGSV; however, suitable resistance genes have not yet been found in natural rice resources. One of the most promising methods to confer resistance against RGSV is the use of RNA interference (RNAi). It is important to target viral genes that play important roles in viral infection and proliferation at an early stage of viral replication. Our recent findings obtained from an RNAi experiment with Rice stripe virus (RSV), a tenuivirus, revealed that the genes for nucleocapsid and movement proteins were appropriate targets for RNAi to confer resistance against RSV. In this study, we transformed rice plants by introducing an RNAi construct of the RGSV genes for the nucelocapsid protein pC5 or movement protein pC6. All progenies from self-fertilized transgenic plants had strong resistance against RGSV infection and did not allow the proliferation of RGSV. Thus, our strategy to target genes for nucleocapsid and movement proteins for conferring viral resistance might be applicable to the plant viruses in the genus Tenuivirus.

Induction of systemic acquired resistance by heat shock treatment in Arabidopsis.

Systemic acquired resistance (SAR) is a potent innate immunity system in plants and has been used in rice fields. Development of SAR, involving priming, is achieved by activation of salicylic acid (SA)-mediated pathway. To determine whether heat shock (HS) treatment can induce SAR, we analyzed the effects of HS on Arabidopsis. HS treatment induced disease resistance, expression of SAR marker genes, and SA accumulation in wild-type but not in SA-deficient sid2 and NahG plants, indicating induction of SAR. Time course analysis of the effects of HS indicated that SAR was activated transiently, differently from biological induction, with a peak at 2-3 d after HS, and that it ceased in several days. Production of reactive oxygen species was observed before SA biosynthesis, which might be a trigger for SAR activation. The data presented here suggest that HS can induce SAR, but there exist unknown regulation mechanisms for the maintenance of SAR.

Efficacy of pneumatic displacement with 40-degree downward gaze positioning for treatment of submacular hemorrhage: report of two cases.

BACKGROUND: Two retrospective case reports are described showing the efficacy of pneumatic displacement with 40-degree downward gaze positioning for treatment of submacular hemorrhage. Case 1 involved an 85-year-old woman who visited our clinic with a complaint of blurred vision in her right eye. Best corrected visual acuity (BCVA) of the right eye was hand motion. Her right eye showed a submacular hemorrhage with retinal macroaneurysm. Prone positioning was difficult because of the patient's age and obesity. The treatment method was pneumatic displacement with 40-degree downward gaze positioning after intravitreal injection of 100% sulfur hexafluoride gas. After the treatment, the hemorrhage moved downward and BCVA of her right eye was 0.1. Case 2 involved a 58-year-old man with a complaint of blurred vision in his left eye. BCVA of his left eye was 0.3 and submacular hemorrhage was observed. He underwent displacement of the hemorrhage as previously described. After treatment, the submacular hemorrhage moved downward. Angiography detected a polypoidal lesion with a vascular network. upon diagnosis of polypoidal choroidal vasculopathy, bevacizumab was injected intravitreally. Two months after gas injection, the submacular hemorrhage disappeared. Optical coherence tomography showed flattening of the submacular retinal pigment epithelium, which was not observed prior to bevacizumab therapy. BCVA of the patient's left eye was 0.9. Pneumatic displacement with 40-degree downward gaze positioning after intravitreal sulfur hexafluoride injection is useful to displace a submacular hemorrhage. Early application of this treatment provides improvement of vision and facilitates fundus examination, which could reveal the cause of the submacular hemorrhage.

Indocyanine green angiographic findings of hypertensive choroidopathy.

PURPOSE: To evaluate fundus lesions in patients with malignant hypertension with indocyanine green angiography (ICGA). METHODS: Cases of hypertensive choroidopathy were followed prospectively with ICGA, fluorescein angiography (FA), and optical coherence tomography (OCT). RESULTS: In Case 1, a 34-year-old man had a 10-day history of blurred vision in both eyes. Visual acuity was 0.2 in the right eye and 0.01 in the left eye. Blood pressure (BP) was 270/178 mmHg, and laboratory tests disclosed severe renal dysfunction. In Case 2, a 31-year-old man had noticed blurred vision in both eyes. Visual acuity was 1.2 in the right eye and 1.0 in the left eye. BP was 272/180 mmHg. Marked optic edema, retinal hemorrhage, cotton-wool patches, and Elschnig spots were seen in both cases. FA showed poorly perfused choroid in the early phase and fluorescein dye leakage from the optic disk. OCT demonstrated serous retinal detachment in both cases. ICGA revealed Elschnig spots corresponding to the patchy choroidal filling defect. ICGA demonstrated a larger area of choroidal filling defect than FA. In Case 1, ICGA revealed dye leakage from large choroidal vessels. These OCT and ICGA findings reduced after early systemic treatment for malignant hypertension. CONCLUSION: Hypofluorescent spots in the choroid thought to be choriocapillaris occlusion and choroidal vessel damage recover as result of early treatment for malignant hypertension. ICGA is useful to reveal the disturbance of choroidal circulation in hypertensive choroidopathy.

