Characterization of the cytochrome [beta] gene in plant pathogenic, basidiomycetes and consequences for QoI resistance

Strobilurins are one of the most important classes of agricultural fungicides, which are also known as Qo inhibitors (QoIs) because they inhibit mitochondrial respiration by binding to the Qo (‘Quinone outside’) site of the cytochrome b. Cytochrome b is part of the cytochrome bc1 complex, located in the inner mitochondrial membrane, and it is encoded by the cytochrome b (cyt b) gene. Since the mode of action of QoI fungicides is highly specific, the risk of resistance is also high. Single amino-acid exchanges in the cytochrome b were found conferring resistance to QoI fungicides in different plant pathogens. The major mechanism of resistance is a point mutation, also called Single Nucleotide Polymorphism (SNP), in the cyt b gene leading to a change at amino acid position 143 from glycine to alanine (G143A) or from phenylalanine to leucine at position 129 (F129L). However, other point mutations were described, all located in two highly conserved cytochrome b regions, so called ‘hot spot’ regions (amino acid residues 120-160 and 250-300). A fragment of the cyt b gene of a range of agronomically important plant pathogenic Basidiomycetes was sequenced at cDNA level, including Puccinia recondita f.sp. tritici, P. graminis f.sp. tritici, P. striiformis f.sp. tritici, P. coronata f.sp. avenae, P. hordei, P. recondita f.sp. secalis, P. sorghi, P. arachidis, P. horiana, Uromyces appendiculatus, Phakopsora pachyrhizi, Hemileia vastatrix and Rhizoctonia solani. The sequence data allowed developing specific primers for the cyt b gene of all these species, which easily amplified the fragment of the gene including the ‘hot spot’ regions. Resistance to QoI fungicides has never been reported until now in plant pathogenic Basidiomycetes, and in all tested isolates included in this study, the sequence of the cyt b gene fragment did not contain any point mutations. The deduced amino acid sequences of the cytochrome b were used to study the relatedness of these pathogens as compared to other species of the Basidiomycetes, Ascomycetes and Oomycetes. The relatedness was also studied at nuclear level using the Internal Transcribed Spacers (ITS) in the ribosomal DNA. Our results demonstrated for the first time that the amino acid sequence of the mitochondrial cytochrome b is a valid tool to study phylogenic relatedness among plant pathogenic Basidiomycetes and supports taxonomic grouping based on morphological structures and host specificity. Moreover, the phylogenic clusters generated by analysing the cytochrome b sequence confirmed previous results based on the analysis of rRNA of the Uredinales and more specifically, on the ITS regions of closely related species of Puccinia. The cytochrome b gene fragment (coding sequence about 950 bp long) was sequenced also at genomic DNA level in different Puccinia species and U. appendiculatus. A shorter fragment (coding sequence about 600-700 bp) was sequenced in P. pachyrhizi, H. vastatrix, Alternaria solani, A. alternata and Plasmopara viticola. The cyt b gene structure of these agronomically important plant pathogens was characterized, especially in the two ‘hot spot’ regions and compared with that of other species. The exon/intron organization was characterized and its possible role investigated for the occurrence of point mutations in the cyt b gene, especially the amino acid substitutions G143A and F129. In all rust species included in this study as well as in A. solani, the mutation G143A was not detected, but a group I intron (self-splicing intron) was observed starting exactly after the codon GGT for glycine at position 143. In pathogens such as A. alternata, Blumeria graminis, Pyricularia grisea, Mycosphaerella graminicola, M. fijiensis, Venturia inaequalis and P. viticola, resistance to QoI fungicides is known and the glycine (triplet GGT) is replaced by alanine (triplet GCT) at position 143 in the resistant genotype. In those pathogen species, no intron was observed after the codon 143. In the rust species as well as in A. solani, the codon GGT at position 143 is located exactly at the exon/intron boundary and is likely part of the signal sequences essential for the recognition of the intronic RNA to be excised. We predicted that a nucleotide substitution in the codon 143, which is two nucleotides upstream from the exon/intron junction, may strongly affect the splicing process, leading to a deficient cytochrome b, which is lethal. Therefore, individuals carrying the G143A substitution and an intron at this position will not survive. As a consequence, the evolution of resistance to QoI fungicides based on G143A is not likely to evolve in plant pathogens such as Puccinia spp., U. appendiculatus, P. pachyrhizi, H. vastatrix and A. solani. It cannot be excluded that mutations other than G143A conferring resistance may arise in upcoming rust populations selected by the use of QoI fungicides, as observed for A. solani, in which the mutation F129L was reported in resistant isolates. Our results cannot predict whether such mutations may evolve at all in rusts, but the sequence data produced during this study and the understanding of the cyt b gene structure represent an essential information for easy isolation of the cyt b gene and detection of point mutations conferring resistance to QoI fungicides that eventually may evolve

