Regulation des Kpp2/Kpp6-MAPK-Moduls in Ustilago maydis durch Phosphatasen

Der Lebenszyklus von Ustilago maydis, dem Erreger des Maisbeulenbrandes, beginnt mit der Ausbildung von Konjugationshyphen und der anschließenden Fusion zweier haploider Sporidien. Die Infektion der Wirtspflanze erfolgt daraufhin nach Ausbildung des Dikaryons durch Appressorien. Diese Entwicklungsprozesse werden durch die beiden Paarungstyploci a und b kontrolliert. Während der bialellische a-Locus Zell-Zellerkennung und Zellfusion reguliert, werden die weiteren Entwicklungsschritte durch den multiallelischen b-Locus kontrolliert. Die Regulation der Expression beider Loci sowie des morphologischen Wechsels erfolgt, neben einem cAMP-Signalweg, über ein konserviertes MAPK-Modul, bestehend aus der MAPKKK Kpp4, der MAPKK Fuz7 und den MAPK Kpp2 und Crk1. Die Aktivierung des MAPK-Moduls erfolgt, nach der Perzeption kompatiblen Pheromons durch anschließende hierarchische Phosphorylierung am konservierten TEY-Motiv. Während in anderen Organismen dual spezifische Protein-Tyrosin-Phosphatasen beschrieben wurden, welche die MAPK durch die Dephosphorylierung des TEY-Motivs negativ regulieren, waren solche Phosphatasen in U. maydis bisher nicht bekannt. In dieser Arbeit wurden mit Rok1 und Mkp2 zwei potentielle Msg5p-homologe dual spezifische Protein-Tyrosin-Phosphatasen identifiziert. rok1 zeigt eine Pheromon-abhängig und mkp2 eine cAMP-abhängig induzierte Expression. Während solopathogene mkp2- Deletionsstämme eine reduzierte Filamentation und eine leicht erniedrigte Virulenz zeigten führte die rok1-Deletion zu einem gegensätzlichen Phänotyp. rok1-Deletionsmutanten zeigen Filamentation, unabhängig von externen Stimuli und kompatiblen b-Loci sowie eine verstärkte transkriptionelle Pheromonantwort. Dementsprechend führt die Überexpression von rok1 zur Inhibition der transkriptionellen und morphologischen Pheromonantwort und verhindert so die Paarung kompatibler Sporidien. Des Weiteren zeigen rok1-Deletionsmutanten eine drastisch erhöhte Virulenz, welche durch vermehrte Appressorienbildung und die effizientere Kolonisierung der Pflanze zu erklären ist. In Wildtyp-Stämmen wird eine effiziente Appressorienbildung nur durch die Kombination der beiden Stimuli „hydrophobe Oberfläche“ und „Fettsäure“ induziert, wohingegen für rok1-Deletionstämme eine hydrophobe Oberfläche ausreicht die Appressorienbildung effizient, sogar über das Maß der maximalen Induktion im Wildtypstamm hinaus, zu induzieren. Somit reguliert Rok1 nicht nur die Pheromonantwort sondern auch die Induktion der Appressorienbildung. Epistasis-Analysen ordnen Rok1 unterhalb der MAPK-Kinase Fuz7 ein. Weiterhin ergaben phosphospezifische Westernanalysen, die Repression der Phosphorylierungsmagnitude der MAP-Kinasen Kpp2 und Kpp6 nach Pheromon-Stimulation durch rok1. Die Repression der MAPKs durch Rok1 spiegelt sich ebenfalls in der Suppression pathogenitätsassoziierter Phänotypen beider MAPKEinzeldeletionsmutanten durch Deletion von rok1 wieder. Somit ist es in der vorliegenden Arbeit erstmals gelungen eine Protein-Phosphatase als negativen Regulator des Kpp2/Kpp6-MAPK-Moduls in U. maydis zu identifizieren und zu charakterisieren. Aufgrund der Eigenschaften der Phosphatase führt die Deletion von rok1 zur genetischen Aktivierung des MAPK-Modul und bietet somit erstmals die Möglichkeit dessen Rolle auch während der biotrophen Entwicklung von U. maydis weiter zu untersuchen

Application of openBIM for the Management of Existing Railway Infrastructure: Case Study of the Cancello - Benevento Railway Line

