A retrospective cohort study comparing a novel, spherical, resorbable particle against five established embolic agents for uterine fibroid embolisation.

AIM: To evaluate the effectiveness of a novel, resorbable, spherical embolic agent compared with other established agents, by studying percentage fibroid infarction (the best indicator of long-term symptom improvement) in patients undergoing uterine fibroid embolisation (UFE). MATERIALS AND METHODS: This retrospective cohort study examined six different embolic agents used for fibroid embolisation, including a new gelatin-based, fully resorbable, spherical agent. The primary effectiveness outcomes were magnetic resonance imaging (MRI)-determined dominant fibroid infarct percentage (DF%) and all fibroid percentage infarct (AF%) at 3 months post-embolisation. MRI-determined uterine artery patency rate was the secondary outcome. Chi-squared test (χ2), relative risk (RR) calculation (primary outcomes), and analysis of variance (ANOVA) (secondary outcome) were the statistical tests employed. RESULTS: One hundred and twenty patients were treated with six embolic agents (20 consecutive patients per group, overall mean age 44.8±6.4, initial uterine volume 570±472 ml, dominant fibroid volume 249±324 ml). Fibroid infarctrates were similar between the cohorts with no significant difference between the new gelatin-based resorbable particle and other embolics in either DF% (χ2=3.92, p=0.56) or AF% (χ2=2.83, p=0.73). Complete DF% RR=1.07 (0.90-1.27) and AF% RR=1.09 (0.85-1.41) suggest non-inferiority of the resorbable particle (d=0.67, p<0.05). A favourable uterine artery patency rate was demonstrated for the resorbable particle compared with gelatin slurry (82.5% versus 27.5%, p<0.001 after Bonferroni adjustment). CONCLUSIONS: This new gelatin-based, fully resorbable particle is an effective embolic agent for fibroid embolisation and achieves an infarct rate non-inferior to established embolics.

Improving trauma imaging in Wales through Kotter's theory of change.

AIM: To improve the practice of trauma computed tomography (CT) within Wales using recognised leadership techniques for change. MATERIALS AND METHODS: Royal College of Radiologists' (RCR) guidance, in addition to other key recent evidence, were used to form an aspirational standard. All centres across Wales with a major emergency department were included. Kotter's theory of change was utilised to facilitate an improvement in practice across the region, with larger units prioritised initially. RESULTS: Of the 13 major emergency units in Wales, eight centres had no formal trauma CT protocol. Only one centre utilised the Bastion protocol (in comparison to 75% of major trauma centres). After the campaign to improve trauma imaging, seven centres now offer the Bastion protocol, with currently only three peripheral centres still without a procedure for whole-body CT. The two largest centres have implemented an emergency department pro forma. CONCLUSION: Trauma CT within Wales has significantly improved as a result of this project. Kotter's theory is demonstrated as an effective tool for facilitating a change in practice on a regional/national scale.

Is the follow-up of small renal angiomyolipomas a necessary precaution?

AIM: To investigate the natural history and rationalize follow-up of renal angiomyolipomas (AMLs). MATERIALS AND METHODS: A prospectively gathered radiology database was scrutinized to identify patients with renal AMLs over a 3 year period (January 2006 to December 2008). Radiological investigations were examined to identify those AMLs exhibiting change during surveillance. RESULTS: A total of 135 patients were identified. Mean age at first detection was 49.6 years and patients were followed up for a median 21.8 months (6-85.3 months). Small AMLs (≤20 mm) were less likely to grow than their larger counterparts [odds ratio 13.3, confidence interval (95% CI) 1.4-123.9, p = 0.02] and exhibited a slower growth rate (0.7 versus 9.2 mm/year). Patients with AMLs that increased in size were significantly younger (median age 43 versus 52 years, p < 0.001). Multiple AMLs or those associated with genetic conditions grew at a significantly greater rate (3 versus 0.1 mm/year, p < 0.001). AMLs with a large extra-renal component are less reliably measured on ultrasound (median error 7 versus 1 mm, p < 0.001). CONCLUSION: This is the first study with the primary purpose to investigate growth of small AMLs (≤20 mm). Small, solitary AMLs (≤20 mm) do not require follow-up due to their low probability of growth. Patients with multiple AMLs and younger patients require closer monitoring due to their comparatively greater AML growth rate. Ultrasound-detected AMLs with an extra-renal component may require computed tomography (CT) to confirm their size.

