Letteratura scientifica selezionata sul tema "Probate Court (Ross County)"
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Articoli di riviste sul tema "Probate Court (Ross County)"
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Tilden, Samuel J. "Ethical and Legal Aspects of Using an Identical Twin as a Skin Transplant Donor for a Severely Burned Minor". American Journal of Law & Medicine 31, n. 1 (marzo 2005): 87–116. http://dx.doi.org/10.1177/009885880503100103.
Testo completoAbstract (sommario):
On January 7, 2003, Sydney Cowan, a healthy six-year-old girl, underwent skin harvesting, specifically to be used for her badly burned identical twin sister, Jennifer. A day earlier, the Probate Court of Jefferson County, Alabama, after considering whether a healthy minor twin sibling could serve as a skin donor for her severely burned sister, authorized parental consent to the surgery. More accurately, the court addressed whether Sydney could undergo surgical procedures that provided her with no physical benefit, but, rather, resulted in harmful effects, such as acute postoperative pain, permanent residua, and potential long-term emotional and psychological dysfunction.Although the transplants were extraordinarily successful, and the newspaper article depicted Sydney's participation in heroic terms, the harvesting of Sydney's skin was ethically problematic. Specifically, I assert that the use of an incompetent minor as a skin transplant donor, even if an identical twin, is not justified unless the transplant will save the recipient's life.
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Howitt, Pamela S., e Eugene Arthur Moore. "Chapter 3: The Efficacy of Intensive Early Intervention An Evaluation of the Oakland County Probate Court Early Offender Program". Juvenile and Family Court Journal 42, n. 3 (agosto 1991): 25–36. http://dx.doi.org/10.1111/j.1755-6988.1991.tb00892.x.
Testo completo
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Ako, Andrew Ako, George Elambo Nkeng e Gloria Eyong Eneke Takem. "Water quality and occurrence of water-borne diseases in the Douala 4th District, Cameroon". Water Science and Technology 59, n. 12 (1 giugno 2009): 2321–29. http://dx.doi.org/10.2166/wst.2009.268.
Testo completoAbstract (sommario):
The monthly occurrence and mean age distribution of water-borne diseases in the Douala 4th District, Cameroon (1995–2006) were studied and probable causes of diseases spread were established. Diseases of interest included gastroenteritis, amoebic dysentery, typhoid fever and cholera. Water-borne disease occurrence was observed to follow a seasonal pattern with peaks occurring between the months of January and May followed by drops between June and October and rose again from November. Children below 5 years were found to be more vulnerable to diarrhoea, gastroenteritis, amoebic dysentery while persons between 15–44 years were more vulnerable to typhoid and cholera. Physico-chemically, water samples had turbidities varying between 5.5–86 NTU, pH values between 4.2 and 7.1 and zero residual chlorine. Bacteriological analysis showed that the total coliform count was averagely 74/100 ml, the faecal colform count was 43/100 ml and the faecal streptococci count was 27/100 ml. Lack of access to potable water, absence of sanitation facilities and environmental factors could be advanced as the probable causes of water-borne disease spread.
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Morton, Jordan M., Evaren E. Page, Nathan W. Liles e James N. George. "Thrombotic Microangiopathy (TMA) and Other Severe Systemic Reactions Caused By Quinine-Containing Beverages: Report of a Patient and Review of Published Reports". Blood 126, n. 23 (3 dicembre 2015): 4651. http://dx.doi.org/10.1182/blood.v126.23.4651.4651.
