A Nucleolar Protein, MoRRP8 Is Required for Development and Pathogenicity in the Rice Blast Fungus

The nucleolus is the largest, membrane-less organelle within the nucleus of eukaryotic cell that plays a critical role in rRNA transcription and assembly of ribosomes. Recently, the nucle olus has been shown to be implicated in an array of processes including the formation of sig nal recognition particles and response to cellular stress. Such diverse functions of nucleolus are mediated by nucleolar proteins. In this study, we characterized a gene coding a putative protein containing a nucleolar localization sequence (NoLS) in the rice blast fungus, Magnaporthe oryzae. Phylogenetic and domain analysis suggested that the protein is ortholo gous to Rrp8 in Saccharomyces cerevisiae. MoRRP8-GFP (translational fusion of MoRRP8 with green fluorescence protein) co-localizes with a nucleolar marker protein, MoNOP1 fused to red fluorescence protein (RFP), indicating that MoRRP8 is a nucleolar protein. Deletion of the MoRRP8 gene caused a reduction in vegetative growth and impinged largely on asexual sporulation. Although the asexual spores of ΔMorrp8 were morphologically indistinguishable from those of wild-type, they showed delay in germination and reduction in appressorium formation. Our pathogenicity assay revealed that the MoRRP8 is required for full virulence and growth within host plants. Taken together, these results suggest that nucleolar processes mediated by MoRRP8 is pivotal for fungal development and pathogenesis.

Effect of Denosumab on the Change of Osteoclast Precursors Compared to Zoledronate Treatment in Postmenopausal Women with Osteoporosis

Background@#A rapid increase in bone turnover and bone loss has been observed in response to the discontinuation of denosumab. It led to an acute increase in the fracture risk, similar to that observed in the untreated patients. We aimed to investigate the effect of denosumab on osteoclast (OC) precursor cells compared to that of zoledronate. @*Methods@#The study compared the effects of denosumab (60 mg/24-week) and zoledronate (5 mg/48-week) over 48 weeks in postmenopausal women with osteoporosis. From patients’ peripheral mononuclear cells, CD14+/CD11b+/vitronectin receptor (VNR)- and CD14+/CD11b+/VNR+ cells were isolated using fluorescent-activated cell sorting, representing early and late OC precursors, respectively. The primary endpoint was the changes in OC precursors after 48 weeks of treatment. @*Results@#Among the 23 patients, 11 were assigned to the denosumab group and 12 to the zoledronate group (mean age, 69 years). After 48 weeks, the changes in OC precursors were similar between and within the groups. Serum C-terminal telopeptide of type I collagen levels were inversely correlated with OC precursor levels after denosumab treatment (r=-0.72, P<0.001). Lumbar spine, femur neck, and total hip bone mineral density (BMD) increased in both groups. Lumbar spine BMD increased more significantly in the denosumab group than in the zoledronate group. @*Conclusions@#Denosumab and zoledronate treatments induced similar changes in OC precursors. During denosumab treatment, old age and suppressed bone turnover were associated with increased OC precursor cell populations. Further validation studies with prospective designs are required.

Molecular Markers for Detecting a Wide Range of Trichoderma spp. that Might Potentially Cause Green Mold in Pleurotus eryngii

In Pleurotus sp., green mold, which is considered a major epidemic, is caused by several Trichoderma species. To develop a rapid molecular marker specific for Trichoderma spp. that potentially cause green mold, eleven Trichoderma species were collected from mushroom farms and the Korean Agricultural Culture Collection (KACC). A dominant fungal isolate from a green mold-infected substrate was identified as Trichoderma pleuroticola based on the sequences of its internal transcribed spacer (ITS) and translation elongation factor 1-a (tef1) genes. In artificial inoculation tests, all Trichoderma spp., including T. atroviride, T. cf. virens, T.citrinoviride, T. harzianum, T. koningii, T. longibrachiatum, T. pleurotum, and T. pleuroticola, showed pathogenicity to some extent, and the observed symptoms were soaked mycelia with a red-brown pigment and retarded mycelium regeneration. A molecular marker was developed for the rapid detection of wide range ofTrichoderma spp. based on the DNA sequence alignment of the ITS1 and ITS2 regions ofTrichoderma spp. The developed primer set detected only Trichoderma spp., and no cross reactivity with edible mushrooms was observed. The detection limits for the PCR assay of T. harzianum (KACC40558), T. pleurotum (KACC44537), and T. pleuroticola (CAF-TP3) were found to be 500, 50, and 5 fg, respectively, and the detection limit for the pathogen-to-host ratio was approximately 1:10,000 (wt/wt).