Machine Learning to Predict Prostate Artery Embolization Outcomes.

PURPOSE: This study leverages pre-procedural data and machine learning (ML) techniques to predict outcomes at one year following prostate artery embolization (PAE). MATERIALS AND METHODS: This retrospective analysis combines data from the UK-ROPE registry and patients that underwent PAE at our institution between 2012 and 2023. Traditional ML approaches, including linear regression, lasso regression, ridge regression, decision trees and random forests, were used with leave-one-out cross-validation to predict international prostate symptom score (IPSS) at baseline and change at 1 year. Predictors included age, prostate volume, Qmax (maximum urinary flow rate), post-void residual volume, Abrams-Griffiths number (urodynamics score) and baseline IPSS (for change at 1 year). We also independently confirmed our findings using a separate dataset. An interactive digital user interface was developed to facilitate real-time outcome prediction. RESULTS: Complete data were available in 128 patients (66.7 ± 6.9 years). All models predicting IPSS demonstrated reasonable performance, with mean absolute error ranging between 4.9-7.3 for baseline IPSS and 5.2-8.2 for change in IPSS. These numbers represent the differences between the patient-reported and model-predicted IPSS scores. Interestingly, the model error in predicting baseline IPSS (based on objective measures alone) significantly correlated with the change in IPSS at 1-year post-PAE (R2 = 0.2, p < 0.001), forming the basis for our digital user interface. CONCLUSION: This study uses ML methods to predict IPSS improvement at 1 year, integrated into a user-friendly interface for real-time prediction. This tool could be used to counsel patients prior to treatment.

Postharvest Disease Development on Southern Highbush Blueberry Fruit in Relation to Berry Flesh Type and Harvest Method.

Postharvest decay, incited by various fungal pathogens, is a major concern in most blueberry production areas of the United States. Because the risk of infection is increased by fruit bruising, which in turn is increased by machine-harvesting, it has been difficult to harvest fruit from the early-maturing but soft-textured southern highbush blueberries (SHB) mechanically for the fresh market. This could change fundamentally with the recent development of SHB genotypes with crisp-textured ("crispy") berries, i.e., fruit with qualitatively firmer flesh and/or more resistant skin. Four replicate row sections of two or three SHB genotypes having crispy fruit and three with conventional fruit were either hand- or machine-harvested at a commercial blueberry farm in northern Florida in April 2009 and May 2010. Harvested fruit were sorted, packed, and placed in cold storage (2°C) for up to 3 weeks. Average counts of aerobic bacteria, total yeasts and molds, coliforms, and Escherichia coli on fruit samples before the cold storage period were below commercial tolerance levels in most cases. In both years, natural disease incidence after cold storage was lowest for hand-harvested crispy fruit and highest for machine-harvested conventional fruit. Interestingly, machine-harvested crispy fruit had the same or lower disease incidence as hand-harvested conventional fruit. Across all treatments, natural postharvest disease incidence was inversely related to fruit firmness, with firmness values >220 g/mm associated with low disease. In separate experiments, samples from the 0-day cold storage period were inoculated at the stem end with Alternaria alternata, Botrytis cinerea, or Colletotrichum acutatum, and disease incidence was assessed after 7 days in a cold room followed by 60 to 72 h at room temperature. In response to artificial inoculation, less disease developed on crispy berries. No significant effect of harvest method was observed, except for A. alternata inoculation in 2009, when hand-harvested fruit developed a lower level of disease than machine-harvested fruit. Taken together, this study suggests that mechanical harvesting of SHB cultivars with crisp-textured berries is feasible from a postharvest pathology perspective.

Robust Image Population Based Stain Color Normalization: How Many Reference Slides Are Enough?