Testo completoAbstract (sommario):
Abstract Case presentation: A 35 year old white female was in excellent health until the sudden onset of fever, chills, headache, myalgias, abdominal and flank pain, nausea and vomiting while driving home from a banquet. She went to an ER the following day; her hematocrit was 39% and platelet count 132,000/µL. She was given IV fluids, antiemetics, and sent home. Two days later, she returned to the ER as her symptoms persisted and she had not urinated since they began; her hematocrit was 32%, platelet count 54,000/µL, creatinine 9.3 mg/dL, AST 350, ALT 274, LDH 2402. Due to anemia, thrombocytopenia, and acute kidney injury (AKI), plasma exchange (PEX) was begun because of suspicion of thrombotic thrombocytopenic purpura; Shiga toxin-induced hemolytic uremic syndrome was also considered. Hemodialysis was begun as her creatinine rose to 10.5 mg/dL and anuria persisted. We saw her the following day; drug-induced TMA was considered to be the most likely etiology because of the very sudden, severe onset with anuric AKI. However her only medicine was Singulair for asthma, which she had taken daily for several years. She had never taken quinine tablets or drunk gin and tonic; then she asked us, "What about vodka and tonic?". She had had "a sip" of vodka and tonic at the banquet; it was a favorite drink. Her previous vodka and tonic was 16 months previously; she had then developed sudden severe headache, neck pain, fever and chills causing suspicion of meningitis. Head CT scan and spinal fluid were normal; blood counts remained normal; creatinine increased transiently to 1.7; an association with vodka and tonic was not suspected. This history confirmed the quinine etiology of her TMA. Subsequently she was demonstrated to have quinine-dependent antibodies reactive with platelets and neutrophils (BloodCenter of Wisconsin). Genetic sequencing (U of Iowa) revealed no mutations associated with complement-mediated TMA. PEX was continued until her platelet count was normal (12 days); hemodialysis was required for 2 months. Five years after TMA, she continues to have hypertension, fatigue and arthralgias. Her creatinine clearance is 54 ml/min/1.73m2, indicating Stage 3A chronic kidney disease. Systematic literature review: A search of 12 databases for reports of adverse reactions to quinine identified 118 patients with definite or probable evidence for a causal association of quinine with idiosyncratic adverse reactions. Most quinine reactions were related to ingestion of tablets, but 24 patients had adverse reactions caused by quinine-containing beverages. Three patients had TMA, similar to our patient; 2 of these 3 patients also had disseminated intravascular coagulation (DIC); one also had liver function abnormalities. Seven patients had isolated severe thrombocytopenia, 1 patient had granulomatous hepatitis, 1 patient had retina toxicity with vision loss; 1 patient had anaphylaxis, and 11 patients had cutaneous reactions (Table). Many patients had multiple recurrent episodes before the quinine etiology was discovered. All patients recovered, but no reports described long-term follow-up. Conclusion: Our patient's experience and our review published reports highlight the potential for severe, systemic adverse reactions caused by quinine-containing beverages. The reactions may involve multiple organ systems. Long-term outcomes are not described in published reports, but our patient has chronic kidney disease 5 years following her acute TMA. The small amounts of quinine in beverages (typically 80 mg/l) are sufficient to cause severe immunologic adverse reactions. Quinine is also added to cocktails to create fluorescence, then described as "Shocktails". A thorough history for exposure to quinine is critical. 24 previously published patients with acute quinine-induced reactions Table. Patients Adverse Events 1 TMA (3 episodes), also DIC 1 TMA (6 episodes), also fever, chills, neutropenia, DIC, liver function abnormalities 1 TMA (1 episode) 7 Thrombocytopenia (3 patients, multiple episodes), nadir platelet count (median) 2000/µL 1 Granulomatous hepatitis, neutropenia (multiple episodes) 1 Retinal toxicity, vision loss 1 Anaphylactic shock (2 episodes) 11 Dermatologic disorders: fixed drug eruptions (7), toxic epidermal necrolysis (1), urticaria (1), photosensitivity, erythematous lesions (1), erythematosus rash (1) Disclosures No relevant conflicts of interest to declare.
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Dutta, Anurag, Liton Chandra Voumik, Athilingam Ramamoorthy, Samrat Ray e Asif Raihan. "Predicting Cryptocurrency Fraud Using ChaosNet: The Ethereum Manifestation". Journal of Risk and Financial Management 16, n. 4 (29 marzo 2023): 216. http://dx.doi.org/10.3390/jrfm16040216.