Changes in plasma lipoxin A4, resolvins and CD59 levels after ischemic and traumatic brain injuries in rats

Ischemic and traumatic brain injuries are the major acute central nervous system disorders that need to be adequately diagnosed and treated. To find biomarkers for these acute brain injuries, plasma levels of some specialized pro-resolving mediators (SPMs, i.e., lipoxin A4 [LXA4], resolvin [Rv] E1, RvE2, RvD1 and RvD2), CD59 and interleukin (IL)-6 were measured at 0, 6, 24, 72, and 168 h after global cerebral ischemic (GCI) and traumatic brain injuries (TBI) in rats. Plasma LXA4 levels tended to increase at 24 and 72 h after GCI. Plasma RvE1, RvE2, RvD1, and RvD2 levels showed a biphasic response to GCI; a significant decrease at 6 h with a return to the levels of the sham group at 24 h, and again a decrease at 72 h. Plasma CD59 levels increased at 6 and 24 h post-GCI, and returned to basal levels at 72 h post-GCI. For TBI, plasma LXA4 levels tended to decrease, while RvE1, RvE2, RvD1, and RvD2 showed barely significant changes. Plasma IL-6 levels were significantly increased after GCI and TBI, but with different time courses. These results show that plasma LXA4, RvE1, RvE2, RvD1, RvD2, and CD59 levels display differential responses to GCI and TBI, and need to be evaluated for their usefulness as biomarkers.

Changes in plasma lipoxin A4, resolvins and CD59 levels after ischemic and traumatic brain injuries in rats

Ischemic and traumatic brain injuries are the major acute central nervous system disorders that need to be adequately diagnosed and treated. To find biomarkers for these acute brain injuries, plasma levels of some specialized pro-resolving mediators (SPMs, i.e., lipoxin A4 [LXA4], resolvin [Rv] E1, RvE2, RvD1 and RvD2), CD59 and interleukin (IL)-6 were measured at 0, 6, 24, 72, and 168 h after global cerebral ischemic (GCI) and traumatic brain injuries (TBI) in rats. Plasma LXA4 levels tended to increase at 24 and 72 h after GCI. Plasma RvE1, RvE2, RvD1, and RvD2 levels showed a biphasic response to GCI; a significant decrease at 6 h with a return to the levels of the sham group at 24 h, and again a decrease at 72 h. Plasma CD59 levels increased at 6 and 24 h post-GCI, and returned to basal levels at 72 h post-GCI. For TBI, plasma LXA4 levels tended to decrease, while RvE1, RvE2, RvD1, and RvD2 showed barely significant changes. Plasma IL-6 levels were significantly increased after GCI and TBI, but with different time courses. These results show that plasma LXA4, RvE1, RvE2, RvD1, RvD2, and CD59 levels display differential responses to GCI and TBI, and need to be evaluated for their usefulness as biomarkers.

Development of a Molecular Marker Linked to the A4 Locus and the Structure of HD Genes in Pleurotus eryngii

Allelic differences in A and B mating-type loci are a prerequisite for the progression of mating in the genus Pleurotus eryngii; thus, the crossing is hampered by this biological barrier in inbreeding. Molecular markers linked to mating types of P. eryngii KNR2312 were investigated with randomly amplified polymorphic DNA to enhance crossing efficiency. An A4-linked sequence was identified and used to find the adjacent genomic region with the entire motif of the A locus from a contig sequenced by PacBio. The sequence-characterized amplified region marker 7-2299 distinguished A4 mating-type monokaryons from KNR2312 and other strains. A BLAST search of flanked sequences revealed that the A4 locus had a general feature consisting of the putative HD1 and HD2 genes. Both putative HD transcription factors contain a homeodomain sequence and a nuclear localization sequence; however, valid dimerization motifs were found only in the HD1 protein. The ACAAT motif, which was reported to have relevance to sex determination, was found in the intergenic region. The SCAR marker could be applicable in the classification of mating types in the P. eryngii breeding program, and the A4 locus could be the basis for a multi-allele detection marker.