Planificación estratégica de recursos humanos para una empresa del rubro consultoría

Incluye BibliografíaEl presente trabajo tiene como objetivo realizar la planificación estratégica del departamento de recursos humanos de la Consultora KAV. Dado que la organización ha transitado por un proceso de fusión con un estudio contable de forma reciente, se propone analizar dicho tema, con el fin de tener un diagnóstico actual de la organización, en el cual se detecten fortalezas y oportunidades de mejora que permitan diseñar un plan de acción. Para ello se analiza en primera instancia el marco teórico, que dará sustento conceptual al trabajo, luego se realiza una instancia de investigación, para la cual se aplica una encuesta a toda la organización y luego se analizan los resultados obtenidos, presentando las dimensiones más relevantes mediante gráficas por tipo de respuesta y categoría. En base a esto, se elabora la planificación estratégica del departamento de recursos humanos de la organización mediante el diagnóstico de los procesos del área y de los resultados obtenidos en la encuesta de clima organizacional. Asimismo, se propone el mapa estratégico de la firma y se define como principal objetivo para el departamento de recursos humanos convertirse en un socio de negocios. A partir del mismo y de los objetivos específicos, se realiza el cronograma de actividades que serán monitoreadas a través de los indicadores planteados en el tablero de gestión. Como cierre, para poder presentar a los socios y evaluar el plan estratégico en su totalidad, se realiza un presupuesto tentativo de las actividades propuestas

Recurrence of Mycosis Fungoides on Multiple Melanocytic Nevi: A Case Report and Review of the Literature

Melanocytic nevi represent a widespread cutaneous finding. Nevertheless, the presence of mycosis fungoides and melanocytic nevi in the same location is an extremely rare event. We report the case of a patient affected by mycosis fungoides and treated with PUVA therapy, with complete remission of the disease. Eight years after therapy discontinuation, he presented epidermal scaling and an erythematous perinevic halo on 3 old melanocytic lesions, the clinical aspect of which was highly suggestive for Meyerson nevi. The histological and immunohistochemical examination of an excised melanocytic lesion revealed histological features consistent with the diagnosis of mycosis fungoides superimposed on junctional melanocytic nevi. The finding of patches of mycosis fungoides superimposed on melanocytic nevi is a rare event; the confounding clinical appearance with eczematous changes around a pre-existing nevus may recall the halo dermatitis known as Meyerson phenomenon; this highlights the importance of clinical and histological examination to make the correct diagnosis of dermatological diseases

Hypokalemia during the early phase of refeeding in patients with cancer

OBJECTIVE: Refeeding syndrome occurs in patients with severe malnutrition when refeeding begins after a long period of starvation. This syndrome increases the risk of clinical complications and mortality. Hypophosphatemia is considered the primary characteristic of the syndrome. The aim of our study was to investigate the presence of other electrolyte alterations in patients with cancer during the early stage of refeeding. METHODS: In this observational study, we enrolled 34 patients with cancer of the upper aerodigestive tract receiving upfront radiotherapy who were also enrolled in a nutrition program. A caloric intake assessment, anthropometric measurements and biochemical laboratory tests were performed. RESULTS: Significant weight loss (∼20%) was found in these patients. In the patients receiving artificial nutrition, we found lower levels of potassium and total protein compared with those who were fed orally (p = 0.03 for potassium and 0.02 for protein, respectively). Patients on enteral tube feeding had a higher caloric intake compared with those who were fed orally (25±5 kcal/kg/day vs. 10±2 kcal/kg/day). CONCLUSION: Hypokalemia, like hypophosphatemia, could be a complication associated with refeeding in patients with cancer. Hypokalemia was present in the early stages of high-calorie refeeding

Accuracy of magnetic resonance imaging to identify pseudocapsule invasion in renal tumors