Testo completoAbstract (sommario):
Cryptocurrencies are in high demand now due to their volatile and untraceable nature. Bitcoin, Ethereum, and Dogecoin are just a few examples. This research seeks to identify deception and probable fraud in Ethereum transactional processes. We have developed this capability via ChaosNet, an Artificial Neural Network constructed using Generalized Luröth Series maps. Chaos has been objectively discovered in the brain at many spatiotemporal scales. Several synthetic neuronal simulations, including the Hindmarsh–Rose model, possess chaos, and individual brain neurons are known to display chaotic bursting phenomena. Although chaos is included in several Artificial Neural Networks (ANNs), for instance, in Recursively Generating Neural Networks, no ANNs exist for classical tasks entirely made up of chaoticity. ChaosNet uses the chaotic GLS neurons’ property of topological transitivity to perform classification problems on pools of data with cutting-edge performance, lowering the necessary training sample count. This synthetic neural network can perform categorization tasks by gathering a definite amount of training data. ChaosNet utilizes some of the best traits of networks composed of biological neurons, which derive from the strong chaotic activity of individual neurons, to solve complex classification tasks on par with or better than standard Artificial Neural Networks. It has been shown to require much fewer training samples. This ability of ChaosNet has been well exploited for the objective of our research. Further, in this article, ChaosNet has been integrated with several well-known ML algorithms to cater to the purposes of this study. The results obtained are better than the generic results.
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Waldstein, Sasha, Sarah E. Arnold-Croop, Laura Carrel e M. Elaine Eyster. "Dehydrated Hereditary Stomatocytosis in a Multi-Generational American Family with a KCNN4 Gardos Channel Mutation". Blood 136, Supplement 1 (5 novembre 2020): 11. http://dx.doi.org/10.1182/blood-2020-137481.
Testo completoAbstract (sommario):
INTRODUCTION: Dehydrated hereditary stomatocytosis (DHSt) is an autosomal dominant red blood cell membrane disorder characterized by hemolytic anemia and splenomegaly. DHSt has an estimated incidence of 1:50,000 births, and the degree of anemia varies within and between families. Although transfusion support during childhood is not uncommon, continued requirement into adulthood is rare. The most frequent cause of DHSt is a gain-of-function mutation of the PIEZO1 gene, leading to delayed channel inactivation that results in a monovalent cation leak and an increase in intracellular calcium (Ca2+). Many of these patients develop recurrent thromboses post splenectomy. Other DHSt patients have mutations in KCNN4, which encodes the Gardos channel, with mutations causing increased Ca2+ sensitivity and potassium efflux. To our knowledge, 42 patients from ten families have been described with four distinct KCNN4 mutations: Arg352His, Val282Met or Val282Glu, and a 28bp deletion encompassing the exon-intron 7 junction. We report herein the eighth family with the Arg352His locus mutation. CASE REPORT: Five subjects from a single family were enrolled in this study (affected proband, unaffected husband, two affected children, and an unaffected grandchild). The proband has had hemolytic anemia since childhood. She had undergone splenectomy at age three and cholecystectomy at eight. Her anemia persisted, with a mean hemoglobin (Hgb) of 10.5g/dL and a reticulocyte count (retic ct) of 12.6%, with no need for transfusion support or iron chelation therapy (Table 1). Extensive testing revealed slightly decreased osmotic fragility and mildly elevated intracellular sodium concentration, of 19.4 mEq/L. Her daughter has mild splenomegaly and anemia, with a mean Hgb of 11.1g/dL and retic ct of 6.7%, whereas her son has more severe disease, with a mean Hgb of 9.3g/dL and a retic ct of >22%. He underwent splenectomy as a teenager for immune thrombocytopenia, and has required chelation therapy with deferasirox since age 31, when his ferritin rose to >1000ng/mL. Similar to results reported by others, splenectomy did not alter the severity of hemolysis in either the proband or her son, and neither developed thrombotic complications 57 and 7 years post splenectomy, respectively. The proband's daughter has a child with a normal Hgb, and is presumed unaffected. Stomatocytes were rarely seen on the peripheral blood smears of the proband and her children. METHODS: A clinically available 39 gene hemolytic anemia panel on the proband failed to identify the mutation underlying this disorder. We therefore performed whole exome sequencing on all five family members. We prioritized the analysis of 23 additional genes that are included in hemolytic anemia panels from two other reference laboratories and are involved in disorders of red blood cell membrane or cytoskeletal proteins of potential clinical relevance to the study population. RESULTS: A single missense mutation, Arg352His within KCNN4, was identified in all three affected individuals. This heterozygous mutation was present in the proband and her affected two children, and absent in her unaffected grandchild and husband. CONCLUSIONS: To our knowledge, this Pennsylvania family is only the eleventh described to have DHSt secondary to a KCNN4 mutation. This disorder is likely much more prevalent than reported, due to the rarity of stomatocytes on peripheral blood smears, the omission of the KCNN4 gene from hemolytic panels offered by some reference laboratories, and variable clinical presentation. KCNN4 mutations should be investigated if other causes are not identified in patients with lifelong hemolytic anemia suspected of having a red cell membrane protein or cytoskeletal disorder. Appropriate diagnosis may allow severely affected patients to be considered for treatment with the experimental Gardos channel inhibitor senicapoc. Additionally, mutational diagnosis is especially important when considering the adverse outcomes post splenectomy in PIEZO1 as compared to KCNN4 mutations. Disclosures Eyster: SPARK:Research Funding;Sanofi:Research Funding;Novo Nordisk:Research Funding;Baxalta/Shire:Research Funding.