Histopathologic evaluation of Hematoxylin & Eosin (H&E) stained slides is essential for disease diagnosis, revealing tissue morphology, structure, and cellular composition. Variations in staining protocols and equipment result in images with color nonconformity. Although pathologists compensate for color variations, these disparities introduce inaccuracies in computational whole slide image (WSI) analysis, accentuating data domain shift and degrading generalization. Current state-of-the-art normalization methods employ a single WSI as reference, but selecting a single WSI representative of a complete WSI-cohort is infeasible, inadvertently introducing normalization bias. We seek the optimal number of slides to construct a more representative reference based on composite/aggregate of multiple H&E density histograms and stain-vectors, obtained from a randomly selected WSI population (WSI-Cohort-Subset). We utilized 1,864 IvyGAP WSIs as a WSI-cohort, and built 200 WSI-Cohort-Subsets varying in size (from 1 to 200 WSI-pairs) using randomly selected WSIs. The WSI-pairs' mean Wasserstein Distances and WSI-Cohort-Subsets' standard deviations were calculated. The Pareto Principle defined the optimal WSI-Cohort-Subset size. The WSI-cohort underwent structure-preserving color normalization using the optimal WSI-Cohort-Subset histogram and stain-vector aggregates. Numerous normalization permutations support WSI-Cohort-Subset aggregates as representative of a WSI-cohort through WSI-cohort CIELAB color space swift convergence, as a result of the law of large numbers and shown as a power law distribution. We show normalization at the optimal (Pareto Principle) WSI-Cohort-Subset size and corresponding CIELAB convergence: a) Quantitatively, using 500 WSI-cohorts; b) Quantitatively, using 8,100 WSI-regions; c) Qualitatively, using 30 cellular tumor normalization permutations. Aggregate-based stain normalization may contribute in increasing computational pathology robustness, reproducibility, and integrity.

Hyaluronidase-induced reductions in myocardial infarct size.

The size of myocardial infarctions following coronary artery occlusion in the rat was determined directly by measurement of creatine phosphokinase activity in homogenized whole left ventricles and by planimetric measurement of the area of the infarctions in histologic sections of serial slices of the left ventricles. Hyaluronidase was shown to produce significant reductions in expected infarct size both 48 hours and 3 weeks after occlusion without impairing fibrosis during the healing phase. Thus, the amount of myocardial necrosis that follows a coronary artery occlusion has been shown directly to be amenable to reduction with a pharmacological intervention.

Factors Predicting a Good Symptomatic Outcome After Prostate Artery Embolisation (PAE).

INTRODUCTION: As prostate artery embolisation (PAE) becomes an established treatment for benign prostatic obstruction, factors predicting good symptomatic outcome remain unclear. Pre-embolisation prostate size as a predictor is controversial with a handful of papers coming to conflicting conclusions. We aimed to investigate if an association existed in our patient cohort between prostate size and clinical benefit, in addition to evaluating percentage volume reduction as a predictor of symptomatic outcome following PAE. MATERIALS OR METHODS: Prospective follow-up of 86 PAE patients at a single institution between June 2012 and January 2016 was conducted (mean age 64.9 years, range 54-80 years). Multiple linear regression analysis was performed to assess strength of association between clinical improvement (change in IPSS) and other variables, of any statistical correlation, through Pearson's bivariate analysis. RESULTS: No major procedural complications were identified and clinical success was achieved in 72.1% (n = 62) at 12 months. Initial prostate size and percentage reduction were found to have a significant association with clinical improvement. Multiple linear regression analysis (r2 = 0.48) demonstrated that percentage volume reduction at 3 months (r = 0.68, p < 0.001) had the strongest correlation with good symptomatic improvement at 12 months after adjusting for confounding factors. CONCLUSION: Both the initial prostate size and percentage volume reduction at 3 months predict good symptomatic outcome at 12 months. These findings therefore aid patient selection and counselling to achieve optimal outcomes for men undergoing prostate artery embolisation.

Long-term preservation of ischemic myocardium after experimental coronary artery occlusion.

