Aluminum accumulation and tolerance in four Amaranthus species

About one-third of the earth’s land area consists of acidic soils. The rhizotoxic Al3+ is one of the primary constraints associated with low soil pH. Various Amaranthus species are important components of the weed flora in tea plantations on acid soils in north Iran. In this study, four Amaranthus species (A. blitoides, A. retroflexus, A. cruentus, and A. tricolor) were grown under hydroponic conditions with total Al concentrations of 0, 20, 50, 200, and 400 µM corresponding to free Al3+ activity of 0, 3.75, 11.97, 60.34, and 125 µM, respectively. Low Al concentrations (20, 50, or 200 µM) stimulated plant growth, A. tricolor demonstrated the highest improvement in shoot growth (93%), whereas A. retroflexus exhibited the greatest improvement in root biomass (367%), total root length (173%), and aproot length (32%). Although the response of shoot biomass to 400 µM Al varied among species, all species were able to accumulate Al in the leaves above the critical level considered for Al hyperaccumulation (1 mg g–1 DW). Our findings revealed Al accumulation in Amaranthus species for the first time at the genus and family levels, suggesting that these species are suitable for the restoration and revegetation of acid-eroded soils

Obtaining emulsions of furanocoumarins from Sosnowsky’s hogweed and <i>in vitro</i> assessment of their phototoxic effect

Background. Furanocoumarin-based drugs are used for photochemotherapy of various diseases. Sosnovsky’s hogweed can be an available source of furanocoumarins for the development of drugs.The aim of the study. To obtain stable emulsions containing furanocoumarins from Sosnowski’s hogweed and to evaluate their photocytotoxicity.Materials and methods. To obtain the emulsions, furanocoumarins were extracted with chloroform from the sap of the aerial part of the Sosnowski’s hogweed. The chloroform extract was clarified by silica gel gradient column chromatography. The extractive fraction containing furanocoumarins was analyzed by high performance liquid chromatography with ultraviolet (UV) detection. An extract with a high content of 8-methoxypsoralen was used to prepare two types of emulsions. The extract was dissolved in peach oil and emulsified in water with tween-80 (emulsion No. 1) and in an aqueous glycerin solution with lecithin (emulsion No. 2).The emulsions were tested for dark and photo-induced toxicity for human lung fibroblasts. The dose of UV radiation for the photoactivation of furanocoumarins was 9 J/cm2. A solution of chlorine e6 was used as a comparison photosensitizer.Results. The obtained emulsions contained 1 mg/ml 8-methoxypsoralen. Both emulsions were homogeneous at macro- and microscopic visualization, remained stable when stored under various temperature conditions for 14 days. Emulsion No. 2 did not show dark toxicity and caused a statistically significant inhibition of cell viability under UV irradiation at a concentration of 12.5–31.3 µg/mL. Emulsion No. 1 had a toxic effect on cells regardless of UV irradiation due to the content of tween-80. According to fluorescent microscopy, the phototoxic effect of emulsion No. 2 was manifested mainly due to apoptosis, in contrast to the effect of chlorine e6, in which there were more pronounced signs of cell necrosis.Conclusion. The developed experimental emulsions of furanocoumarins from Sosnovski’s hogweed are an example of promising medicinal photosensitizers of plant origin for phototherapy of various dermatological and oncological diseases

Tracking Endogenous Amelogenin and Ameloblastin In Vivo

Research on enamel matrix proteins (EMPs) is centered on understanding their role in enamel biomineralization and their bioactivity for tissue engineering. While therapeutic application of EMPs has been widely documented, their expression and biological function in non-enamel tissues is unclear. Our first aim was to screen for amelogenin (AMELX) and ameloblastin (AMBN) gene expression in mandibular bones and soft tissues isolated from adult mice (15 weeks old). Using RT-PCR, we showed mRNA expression of AMELX and AMBN in mandibular alveolar and basal bones and, at low levels, in several soft tissues; eyes and ovaries were RNA-positive for AMELX and eyes, tongues and testicles for AMBN. Moreover, in mandibular tissues AMELX and AMBN mRNA levels varied according to two parameters: 1) ontogenic stage (decreasing with age), and 2) tissue-type (e.g. higher level in dental epithelial cells and alveolar bone when compared to basal bone and dental mesenchymal cells in 1 week old mice). In situ hybridization and immunohistodetection were performed in mandibular tissues using AMELX KO mice as controls. We identified AMELX-producing (RNA-positive) cells lining the adjacent alveolar bone and AMBN and AMELX proteins in the microenvironment surrounding EMPs-producing cells. Western blotting of proteins extracted by non-dissociative means revealed that AMELX and AMBN are not exclusive to mineralized matrix; they are present to some degree in a solubilized state in mandibular bone and presumably have some capacity to diffuse. Our data support the notion that AMELX and AMBN may function as growth factor-like molecules solubilized in the aqueous microenvironment. In jaws, they might play some role in bone physiology through autocrine/paracrine pathways, particularly during development and stress-induced remodeling

