Minor head injury in anticoagulated patients: Outcomes and analysis of clinical predictors. A prospective study.

BACKGROUND: The optimal management of patients taking oral anticoagulants who experience minor head injury (MHI) is unclear. The availability of validated protocols and reliable predictors of prognosis would be of great benefit. We investigated clinical factors as predictors of clinical outcomes and intracranial injury (ICI). METHODS: We conducted a single-cohort, prospective, observational study in an ED. Our structured clinical pathway included a first head CT scan, 24 h observation and a second CT scan. The primary outcome was the occurrence of MHI-related death or re-admission to ED at day +30. The secondary outcome was the rate of delayed ICI (dICI), defined as second positive CT scan after a first negative CT scan. We assessed some clinical predictors derived from guidelines and clinical prediction rules as potential risk factors for the outcomes. RESULTS: 450 patients with a negative first CT scan who underwent a second CT scan composed our 'study population'. The rate of the primary outcome was 4%. The rate of the secondary outcome was 4.7%. Upon univariate and multivariate analysis no statistically significant predictors for the outcomes were found. CONCLUSIONS: Previous retrospective studies showed a lot of negative predictive factors for anticoagulated patients suffering a minor head injury. In our prospective study no clinical factors emerged as predictors of poor clinical outcomes and dICI. So, even if we confirmed a low rate of adverse outcomes, the best management of these patients in ED remains not so clear and future trials are needed.

DNA damage in workers exposed to pigment grade titanium dioxide (TiO2) and association with biomarkers of oxidative stress and inflammation.

The present study was aimed at investigating DNA damage, micronuclei frequency and meta-nuclear alterations in buccal cells of workers involved in pigment-grade TiO2 production (15 exposed and 20 not-exposed). We also assessed associations of genotoxicity biomarkers with oxidative stress/inflammatory biomarkers in urine and exhaled breath condensate (EBC), as well as possible associations between biomarkers and reported respiratory symptoms. In spite of compliance with TiO2 Occupational Exposure Limits, results showed increased direct/oxidative DNA damage and micronuclei frequency in exposed workers. Genotoxicity parameters were associated with oxidative stress/inflammation biomarkers in urine and EBC, thus confirming that TiO2 exposure can affect the oxidative balance. Workers with higher genotoxic/oxidative stress biomarkers levels reported early respiratory symptoms suggesting that molecular alterations can be predictive of early health dysfunctions. These findings suggest the need to assess early health impairment in health surveillance programs and to address properly safety issues in workplaces where TiO2 is handled.

KCTD1 is a new modulator of the KCASH family of Hedgehog suppressors.

The Sonic Hedgehog (Hh) signal transduction pathway plays a critical role in many developmental processes and, when deregulated, may contribute to several cancers, including basal cell carcinoma, medulloblastoma, colorectal, prostate, and pancreatic cancer. In recent years, several Hh inhibitors have been developed, mainly acting on the Smo receptor. However, drug resistance due to Smo mutations or non-canonical Hh pathway activation highlights the need to identify further mechanisms of Hh pathway modulation. Among these, deacetylation of the Hh transcription factor Gli1 by the histone deacetylase HDAC1 increases Hh activity. On the other end, the KCASH family of oncosuppressors binds HDAC1, leading to its ubiquitination and subsequent proteasomal degradation, leaving Gli1 acetylated and not active. It was recently demonstrated that the potassium channel containing protein KCTD15 is able to interact with KCASH2 protein and stabilize it, enhancing its effect on HDAC1 and Hh pathway. KCTD15 and KCTD1 proteins share a high homology and are clustered in a specific KCTD subfamily. We characterize here KCTD1 role on the Hh pathway. Therefore, we demonstrated KCTD1 interaction with KCASH1 and KCASH2 proteins, and its role in their stabilization by reducing their ubiquitination and proteasome-mediated degradation. Consequently, KCTD1 expression reduces HDAC1 protein levels and Hh/Gli1 activity, inhibiting Hh dependent cell proliferation in Hh tumour cells. Furthermore, analysis of expression data on publicly available databases indicates that KCTD1 expression is reduced in Hh dependent MB samples, compared to normal cerebella, suggesting that KCTD1 may represent a new putative target for therapeutic approaches against Hh-dependent tumour.

Sedimentological features of asbestos cement fragments in coastal environments (Taranto, southern Italy).

