[Aneurysmal bone cyst of the ethmoid sinus and skull base in a 3-year old child: a rare location of a benign bone lesion]. / Aneurysmatische Knochenzyste der Rhinobasis bei einem 3-jährigen Kind: Seltene Lokalisation einer benignen Knochenläsion.

Aneurysmal bone cyst is a benign osteolytic lesion in childhood and adolescence which primarily arises in metaphyseal long bones. Its presence in bones of the skull base is very rare. In a 3-year old girl presenting with proptosis, MRI demonstrated a well-defined displacing growing mass in the ethmoid sinus, orbita and anterior fossa. The histopathologic examination of biopsy specimens confirmed an aneurysmal bone cyst. Despite radical surgery the child suffered from two recurrences of the lesion in the first year after initial diagnosis. There has been no subsequent recurrence during the last 3 years. Since this lesion is rarely seen at the skull base, is difficult to differentiate clinically and by histopathology and may take an abnormal course, it is described in this case report to emphasize that it should be included in the differential diagnosis of ENT tumors at this location.

Signaling for growth orientation and cell differentiation by surface topography in uromyces.

The dimensions of the topographical signals for growth orientation and infection structure formation, a cell differentiation event that includes nuclear division, were determined for the stomatal penetrating rust fungus Uromyces appendiculatus. The differentiation signal was found to be a simple ridge on the substrate surface that had a markedly optimum height of 0.5 micrometer. Such ridges were microfabricated on silicon wafers by using electron-beam lithography. A similar ridge, in the form of a stomatal lip, was found associated with the stomatal guard cells of the bean (Phaseolus vulgaris) leaf. Ridge elevations greater than 1.0 micrometer or less than 0.25 micrometer did not serve as effective signals. Germ tubes of the fungus were highly oriented by ridge spacings of 0.5 to 6.7 micrometers. The data indicate that the fungus is able to distinguish uniquely minute differences in leaf surface topography in order to infect the host plant.

Thallphytic alleopathy: isolation and identification of laetisaric Acid.

Laetisaria arvalis, a soil-dwelling basidiomycete fungus, secretes an allelopathic agent that induces rapid hyphal lysis in several phytopathogenic fungi. The active compound was isolated from chloroform:methanol extracts of L. arvalis mycelia and characterized as a previously unknown hydroxy fatty acid, (Z,Z-9,12-8-hydroxyoctadecadienoic acid.

A mechanosensitive channel in whole cells and in membrane patches of the fungus Uromyces.

Bean leaf stomata provide a topographical signal that induces germlings of the phytopathogen Uromyces appendiculatus to develop specialized infection structures. Protoplasts from germ tubes of this fungus, when examined with patch-clamp electrodes, displayed the activities of a 600-picosiemen mechanosensitive ion channel. This channel passes a variety of cations, including Ca2+, and is blocked by Gd3+ at 50 micromolar. This channel could transduce the membrane stress induced by the leaf topography into an influx of ions, including Ca2+, that may trigger differentiation.

Diversity of ampeloviruses in mealybug and soft scale vectors and in grapevine hosts from leafroll-affected vineyards.

The occurrence and diversity of Grapevine leafroll-associated virus 1 (GLRaV-1) and Grapevine leafroll-associated virus 3 (GLRaV-3) in the soft scales Parthenolecanium corni and Pulvinaria innumerabilis and in the mealybug Pseudococcus maritimus was determined in leafroll-affected vineyards in the Finger Lakes region of New York. Groups of 1 to 4 specimens were collected under loose grapevine bark and tested by reverse-transcription polymerase chain reaction (RT-PCR) for segments of the second diverged copy of the GLRaV-1 coat protein gene or GLRaV-3 heat-shock protein 70-homologue gene. Virus-specific RT-PCR products were amplified from immature insect vectors and adult mealybugs. Single viral amplicons were obtained mostly from immature vectors (35%, 30 of 85) and dual viral amplicons from immature (16%, 10 of 61) and adult (100%, 14 of 14) mealybugs, including individuals. These observations suggested a simultaneous uptake of GLRaV-1 and GLRaV-3 by individual mealybugs. Furthermore, a comparative nucleotide sequence analysis of viral amplicons from soft scales, mealybugs, and grapevines from which vectors were collected showed identical or highly similar haplotypes, indicating that uptake of GLRaV-1 and GLRaV-3 likely occurred by direct feeding of vectors on their host plants.