T Cell-Specific Knockout of STAT3 Ameliorates Dextran Sulfate Sodium-Induced Colitis by Reducing the Inflammatory Response

Signal transducer and activator of transcription 3 (STAT3) has a crucial role in various autoimmune disorders including, inflammatory bowel disease (IBD). Our previous study demonstrated that STAT3 activation by IL-6 in colonic epithelial cells exacerbates experimental ulcerative colitis. Activated T lymphocytes are also found in ulcerative colitis patients with intestinal inflammation, but the role of STAT3 in T cells remains elusive. To determine the STAT3 function of T cells in intestinal inflammation, we generated T cell-specific STAT3 knockout (KO) mice and used dextran sulfate sodium (DSS) to induce colitis. In this study, we demonstrated that T cell-specific STAT3 deletion alleviated DSS-induced colitis in mice, resulting in reduced histological scores and myeloperoxidase (MPO) activity. Importantly, the population of T cells in the spleen and lymph nodes was significantly decreased in the control and DSS-induced groups of STAT3 KO mice. In addition, STAT3 deficiency in T cells markedly reduced the production of interferon (IFN)-γ, IL-6, and IL-17A, whereas IL-10 secretion was increased. Collectively, the results suggest that STAT3 in T cells may be a therapeutic target in ulcerative colitis by balancing the immune response through T cell homeostasis.

The Clinical Results of 6-mm Iris-Fixated Phakic IOL

PURPOSE: To evaluate the clinical results of 6 mm iris-fixated phakic intraocular lens (Artisan(R) lens) implantation in myopic patients. METHODS: Forty eyes of 23 myopic patients underwent 6-mm Artisan(R) phakic IOL implantation and were followed-up for over 6 months. We prospectively analyzed the efficacy, stability, predictability, change of astigmatism, endothelial cell count, pupil diameter, degree of decentration, subjective satisfaction and complications. RESULTS: The preoperative mean spherical equivalent was -9.46 D, and postoperative spherical equivalent was -0.70D at 1 month, -0.55D at 3 months, -0.54D at 6 months, -0.78D at 12 months and remained stable during the follow-up period. The preoperative mean astigmatism was -1.88D and postoperative astigmatism was -0.87D at 6 months. Postoperative uncorrected visual acuity (UCVA) was more than 0.8 in 85% of the eyes at 1 month, 91% at 3 months, 95% at 6 months, and 89% at 12 months. The spherical equivalent refraction after surgery was within 1.0D of emmetropia in 78% of eyes at 1 month, 80% at 3 months, 88% at 6 months, and 64% at 12 months. There was no significant decrease in the endothelial cell count during the follow-up period. Preoperative scotopic pupil diameters were significantly decreased at 1 month and the mean decentration of the lens was 0.38 mm. Patient satisfaction was generally high. Complications included the transient elevation of intraocular pressure in 1 eye, corneal edema in 4 eyes, iritis in 1 eye, traumatic dislocation in 1 eye and iris atrophy in 2 eyes. CONCLUSIONS: The 6-mm Artisan(R)phakic IOL implantation may be an effective surgical procedure for myopic patients who cannot undergo corneal refractive surgery, as it provided good visual results, predictability, patient satisfaction, and short-term safety.