Purpose: To evaluate accuracy of MRI in detecting renal tumor pseudocapsule (PC) invasion and to propose a classification based on imaging of PC status in patients with renal cell carcinoma. Methods: From January 2017 to June 2018, 58 consecutive patients with localized renal cell carcinoma were prospectively enrolled. MRI was performed preoperatively and PC was classified, according to its features, as follows: MRI-Cap 0 (absence of PC), MRI-Cap 1 (presence of a clearly identifiable PC), MRI-Cap 2 (focally interrupted PC), and MRI-Cap 3 (clearly interrupted and infiltrated PC). A 3D image reconstruction showing MRI-Cap score was provided to both surgeon and pathologist to obtain complete preoperative evaluation and to compare imaging and pathology reports. All patients underwent laparoscopic partial nephrectomy. In surgical specimens, PC was classified according to the renal tumor capsule invasion scoring system (i-Cap). Results: A concordance between MRI-Cap and i-Cap was found in 50/58 (86%) cases. ρ coefficient for each MRI-cap and iCap categories was: MRI-Cap 0: 0.89 (p < 0.0001), MRI-Cap1: 0.75 (p < 0.0001), MRI-Cap 2: 0.76 (p < 0.0001), and MRI-Cap3: 0.87 (p < 0.0001). Sensitivity, specificity, positive predictive value, negative predictive value, and AUC were: MRI-Cap 0: Se 97.87% Spec 83.3%, PPV 95.8%, NPV 90.9%, and AUC 90.9; MRI-Cap 1: Se 77% Spec 95.5%, PPV 83.3%, NPV 93.5%, and AUC 0.86; MRI-Cap 2- iCap 2: Se 88% Spec 90%, PPV 79%, NPV 95%, and AUC 0.89; MRI-Cap 3: Se 94% Spec 95%, PPV 88%, NPV 97%, and AUC 0.94. Conclusions: MRI-Cap classification is accurate in evaluating renal tumor PC features. PC features can provide an imaging-guided landmark to figure out where a minimal margin could be preferable during nephron-sparing surgery

Social Parasite Ants in the Alps: a New Site of the Vulnerable Myrmica myrmicoxena and New Uppermost Altitudinal Limit for M. microrubra

We conducted a survey on the Alpine fauna of one of the largest Natural Park of the Italian Alps (Stelvio National Park) in the framework of a broad ecological monitoring of Alpine biodiversity. A two-years standardized sampling employing pitfall traps along a 1200 m altitudinal gradient led to the discovery of two interesting inquiline social parasite ants of the genus Myrmica: M. myrmicoxena Forel, 1895 and M. microrubra Seifert, 1993. Myrmica myrmicoxena, which is classified as Vulnerable according to the IUCN Red List, was so far known from only three sites across a narrow geographic range between Italy and Switzerland. Our data support the previous hypothesis over its ecology and host association. Myrmica microrubra is considered an incipient species of high evolutionary interest, sometimes regarded as an intraspecific form of M. rubra. While having a wide distribution in Europe, its presence in Italy was hitherto known only from a single site, and our record extends its altitudinal distribution limit in Europe upwards by about 600 m

Flow Index accurately identifies breaths with low or high inspiratory effort during pressure support ventilation

Background Flow Index, a numerical expression of the shape of the inspiratory flow-time waveform recorded during pressure support ventilation, is associated with patient inspiratory effort. The aim of this study was to assess the accuracy of Flow Index in detecting high or low inspiratory effort during pressure support ventilation and to establish cutoff values for the Flow index to identify these conditions. The secondary aim was to compare the performance of Flow index,of breathing pattern parameters and of airway occlusion pressure (P0.1) in detecting high or low inspiratory effort during pressure support ventilation. Methods Data from 24 subjects was included in the analysis, accounting for a total of 702 breaths. Breaths with high inspiratory effort were defined by a pressure developed by inspiratory muscles (Pmusc) greater than 10 cmH2O while breaths with low inspiratory effort were defined by a Pmusc lower than 5 cmH2O. The areas under the receiver operating characteristic curves of Flow Index and respiratory rate, tidal volume,respiratory rate over tidal volume and P0.1 were analyzed and compared to identify breaths with low or high inspiratory effort. Results Pmusc, P0.1, Pressure Time Product and Flow Index differed between breaths with high, low and intermediate inspiratory effort, while RR, RR/VT and VT/kg of IBW did not differ in a statistically significant way. A Flow index higher than 4.5 identified breaths with high inspiratory effort [AUC 0.89 (CI 95% 0.85–0.93)], a Flow Index lower than 2.6 identified breaths with low inspiratory effort [AUC 0.80 (CI 95% 0.76–0.83)]. Conclusions Flow Index is accurate in detecting high and low spontaneous inspiratory effort during pressure support ventilation