Dispersal and movement mechanisms of Phytophthora capsici sporangia.

Understanding the mechanisms of Phytophthora capsici sporangial dissemination is paramount to understanding epidemic initiation and development. Direct laboratory observations showed P. capsici sporangial dispersal occurred in water with capillary force, but did not occur in response to wind or a reduction in relative humidity. Atmospheric sporangial concentrations were monitored under field conditions using a volumetric spore sampler in a commercial cucurbit field and in an experimental setting where copious sporangia were continuously available in close proximity to the spore trap. Dispersal was infrequent (0.7% of total hours monitored) during sampling in a commercial field; 14 sporangia were detected during a 7.5-week sampling period. In the experimental field situation, dispersal occurred in 4.6% of the hours sampled and 438 sporangia were impacted onto tapes during a 7-week sampling period. Airborne sporangial concentrations were positively associated with rainfall at both sites, but not vapor pressure deficit. Furthermore, in the experimental field situation, wind speed was not significant in regression analysis. Wind speed was not measured in the commercial field. Hence, both direct laboratory observations and volumetric spore sampling indicate that dispersal of sporangia via wind currents is infrequent, and sporangia are unlikely to be naturally dispersed among fields by wind alone.

Surface motility and associated surfactant production in Agrobacterium vitis.

AIMS: Agrobacterium vitis is the causal agent of crown gall of grapevine. Surface motility (swarming), an important mechanism for bacterial colonization of new environments and a previously unknown behaviour of Ag. vitis, was demonstrated. METHODS: Surface motility assays were performed on half-strength potato dextrose agar (Difco) containing 0.75% agar. To test for surfactant production, a drop-collapse test was used. Quorum-sensing (QS) negative and complemented mutants were tested for swarming activity. RESULTS: Ninety-one Agrobacterium strains representing -Agrobacterium tumefaciens (17 strains), Agrobacterium rhizogenes (14 strains) and Ag. vitis (60 strains) were tested for swarming and production of surfactant. All Ag. vitis strains expressed a surface-related motility. In contrast, none of 17 strains of Ag. tumefaciens or 14 strains of Ag. rhizogenes exhibited this behaviour. Surface motility in Ag. vitis was associated with surfactant secretion; both of which are regulated by a QS system previously associated with induction of a hypersensitive response on tobacco and necrosis on grape. An aviR (belongs to luxR family) mutant was surface motility negative and did not produce surfactant. An avsI mutant (autoinducer synthase) was also surface motility negative and was complemented with an Ag. tumefaciens clone expressing avsI. CONCLUSIONS: Agrobacterium vitis is able to produce a characteristic swarming phenotype that is regulated by a complex QS system. SIGNIFICANCE AND IMPACT OF THE STUDY: Swarming activity is unique to Ag. vitis among Agrobacterium sp. and may be associated with the ability of the pathogen to colonize grapevines.

Survey for the Three Major Leafroll Disease-Associated Viruses in Finger Lakes Vineyards in New York.

Vineyards in the Finger Lakes region in New York were surveyed for the three major viruses associated with leafroll disease, i.e., Grapevine leafroll-associated virus 1 (GLRaV-1), Grapevine leafroll-associated virus 2 (GLRaV-2), and Grapevine leafroll-associated virus 3 (GLRaV-3). Target viruses were detected in nearly two-thirds (68%, 65 of 95) of the vineyard blocks surveyed by enzyme-linked immunosorbent assay. Single infections by GLRaV-1, GLRaV-2, and GLRaV-3 occurred in 10% (113 of 1,124), 3% (36 of 1,124), and 15% (173 of 1,124) of the samples tested, respectively, whereas mixed infections affected 3.6% (40 of 1,124) of them, essentially with GLRaV-1 and GLRaV-3 (2.5%, 28 of 1,124). Presence of the target viruses was confirmed in selected samples by reverse transcription-polymerase chain reaction and sequencing. Comparative analysis indicated moderate to high nucleotide sequence identities in the second diverged copy of the GLRaV-1 coat protein gene (81.0 to 86.7%), GLRaV-2 coat protein gene (87.6 to 99.2%), and GLRaV-3 heat shock protein 70 homologue gene (91.5 to 98.3%) of New York isolates with corresponding virus reference strains. The prevalence of the three major leafroll disease-associated viruses in Finger Lakes vineyards results likely from poor sanitary status of planting materials, stressing the need to reinstate a certification program in New York.