Asbestos cement materials (ACMs) are widespread in coastal environments as result of illegal dumping activities. This study focuses on the Taranto area (Italy) in the Mar Grande basin within the northern sector of the Ionian Sea. The complex history of dumping building materials containing high amounts of ACM into the coastal zone, and the erosion, transport and deposition in Marechiaro Bay is a serious environmental hazard. An interdisciplinary research methodology defines the temporal dumping succession, and the erosional processes and phases, the diffusion of ACM, the mineralogical characteristics, and existing physical status of the ACM. A multiscale investigation was conducted. Results show that from 1992 to 2000 a significant increment of dumping operations have occurred. The current cliff has been subject to erosion and redeposition phases, developing a new beach composed of these polluted man-made sediments. The findings persuade the local authorities to close the beach requiring remediation interventions.

Autologous chondrocyte implantation provides good long-term clinical results in the treatment of knee osteoarthritis: a systematic review.

PURPOSE: To evaluate the mid- and long-term efficacy of autologous chondrocyte implantation (ACI) and matrix-assisted chondrocyte implantation (MACI) to treat patients with knee cartilage defects in the presence of osteoarthritis (OA). METHODS: PubMed and Cochrane databases were systematically searched for studies describing the treatment of knee OA with ACI or MACI (Kellgren-Lawrence (KL) ≥ 1, minimum follow-up 36 months). Results were reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines and included Lysholm, Western Ontario McMaster University and International Knee Documentation Committee scores. RESULTS: Of the 127 full-text articles assessed for eligibility, only five studies were selected based on inclusion/exclusion criteria (2 on ACI and 3 on MACI). In both groups, the defects were mainly located at femoral level, size 2.2-15.1 cm2 in the ACI and 2.0-7.6 cm2 in the MACI group. ACI was mostly used for patients affected by KL I, whereas MACI for patients with KL II-IV. The data obtained from 235 patients (161 ACI, 74 MACI) showed that ACI and MACI sustained stable clinical improvements up to 11 and 15 years, respectively, with a failure rate of about 10% up to 11 years. Scarce biological details regarding chondrocyte implantation were reported. CONCLUSIONS: ACI and MACI procedures for the treatment of knee cartilage lesions associated to OA showed long-term success and allowed delaying arthroplasty. Additional trials reporting homogenous data and precise patient characterization are needed to conduct an effective literature meta-analysis and identify the clinical relevance of these procedures. LEVEL OF EVIDENCE: IV.

Culture of 3D bioprinted bone constructs requires an increased fluid dynamic stimulation.

In vitro flow-induced mechanical stimulation of developing bone tissue constructs has been shown to favor mineral deposition in scaffolds seeded with cells directly exposed to the fluid flow. However, the effect of fluid dynamic parameters, such as shear stress (SS), within 3D bioprinted constructs is still unclear. Thus, this study aimed at correlating the SS levels and the mineral deposition in 3D bioprinted constructs, evaluating the possible dampening effect of the hydrogel. Human mesenchymal stem cells (hMSCs) were embedded in 3D bioprinted porous structures made of alginate and gelatin. 3D bioprinted constructs were cultured in an osteogenic medium assessing the influence of different flow rates (0, 0.7 and 7 ml/min) on calcium and collagen deposition through histology, and bone volume (BV) through micro-computed tomography. Uniform distribution of calcium and collagen was observed in all groups. Nevertheless, BV significantly increased in perfused groups as compared to static control, ranging from 0.35±0.28 mm3, 11.90±8.74 mm3 and 25.81±5.02 mm3 at week 3 to 2.28±0.78 mm3, 22.55±2.45 mm3 and 46.05±5.95 mm3 at week 6 in static, 0.7 and 7 ml/min groups, respectively. SS values on construct fibers in the range 10-100 mPa in 7 ml/min samples were twice as high as those in 0.7 ml/min samples showing the same trend of BV. The obtained results suggest that it is necessary to enhance the flow-induced mechanical stimulation of cell-embedding hydrogels to increase the amount of mineral deposited by hMSCs, compared to what is generally reported for the development of in vitro bone constructs. STATEMENT OF SIGNIFICANCE: In this study, we evaluated for the first time how the hydrogel structure dampens the effect of flow-induced mechanical stimulation during the culture of 3D bioprinted bone tissue constructs. By combining computational and experimental techniques we demonstrated that those shear stress thresholds generally considered for culturing cells seeded on scaffold surface, are no longer applicable when cells are embedded in 3D bioprinted constructs. Significantly, more bone volume was formed in constructs exposed to shear stress values generally considered as detrimental than in constructs exposed shear stress values generally considered as beneficial after 3 weeks and 6 weeks of dynamic culture using a perfusion bioreactor.