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"The Sourcebook of county court records: a national guide to civil, criminal, and probate records at the county and municipal levels within the state court systems". Choice Reviews Online 35, n. 01 (1 settembre 1997): 35–0010. http://dx.doi.org/10.5860/choice.35-0010.
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Chan, Antonia, Yuhsin Huang, Maja Ivanovic, Maja Ivanovic, Sarah Waliany, Ryan Batchelder, Joel Neal et al. "Abstract 16450: Increased Monocyte Count and Neutrophil-to-Lymphocyte Ratio With ICI Myocarditis". Circulation 148, Suppl_1 (7 novembre 2023). http://dx.doi.org/10.1161/circ.148.suppl_1.16450.
Testo completoAbstract (sommario):
Introduction: Diagnosing immune checkpoint inhibitor (ICI) myocarditis remains complex in practice, often requiring advanced imaging or invasive endomyocardial biopsy. We constructed an institutional cohort of patients who received at least 1 dose of ICI and developed probable or definite ICI myocarditis. Hypothesis: We hypothesized that routine serum biomarkers, including troponin I, neutrophil-to-lymphocyte ratio (NLR), monocytes, and AST/ALT, would be elevated and peak at diagnosis. Methods: This retrospective cohort study used EHR data from patients on ICI at a single academic institution who developed probable or definite myocarditis from 7/2015 - 1/2023. Inclusion criteria and diagnosis date were defined using Bonaca criteria and/or documented clinical consensus, e.g. initiation of steroids for presumed myocarditis. Primary outcomes were levels of troponin I/high sensitivity troponin I, monocytes, NLR, and AST/ALT. LOESS regression with default smoothing parameter α = 0.75 was used to evaluate biomarker patterns 7 days pre- and post- diagnosis. Results: We included 16 cases of myocarditis. At diagnosis, most patients had at least 1 significant elevation in troponin I (100%), AST (81%), ALT (63%), or monocytes (69%) while on ICI (Figure 1). After diagnosis, absolute monocyte and lymphocyte count initially decreased before rebounding. Serum NLR rose prior to and briefly after diagnosis (Figure 2). Conclusions: Peripheral cardiac and non-cardiac biomarkers change with ICI myocarditis. Further research on routine non-cardiac biomarkers associated with myocarditis and other ICI toxicities may help prevent high-grade adverse events.
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Boudaghpour, Siamak, e Saeideh Sabooteh. "ENVIRONMENTAL LEAKAGE POLLUTIONS EVALUATIONS IN URBAN WATER DISTRIBUTION NETWORK USING UNACCOUNTED WATER PRINCIPLES (SHOKUHIEH INDUSTRIAL TOWN IN IRAN)". Stavební obzor - Civil Engineering Journal 29, n. 2 (31 luglio 2020). http://dx.doi.org/10.14311/cej.2020.01.0019.
