Vegetative incompatibility and potential involvement of a mycovirus in the Italian population of Geosmithia morbida

Studies carried out during an Italian outbreak of the Thousand Cankers Disease of walnut, demonstrated that non-coalescing cankers on host plants, separated by equidistant uninfected zones, were associated with incompatible strains of Geosmithia morbida. Confirmation of the vegetative incompatibility of paired fungal isolates, randomly collected from black walnuts, was obtained from observations of a clear separation zones and the absence of anastomoses. Pairing tests with two incompatible monoconidial strains indicated differences in morphology and growth rates. Electron microscopy revealed the presence of icosahedral mycovirus-like particles in one of the monoconidial strains that demonstrated low degrees of virulence in planta compared with a particlefree monoconidial strain. The occurrence of a vegetative incompatibility system in recently introduced populations of G. morbida has considerable implications for fungal biology. Incompatibility in G. morbida and potential direct or indirect roles of the observed virus-like particles have potential ecological and epidemiological consequences. \ua9 Firenze University Press

Vegetative incompatibility and potential involvement of a mycovirus in the Italian population of Geosmithia morbida

Studies carried out during an Italian outbreak of the Thousand Cankers Disease of walnut, demonstrated that non-coalescing cankers on host plants, separated by equidistant uninfected zones, were associated with incompatible strains of Geosmithia morbida. Confirmation of the vegetative incompatibility of paired fungal isolates, randomly collected from black walnuts, was obtained from observations of a clear separation zones and the absence of anastomoses. Pairing tests with two incompatible monoconidial strains indicated differences in morphology and growth rates. Electron microscopy revealed the presence of icosahedral mycovirus-like particles in one of the monoconidial strains that demonstrated low degrees of virulence in planta compared with a particle-free monoconidial strain. The occurrence of a vegetative incompatibility system in recently introduced populations of G. morbida has considerable implications for fungal biology. Incompatibility in G. morbida and potential direct or indirect roles of the observed virus-like particles have potential ecological and epidemiological consequences

Clinical and dermal thickness assessment one year after hyaluronic acid filler treatment

INTRODUÇÃO: Poucos estudos avaliam a durabilidade dos produtos de preenchimento facial de forma objetiva e não-invasiva. O objetivo deste estudo foi avaliar a durabilidade de formulação de ácido hialurônico (Derma Hyal ®) no preenchimento de sulco nasogeniano, comparando percepção do efeito clínico e espessura de partes moles medida por ultrassonografia. MÉTODO: Dez mulheres foram tratadas. Avaliação foi realizada após um, três, seis, nove e doze meses, clinicamente e por ultrassonografia. RESULTADOS: A média da espessura à ultrassonografia foi 0,38 ± 0,14 no pré, 0,69 ± 0,19 após 1 mês, 0,65 ± 0,17 após 3 meses, 0,61 ± 0,22 após 6 meses, 0,57 ± 0,23 após 9 meses e 0,55 ± 0,14 após 12 meses. Os dados analisados pelo teste Friedman não se mostraram estatisticamente significativos. Em relação à satisfação, aos 6 meses, 3 pacientes consideraram o resultado insatisfatório, 5, pouco satisfatório e 2, satisfatório. Apenas uma paciente considerou o resultado satisfatório aos 9 meses. Aos 12 meses, uma considerou pouco satisfatório e as demais, insatisfatório. Todas responderam que realizariam novamente o procedimento. CONCLUSÕES: A ultrassonografia demonstrou ser método objetivo e não-invasivo na avaliação da durabilidade de preenchimento, desde que avaliado com correlação clínica. Demonstrou-se aumento da espessura cutânea até 12 meses após injeção em sulco nasogeniano (SNG), com decréscimo progressivo. Embora exista diferença dos valores da ultrassonografia ao longo do tempo, essas não foram estatisticamente significantes. Clinicamente, o efeito percebido pelas pacientes se deu até o nono mês após aplicação do produto avaliado.BACKGROUND: Few scientific studies evaluate the durability of existent fillers with objective parameters and non invasive methods. The objective of this study was to evaluate durability of a hyaluronic acid formula (Derma Hyal ®) in the nasolabial fold treatment, comparing the perception of clinical effect and soft tissue thickness measured by ultrasound. METHODS: Ten women were treated. Evaluation was made before, after 1, 3, 6, 9 and 12 months, clinically and by ultrasound. RESULTS: Median ultrasound thickness was 0.38 ± 0.14 pre, 0.69 ± 0.19 after 1 month, 0.65 ± 0.17 after 3 months, 0.61 ± 0.22 after 6 months, 0, 57 ± 0.23 after 9 months and 0.55 ± 0.14 after 12 months. Data analyzed by Friedman's test were not statistically significant. Regarding patient satisfaction after 6 months, 3 considered the results unsatisfactory, 5 fairly satisfactory and 2 satisfactory. Only one patient considered the treatment satisfactory at 9 months. By 12 months one considered it fairly satisfactory and nine unsatisfactory. All patients said they would do treatment again in another opportunity. CONCLUSIONS: Ultrasound evaluation of dermal thickness after filler treatment proved to be an objective and non invasive method, since when associated with clinical examination. Increase in soft tissue thickness was demonstrated until 12 months after nasolabial fold injection, with progressive decrease after this period. Despite the increase in soft tissue thickness at ultrasound, it was not statistically significant. Clinical effect was noticed by patients until nine months after treatment with this product