The Adipose Organ Is a Unitary Structure in Mice and Humans.

Obesity is the fifth leading cause of death worldwide. In mice and humans with obesity, the adipose organ undergoes remarkable morpho-functional alterations. The comprehension of the adipose organ function and organization is of paramount importance to understand its pathology and formulate future therapeutic strategies. In the present study, we performed anatomical dissections, magnetic resonance imaging, computed axial tomography and histological and immunohistochemical assessments of humans and mouse adipose tissues. We demonstrate that most of the two types of adipose tissues (white, WAT and brown, BAT) form a large unitary structure fulfilling all the requirements necessary to be considered as a true organ in both species. A detailed analysis of the gross anatomy of mouse adipose organs in different pathophysiological conditions (normal, cold, pregnancy, obesity) shows that the organ consists of a unitary structure composed of different tissues: WAT, BAT, and glands (pregnancy). Data from autoptic dissection of 8 cadavers, 2 females and 6 males (Age: 37.5 ± 9.7, BMI: 23 ± 2.7 kg/m2) and from detailed digital dissection of 4 digitalized cadavers, 2 females and 2 males (Age: 39 ± 14.2 years, BMI: 22.8 ± 4.3 kg/m2) confirmed the mixed (WAT and BAT) composition and the unitary structure of the adipose organ also in humans. Considering the remarkable endocrine roles of WAT and BAT, the definition of the endocrine adipose organ would be even more appropriate in mice and humans.

Musculoskeletal tissues-on-a-chip: role of natural polymers in reproducing tissue-specific microenvironments.

Over the past years, 3Din vitromodels have been widely employed in the regenerative medicine field. Among them, organ-on-a-chip technology has the potential to elucidate cellular mechanism exploiting multichannel microfluidic devices to establish 3D co-culture systems that offer control over the cellular, physico-chemical and biochemical microenvironments. To deliver the most relevant cues to cells, it is of paramount importance to select the most appropriate matrix for mimicking the extracellular matrix of the native tissue. Natural polymers-based hydrogels are the elected candidates for reproducing tissue-specific microenvironments in musculoskeletal tissue-on-a-chip models owning to their interesting and peculiar physico-chemical, mechanical and biological properties. Despite these advantages, there is still a gap between the biomaterials complexity in conventional tissue engineering and the application of these biomaterials in 3Din vitromicrofluidic models. In this review, the aim is to suggest the adoption of more suitable biomaterials, alternative crosslinking strategies and tissue engineered-inspired approaches in organ-on-a-chip to better mimic the complexity of physiological musculoskeletal tissues. Accordingly, after giving an overview of the musculoskeletal tissue compositions, the properties of the main natural polymers employed in microfluidic systems are investigated, together with the main musculoskeletal tissues-on-a-chip devices.

Antibiotic prescriptions in the context of suspected bacterial respiratory tract superinfections in the COVID-19 era: a retrospective quantitative analysis of antibiotic consumption and identification of antibiotic prescription drivers.

This study aims to quantify antibiotic consumption for suspected respiratory tract superinfections in COVID-19 patients, while investigating the associated drivers of antibiotic prescribing in light of the current signs of antibiotic overuse. Adult patients with a positive COVID-19 diagnosis admitted to a Belgian 721-bed university hospital were analyzed retrospectively (March 11th-May 4th, 2020), excluding short-term admissions (< 24 h). Antibiotic prescriptions were analyzed and quantified, using Defined Daily Doses (DDD) per admission and per 100 bed days. Possible drivers of antibiotic prescribing were identified by means of mixed effects logistic modelling analysis with backwards selection. Of all included admissions (n = 429), 39% (n = 171) were prescribed antibiotics for (presumed) respiratory tract superinfection (3.6 DDD/admission; 31.5 DDD/100 bed days). Consumption of beta-lactamase inhibitor-penicillin combinations was the highest (2.55 DDD/admission; 23.3 DDD/100 bed days). Four drivers were identified: fever on admission (OR 2.97; 95% CI 1.42-6.22), lower SpO2/FiO2 ratio on admission (OR 0.96; 95% CI 0.92-0.99), underlying pulmonary disease (OR 3.04; 95% CI 1.12-8.27) and longer hospital stay (OR 1.09; 95% CI 1.03-1.16). We present detailed quantitative antibiotic data for presumed respiratory tract superinfections in hospitalized COVID-19 patients. In addition to knowledge on antibiotic consumption, we hope antimicrobial stewardship programs will be able to use the drivers identified in this study to optimize their interventions in COVID-19 wards.