Soluble IL-6 receptor in vitreous fluid of patients with proliferative diabetic retinopathy.

PURPOSE: To identify the biological reaction of soluble interleukin-6 receptor (sIL-6R) in the vitreous of patients with proliferative diabetic retinopathy (PDR). METHODS: The subjects were 45 patients (45 eyes) with vitreoretinal diseases. The patients were divided into three groups: the PDR group comprised 28 patients (28 eyes) with PDR; the pre-proliferative diabetic retinopathy (PPDR) group comprised seven patients (seven eyes) with PPDR combined with diabetic macular edema; and the nondiabetic group comprised ten patients (ten eyes) with idiopathic macular hole or idiopathic epiretinal membrane. Vitreous samples were obtained at vitrectomy. sIL-6R, vascular endothelial growth factor (VEGF), and protein concentration in vitreous samples were determined by enzyme-linked immunosorbent assay (ELISA). sIL-6R levels in serum were also determined by ELISA in nine of the 28 patients with PDR and in six healthy volunteers as controls. RESULTS: In vitreous fluid, the levels of sIL-6R in the PDR group, PPDR group, and nondiabetic group were 612.7 +/- 233.8 (mean +/- SD), 746.3 +/- 523.1, and 215.4 +/- 98.3 pg/ml, respectively. Vitreous levels of sIL-6R in the PDR and PPDR groups were significantly higher than those in the nondiabetic group (PDR group, P < 0.0001; PPDR group, P < 0.01). In serum, the levels of sIL-6R were 39.38 +/- 9.43 ng/ml in the PDR group and 22.84 +/- 5.32 ng/ml in the control group. sIL-6R levels in serum in the PDR group were significantly higher than those in the control group (P < 0.01). A partial correlation analysis showed a significant correlation between the levels of sL-6R and VEGF in the vitreous in the PDR group (r = 0.34, P < 0.05). CONCLUSIONS: We conclude that the level of sIL-6R in vitreous fluid can be considered as a biomarker of PDR.

[Effect of photodynamic therapy with ATX-S 10 (Na) on experimental choroidal neovascularization].

PURPOSE: To evaluate an appropriate irradiative condition for selective occlusion of experimental choroidal neovascularization(CNV) with photodynamic therapy (PDT) using ATX-S 10 (Na). METHODS: Experimental CNV was induced in monkey eyes by laser photocoagulation. PDT(dose of irradiative energy 40 to 80J/cm2) was performed after 3.5 mg/kg of body weight intravenous injections of ATX-S 10(Na). CNV and retinal vessel occlusion induced by PDT was evaluated by fluorescein angiography (FA) at 1 and 7 days after irradiation. If FA showed no fluorescein dye leakage from CNV at 1 and 7 days after irradiation, CNV was evaluated by histopathological analysis at 7 days after irradiation. RESULTS: Within 30 to 33 minutes after ATX-S 10(Na) injection and irradiation with 50 to 60 J/ cm2, FA showed no fluorescein dye leakage from CNV and no closure of retinal vessels at 1 and 7 days after irradiation. Light micrographs showed occluded CNV, and retinal vessels remained patent and there was no apparent change in the inner layer of the retina. CONCLUSIONS: Irradiative condition of ATX-S10 (Na) 3.5 mg/kg was appropriate 30 to 33 minutes after ATX-S 10(Na) injection and irradiation with 50 to 60 J/cm2.

Twenty-gauge Transconjunctival Vitrectomy.

PURPOSE: To describe a new 20-gauge transconjunctival vitrectomy procedure that we designed, to evaluate its usefulness, and to compare two different methods of use. METHODS: The records of 431 patients (433 eyes) who underwent 20-gauge transconjunctival vitrectomy at the Surugadai Hospital of Nihon University between March 2003 and January 2004 were studied retrospectively. In surgical method 1, 20-gauge wounds were made through the conjunctiva and sclera together. In surgical method 2, 20-gauge conjunctival openings were made 2 mm posterior to the sclerotomies. Absorbable sutures were used to stitch the scleral and conjunctival openings simultaneously in both methods. RESULTS: A 20-gauge transconjunctival vitrectomy has various benefits and is indicated for nearly all ocular diseases. Moreover, it is not limited by the intraocular instruments required. However, since cannulas were not placed in all ports, conjunctival edema occurred more easily with surgical method 2 owing to the leakage of perfusion fluid. CONCLUSIONS: This new 20-gauge transconjunctival vitrectomy procedure has various benefits and is indicated for nearly all ocular diseases. It is not limited by the intraocular instruments required.