The results of experiments with indirect methods have suggested that various interventions reduce infarct size after coronary artery occlusion. To determine and quantify directly both the short- and long-term effects of several interventions on myocardial salvage without relying on indirect methods, the left coronary artery was occluded in 880 rats; they were then given either no treatment or one of the following interventions: (a) hyaluronidase, an enzyme that hydrolyzes interstitial glycoproteins, 1,500 National Formulary (NF) U/kg i.v. 5 min and 24 h after occlusion; (b) cobra venom factor, a protein that depletes the third component of complement, 20 U/kg i.v. 5 min after occlusion; (c) a glucocorticoid: hydrocortisone, 50 mg/kg i.v. 5 min after occlusion; or the five-fold more potent methylprednisolone (MP): (i) 50 mg/kg i.v. 5 min after occlusion or (ii) 50 mg/kg i.v. 5 min after occlusion followed by 50 mg/kg i.m. 3, 6, and 24 h after occlusion; or (d) reserpine, an agent that depletes the heart of catecholamines, 0.5 mg/kg i.m. once on each of the 3 days before occlusion. The animals were sacrificed either 2 days after occlusion, i.e., at the time of peak necrosis, or after 3 wk, i.e., after the infarct was completely healed. The amount of preserved myocardium was then assessed by two independent techniques: planimetric measurement of serial histologic sections and creatine kinase activity of the whole left ventricle. The amount of normal myocardium preserved at 21 days postocclusion was significantly increased, by 22.3+/-7.8% (P < 0.025) after the administration of hyaluronidase, by 25.3+/-5.8% (P < 0.005) after cobra venom factor, by 14.5+/-6.9% (P < 0.05) after hydrocortisone, by 20.8+/-8.2% (P < 0.025) after the single dose of MP, by 20.9+/-3.9% (P < 0.001) after the four doses of MP, and by 10.2+/-3.7% (P < 0.05) as a result of pretreatment with reserpine. The four doses of MP significantly thinned the infarct-by 25.6+/-2.9% (P < 0.001)-and although ventricular rupture did not occur, the intervention caused distension of the left ventricle as a result of stretching of the infarcted tissue during scar formation. Thus, myocardium acutely jeopardized by ischemia can be preserved on a long-term basis.

Sympathetic activation in exercise is not dependent on muscle acidosis. Direct evidence from studies in metabolic myopathies.

Muscle acidosis has been implicated as a major determinant of reflex sympathetic activation during exercise. To test this hypothesis we studied sympathetic exercise responses in metabolic myopathies in which muscle acidosis is impaired or augmented during exercise. As an index of reflex sympathetic activation to muscle, microneurographic measurements of muscle sympathetic nerve activity (MSNA) were obtained from the peroneal nerve. MSNA was measured during static handgrip exercise at 30% of maximal voluntary contraction force to exhaustion in patients in whom exercise-induced muscle acidosis is absent (seven myophosphorylase deficient patients; MD [McArdle's disease], and one patient with muscle phosphofructokinase deficiency [PFKD]), augmented (one patient with mitochondrial myopathy [MM]), or normal (five healthy controls). Muscle pH was monitored by 31P-magnetic resonance spectroscopy during handgrip exercise in the five control subjects, four MD patients, and the MM and PFKD patients. With handgrip to exhaustion, the increase in MSNA over baseline (bursts per minute [bpm] and total activity [%]) was not impaired in patients with MD (17+/-2 bpm, 124+/-42%) or PFKD (65 bpm, 307%), and was not enhanced in the MM patient (24 bpm, 131%) compared with controls (17+/-4 bpm, 115+/-17%). Post-handgrip ischemia studied in one McArdle patient, caused sustained elevation of MSNA above basal suggesting a chemoreflex activation of MSNA. Handgrip exercise elicited an enhanced drop in muscle pH of 0.51 U in the MM patient compared with the decrease in controls of 0.13+/-0.02 U. In contrast, muscle pH increased with exercise in MD by 0.12+/-0.05 U and in PFKD by 0.01 U. In conclusion, patients with glycogenolytic, glycolytic, and oxidative phosphorylation defects show normal muscle sympathetic nerve responses to static exercise. These findings indicate that muscle acidosis is not a prerequisite for sympathetic activation in exercise.

Sequence specificity in the recognition of the epidermal growth factor receptor by the abl Src homology 2 domain.

The transforming activity of the abl gene product requires a functional src homology 2 (SH2) domain. An assay was developed to evaluate this function by examining binding of a bacterially-expressed abl SH2 domain to the activated EGF receptor, used as a surrogate tyrosine phosphorylated protein. The sequence specificity of this interaction has been explored with a series of point mutants of EGF receptor. Analysis of equilibrium binding reveals that substitution of Tyr1086 for Phe in the EGF receptor produced a 10-fold reduced affinity for abl SH2 domain binding as compared to the wildtype receptor. Moreover, a phosphorylated peptide modeled on the sequences surrounding Tyr1086 specifically inhibits abl SH2 binding, with an IC50 of approximately 10 microM. Evaluation of a series of additional peptides, modeled on the Tyr1086 sequence, revealed that the carboxy terminal residues directly next to the phosphotyrosine were particularly critical to this binding. Molecular modeling studies of the pTyr1086 peptide revealed the potential hydrophobic, ionic and hydrogen bonding interactions involved in the functions of the abl SH2 domain.