Adaptación y validación preliminar argentina del cuestionario de evaluación del apego (CaMir) / Adaptació i validació preliminar argentina del qüestionari d'avaluació de l’aferrament (CaMir) / Preliminary Argentinean adaptation and validation of the bonding assessment questionnaire (CaMir)

El propósitode este estudio fue adaptar lingüísticamente y evaluar validez y fiabilidad del instrumento de evaluación demodelos individuales de relación del estilo de apego, CaMir (Cartes: Modèles Individuels de Relation), en unamuestra argentina de adolescentes y adultos. Se trabajó con una muestra de 549 sujetos de ambos sexos deentre 14 y 80 años. El análisis factorial exploratorio develó una estructura de cuatro factores y el análisis fac-torial confirmatorio corroboró la misma, así como los índices de fiabilidad de los factores. Se confirma que esun instrumento válido y confiable para evaluar los modelos individuales de relación en adolescentes y adultosargentinos. Finalmente, se señalan limitaciones y contribuciones.(AU)

The pur-pose of this study was to linguistically adapt and evaluate the validity and reliability of the individual modelsof attachment style relationship assessment instrument, CaMir (Cartes: Modèles Individuels de Relation), in anArgentinean sample of adolescents and adults. A sample of 549 subjects of both sexes aged between 14 and80 years was used. The exploratory factor analysis revealed a four-factor structure and the confirmatory factoranalysis corroborated it, as well as the reliability indices of the factors. It is confirmed that it is a valid and reliableinstrument to assess individual relationship patterns in Argentinean adolescents and adults. Finally, limitations andcontributions are pointed out.(AU)

El propòsitd'aquest estudi va ser adaptar lingüísticament i avaluar la validesa i la fiabilitat de l'instrument d'avaluació demodels individuals de relació de l'estil d'aferrament, CaMir (Cartes: Modèles Individuels de Relation), en una mos-tra argentina d'adolescents i adults. Es va treballar amb una mostra de 549 subjectes d'ambdós sexes d'entre14 i 80 anys. L'anàlisi factorial exploratòria va desvetllar una estructura de quatre factors i l'anàlisi factorial con-firmatori la va corroborar, així com els índexs de fiabilitat dels factors. Es confirma que és un instrument vàlidi fiable per avaluar els models individuals de relació en adolescents i adults argentins. Finalment, s'assenyalenlimitacions i contribucions.(AU)

A geo-chemo-mechanical study of a highly polluted marine system (Taranto, Italy) for the enhancement of the conceptual site model.

The paper presents the results of the analysis of the geo-chemo-mechanical data gathered through an innovative multidisciplinary investigation campaign in the Mar Piccolo basin, a heavily polluted marine bay aside the town of Taranto (Southern Italy). The basin is part of an area declared at high environmental risk by the Italian government. The cutting-edge approach to the environmental characterization of the site was promoted by the Special Commissioner for urgent measures of reclamation, environmental improvements and redevelopment of Taranto and involved experts from several research fields, who cooperated to gather a new insight into the origin, distribution, mobility and fate of the contaminants within the basin. The investigation campaign was designed to implement advanced research methodologies and testing strategies. Differently from traditional investigation campaigns, aimed solely at the assessment of the contamination state within sediments lying in the top layers, the new campaign provided an interpretation of the geo-chemo-mechanical properties and state of the sediments forming the deposit at the seafloor. The integrated, multidisciplinary and holistic approach, that considered geotechnical engineering, electrical and electronical engineering, geological, sedimentological, mineralogical, hydraulic engineering, hydrological, chemical, geochemical, biological fields, supported a comprehensive understanding of the influence of the contamination on the hydro-mechanical properties of the sediments, which need to be accounted for in the selection and design of the risk mitigation measures. The findings of the research represent the input ingredients of the conceptual model of the site, premise to model the evolutionary contamination scenarios within the basin, of guidance for the environmental risk management. The study testifies the importance of the cooperative approach among researchers of different fields to fulfil the interpretation of complex polluted eco-systems.

Tocilizumab in COVID-19 interstitial pneumonia.