High sodium intake, glomerular hyperfiltration and protein catabolism in patients with essential hypertension

Aims: A blood pressure-independent metabolic shift toward a catabolic state upon high sodium (Na+) diet, ultimately favouring body fluid preservation, has recently been described in pre-clinical controlled settings. We sought to investigate the real-life impact of high Na+ intake on measures of renal Na+/water handling and metabolic signatures, as surrogates for cardiovascular risk, in hypertensive patients. Methods and results: We analysed clinical and biochemical data from 766 consecutive patients with essential hypertension, collected at the time of screening for secondary causes. The systematic screening protocol included 24h urine collection on usual diet and avoidance of renin-angiotensin-aldosterone system-confounding medications. Urinary 24h-Na+ excretion, used to define classes of Na+ intake (Low ≤2.3g/d; Medium 2.3-5g/d; High >5g/d), was an independent predictor of glomerular filtration rate after correction for age, sex, blood pressure, BMI, aldosterone and potassium excretion (p = 0.001; Low: 94.1 [69.9-118.8] vs High: 127.5 [108.3-147.8] ml/min/1.73m2). Renal Na+ and water handling diverged, with higher fractional excretion of Na+ and lower fractional excretion of water in those with evidence of High Na+ intake (FENa: Low 0.39% [0.30-0.47] vs. High 0.81% [0.73-0.98], p < 0.001; FEwater: Low 1.13% [0.73-1.72] vs. High 0.89% [0.69-1.12], p = 0.015). Despite higher FENa, these patients showed higher absolute 24h Na+ reabsorption and higher associated tubular energy expenditure, estimated by tubular Na+/ATP stoichiometry, accordingly (ΔHigh-Low = 18 [12-24] kcal/d, p < 0.001). At non-targeted LC/MS plasma metabolomics in an unselected subcohort (n = 67), metabolites which were more abundant in High vs. Low Na+ intake (p < 0.05) mostly entailed intermediates or end products of protein catabolism/urea cycle. Conclusions: When exposed to high Na+ intake, kidneys dissociate Na+ and water handling. In hypertensive patients, this comes at the cost of higher glomerular filtration rate, increased tubular energy expenditure and protein catabolism from endogenous (muscle) or excess exogenous (dietary) sources. Glomerular hyperfiltration and the metabolic shift may have broad implications on global cardiovascular risk independent of blood pressure

Intracellular Trafficking of Guanylate-Binding Proteins Is Regulated by Heterodimerization in a Hierarchical Manner