In the field of infrastructure, the development and application of the openBIM (open Building Information Modeling) approach and related standards (principally Industry Foundation Classes) remain limited with regard to processes in O&M (Operation and Maintenance) phases, as well as the broader context of AM (Asset Management). This article deals with the activities carried out as part of a pilot project based on the need to manage the operation and assess the condition and value of existing infrastructure along the Cancello–Benevento railway line. The principal goal was to systematize information by digitalizing the infrastructure, in order to enable the assessment of possible performance gaps (compared to national railway standards) in the event of integration within the national infrastructure. In compliance with the project requirements, a digitalization strategy was designed for the definition of surveying activities and the implementation of openBIM systems for the development of an object library and a federated digital model, structured within the collaborative platform that was used, and allowing management, maintenance, and subsequent financial evaluation in the broader context of asset management. The project involved the collaboration of railway operators, a university, and a software company that implemented innovative concepts concerning IFC (specifically, IFC4x2 was used) through the development of dedicated software solutions. The digital solution we proposed enabled the use of digital models as access keys to survey and maintenance information (ERP platforms used by the railway operators) that was available in real time. This project was nominated at the buildingSMART awards 2021 and was one of three finalists in the “Asset Management Using openBIM” category

Pretreatment with verapamil in patients with persistent or chronic atrial fibrillation who underwent electrical cardioversion

AbstractOBJECTIVESTo evaluate, in a prospective and randomized fashion, the efficacy of a pretreatment with verapamil (V) in reducing recurrences of atrial fibrillation (AF) after electrical cardioversion (C).BACKGROUNDThe increased vulnerability for AF recurrence is probably due to AF-induced changes in the electrophysiologic properties of the atria. This electrical remodeling seems to be due to intracellular calcium overload.METHODSOne hundred seven patients with persistent or chronic AF underwent external and/or internal C. All patients received oral propafenone (P) (900 mg/day) three days before and during the entire period of follow-up (three months). In the first group, patients received only the P. In the second group, in adjunct to P, oral V (240 mg/day) was initiated three days before C and continued during the follow-up. Finally, in the third group, oral V was administered three days before and continued only for three days after electrical C.RESULTSDuring the three months of follow-up, 23 patients (23.7%) had AF recurrence. Mantel-Haenszel cumulative chi-square reached a significant level only when comparing AF free survival curves of group I versus group II and group III (chi-square = 5.2 and 4, respectively; p < 0.05). Significantly, 15 (65.2%) AF relapses occurred during the first week after cardioversion with a higher incidence in group I (10/33 patients, 30.3%) than group II (2/34 patients, 5.9%; p = 0.01) and group III (3/30 patients, 10%; p = 0.04).CONCLUSIONSSix days of oral V administration centered on the C day, combined with P, significantly reduce the incidence of early recurrences of AF compared with P alone

Functional impairment is associated with an increased risk of mortality in patients on chronic hemodialysis

Background: Functional impairment is associated with adverse outcomes in older people, as well as in patients on chronic hemodialysis. The aim of the present study was to determine the characteristics associated with functional impairment in chronic hemodialysis, and to evaluate if functional impairment represents a risk factor for reduced survival in chronic hemodialysis. Methods: All 132 chronic hemodialysis referring to the Hemodialysis Service of the Catholic University, Rome, Italy between November 2007 and May 2015 were included. All patients underwent comprehensive geriatric assessment; functional ability was estimated using two questionnaires exploring independency in bathing, dressing, toileting, transferring, continence, feeding (ADLs), and independency in using the telephone, shopping, food preparation, housekeeping, laundering, traveling, taking medications, and handling finances (IADLs). Functional impairment was diagnosed in presence of dependence in one or more ADLs/IADLs. Mood was assessed using the 30-item Geriatric Depression Scale. Logistic regression was used to evaluate factors associated with functional impairment. The association between functional impairment and survival was assessed by Cox regression. Results: ADLs impairment was present in 34 (26 %) participants, while IADLs impairment was detected in 64 (48 %) subjects. After a follow up of 90 months, 55 (42 %) patients died. In logistic regression, depressive symptoms were associated with ADLs and IADLs impairment (OR 1.12; 95 % CI = 1.02-1.23; OR 1.16; 95 % CI = 1.02-1.33; respectively). In Cox regression, ADLs impairment was associated with mortality (HR 2.47; 95 % CI-1.07-5.67) while IADLs impairment was not associated with reduced survival (HR .80; 95 % CI-.36-1.76). Conclusions: Functional impairment is associated with depressive symptoms; also, impairment in the ADLs represents a risk factor of reduced survival in chronic hemodialysis. These associations and their potential implication should be assessed in dedicated studies

Subclinical atherosclerosis is linked to small intestinal bacterial overgrowth via vitamin K2-dependent mechanisms