Correlation between angiographic and pathological findings in experimental choroidal neovascular membranes after transpupillary thermotherapy.

PURPOSE: To evaluate the correlation between angiographic and pathological findings in experimental choroidal neovascular membranes after transpupillary thermotherapy (TTT). METHODS: Experimental subretinal choroidal neovascular membranes were produced in the monkey eye macula by intense photocoagulation. TTT was delivered, targeting the choroidal neovascular membranes, using a diode laser at 810 nm. One week and 4 weeks after TTT, indocyanine green angiography (IA), fluorescein angiography (FA), optical coherence tomography (OCT), and histological examinations were performed. RESULTS: One week after TTT, indocyanine green dye leakage from choroidal neovascular membranes was reduced on IA, and FA showed staining in the late phase. Light microscopic examination showed numerous small-caliber blood vessels and persistent subretinal exudation. Four weeks after TTT, there was no indocyanine green dye leakage on IA, and FA showed staining. OCT showed a decrease in retinal detachment. Light microscopic examination showed fewer blood vessels and subretinal exudation had disappeared. Choroidal neovascular membranes were composed of fibrous tissue. CONCLUSIONS: Changes in angiographic and OCT findings observed after TTT were inferred to be the result of accelerated fibrosis due to TTT and the disappearance of exudation.

Correlation between indocyanine green angiographic findings and histopathology of polypoidal choroidal vasculopathy.

PURPOSE: To analyze the histopathology of polypoidal choroidal vasculopathy (PCV) and choroidal neovascularization (CNV) developing from PCV, the authors evaluated correlations between pathological findings and the findings of preoperative indocyanine green angiography (IA). METHODS: Two specimens were obtained during CNV excision associated with PCV. PCV tissue was excised with the CNV. The specimens were examined by light microscopy. RESULTS: In one case, IA revealed polypoidal lesions exhibiting hyperfluorescence in both the early and the late phase, and in the affected area, abnormally dilated vessels were identified histologically underneath relatively healthy retinal pigment epithelium (RPE). In the other case, the polypoidal lesions seen on IA showed early hyperfluorescence and late isofluorescence, and dilated vessels were observed under the RPE; perivascular amorphous material was present. The RPE adhered to the side of the choroid, and there was CNV under the neurosensory retina in both cases. The CNV had numerous vascular lumens, was not surrounded by the RPE, and exhibited few fibrous components. CONCLUSIONS: IA findings vary depending on the condition of the RPE located above the PCV and the extent of amorphous material around the PCV.

[Surgical removal of choroidal neovascularization in angioid streaks].

PURPOSE: All the studies so far on surgical removal of choroidal neovascularization (CNV) in angioid streaks(AS) were conducted on a small number of cases. Therefore, a definitive evaluation of the surgical method was not available. The present study aimed to evaluate this surgical modality. SUBJECTS AND METHOD: We performed surgical removal of foveal CNV accompanied by AS. Eighteen eyes of seventeen patients were available for follow-up of over 12 months. Surgical indications included foveal CNV, fluorescence leakage from the CNV in late-phase fluorescein angiography, and visual acuity of 0.3 or lower. RESULTS: The best visual acuity was improved in 44% and unchanged in 44%. The final visual acuity was improved in 33% and unchanged in 39%. 22% had a preoperative visual acuity of 0.2 or above; and 44% and 17% achieved best and final visual acuity, respectively, of 0.2 or above. Since all cases developed atrophy of the choriocapillaris in the fovea, none of the cases were capable of fixation within atrophy. The fixation point was localized outside atrophy in 56% and fixation was poor in 44%. The diameter of postoperative atrophy of choriocapillaris was significantly greater (p < 0.05) than the preoperative CNV diameter. CNV recurred in 8 eyes(44%), 88% of which occurred within one year. CONCLUSION: Surgical removal of CNV in AS is an effective method to maintain preoperative visual acuity.

Glottal configuration, acoustic, and aerodynamic changes induced by variation in suture direction in arytenoid adduction procedures.

Arytenoid adduction is a phonosurgical procedure in which the arytenoid cartilages are approximated to reduce posterior glottal gap size and improve voice. Voice outcomes following arytenoid adduction are not always optimal. The goal of this study was to systematically vary suture direction and force of pull on the arytenoid cartilages in a human excised laryngeal model to determine the optimal combination of factors for reducing glottal gap and improving voice. Several factors demonstrated significant effects. Changes in suture direction and force of pull affected glottal configuration in both the horizontal and vertical planes. Increased force of pull on the muscular process resulted in increased adduction of the vocal process for all suture directions. Changes in suture direction and force of pull also affected acoustic and aerodynamic measures of induced voice. Therefore, voice outcomes can be optimized with arytenoid adduction if the vocal fold plane is accurately adjusted.