Testo completoAbstract (sommario):
Consideration of non-revenue water in urban water distribution networks is of special importance in planning urban water distribution network and in water consumption. Water loss, especially in the distribution networks, forms a large part of the unaccounted for water (UFW). Considering the serious crisis in water resources, and taking into account the existing limitations of supplying hygienic drinking water, increasing attempts must be made at reducing the volume of water loss. The present study evaluated the environmental effects of pollution leakage from broken pipes into the water distribution network in the Shokuhieh. General principles and concepts of UFW, apparent loss was reviewed. Based on these concepts, probable pipe break points, and the resulting contaminants entering the network were considered. A software model of the water distribution network in the study area was then used to investigate qualitative and quantitative results related to intrusion of contaminants into the water distribution network and diffusion of pollutants into the network. Results of the present study and physicochemical tests performed on water revealed that BOD and COD concentrations increased from 5-10 to 10-20 mg/L and the coliform count rose from zero to 20 MPN as a result of pipe breaks in the distribution network.
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Obinze, Sandra, Philippa C. Ojimelukwe e Ben A. Eke. "Box fermentation and solar drying improve the nutrient composition and organoleptic quality of chocolate from cocoa beans". Frontiers in Sustainable Food Systems 6 (22 novembre 2022). http://dx.doi.org/10.3389/fsufs.2022.1023123.
Testo completoAbstract (sommario):
Chocolate is a well-liked and popular food product made from the cocoa bean. The objective of this research was to evaluate the effects of box fermentation and solar drying of cocoa bean on chocolate quality. Fermentation was carried out in a perforated wooden box for 168 h with periodic turning after every 48 h. The succession of microorganisms during fermentation and total microbial count were monitored. Both the fermented and unfermented samples were solar dried. During drying the change in weight of the beans, amount of solar radiation, and ambient wind speed of the atmosphere were measured. The approximate nutrient (crude protein, crude fat, total ash, crude fiber, and carbohydrate) and phytochemical (phenol) contents of the dried cocoa beans were evaluated. Sensory properties of chocolate, produced from the two samples (box fermented and dried as well as unfermented and dried) were compared. The initial temperature of the fermenting cocoa mash was 30°C. It rose to 46°C by the 96th h of fermentation and sharply declined to 38°C by the 120th h. Candida, Pseudomonas and Staphylococcus spp were probable organisms identified with the fermenting mass at the initial stage of the fermentation. Lactic acid bacteria dominated after 72 h. Bacillus spp was also observed until the 168th h. Solar drying of unfermented bean lasted for 4 days, while that of fermented bean lasted for 3 days. Fermentation increased the crude protein, total ash and carbohydrate contents of the cocoa beans. It also improved the appearance, and aroma of chocolate from the cocoa beans.
Tesi sul tema "Probate Court (Ross County)"
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Dabner, Carol Patrice. "Legal Compliance in Guardianship Cases An Exploratory Study: Investigating Denton County Probate Court Visitors' Program Success with Legal Compliance in Guardianship Cases in 2013". Thesis, University of North Texas, 2016. https://digital.library.unt.edu/ark:/67531/metadc955079/.
Testo completoAbstract (sommario):
Dabner, Carol P. Legal Compliance in Guardianship Cases. An Exploratory Study: Investigating Denton County Probate Court Visitors' Program Success with Legal Compliance in Guardianship Cases. Doctor of Philosophy (Applied Gerontology), December 2016, 140 pp., 18 tables, references, 20 titles.
Aim: To evaluate the effectiveness of the legal compliance of the Denton County Probate Court Visitor's program in the year 2013.
Rationale: Guardianship case management success is based on the presence of legal compliance of both guardians and the Court. When a guardian is legally compliant, a ward is receiving the statutorily minimum standards of care. Legal compliance equates (evidence of) the Ward receiving legally sufficient care. Research has not been vast; it has been consistent as to necessity of guardianship training, monitoring, and narrow focus of research. Evidence based research will assist in defining and developing appropriate court monitoring programs, which can add to the quality of care for elderly and disabled adults.
Methods: 1,300 guardianship cases in the probate court. Of these cases, 910 had annual reports of the person filed, which 304 were reviewed using the Legal Compliance Audit. Eight (8) factors of compliance were reviewed with three (3) being Court actions and five (5) being guardian actions.
Results: Exploratory study provides evidence based research of the necessary changes to develop the Denton County Probate Court Visitor's program. The guardians are more legal compliant than the Court.
Libri sul tema "Probate Court (Ross County)"
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Janet, Ackley, Ohio. Probate Court (Ross County) e Ross County Genealogical Society, a cura di. Abstracts from the Mothers' pension records, Ross County, Ohio Probate Court: December 9, 1915 to June 13, 1936, volumes 1, 2, & 3. Chillicothe, Ohio]: [Ross County Genealogical Society], 2009.