Study on the interface between light-cured glass ionomer base and indirect composite resin inlay and dentin / 대한치과보존학회지

This study was done to evaluate the shear bond strength between light-cured glass ionomer cement (GIC) base and resin cement for luting indirect resin inlay and to observe bonding aspects which is produced at the interface between them by SEM. Two types of light cured GIC (Fuji II LC Improved, GC Co. Tokyo, Japan and Vitrebond(TM), 3M, Paul, Minnesota, U.S.A) were used in this study. For shear bond test, GIC specimens were made and immersed in 37degrees C distilled water for 1 hour, 24 hours, 1 week and 2 weeks. Eighty resin inlays were prepared with Artglass(R) (Heraeus Kultzer, Germany) and luted with Variolink(R) II (Ivoclar Vivadent, Liechtenstein). Shear bond strength of each specimen was measured and fractured surface were examined. Statistical analysis was done with one-way ANOVA. Twenty four extracted human third molars were selected and Class II cavities were prepared and GIC based at axiopulpal lineangle. The specimens were immersed in 37degrees C distilled water for 1 hour, 24 hours, 1 week and 2 weeks. And then the resin inlays were luted to prepared teeth. The specimens were sectioned vertically with low speed saw. The bonding aspect of the specimens were observed by SEM (JSM-5400(R), Jeol, Tokyo, Japan). There was no significant difference between the shear bond strength according to storage periods of light cured GIC base. And cohesive failure was mostly appeared in GIC. On scanning electron micrograph, about 30 - 120 microm of the gaps were observed on the interface between GIC base and dentin. No gaps were observed on the interface between GIC and resin inlay.

Retinal Vascular Patterns: Part I. A/V crossings, argyrophilic perivascular network, exogenous retinal vasculitis and arterial annulus

The present paper will concern the venous changes in the arteriovenous crossings, argyrophilic perivasccular network, exogenous retinal vasculitis and arteriolar annulus. Material for the study of arteriovenous crossings was obtained post mortem (19 eyes), which was offered from department of ophthalmology, medical faculty, Kyushu University in Japan, while an eye for the study of vascular changes with traumatic global rupture foHowing to exogenous purulent retinal vasculitis was obtained from eyes enucleated surgically. Material (10 eyes) for the study of argyrophilic perivascular network and arteriolar annuIi was obtained from dogs (5 eyes) and bovines (5 eyes). The technique of perparation of the slides was made of flat preparations of the retina that all nonvascular components were digested with tris- buffered 3%-trypsin solution and stained with PAS (periodic-acid Schiff), hematoxylin, eosin and Vilder's reticulin stain. Results obtained as follows. 1. Various degree of narrowing of the vein in arteriovenous crossing sites was observed in flat preparation of retinal vessels without any staining technique, which might be related to arteriolar sclerotic changes. 2. An encircling perivascular argyrophilic network consisting of wavy fibers and some externallyr interlacing argyrophilic longitudinal fibers were observed in dog retinal arterioles but coarser fibers in venules. Some strand-like intercapillary bridge was argyrophilic in Wilder's reticulin stain of the human retinal vessels. 3. Retinal venules and its capillaries in a 40-year-old man with traumatic global rupture following to purulent exogenous retinal vasculitis predominantly affected. The walls of capillaries had multiple aneurysmal dilatations, and in some ruptured area numerous basophilic bodies in H-E. stain were observed. The cells of abnormally dilated capillaries were missing over wide areas, and in some capillaries predominated loss of endothelial cells. 4. Arteriolar Annulus in positive for periodic-acid Schiff technique was hyper-cellularities in H-E. staining, which did not appear to be different from surrounding cells in arterioles.

Studies on the Phosphorylase Activity in Ciliary Muscle

The author studied the effects of adrenergic and its blocking agents, ACTH, methylprednisolone, 5-hydroxytryptamine, and prostaglandin E1 and E2 on the phosphorylase activity in bovine ciliary muscle in vitro and obtained the following results. 1. The phosphorylase activity in bovine ciliary muscle was enhanced by the adrenergic agents. such as epinephrine. norepinephrine and isoproterenol, but was not affected by adrenergic alpha receptor blocking agents, such as phenoxybenzamine and phentolamine, and adrenergic beta receptor blockades. such as propranolol and pronethalol. Pretreatment with propranolol prevented partly the epinephrine-induced increase of the phosphorylase activity in bovine ciliary muscle. 2. The phosphorylase activity in bovine ciliary muscle was shown to be stimulated by ACTH, methylprednisolone and 5-hydroxytryptamine but not by prostaglandin E1 and E2. 3. The phosphorylase activity in bovine ciliary muscle which was stimulated by epinephrine, ACTH, methylprednisolone or 5-hydroxytryptamine was markedly blocked by pretreatment with prost aglandin E1 but less by prostaglandin E2. 4. Prostaglandin E1 exerted no effects on the phosphorylase activity in bovine ciliary muscle following the addition of cyclic AMP or MgH++-ATP, but markedly depressed that stimulated by epinephrine.