Generality of the prerequisite of conidium attachment to a hydrophobic substratum as a signal for germination among Phyllosticta species.

It has been shown that conidia of Phyllosticta ampelicida require attachment to a substratum to initiate germination. Furthermore this attachment occurs only on hydrophobic surfaces. This study was initiated to ascertain the breadth of this phenomenon among other species of the genus Phyllosticta. We tested 23 isolates of Phyllosticta representing at least 14 named species. These isolates were collected from North America, Asia and Africa. For 22 of the 23 isolates tested spore attachment occurred at a rate of 60-100% on hydrophobic polystyrene but at 0-5% on hydrophilic polystyrene. The one exception to the preference for a hydrophobic substratum for attachment was an unnamed species of Phyllosticta from Rhus glauca that attached less than 10% on either surface. A similar response was observed when assaying germination and appressorium formation for 17 isolates. Germination and appressorium formation for these isolates proceeded on hydrophobic polystyrene but not on nutrient agar, which is hydrophilic. In five of the tested isolates germination was high on both hydrophobic polystyrene and hydrophilic nutrient media. The isolate from Rhus glauca did not germinate appreciably on either surface. Taken together these results suggest that the requirement for conidium contact/attachment to trigger germination is pervasive to the genus Phyllosticta.

Two new fluorescent dyes applicable for visualization of fungal cell walls.

Two fluorophores, Solophenyl Flavine 7GFE 500 and Pontamine Fast Scarlet 4B, not heretofore reported upon are described as useful dyes of fungal cell walls, septa and bud scars examined microscopically. The dyes, depending on the filter sets used, yield fluorescently stained material generally in the blue to green and yellow to red wavelengths for Solophenyl Flavine 7GFE 500 and Pontamine Fast Scarlet 4B, respectively. They provide an excellent alternative to the more commonly used fluorophore, Calcofluor White M2R. The two fluorophores, in addition to being used at various spectral wavelengths from mercury arc sources, can be used with laser sources providing 488 nm and 543 nm line wavelengths, common to most scanning confocal microscopes. Unlike Calcofluor, Solophenyl Flavine 7GFE 500 and Pontamine Fast Scarlet 4B do not fade quickly when exposed to selected light wavelengths; however, like Calcofluors they are compatible with living fungal cells.

Visualization of actin in situ by rhodamine-conjugated phalloin in the fungus Uromyces phaseoli.

Rhodamine-conjugated phalloin, a derivative of phalloidin which binds to F-actin, was associated with three types of structures in uredospore germlings of the bean rust fungus, Uromyces phaseoli. The structures were filaments, peripheral plaques, and intranuclear inclusions. The filaments, located throughout the germ tube but especially in the more basipetal regions, were observed as either barely perceivable, fine elements or as easily detectable, coarser structures. The plaques, which we suggest to be equivalent to filasomes, occurred near the periphery of the cell's cytoplasm. They were most numerous in the hyphal tip regions. Nuclear inclusions occurred within the nucleoplasm subjacent to the spindle pole body. Treatments with KI and phalloidin substantiated that the fluorescently labelled sites were F-actin. Treatment of the germlings with cytochalasin E caused the intranuclear inclusions to become extended often with branched, fine filaments. Similar treatments led to a disappearance of cytoplasmic filaments, but had no perceivable effect on the peripheral plaques.

Isolation and identification of actin from the phytopathogenic filamentous fungus uromyces appendiculatus.