Guanylate-binding proteins (GBPs) belong to the dynamin family of large GTPases and represent the major IFN-γ-induced proteins. Here we systematically investigated the mechanisms regulating the subcellular localization of GBPs. Three GBPs (GBP-1, GBP-2 and GBP-5) carry a C-terminal CaaX-prenylation signal, which is typical for small GTPases of the Ras family, and increases the membrane affinity of proteins. In this study, we demonstrated that GBP-1, GBP-2 and GBP-5 are prenylated in vivo and that prenylation is required for the membrane association of GBP-1, GBP-2 and GBP-5. Using co-immunoprecipitation, yeast-two-hybrid analysis and fluorescence complementation assays, we showed for the first time that GBPs are able to homodimerize in vivo and that the membrane association of GBPs is regulated by dimerization similarly to dynamin. Interestingly, GBPs could also heterodimerize. This resulted in hierarchical positioning effects on the intracellular localization of the proteins. Specifically, GBP-1 recruited GBP-5 and GBP-2 into its own cellular compartment and GBP-5 repositioned GBP-2. In addition, GBP-1, GBP-2 and GBP-5 were able to redirect non-prenylated GBPs to their compartment in a prenylation-dependent manner. Overall, these findings prove in vivo the ability of GBPs to dimerize, indicate that heterodimerization regulates sub-cellular localization of GBPs and underscore putative membrane-associated functions of this family of proteins

How future surgery will benefit from SARS-COV-2-related measures: a SPIGC survey conveying the perspective of Italian surgeons

COVID-19 negatively affected surgical activity, but the potential benefits resulting from adopted measures remain unclear. The aim of this study was to evaluate the change in surgical activity and potential benefit from COVID-19 measures in perspective of Italian surgeons on behalf of SPIGC. A nationwide online survey on surgical practice before, during, and after COVID-19 pandemic was conducted in March-April 2022 (NCT:05323851). Effects of COVID-19 hospital-related measures on surgical patients' management and personal professional development across surgical specialties were explored. Data on demographics, pre-operative/peri-operative/post-operative management, and professional development were collected. Outcomes were matched with the corresponding volume. Four hundred and seventy-three respondents were included in final analysis across 14 surgical specialties. Since SARS-CoV-2 pandemic, application of telematic consultations (4.1% vs. 21.6%; p < 0.0001) and diagnostic evaluations (16.4% vs. 42.2%; p < 0.0001) increased. Elective surgical activities significantly reduced and surgeons opted more frequently for conservative management with a possible indication for elective (26.3% vs. 35.7%; p < 0.0001) or urgent (20.4% vs. 38.5%; p < 0.0001) surgery. All new COVID-related measures are perceived to be maintained in the future. Surgeons' personal education online increased from 12.6% (pre-COVID) to 86.6% (post-COVID; p < 0.0001). Online educational activities are considered a beneficial effect from COVID pandemic (56.4%). COVID-19 had a great impact on surgical specialties, with significant reduction of operation volume. However, some forced changes turned out to be benefits. Isolation measures pushed the use of telemedicine and telemetric devices for outpatient practice and favored communication for educational purposes and surgeon-patient/family communication. From the Italian surgeons' perspective, COVID-related measures will continue to influence future surgical clinical practice