Pathogenetic Mechanisms of Corneal Inflammation

This paper is to review pathogenetic mechanisms in inflammatory diseases of the cornea. After demonstration of immune corneal reactions were first undertaken in the early 20th century, the considerable changes in concept of corneal inflammation were taken. It has been the focus of immune mechanism to introduce the various aspects of corneal inflammation, especially definition of inflammation, inflammatory stimulus, normal host response, and histopathalogical evolutionof simple corneal ulcer and repair of corneal tissues.

Effect of Practolol on Ocular Tension in Rabbits

A trial with eye drops of practolol, 0.2% and propranolol, 0.1%, and intravenous injection of practolol, 1-4mg/kg. and propranolol, 0.5-2mg/kg. in rabbits was done for the changes of ocular tension. Ocular tension was estimated by Schotz tonometer. The E.C.G. was recorded before and after treatment of the above drugs. In the rabbits all of doses of practolol, topically and systemically applied failed to reduce ocular tension, but propranolol reduced slightly ocular tension which was suggested that the lowering of ocular tension was mediated by its local anesthetic activity.

Tensile Strength of the Fibrous Coats of Rabbit Eye

This report describes the elongation and rupture pattern studied on 42 tissue slices of the fibrous coats (Cornea, Sclera and Limbus) of the fresh rabbit eye. The size of slices was 1.OOO +/- O.667 long by 0.900 +/- 0.667mm. wide. For each slice elongation was measured under the microscope in a magnitude of 30 X. The elongation, micron per each 10 gram load added, was taken as an elongation ratio. The results were as follows: 1. The shapes of elongation curve of all slices showed two different patterns between 1st limit (20~30gm.) and 2nd limit (150-180gm.), that was markedly greater elongation ratio in 1st limit than that in 2nd limit. 2. Elongation curves showed increased magnitude from central portion toward limbus in the cornea and from limbus to posterior pole in sclera. 3. Mean elongation ratio was greater in latitudinal than meridional of all slices. 4. Mean elongation ratio of the macular area was similar to that of the posterior portion of sclera. 5. The range of weight required to make rupturing of each slice was 150~230gm., though the slices were taken in same portion.

Dye Test for Toxoplasmosis to normal population in Pusan

Serological survey, using methylene blue dye test for toxoplasmosis was carried out upon 395 normal population, 21 cases of uveal lesions, and 95 cases of acute febrile and chronic systemic diseases. Toxoplasma gondii preferred for this test was "Anyang straio" from Veterinary Researh Anynag, Korea in March 1968. The results were as follows: 1. In normal population 9.62 per cent of the sera examined showed positive titers of 1:16 or above in adult and 1:8 or above in children. 2. The positive in serologic survey was prevalent 3rd to 5th decades of both sex. 3. 4 out of 21 cases of uveal lesions were positive of which titers showed 1:64 or above in adults and 1:32 in a girl. 4. In 95 cases of acute febrile and chronic systemic diseases 4.2 per cent of the sera examined showed positive titers of 1:16 or above.

Migration of Photoreceptor Cells Into the Subretinal Space in Rabbit Retina

The migration of photoreceptors into the subretinal space has been reported to occur in developing and aged rat retina, in aged human retina, in detached owl monkey, and in detached cat retina. Subretinal photoreceptor cells have been observed in aphakic-vitrectomized rabbit eyes with or without intravitreal saline or antibiotic injection. Variety of photoreceptor cell nuclei can be distinguished. The nuclei of photoreceptor cells change their shape so as to pass easily through the juncition and thereafter they returned to their original shape in sub retinal space. The mechanisms and biological significance of this phenomenon are not yet certain, but it may by one of the important factors contributing to decreased vision with aging.