Crude protein extracts of Uromyces appendiculatus contain a polypeptide that resembles actin in several ways. This protein eluates from DEAE-cellulose with concentrations of KCl known to release actin of other species from the cation. The polypeptide is recognized by polyclonal antibodies directed to sodium dodecyl sulfate-denatured actin of chicken gizzard as well as by a monoclonal antibody also made to gizzard actin from chicken, but not by antibodies made against rabbit skeletal muscle actin. Western blot analysis after electrophoresis of the protein on polyacrylamide revealed that the protein has an electrophoretic mobility very similar to that of rabbit skeletal muscle actin. We were unable either to isolate actin by affinity chromatography using immobilized DNase-I, or to identify bean rust actin using DNase-I inhibition assays. Nevertheless, large quantities of the protein sedimented by high speed centrifugation. The sedimented protein resisted attempts to solubilize it under conditions normally used to depolymerize actin filaments. Both of the latter findings indicate unusual features of bean rust actin. Immunocytochemical studies of actin localization in germlings of the fungus using two chicken gizzard actin antibodies revealed actin-containing sites which were similar to those previously observed with fluorescently tagged phallotoxin derivatives.

Isolation and characterization of coated vesicles from filamentous fungi.

Coated vesicles have been shown to exist in Neurospora crassa (Ascomycetes) and Uromyces phaseoli (Basidiomycetes) growing germlings. Separation of coated vesicles in both fungi was obtained when the high-speed (100,000g) pellet was fractioned on a Sephacryl S-1000 gel filtration column, according to the procedure of Mueller and Branton. Electron micrographs of negatively stained coated vesicles from fractions of gel filtration show the same striking lattice coated vesicles similar to vertebrate coated vesicles. We observe two major size classes of coated vesicles in both fungi: the larger class (100-180 nm) is similar in size to vertebrate coated vesicles; the smaller class (50-80 nm) is mostly found in both fungi. When examined by SDS-PAGE, the Sephacryl column fractions containing the maximum concentration of electron microscopically visible coated vesicles coincide with the bands of the protein coat reported as clathrin. The protein composition on SDS-PAGE of the coated vesicles indicates a major polypeptide species of 180 kDa and minor 30 to 36 kDa species. Polypeptides of 100 kDa and 64 kDa are also found in the fractions containing coated vesicles.

Escape behavior during academic tasks: a preliminary analysis of idiosyncratic establishing operations.

The presence or absence of idiosyncratic stimuli has been demonstrated to predictably alter the occurrence of problem behavior. By specifying stimuli related to negatively reinforced behavior during academic tasks, it may be possible to identify methods of instruction that decrease the occurrence of problem behavior. The current study used a four-step procedure that involved a functional analysis, descriptive assessment, establishing operations (EO) analysis, and follow-up evaluation (a) to identify the operant function of destructive behavior and (b) to evaluate the effects of idiosyncratic features of academic task demands and related methods of instruction on the occurrence of negatively reinforced destructive behavior of 3 boys with developmental disabilities and autism in a classroom setting. The data suggest that the four-step procedure was effective in identifying methods of instruction that decreased the likelihood of destructive behavior without disrupting the maintaining contingencies for destructive behavior. Results are discussed in terms of establishing operations for negatively reinforced destructive behavior during academic tasks and related methods of instruction in classroom settings.

Synchrotron X-ray micro-computed tomography as a tool for in situ elucidation of insect bacteriomes.

Obligate bacterial endosymbionts are common, influential associates of arthropods, and are often found in specific organs termed bacteriomes. Three dimensional images of bacteriomes of the leafhopper Orosius albicinctus (Hemiptera: Cicadellidae) were reconstructed from synchrotron-based X-ray micro-computed tomography (CT). Results show that bilateral bacteriomes are located between the first and second abdominal tergites, are mushroom-shaped and consist two different types of tissue. Fluorescence in situ hybridization reveals that the primary bacterial symbiont Sulcia muelleri is in the 'cap' part of the of organ. The technique allows a noninvasive, in situ, means of visualizing bacteriomes and will facilitate understanding their form and function.

Evaluation of complications and short-term outcome after unilateral or single-session bilateral tibial tuberosity advancement for cranial cruciate rupture in dogs.