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Ohio Genealogical Society. Lorain County Chapter. Lorain County Probate Court deaths. Elyria, Ohio: Lorain County Chapter of the Ohio Genealogical Society, 2006.
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Ohio Genealogical Society. Lorain County Chapter. Lorain County Probate Court births. Elyria, Ohio: Lorain County Chapter of the Ohio Genealogical Society, 2005.
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Pickaway County Historical Society (Ohio). Pickaway County probate court index. Circleville, Ohio: Pickaway County Historical Society, 2008.
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County), Missouri Probate Court (Butler. Butler County, Missouri, Probate Court minutes. Broseley, MO: T. Glass, 1988.
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Ohio Genealogical Society. Auglaize County Chapter. Auglaize County, Ohio probate court birth records. Wapakoneta, Ohio: The Chapter, 1993.
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Ohio Genealogical Society. Mercer County Chapter. Probate court death records, Mercer County, Ohio. Celina, Ohio: Ohio Genealogical Society, Mercer County Chapter, 1998.
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Ohio Genealogical Society. Mercer County Chapter. Probate court birth records, Mercer County, Ohio. Celina, Ohio: The Mercer County Chapter of the Ohio Genealogical Society, 2001.
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Ohio Genealogical Society. Mercer County Chapter., a cura di. Probate court death records: Mercer County, Ohio. Celina, Ohio (P.O. Box 437, Celina 45822-0437): The Society, 1998.
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Ohio Genealogical Society. Perry County Chapter., a cura di. Perry County, Ohio Probate Court birth records. [Junction City, Ohio]: Perry County Chapter of the Ohio Genealogical Society, 2000.
Cerca il testo completoCapitoli di libri sul tema "Probate Court (Ross County)"
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Davies, Mark R. "Wills And Probate". In Solicitors’ Negligence And Liability, 353–76. Oxford University PressOxford, 2008. http://dx.doi.org/10.1093/oso/9780199284399.003.0014.
Testo completoAbstract (sommario):
Abstract The statistical picture regarding solicitors’ negligence in the area of wills and probate is less marked than the other important private client area, conveyancing. The pertinent case law relating to wills and probate also has a relatively short history. The reason for this is that if a solicitor is negligent in preparing a will, the error should usually be cheap and simple to remedy if discovered during the prospective testator’s lifetime. If not discovered until after the testator’s death, the potential claimants, often intended beneficiaries, are third parties who lack a contractual relationship with the solicitor. Only with the decision in Ross v Caunters, did the courts fully begin to address the capacity of such third parties to bring actions in tort.
Atti di convegni sul tema "Probate Court (Ross County)"
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Tashima, Tsuguhisa, Takashi Sasaki, Anestis I. Kalfas e Reza S. Abhari. "Blade Loading Influence on Unsteady Flow Interactions in Axial Steam Turbines". In ASME Turbo Expo 2007: Power for Land, Sea, and Air. ASMEDC, 2007. http://dx.doi.org/10.1115/gt2007-27452.
Testo completoAbstract (sommario):
This paper presents the results of a comparative experimental investigation of two turbine test cases with different second stage stator count. The objective is to investigate the possibility of reducing the product cost and weights by reducing stator count while improving overall turbine performance. The experiment was performed in a two-stage axial research turbine facility. The blade geometries under investigations are of typical of the high-pressure section of steam-turbines and the blade rows except the second stage stator are identical between the two test cases. The low solidity stator has 3/4 blades count of the high solidity stator while keeping the same axial chord length. Unsteady and steady data were obtained using the fast response probe and the miniature pneumatic probes at the exit of the first rotor, at the exit of the second stator and the second rotor. The prime interest is focused on the way that performance is changed by the different number of wakes and vortices transporting through the downstream blade rows. In the reduced blade count case, the unsteady flow interactions causes the larger distributions of all flow properties measured. A marked difference is observed near 10 to 35% span where the hub passage vortex is dominant. In the low solidity stator case, the time dependent vorticity variation was found to be 40% larger compared to the high solidity stator case. In correspondence to the vorticity variation, the loss coefficient also varies depending on time. Pneumatic measurements show that the difference of second rotor loss coefficient between the cases is within 1.5% except the tip area. However, unsteady loss variation differs between the test cases depending on the change of the flow structures. The mechanism of this unsteady loss variation is analyzed in detail.