Aim To assess the rate of matrix Gla-protein carboxylation in patients with small intestinal bacterial overgrowth (SIBO) and to decipher its association with subclinical atherosclerosis. METHODS Patients with suspected SIBO who presented with a low risk for cardiovascular disease and showed no evidence of atherosclerotic plaques were included in the study. A glucose breath test was performed in order to confirm the diagnosis of SIBO and vascular assessment was carried out by ultrasound examination. Plasma levels of the inactive form of MGP (dephosphorylateduncarboxylated matrix Gla-protein) were quantified by ELISA and vitamin K2 intake was estimated using a food frequency questionnaire. RESULTS Thirty-nine patients were included in the study. SIBO was confirmed in 12/39 (30.8%) patients who also presented with a higher concentration of dephosphorylated-uncarboxylated matrix Gla-protein (9.5 \u3bc g/L vs 4.2 \u3bc g/L; P = 0.004). Arterial stiffness was elevated in the SIBO group (pulse-wave velocity 10.25 m/s vs 7.68 m/s; P = 0.002) and this phenomenon was observed to correlate linearly with the levels of dephosphorylated-uncarboxylated matrix Gla-protein ( f = 0.220, R 2 = 0.366, P = 0.03). Carotid intima-media thickness and arterial calcifications were not observed to be significantly elevated as compared to controls. CONCLUSION SIBO is associated with reduced matrix Gla-protein activation as well as arterial stiffening. Both these observations are regarded as important indicators of subclinical atherosclerosis. Hence, screening for SIBO, intestinal decontamination and supplementation with vitamin K2 has the potential to be incorporated into clinical practice as additional preventive measures

Long-Lasting Efficacy of Radio Electric Asymmetric Conveyer Neuromodulation Treatment on Functional Dysmetria, an Adaptive Motor Behavior

BackgroundFluctuating asymmetry (FA) is widely defined as the deviation from perfect bilateral symmetry and is considered an epigenetic measure of environmental stress. Rinaldi and Fontani hypothesized that the FA morpho-functional changes originate from an adaptive motor behavior determined by functional alterations in the cerebellum and neural circuits, not caused by a lesion, but induced by environmental stress. They called this phenomenon functional dysmetria (FD). On this premise, they developed the radio electric asymmetric conveyer (REAC) technology, a neuromodulation technology aimed at optimizing the best neuro-psycho-motor strategies in relation to environmental interaction.AimsPrevious studies showed that specific REAC neuro postural optimization (NPO) treatment can induce stable FD recovery. This study aimed to verify the duration of the NPO effect in inducing the stable FD recovery over timeMaterials and methodsData were retrospectively collected from a population of 29,794 subjects who underwent a specific semiological FD assessment and received the NPO treatment, regardless of the pathology referred.ResultsThe analysis of the data collected by the various participants in the study led us to ascertain the disappearance of FD in 100% of the cases treated, with a stability of the result detected up to 18 years after the single administration of the REAC NPO treatment.ConclusionsThe REAC NPO neurobiological modulation treatment consisting of a single administration surprisingly maintains a very long efficacy in the correction of FD. This effect can be explained as the long-lasting capacity of the NPO treatment to induce greater functional efficiency of the brain dynamics as proven in previous studies

Disease-specific and general health-related quality of life in newly diagnosed prostate cancer patients: The Pros-IT CNR study

Background: The National Research Council (CNR) prostate cancer monitoring project in Italy (Pros-IT CNR) is an observational, prospective, ongoing, multicentre study aiming to monitor a sample of Italian males diagnosed as new cases of prostate cancer. The present study aims to present data on the quality of life at time prostate cancer is diagnosed. Methods: One thousand seven hundred five patients were enrolled. Quality of life is evaluated at the time cancer was diagnosed and at subsequent assessments via the Italian version of the University of California Los Angeles-Prostate Cancer Index (UCLA-PCI) and the Short Form Health Survey (SF-12). Results: At diagnosis, lower scores on the physical component of the SF-12 were associated to older ages, obesity and the presence of 3+ moderate/severe comorbidities. Lower scores on the mental component were associated to younger ages, the presence of 3+ moderate/severe comorbidities and a T-score higher than one. Urinary and bowel functions according to UCLA-PCI were generally good. Almost 5% of the sample reported using at least one safety pad daily to control urinary loss; less than 3% reported moderate/severe problems attributable to bowel functions, and sexual function was a moderate/severe problem for 26.7%. Diabetes, 3+ moderate/severe comorbidities, T2 or T3-T4 categories and a Gleason score of eight or more were significantly associated with lower sexual function scores at diagnosis. Conclusions: Data collected by the Pros-IT CNR study have clarified the baseline status of newly diagnosed prostate cancer patients. A comprehensive assessment of quality of life will allow to objectively evaluate outcomes of different profile of care