Molecular cloning of troponin I expressed in fast white muscle of a teleost fish, the Atlantic herring (Clupea harengus L.).

By cDNA cloning and 5'-RACE analysis we characterised a Clupea harengus troponin I mRNA expressed in larvae and in adult white (fast) muscle, but not in red (slow) or cardiac muscle. The mRNA encodes a TnI protein of the short chain length (176 residues) N-terminally truncated type previously observed only in tetrapod skeletal muscles. Despite its expression specificity the herring TnI does not particularly resemble the tetrapod fast skeletal muscle isoform in sequence but appears to be outside of the tetrapod TnIfast/Tnslow/TnIcardiac isoform family. Surprisingly, the actin/TnC-binding sequence resembles that of arthropod, rather than tetrapod vertebrate, troponin I's and has, besides, unique features not seen in any other troponin I's, vertebrate or invertebrate.

Substance P and somatostatin content and transport in vagus and sciatic nerves of the streptozocin-induced diabetic rat.

Substance P (SP) and somatostatin (SS) are two widely distributed neuropeptides that within the vagus and sciatic nerves are localized predominantly in sensory fibers. The effect of diabetes mellitus on their content or transport in sensory nerves is unknown. With the nerve ligation technique, the peripheral orthograde 24-h transport of both peptides was quantified in the vagus nerve 3 days or 1 mo after induction of streptozocin (STZ) diabetes and in both the vagus and sciatic nerves after diabetes of 3 mo duration. In acute (3-day) diabetics, neuropeptide transport in the vagus was unaltered. After 1 mo, SP transport was significantly increased; content in unligated contralateral nerve was unaltered. Transport of SS was unchanged, and content in contralateral nerve was too low to reliably quantitate. After diabetes of 3-mo duration, transport of both peptides in the vagus nerve was increased in STZ-induced diabetic (STZ-D) rats versus both weight- and age-matched controls: SP 474 +/- 17 (N = 10) vs. 358 +/- 32 (N = 13) pg/24 h, STZ-D rats vs. controls, mean +/- SE, P less than .03; SS 29 +/- 4 vs. 20 +/- 3 pg/24 h, STZ-D rats vs. controls, P less than .02. In the sciatic nerve, SP transport and content were unaltered. SS content was significantly reduced: 17 +/- 3 vs. 30 +/- 3 pg/3-mm nerve segment, STZ-D rats vs. controls, P less than .01. SS transport in the sciatic nerve of diabetic rats was variably reduced (P less than .07), and transport rates were increased (1.41 +/- 0.13 vs. 0.96 +/- 0.10 mm/h.(ABSTRACT TRUNCATED AT 250 WORDS)

Differentiation of peptide molecular recognition by phospholipase C gamma-1 Src homology-2 domain and a mutant Tyr phosphatase PTP1bC215S.

Activated epidermal growth factor receptor (EGFR) undergoes autophosphorylation on several cytoplasmic tyrosine residues, which may then associate with the src homology-2 (SH2) domains of effector proteins such as phospholipase C gamma-1 (PLC gamma-1). Specific phosphotyrosine (pTyr)-modified EGFR fragment peptides can inhibit this intermolecular binding between activated EGFR and a tandem amino- and carboxy-terminal (N/C) SH2 protein construct derived from PLC gamma-1. In this study, we further explored the molecular recognition of phosphorylated EGFR988-998 (Asp-Ala-Asp-Glu-pTyr-Leu-Ile-Pro-Gln-Gln-Gly, I) by PLC gamma-1 N/C SH2 in terms of singular Ala substitutions for amino acid residues N- and C-terminal to the pTyr (P site) of phosphopeptide I. Comparison of the extent to which these phosphopeptides inhibited binding of PLC gamma-1 N/C SH2 to activated EGFR showed the critical importance of amino acid side chains at positions P+2 (Ile994), P+3 (Pro995), and P+4 (Gln996). Relative to phosphopeptide I, multiple Ala substitution throughout the N-terminal sequence, N-terminal sequence, N-terminal truncation, or dephosphorylation of pTyr each resulted in significantly decreased binding to PLC gamma-1 N/C SH2. These structure-activity results were analyzed by molecular modeling studies of the predicted binding of phosphopeptide I to each the N- and C-terminal SH2 domains of PLC gamma-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Phytophthora sojae avirulence genes, RAPD, and RFLP markers used to construct a detailed genetic linkage map.