OLIGODENDROCYTES FROM SPINAL CORD MENINGES: AMPLIFICATION, CHARACTERIZATION AND TRANSPLANTATION IN CONTUSIVE INJURY

Nonostante le lesioni traumatiche del midollo spinale siano abbastanza frequenti e comportino una grave sofferenza per l\u2019individuo (sia livello fisico che psicologico) e pesanti costi di assistenza diretta ed indiretta, per questa patologia la terapia \ue8 ancora limitata ad interventi di minimizzazione del danno in fase acuta e di supporto e sostegno fisico nella fase cronica. Lo sviluppo della medicina rigenerativa ha ovviamente prodotto una grande aspettativa in questo settore; la scoperta della presenza di cellule neurali staminali nel sistema nervoso centrale dell\u2019adulto e l\u2019ampliarsi della conoscenza dei meccanismi che ne regolano il destino hanno infatti fatto intravedere la possibilit\ue0 di applicazioni terapeutiche di queste cellule nel danno al midollo spinale. Di conseguenza, un buon numero di trials clinici basati sul trapianto di cellule staminali di diversa origine \ue8 stato attivato anche se, al momento, non \ue8 ancora emersa una soluzione definitiva. La maggior parte dei trapianti sperimentati \ue8 stata condotta sinora con cellule staminali non neurali in quanto queste sono di facile reperimento anche da individui adulti; per quanto infatti le cellule staminali neurali siano efficientemente prelevabili da blastocisti e da embrioni, nell'adulto le uniche fonti consistenti sono rappresentate solo da alcune nicchie localizzate in vicinanza dell'ippocampo e dei ventricoli, quindi in zone di difficile accesso per un prelievo autologo o in donatore vivente. Il nostro gruppo ha recentemente dimostrato che le leptomeningi ospitano una popolazione di cellule con propriet\ue0 staminali neurali presenti nel roditore anche in et\ue0 adulta. Queste cellule possono essere coltivate ed espanse in vitro come neurosfere e possono essere indotte a differenziare in neuroni ed oligodendrociti. Se trapiantate in area ippocampale o ventricolare, queste cellule si integrano con il tessuto normale, entrando apparentemente a far parte dell'esistente rete neuronale. Inoltre, a seguito di danno traumatico del midollo spinale, le cellule delle leptomeningi si attivano e migrano all\u2019interno del parenchima, dove partecipano alla reazione al trauma. Considerata la facile accessibilit\ue0 chirurgica delle meningi e la loro presenza in et\ue0 adulta, le cellule staminali delle leptomeningi (LeSCs) rappresentano un potenziale candidato per la terapia rigenerativa del danno al midollo spinale; altrettanto importante \ue8 l'osservazione che le LeSCs possono essere isolate anche da biopsie di meningi umane prelevate nel corso di interventi neurochirurgici (asportazione di tumori). Questo progetto di tesi indaga in profondit\ue0 la possibile applicazione di LeSCs per terapie rigenerative di traumi del midollo spinale; considerato il fatto che i fenomeni di demielinizzazione post-traumatica giocano un ruolo fondamentale nella patogenesi del danno al midollo spinale e che studi condotti con cellule di diversa origine hanno dimostrato come un approccio di medicina rigenerativa basato sulla stimolazione dei processi di rimielinizzazione possa portare a risultati promettenti, nel mio lavoro ho innanzitutto sviluppato ed ottimizzato un metodo in grado di amplificare le LeSCs in vitro e di differenziarle efficacemente in oligodendrociti. \uc9 stato quindi messo a punto un protocollo innovativo di crescita e differenziamento delle LeSCs e, mediante l\u2019analisi dell\u2019espressione proteica e genica, \ue8 stato analizzato e dimostrato come queste cellule acquisiscano sia la tipica morfologia degli oligodendrociti, sia un\u2019elevata espressione di diversi geni mielina-specifici. Il potenziale rigenerativo degli oligodendrociti derivati dalle LeSCs \ue8 stato quindi verificato in vivo in un modello animale di danno contusivo al midollo spinale. Nelle nostre condizioni sperimentali il trapianto degli oligodendrociti \ue8 associato ad un significativo aumento del recupero di alcune funzioni motorie, come determinato dalla valutazione con BBB score e analisi CatWalk. In conclusione, questo lavoro suggerisce per la prima volta che le cellule staminali delle leptomeningi possono rappresentare una risorsa nella terapia cellulare del danno del midollo spinale e apre la strada per futuri studi di medicina rigenerativa applicabili all\u2019uomo.Spinal cord injury (SCI) is a single event with devastating effects on the life of patients both in physiological and psychological terms and for which only supportive and damage-limiting interventions are available at the moment. In the last decades, regenerative therapies based on cell transplantation have generated increasing attention as a potential therapeutic approach for degenerative diseases such as spinal cord injury. In addition, the discovery of neural stem cells in the adult central nervous system and the expansion of the knowledge of the mechanisms regulating their fate have increased the expectations for therapeutic application of these cells to spinal cord injury. Indeed, a considerable number of potential cell-based regenerative therapies have reached the stage of clinical trial, but a clear solution has not emerged yet. We have recently shown that the leptomeninges host a cell population with neural stem/progenitor properties both in vitro and in vivo: isolated leptomeningeal cells can be propagated in vitro as neurospheres and induced to differentiate into neurons and oligodendrocytes. Moreover, they have been shown to become activated by injury to both the brain and the spinal cord and to migrate in the parenchyma, where they participate in the reaction to the injury. Considering the easily accessible anatomical location of the meninges, leptomeningeal stem/progenitor cells (LeSCs) represent a potential candidate for regenerative cell therapy for spinal cord injury. With this work, we provide a first evidence that leptomeningeal cells might indeed play a role in regenerative therapies applied to SCI. Considering the pathogenetic role of demyelination in SCI and that remyelination is a promising therapeutic approach, we first developed and optimized a method for efficient in vitro production of LeSCs and differentiation into mature oligodendrocytes; protein and gene expression analysis showed that by the end of the protocol cultured LeSCs acquired both the typical morphology of mature oligodendrocytes and the elevated expression of different myelin-specific genes. In addition, we performed a pilot study of the regenerative potential of LeSCs-derived oligodendrocyte precursors in an animal model of contusive spinal cord injury. In our conditions, cells transplantation was associated with a significant improvement of some of the motor functions, as determined by behavioural evaluation through BBB score and CatWalk gait analysis. This work indicates for the first time that leptomeningeal stem/progenitor cells could represent an asset in both transplantational and pharmacological therapy for spinal cord injury and paves the way to further studies of regenerative medicine in human SCI