Identification of miRNAs regulating MAPT expression and their analysis in plasma of patients with dementia

Background: Dementia is one of the most common diseases in elderly people and hundreds of thousand new cases per year of Alzheimer’s disease (AD) are estimated. While the recent decade has seen significant advances in the development of novel biomarkers to identify dementias at their early stage, a great effort has been recently made to identify biomarkers able to improve differential diagnosis. However, only few potential candidates, mainly detectable in cerebrospinal fluid (CSF), have been described so far. Methods: We searched for miRNAs regulating MAPT translation. We employed a capture technology able to find the miRNAs directly bound to the MAPT transcript in cell lines. Afterwards, we evaluated the levels of these miRNAs in plasma samples from FTD (n = 42) and AD patients (n = 33) and relative healthy controls (HCs) (n = 42) by using qRT-PCR. Results: Firstly, we found all miRNAs that interact with the MAPT transcript. Ten miRNAs have been selected to verify their effect on Tau levels increasing or reducing miRNA levels by using cell transfections with plasmids expressing the miRNAs genes or LNA antagomiRs. Following the results obtained, miR-92a-3p, miR-320a and miR-320b were selected to analyse their levels in plasma samples of patients with FTD and AD respect to HCs. The analysis showed that the miR-92a-1-3p was under-expressed in both AD and FTD compared to HCs. Moreover, miR-320a was upregulated in FTD vs. AD patients, particularly in men when we stratified by sex. Respect to HC, the only difference is showed in men with AD who have reduced levels of this miRNA. Instead, miR-320b is up-regulated in both dementias, but only patients with FTD maintain this trend in both genders. Conclusions: Our results seem to identify miR-92a-3p and miR-320a as possible good biomarkers to discriminate AD from HC, while miR-320b to discriminate FTD from HC, particularly in males. Combining three miRNAs improves the accuracy only in females, particularly for differential diagnosis (FTD vs. AD) and to distinguish FTD from H

A Feasible Methodological Approach to Estimate the Burden of Autism Spectrum Disorder: Results from the EPI-ASD Study in the Province of Lecce (Southern Italy)

Diagnoses of Autism Spectrum Disorder (ASD) have rapidly increased globally. However, the lack of comprehensive epidemiological surveys and surveillance systems, able to provide official data at a national or European level is one of the main issues in the monitoring of this condition. The present study aimed to estimate the prevalence of ASD in children and adolescents aged 3-18 years old living in the province of Lecce (Southern Italy) through official data provided by the Local Health Authority of Lecce (ASL/LE) up to 31 October 2020, and compare it with school-based data concerning the number of students needing support for ASD. Based on data provided by the ASL/LE, in 2020 there were 509 cases of ASD among children and adolescents aged 3-18 years old, corresponding to a prevalence of 0.46%. A total of 408 (80.2%) were boys and 101 (19.8%) were girls. In relation to their age, 155 ASD cases (0.90%) were diagnosed in the 3-5 age group, while 222 (0.55%) in the 6-11 age group and 132 (0.25%) in the 12-18 age group. Prevalence of ASD assessed by school-based dataset was underestimated in the 3-5 age group, while the 6-11 and 12-18 age groups were consistent with the official data provided by the ASL/LE

Diabete Tipo 1, Tipo 2 e Tipo X

Il muro concettuale secondo il quale il diabete in età pediatrica ha preferibilmente una patogenesi autoimmune sta ormai definitivamente crollando. Il diabete in età infantile e adolescenziale è molto più eterogeneo dal punto di vista eziopatogenetico di quanto si pensasse. In presenza di una qualsiasi iperglicemia è ormai diventato importantissimo chiedersi la patogenesi di questo sintomo utilizzando tutti gli strumenti che abbiamo oggi a disposizione