Two crosses between different races of Phytophthora sojae were established using one race as a common parent in both crosses. F2 populations comprising over 200 individuals were generated for each cross. A subset of 53 F2 individuals from each cross was selected at random for genetic analysis of virulence/avirulence and molecular markers, and finally the construction of a detailed genetic linkage map. The linkage map developed for P. sojae is based on a total of 257 markers (22 RFLP, 228 RAPD, and 7 avirulence genes). The linkage map comprises 10 major and 12 minor linkage groups covering a total of 830.5 cM. Close linkage was observed between Avr4 and Avr6 (0.0 cM), Avr1b and Avr1k (0.0 cM), and Avr3a and Avr5 (4.6 cM). Coupling phase linkage of RFLP and RAPD markers to all seven avirulence genes was identified at the minimum and maximum distances of 0.0 and 14.7 cM, respectively.

Differential expression of peroxidase isogenes during the early stages of infection of the tropical forage legume Stylosanthes humilis by Colletotrichum gloeosporioides.

Infection of Stylosanthes humilis by the fungal phytopathogen Colletotrichum gloeosporioides is associated with an increase in peroxidase enzyme activity within 24 h postinoculation. Peroxidase gene expression was investigated as a first step towards understanding the regulation and functional importance of this host response to fungal attack. Four distinct cDNAs Shpx 2, 5, 6, and 12, isolated from a cDNA library of S. humilis contained deduced amino acid (aa) sequence motifs characteristic of peroxidases. Three of these (Shpx 2, 5, and 6) were full-length and their deduced proteins each fell into a different homology group based on comparisons with other plant peroxidases. Each cDNA appeared to hybridize to only one or two genes in S. humilis. mRNAs corresponding to Shpx2, Shpx6, and Shpx12 were expressed relatively abundantly in young leaves, with lesser expression of Shpx2 and Shpx6 and no expression of Shpx12 detected in roots. No expression of these genes was detected in stems or old leaves. The mRNA of Shpx5 was relatively abundant in stems and to a lesser extent in young leaves. However, infection of young leaves with C. gloeosporioides greatly increased expression of the mRNAs of Shpx2 and Shpx6 but not Shpx5 nor Shpx12 compared to mock-inoculated controls. The mRNA of Shpx6 was strongly induced by the pathogen 4 h postinoculation, a time which precedes fungal penetration, while Shpx2 was induced to higher levels than controls at 24 h after inoculation. The mRNAs of both Shpx2 and Shpx6 but not Shpx5 and Shpx12 were also induced by wounding.(ABSTRACT TRUNCATED AT 250 WORDS)

CgDN3: an essential pathogenicity gene of colletotrichum gloeosporioides necessary to avert a hypersensitive-like response in the host Stylosanthes guianensis.

A gene of Colletotrichum gloeosporioides that is induced by nitrogen starvation in axenic culture and is expressed at the early stages of infection of the host Stylosanthes guianensis has been identified and its role in pathogenicity tested. The sequence of this gene, named CgDN3, indicated that it encodes a protein of 74 amino acids that contains a predicted 18 amino acid signal sequence for secretion of a basic 54 amino acid mature protein with weak homology to an internal region of plant wall-associated receptor kinases. Mutants of C. gloeosporioides were produced by homologous recombination in which part of the coding sequence and promoter region of the CgDN3 gene was replaced with a hygromycin-resistance gene cassette. Mutations in the CgDN3 gene were confirmed in two independent transformants and Northern (RNA) analysis demonstrated the disrupted CgDN3 gene was not expressed. The mutants had faster mycelial growth rates in vitro but produced spores that germinated to form appressoria normally on the leaf surface. However, the CgDN3 mutants were unable to infect and reproduce on intact host leaves. Microscopic analysis revealed small clusters of necrotic host cells at inoculation sites on leaves, suggesting that these mutants elicited a localized, host hypersensitive-like response. The mutants were able to grow necrotrophically and reproduce on leaves when conidia were inoculated directly onto wound sites. The putative promoter region of the CgDN3 gene was fused to a gene encoding a modified jellyfish green fluorescent protein and introduced into the fungus. Following inoculation, strong expression of green fluorescent protein was observed in primary infection vesicles in infected epidermal cells with weaker expression evident in hyphae growing within infected leaf tissue. These findings indicate that CgDN3 encodes a novel pathogenicity determinant associated with the biotrophic phase of primary infection and required to avert a hypersensitive-like response by a compatible host.