The effects of nanoparticle properties on biological imaging and photothermal cancer treatment

Over the past two decades, gold nanoparticles (AuNPs) have emerged as promising tools for biomedical applications. Their unique optical properties enable sensitive detection and effective treatment strategies. Additionally, the expanding toolkit of AuNP colloidal synthesis, combined with their straightforward surface functionalization, allowing for their conjugation with a variety of targeting and / or therapeutic ligands, contribute to their increasing use. This thesis explores the effects of nanoparticle size, shape, composition, and surface chemistry in the design and application of AuNPs for biological imaging and cancer treatment applications. The 1st chapter introduces various AuNP synthesis, characterization, and conjugation strategies, and presents an overview of their tunable optical properties. Recent AuNP applications such as biological imaging, diagnostics, and cancer treatments (Chapter 1) are reviewed to prepare the reader for the remaining chapters. Then, Chapter 2 discusses the effect of varying surface chemistries on nanoparticle localization within living cells. Using different targeting ligands, a dynamic profile of AuNP localization was obtained. Cellular localization was found to critically affect AuNP scattering properties, a crucial component of biological imaging. Increased subcellular targeting was found to result in greater and more rapid localization, resulting in increased light scattering and enhanced imaging (Chapter 2). Subsequently, the nuclear-targeted AuNPs (NT-AuNPs) previously found to give the greatest imaging enhancement were employed as probes to increase the inherent light scattering from cells. Chapter 3 describes a technique to use these NT-AuNPs to compare the relative efficacies of three clinically relevant chemotherapeutic drugs. This allows the use of a single sample of cells in real-time using inexpensive lab equipment, saving time and material costs while imparting the potential to rapidly screen drugs or analogs to determine the most effective option. The remainder of this thesis focuses on plasmonic photothermal therapy (PPT), an emerging treatment where AuNPs convert light into heat, causing cell death specifically in the vicinity of the targeted AuNPs. Chapter 4 discusses the use of NT-AuNPs to induce PPT cell death while simultaneously serving as scattering probes to monitor the associated molecular changes through time-dependent surface-enhanced Raman spectroscopy of single cells. The same molecular changes were observed using different AuNP sizes, concentrations, and laser intensities, indicating the consistency mechanism of action of PPT. Finally, the use of platinum-coated gold nanorods (PtAuNRs) is introduced in Chapter 5 to mitigate the side effects of PPT. Platinum, commonly used for oxygen reduction in catalysis, is incorporated to scavenge reactive oxygen species (ROS), allowing the decoupling of thermal and chemical effects during PPT. The PtAuNRs protected untreated cells from the ROS byproducts of PPT, making them ideal candidates to advance the treatment while reducing deleterious side effects. This thesis presents a fundamental investigation of the influence of AuNP properties on imaging and cancer treatment, which can be used to continue advancing their utility and applications.Ph.D

Il valore diagnostico incrementale del test di Rorschach rispetto al PDM nell'assessment multimetodo dei disturbi di personalità