Rapporti di organizzazioni sul tema "Probate Court (Ross County)"
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Horwitz, Benjamin A., e Barbara Gillian Turgeon. Fungal Iron Acquisition, Oxidative Stress and Virulence in the Cochliobolus-maize Interaction. United States Department of Agriculture, marzo 2012. http://dx.doi.org/10.32747/2012.7709885.bard.
Testo completoAbstract (sommario):
Our project focused on genes for high affinity iron acquisition in Cochliobolus heterostrophus, a necrotrophic pathogen of maize, and their intertwined relationship to oxidative stress status and virulence of the fungus on the host. An intriguing question was why mutants lacking the nonribosomal peptide synthetase (NRPS) gene (NPS6) responsible for synthesis of the extracellular siderophore, coprogen, are sensitive to oxidative stress. Our overall objective was to understand the mechanistic connection between iron stress and oxidative stress as related to virulence of a plant pathogen to its host. The first objective was to examine the interface where small molecule peptide and reactive oxygen species (ROS) mechanisms overlap. The second objective was to determine if the molecular explanation for common function is common signal transduction pathways. These pathways, built around sensor kinases, response regulators, and transcription factors may link sequestering of iron, production of antioxidants, resistance to oxidative stress, and virulence. We tested these hypotheses by genetic manipulation of the pathogen, virulence assays on the host plant, and by following the expression of key fungal genes. An addition to the original program, made in the first year, was to develop, for fungi, a genetically encoded indicator of redox state based on the commercially available Gfp-based probe pHyper, designed for animal cell biology. We implemented several tools including a genetically encoded indicator of redox state, a procedure to grow iron-depleted plants, and constructed a number of new mutants in regulatory genes. Lack of the major Fe acquisition pathways results in an almost completely avirulent phenotype, showing how critical Fe acquisition is for the pathogen to cause disease. Mutants in conserved signaling pathways have normal ability to regulate NPS6 in response to Fe levels, as do mutants in Lae1 and Vel1, two master regulators of gene expression. Vel1 mutants are sensitive to oxidative stress, and the reason may be underexpression of a catalase gene. In nps6 mutants, CAT3 is also underexpressed, perhaps explaining the sensitivity to oxidative stress. We constructed a deletion mutant for the Fe sensor-regulator SreA and found that it is required for down regulation of NPS6 under Fe-replete conditions. Lack of SreA, though, did not make the fungus over-sensitive to ROS, though the mutant had a slow growth rate. This suggests that overproduction of siderophore under Fe-replete conditions is not very damaging. On the other hand, increasing Fe levels protected nps6 mutants from inhibition by ROS, implying that Fe-catalyzed Fenton reactions are not the main factor in its sensitivity to ROS. We have made some progress in understanding why siderophore mutants are sensitive to oxidative stress, and in doing so, defined some novel regulatory relationships. Catalase genes, which are not directly related to siderophore biosynthesis, are underexpressed in nps6 mutants, suggesting that the siderophore product (with or without bound Fe) may act as a signal. Siderophores, therefore, could be a target for intervention in the field, either by supplying an incorrect signal or blocking a signal normally provided during infection. We already know that nps6 mutants cause smaller lesions and have difficulty establishing invasive growth in the host. Lae1 and Vel1 are the first factors shown to regulate both super virulence conferred by T-toxin, and basic pathogenicity, due to unknown factors. The mutants are also altered in oxidative stress responses, key to success in the infection court, asexual and sexual development, essential for fungal dissemination in the field, aerial hyphal growth, and pigment biosynthesis, essential for survival in the field. Mutants in genes encoding NADPH oxidase (Nox) are compromised in development and virulence. Indeed the triple mutant, which should lack all Nox activity, was nearly avirulent. Again, gene expression experiments provided us with initial evidence that superoxide produced by the fungus may be most important as a signal. Blocking oxidant production by the pathogen may be a way to protect the plant host, in interactions with necrotrophs such as C. heterostrophus which seem to thrive in an oxidant environment.