Adrenocorticotropin-adrenal regulation of transported substance P in the vagus nerve of the rat.

The factors regulating substance P (SP) synthesis and quantity of transport in the sensory vagus nerve are unknown. To examine this issue rats were administered ACTH or corticosterone or subjected to adrenalectomy, and the quantity of peripherally directed transported SP was measured in those animals as an indication of neuropeptide synthesis. ACTH treatment (12 U/day, sc, for 14 days) resulted in significant adrenal hypertrophy and increased corticosterone levels. The 24-h accumulation of SP proximal to ligature in the cervical vagus was significantly reduced [mean net proximal segment content: controls, 529 +/- 42 (+/- SEM) pg/3 mm segment; ACTH, 282 +/- 44]. The content in the unligated nerve, one sixth or less than that proximal to ligature, was not different in the two groups. In a separate experiment, ACTH (6 U/day for 14 days) had no effect compared to controls, whereas 16 U/day reduced transported SP. The content in the unligated nerve was again not different in the two groups. In the same experiment, corticosterone (2.5 mg/100 g BW, sc, for 14 days) reduced the quantity of transported SP. Total protein content in proximal segments was reduced only in the corticosterone group and was identical in all groups in unligated nerve. Adrenalectomy modestly increased transport by 20% and contralateral unligated nerve content by a similar percentage. The quantity of transported somatostatin, another vagal neuropeptide partly derived from sensory cell bodies, was either increased or unaltered by the experimental manipulations. In summary, these studies demonstrate that the chronic administration of ACTH or corticosterone significantly decreases the quantity of peripherally transported SP in the sensory vagus nerve and, presumably, synthesis within the vagal sensory ganglia. Down-regulation of synthesized/transported neuropeptide suggests a mechanism by which the ACTH-adrenal axis, acting through visceral sensory nerves, may modulate autonomic or central nervous system vagally mediated reflex arcs.

Studies of the axoplasmic transport of somatostatin in the vagus nerve of the rat.

These studies were performed to examine the axoplasmic transport of somatostatin (SS) in the cervical vagus nerve of the rat. As a preliminary step, the immunoreactive SS (IR-SS) obtained from extracts of the vagal nodose ganglion and the vagus nerve was subjected to chromatographic analysis. On a Bio-Gel P-10 column, 92% of the nodose ganglion IR-SS and 98% of the vagal IR-SS coeluted with synthetic SS-14. The remaining immunoreactivity in both areas coeluted with synthetic SS-28. Vagal IR-SS demonstrated migratory characteristics identical to those of synthetic SS-14 on high performance liquid chromatography. A larger molecular weight form of IR-SS, which may correspond to prosomatostatin, was not identified in either site. When the vagus nerve was ligated distal to the nodose ganglion, the content of IR-SS increased in a time-dependent manner in the 3-mm segment of nerve proximal to the ligature. No increase in IR-SS was observed in an equal segment of nerve distal to the ligation or in the unligated contralateral nerve. Twenty-four hours after the ligation, the content of IR-SS (picograms per 3 mm; mean +/- SD) was: proximal segment, 33.9 +/- 9.6; distal segment, 3.4 +/- 3.0; and contralateral nerve, 1.7 +/- 0.7. The apparent transport velocity of IR-SS was estimated to be 2.1 +/- 1.5 mm/h. A variety of experimental approaches were used to characterize the mechanisms underlying the transport process and to define the anatomical sites of origin of the transported peptide. The application of colchicine to the vagus nerve resulted in an accumulation of IR-SS above the area which was not significantly different from that obtained after nerve ligation. When the vagus nerve was crushed above the nodose ganglion, the accumulation of IR-SS in the proximal segment was reduced by 50%, although the IR-SS content in the nodose ganglion and in the intervening nerve segments was unchanged by this procedure. The induction of a chemical sympathectomy with guanethidine had no effect on the accumulation of IR-SS. The administration of capsaicin during the neonatal period or in adult life had no effect on the transport of IR-SS, but greatly decreased the transport of substance P.(ABSTRACT TRUNCATED AT 400 WORDS)