La ricerca si propone di verificare il “valore diagnostico incrementale” del test di Rorschach nell’implementazione di una procedura di assessment multimetodo per i disturbi di personalità che includa anche gli assi P ed M del PDM (Manuale Diagnostico Psicodinamico; Task Force, 2006), e che alla luce dei limiti della tassonomia categoriale DSM-based e degli strumenti self-report, li integri con misure clinician-report (SWAP-200, Prototipi Diagnostici Psicodinamici, Questionario sul Funzionamento Mentale) e performance-based. Il “problema della convergenza eterometodo”(Bornstein, 2009) è infatti determinato dai differenti livelli del funzionamento individuale che i diversi strumenti sono in grado di cogliere. La scelta del metodo di siglatura di Klopfer si propone di fornire un primo contributo verso la validazione, alla luce dei vantaggi per una descrizione psicodinamica “multilivello” della personalità che esso fornisce (oltre ad es. alla maggiore economicità clinica e alla quasi sempre più soddisfacente IRR), e delle criticità, poco indagate, del CS di Exner. Saranno illustrati i dati preliminari relativi a un campione di pazienti con DP secondo il DSM-5 afferenti a setting pubblici e privati e valutati con gli strumenti suddetti in maniera indipendente, per verificare empiricamente l’ipotesi dell’esistenza di clusters o misure composite, costituiti da un insieme predefinito di variabili Rorschach, corrispondenti a ciascun disturbo di personalità DSM e alle funzioni mentali dell'Asse M del PDM, e valutarne il “valore diagnostico incrementale” rispetto ai PDP. Sarà discussa l'utilità delle variabili Rorschach nel cogliere le componenti più profonde dell’assetto psichico dei DP per la diagnosi differenziale di disturbi che presentano dimensioni di funzionamento almeno in parte sovrapponibili, per i DP su cui ci sono pochi o nessuno studio, e alla luce delle nuove conoscenze circa caratteristiche controverse di alcuni DP

Using cement as filler to enhance asphalt mixes performance in hot climate regions

This paper investigates the addition of different percentages of ordinary Portland cement as a filler in conventional asphalt concrete for a range of heavy traffic. Road pavement agencies in hot areas face the daunting challenge of preserving their pavements in a fair to good condition to increase their lifespan. This challenge is due to the high occurrence of permanent pavement deformation via rutting, which is one of the major distress factors influencing pavements. This is a particularly serious issue in hot and arid countries which are closely associated with various aggravating factors. These aggravating factors include the choice of bitumen binder viscosity, the type of bitumen, the available low-quality materials, and the high environmental temperatures. Ultimately, poor performance will show within the first few years of service as permanent deformations such as rutting, shoving, and depressions. The examined properties include the resilient modulus and the resistance to rutting. Findings indicate that the resistance to rutting and the rigidity of the asphalt concrete are both substantially increased as the cement content is increased. Moreover, to meet the heavy traffic spectrum requirements, increasing the embedded cement content in the asphalt concrete improves pavement structural capacity. Finally, based on the rigidity expected for different cement levels, design curves are provided for pavement design in hot climates using low quality aggregate materials

Rischio psicopatologico nei pazienti con malattia rara. Uno studio multicentrico

Nonostante la bassa prevalenza delle singole patologie, le Malattie Rare (MR) rappresentano un importante problema di sanità pubblica a causa dell’elevata numerosità, cronicità, grado di invalidità e spesso di mortalità precoce che comportano (Taruscio, 2011). Numerose sono inoltre le problematiche relative alla diagnosi, spesso tardiva (per l’eterogenità nell’insorgenza, nell’eziopatogenesi e nella sintomatologia), al trattamento (un basso numero di esse può contare su terapie risolutive) e alla raccolta di dati epidemiologici attendibili e omogenei tra i diversi Paesi –nell'ICD-10 molte MR non sono nemmeno contemplate-. Nel complesso costituiscono dunque un carico significativo per un consistente numero di pazienti e loro familiari, che risultano verosimilmente a rischio per l'insorgenza di psicopatologia comorbida. Sarà presentata parte dei risultati di uno studio nazionale multicentrico condotto dal Centro Nazionale Malattie Rare (CNMR) dell’Istituto Superiore di Sanità (ISS), relativi a emozioni espresse, vissuti personali, bisogno di sostegno psicologico nelle varie fasi di malattia (prima e dopo la diagnosi, nell'adattamento), aspettative sulla natura del servizio erogato e temi espressi nelle narrazioni dell'utenza dei Centri Regionali/Provinciali di Coordinamento e degli sportelli di ascolto per le MR. I dati, che rappresentano la prima indagine sistematica in Italia dei bisogni psicologici di questi pazienti, saranno discussi rispetto alla possibilità di rischio psicopatologico, anche alla luce del confronto con altre patologie mediche in termini di similitudini e peculiarità, e al fine duplice di documentare la necessità di formazione specifica degli operatori della salute mentale da un lato, e di implementare percorsi reali di invio a consultazione dell'utenza considerata a rischio all'interno dei presidi di cura specialistici, entrambi praticamente inesistenti nel SSN