Hikui disease in nine koi carp (Cyprinus carpio): First description of a cutaneous perivascular wall tumour

Background: Hikui disease is a well known disfiguring disease of koi carp (Cyprinus carpio) primarily affecting fish with red pigmentation. It causes light orange to golden yellow, multifocal to coalescing raised patches, starting from the red cutaneous areas. Some cases respond to surgery or topical treatment, but recurrence is common. Objectives: To describe the clinical and pathological presentation of Hikui disease and its cause. Animals: Nine affected koi carp belonging to private hobbyists. Methods: Eight fish underwent surgery or biopsy; one was euthanized. Tissues were submitted for histology, immunohistochemistry and transmission electron microscopy. Results: Five fish showed typical lesions of Hikui disease, whereas four fish showed an atypical presentation characterized by focal or multifocal, oedematous, dark red cutaneous plaques or nodules. Histology showed unencapsulated, infiltrating and densely cellular neoplasms composed of spindle cells arranged in bundles, rows and whorls frequently centred on capillaries. Immunohistochemistry for smooth muscle actin labelled neoplastic cells in all cases. Ultrastructure showed neoplastic cells with slender cytoplasmic processes encircling the capillaries, a thin basal membrane and occasional plasmalemmal vesicles. Conclusions and clinical importance: All of the data supported a neoplastic process producing perivascular wall tumours. Immunoreactivity to smooth muscle actin and the ultrastructural features were indicative of a pericyte origin (haemangiopericytoma). This is the first report dealing with Hikui disease that has achieved a conclusive diagnosis. The neoplastic nature of this condition suggests the potential usefulness of a surgical approach in the clinical management of less severe cases

In vitro characterization of the differentiation of meningeal Stem/precursor cells into the oligodendrocyte lineage

In most diseases of the spinal cord, demyelination plays an important role in the generation and progression of the neurodegenerative lesion. Despite the fact that the spinal cord shows potential for regeneration, this is limited and apparently insufficient for repair and attempts to establish regenerative approaches by neural stem cell transplantation have been so far only partially successful. We have recently described that Leptomeninges of the spinal cord are a niche for neural stem cells (Decimo et al, 2011). These Leptomeningeal Stem Cells (LeSCs) are endowed with self-renewal properties and can differentiate into neurons and oligodendrocytes when stimulated with the appropriate factors. Considering that remyelinization may be a realistic and efficient goal for transplantation-based regenerative therapies of the spinal cord, we decided to define conditions for efficient and high-yield generation of precursor and differentiated cells of the oligodendrocyte lineage. In conclusion, we have established an efficient and reproducible method to generate large numbers of cells of oligodendrocyte-lineage at different and controlled levels of differentiation. Testing of remyelinization efficiency by different sets of cells will be performed in vitro and by transplantation experiments in vivo using animal models of traumatic lesion of the spinal cord and of multiple sclerosis