OBJECTIVE: To compare the resulting complications, short-term results, and client satisfaction for treatment of cranial cruciate ligament rupture using either unilateral or bilateral single-session tibial tuberosity advancement (TTA) in dogs. METHODS: Medical records of 68 dogs (101 stifles) undergoing unilateral or bilateral single-session TTA were evaluated. Data gathered included signalment, history, physical examination findings, anaesthesia and surgical time, type of cranial cruciate ligament rupture and meniscal injury, implants, and intra-operative and postoperative complications. A mixed effect logistic regression analysis was performed to determine if complications were grouped by surgical procedure. Linear regression was performed to determine the influence of the variables on the occurrence of complications. Values of p <0.05 were considered significant. RESULTS: No major intra-operative complications occurred. Twenty stifles (20%) developed a complication after surgery (11 minor, 9 major). There was no significant difference in occurrence of complications between dogs undergoing unilateral (n = 8) or bilateral single-session (n = 12) TTA (p = 0.69). The only risk factor found to be associated with complication occurrence was age. CLINICAL SIGNIFICANCE: This is the first report evaluating the use of bilateral simultaneous TTA. There was no significant difference in complication rates between unilateral and bilateral single-session TTA. Additional evaluation is needed to fully determine the extent of complications and long-term outcome of bilateral single-session TTA.

PTH-receptors regulate norepinephrine release in human heart and kidney.

Recent data suggests that chronic renal failure and hyperparathyroidism are associated with sympathetic overactivity. Since peptide hormones are known to modulate norepinephrine (NE) release by activating prejunctional receptors, this study investigates whether parathyroid hormone fragment (1-34) (hPTH(1-34)) increases neuronal NE release in human heart and kidney. Using specific PTH-receptor agonists and antagonists, this study furthermore highlights functional differences between PTH1 and PTH2 receptors. Human atrial and renal tissues were incubated with [(3)H]-NE and superfused. Three electrical stimulations (5Hz, 1min) induced a stable [(3)H]-NE release which was taken as an index of endogenous NE release. RT-PCR with specific primers for PTH1- and PTH2-receptor was performed in heart and kidney. hPTH(1-34) (0.01-0.1µmol/L) and a stable analog of its second messenger cAMP (8-bromo-cAMP) increased [(3)H]-NE release in human atria. This facilitatory effect of PTH was also observed in human renal cortex. The PTH1-receptor antagonist (D-Trp(12), Tyr(34))-pTH-(7-34) (0.5µmol/L) abolished the effect of hPTH(1-34). This data was verified using isolated perfused mouse kidneys. Tuberoinfundibular peptide of 39 residues (TIP-39) (0.1nmol/L-0.1µmol/L) decreased [(3)H]-NE release in atria. PTH1- and PTH2-receptor expressions were demonstrated in human heart and kidney. Moreover, a splice variant of the PTH2-receptor was detected in human kidney. In conclusion, PTH is able to facilitate NE release in human atria and renal cortex by activation of PTH1-receptors. The highly increased PTH levels that can be observed in chronic renal failure might be one contributor for the elevated sympathetic nerve activity and the associated cardiovascular mortality in patients with end stage renal disease.

In vitro study of near-wall flow in a cerebral aneurysm model with and without coils.

BACKGROUND AND PURPOSE: Coil embolization procedures change the flow conditions in the cerebral aneurysm and, therefore, in the near-wall region. Knowledge of these flow changes may be helpful to optimize therapy. The goal of this study was to investigate the effect of the coil-packing attenuation on the near-wall flow and its variability due to differences in the coil structure. MATERIALS AND METHODS: An enlarged transparent model of an ACA aneurysm was fabricated on the basis of CT angiography. The near-wall flow was visualized by using a recently proposed technique called Wall-PIV. Coil-packing attenuation of 10%, 15%, and 20% were investigated and compared with an aneurysmal flow without coils. Then the flow variability due to the coil introduction was analyzed in 10 experiments by using a packing attenuation of 15%. RESULTS: A small packing attenuation of 10% already alters the near-wall flow significantly in a large part of the aneurysmal sac. These flow changes are characterized by a slow flow with short (interrupted) path lines. An increased packing attenuation expands the wall area exposed to the altered flow conditions. This area, however, depends on the coil position and/or on the 3D coil structure in the aneurysm. CONCLUSIONS: To our knowledge, this is the first time the near-wall flow changes caused by coils in an aneurysm model have been visualized. It can be concluded that future hydrodynamic studies of coil therapy should include an investigation of the coil structure in addition to the coil-packing attenuation.