On the Ratio to the Physiologic Cupping and Optic Disc. and Optic Disc

A method of charting and scoring the optic disc and cupping from fundus color photographs is presented. The imase that the color slides were projected on the graphic sheet was traced and calculated C/D ratio as horizontal diameter and mensuration of division by measure. The result showed statiscal significancy in calculation of the C/D ratio as mensuration of division in cases which the cupping showed irregular form.

Dilated and Fixed Pupil

A Dilated pupil with a loss of reactions to light and accommodation convergence exhibit these defects in varying degree, and occur under a variety of circumstances. These conditions include unilateral dilatation and fixity of the pupil as a result of injury to the eye and as the result of ipsilateral cerebral compression and bilaterally dilated and fixed pupils seen commonly in juvenile paresis and tabo-paresis. There are another unknown circumstances as tonic pupil. The authors presented a case of 22 years old Korean female under the diagnosis of dilated and fixed pupil which possibly originated from syphilis in as much as she was strongly positive reaction in serum S.T.S. and her abnormal pupil was recovered after anti-syphilitic therapy. The authors reviewed the literatures concerning these conditions and discussed about the possibility of early involvement of syphilis in the nervous system and the possible site of lesion which interefere pupillary reflex to light and accommodation-convergence. Finally authors commented the possible site of lesion is efferent pupilloconstrictor fibers which pass through the tentorial gap. In this region para-sympathetic pupilloconstrictor fibers, concentrated over the superior are of the oculomotor nerve, may be compressed upon its superior surface by inflammatory swelling of syphilitic involvement of adjacent structures.

Follow-up Study of Trabeculectomy

In 1968, Cairns introduced the trabeculectomy operation and reported it to be a highly succesful form of surgery for the reduction of intraocular pressure in primary open-angle glaucoma. At the present time trabeculectomy is the treatment of choice for patients with primary open-angle glaucoma requiring surgery. In the period January, 1975. to March, 1978, 72 eyes of chronic simple, chronic angle-closure, secondary, and developmental glaucoma were operated by trabeculectomy and/or trabeculotomy in Busan National University Hospital. Follow-up study was achieved in 28 eyes(chronic simple glaucoms 17, chronic angle-closure glaucoma 4, secondary glaucoma 3, and developmental 4) of 72 eyes, and the duration of follow-up ranged from 1 year to 2 and 1/2 years(average 18 months). Our operative procedure, which was different from others, was made with 1/3 thickness of scleral flap, and made 2 knots with 8-0 nylon placed in free edge of scleral flap and 1/3 of all cases was not placed scleral flap sutures. We achieved success in 23 eyes (82.1% )of 28 eyes of all types of glaucoma and 14 eyes (82.4%) of 17 eyes of openangle glaucoma. The control of intraocular pressure in interrelationship to formation of filtering bleb was statistically significance(x2(0.05, 1)=3.84>5.46), and to scleral flap suture was not significance(x2(0,05, 1)=3.84>0.39). The complications during and after operation were delayed formation of anterior chamber, hyphema, posterior synechia, vitreous prolapse, choroidal detachment, and lens and vitreous opacity in order.

The Malignant Tumors of the Eye

We reported the clinical and histopathologic review of 71 cases of primary and metastk malignant tumors of the eye and its adnexa for last 15 years at Department of Ophthalmology, Busan National University Hospital. The followings were summarized. 1. Malignant tumors of the eye and its adnexa were more frequent in man, their ratio of male and female revealed 47:25. Their occurrence by age were under 10 year-old, 5th, 6th, and 4th decades in order. 2. The frequency of occurrence of malignant tumors in the eye and its adnexa was retinobla stoma, primary squamous and basal cell carcinoma and secondary squamous cell carcinoma in order. 3. In primary malignant tumors of eyelid, it revealed same incidence of squamous and basal cell carcinoma, and adenocarcinoma was also relatively high incidence.