Interventi del terapeuta e alleanza: un’indagine empirica

L’alleanza terapeutica è stata riconosciuta come quel fattore comune, trasversale ai diversi approcci terapeutici, in grado di spiegare buona parte dell’efficacia delle psicoterapie (Horvath, Bedi, 2002). Se l’effetto dell’alleanza rispetto all’outcome resta un fatto assodato, appare tuttavia più convincente un modello di azione terapeutica che non contrappone aspetti “tecnici” e genericamente “relazionali” in psicoterapia. Questo studio si propone pertanto di: 1) indagare la relazione tra interventi del terapeuta e alleanza; 2) identificare gli elementi del processo terapeutico che sono associati all’alleanza tra paziente e terapeuta; 3) verificare se gli interventi del terapeuta maggiormente correlati all’alleanza aderiscono a specifici modelli di trattamento. 30 sedute di psicoterapia di 15 coppie paziente/terapeuta sono state selezionate in modo casuale e valutate con una batteria di strumenti inclusi: lo Psychotherapy Process Q-set (PQS; Jones, 2000), uno strumento Q-sort costituito da 100 item che descrivono atteggiamenti, comportamenti o esperienze del paziente, azioni, interventi o atteggiamenti del terapeuta e la natura dell’interazione diadica; il Working Alliance Inventory - Observer Version (WAI-O; Horvath, 1981, 1982), uno strumento costituito da 36 item e formato da tre scale che si riferiscono alle tre componenti dell’alleanza individuate da Bordin (1979) – obiettivi (Goal), compiti (Task), legami (Bond); e il Comparative Psychotherapy Process Scale (CPPS; Hilsenroth et al., 2005), uno strumento costituito da 20 item e utilizzato per valutare le tecniche di intervento di tipo psicodinamico o cognitivo-comportamentale adottate dal terapeuta. Le correlazioni tra interventi del terapeuta e alleanza sono risultate significative e positive. Rispetto agli item del PQS associati all’alleanza, molti corrispondono o sono per lo più riconducibili a interventi tecnici di tipo psicodinamico, altri a determinati contributi del paziente, e delineano specifiche modalità di interazione terapeutica. Tali risultati sembrerebbero confermare il carattere bipersonale, di co-costruzione intersoggettiva della relazione terapeutica

A Real-Time Surface Enhanced Raman Spectroscopy Study of Plasmonic Photothermal Cell Death Using Targeted Gold Nanoparticles

Plasmonic nanoparticles are increasingly utilized in biomedical applications including imaging, diagnostics, drug delivery, and plasmonic photothermal therapy (PPT). PPT involves the rapid conversion of light into heat by plasmonic nanoparticles targeted to a tumor, causing hyperthermia-induced cell death. These nanoparticles can be passively targeted utilizing the enhanced permeability and retention effect, or actively targeted using proteins, peptides, or other small molecules. Here, we report the use of actively targeted spherical gold nanoparticles (AuNPs), both to induce PPT cell death, and to monitor the associated molecular changes through time-dependent surface enhanced Raman spectroscopy within a single cell. We monitored these changes in real-time and found that heat generated from the aggregated nanoparticles absorbing near-infrared (NIR) laser light of sufficient powers caused modifications in the protein and lipid structures within the cell and ultimately led to cell death. The same molecular changes were observed using different nanoparticle sizes and laser intensities, indicating the consistency of the molecular changes throughout PPT-induced cell death from actively targeted AuNPs. We also confirmed these observations by comparing them to reference spectra obtained by cell death induced by oven heating at 100 °C. The ability to monitor PPT-induced cell death in real-time will help understand the changes on a molecular level and offers us a basis to understand the molecular mechanisms involved in photothermal cancer cell death