BACKGROUND: Published reports on tocilizumab in COVID-19 pneumonitis show conflicting results due to weak designs or heterogeneity in critical methodological issues. METHODS: This open-label trial, structured according to Simon's optimal design, aims to identify factors predicting which patients could benefit from anti-IL6 strategies and to enhance the design of unequivocal and reliable future randomized trials. A total of 46 patients with COVID-19 pneumonia needing of oxygen therapy to maintain SO2 > 93% and with recent worsening of lung function received a single infusion of tocilizumab. Clinical and biological markers were measured to test their predictive values. Primary end point was early and sustained clinical response. RESULTS: Twenty-one patients fulfilled pre-defined response criteria. Lower levels of IL-6 at 24 h after tocilizumab infusion (P = 0.049) and higher baseline values of PaO2/FiO2 (P = 0.008) predicted a favourable response. CONCLUSIONS: Objective clinical response rate overcame the pre-defined threshold of 30%. Efficacy of tocilizumab to improve respiratory function in patients selected according to our inclusion criteria warrants investigations in randomized trials.

Cytogenetic biomonitoring of road tunnel construction workers: buccal micronucleus cytome assay.

Results: Road tunnel construction workers revealed higher frequencies of cells with genotoxic damage (i.e., MN and NBUD). MN and NBUD resulted to be Poisson distributed and counts of these genotoxicity biomarkers were then analysed by Poisson regression. The frequency ratio (FR) for MN was 1.31 (95% CI: 0.84-2.04), with an increase in the exposed subjects; this finding, though indicating a higher genotoxic risk in the exposed subjects, did not reach statistical significance. On the other hand, the FR for NBUD was 3.49 (95% CI: 1.86-6.56), with a statistically significant increased risk of chromosomal damage. Even the frequencies of binucleated cells (a marker of cell proliferation) and pyknotic cells (a cell death biomarker) were significantly higher in tunnel workers. Introduction: Tunnel construction workers are exposed to complex mixtures of toxic agents, some of which are known to be genotoxic. The aim of this study was to evaluate the genotoxic risk in this occupational setting by comparing tunnel workers with a control group for frequencies of nuclear aberrations in oral exfoliated cells. Methods: To evaluate the genotoxic effects of tunnel air pollutants, we conducted a cross-sectional, molecular epidemiological study (35 tunnel workers and 35 healthy controls) using the buccal micronucleus cytome assay. A questionnaire was administered to obtain information about demographic variables, lifestyle, dietary habits, anthropometric data, and occupational history. Buccal mucosa cells were collected by scraping the buccal mucosa with a small-headed toothbrush. Coded slides were examined blind by trained scorers for micronuclei (MN), nuclear buds (NBUD), and other nuclear abnormalities. Conclusions: Our observations provide further knowledge and understanding of the occupational hazards of tunnel workers and confirm the complexity of effects (cytotoxic and genotoxic) probably induced by fumes and dust produced in underground operations.

A Systematic Review of Primary and Secondary Callous-Unemotional Traits and Psychopathy Variants in Youth.

Two variants of callous-unemotional (CU) traits and psychopathy have been proposed, referred to as primary and secondary. Whereas primary variants are thought to be underpinned by insufficient arousal to emotional cues, secondary variants are thought to develop as a coping mechanism in response to trauma exposure. Compared with adult samples, research on primary and secondary variants in children and adolescents under the age of 18 has only emerged in the past decade, and there is ongoing debate with regards to the identification, defining characteristics, and distinct correlates of these variants. The present systematic review synthesizes the current literature on primary and secondary variants in relation to: (1) constructs used to distinguish and define primary and secondary variants; (2) study population characteristics; (3) data analytic techniques to differentiate variants; and (4) differential associations with theoretically relevant indices related to emotional processing, maltreatment, biomarkers, and behavioral outcomes (e.g., substance use, aggression). This is the first systematic review to examine the growing literature on primary and secondary CU and psychopathy variants among youth. Findings support the distinction between youth with primary versus secondary variants and demonstrate that this distinction is related to unique clinical correlates. Recommendations are made for future research in the field.

Growth kinetics of Myceliophthora thermophila M.7·7 in solid-state cultivation.

AIMS: This work aimed to estimate the growth of Myceliophthora thermophila M.7·7 in solid-state cultivation (SSC) through quantification of N-acetyl-d-glucosamine (NAG) and enzyme activity. METHODS AND RESULTS: The fungus was cultivated in sugarcane bagasse and wheat bran. A consistent statistical analysis was done to assess the reliability of experimental data. Logistic model equation was fitted to experimental data and growth parameters were estimated. The results showed strong influence of the sample size on NAG and a minimum recommended sample size was identified. Scanning electron microscopy (SEM) was used to identify the strategy of substrate colonization. Wheat bran was attacked firstly, while sugarcane bagasse was consumed after wheat bran depletion. The biomass growth was poorly estimated by secretion kinetics of α-amylase, endoglucanase, protease and xylanase, but enzyme kinetics were important for understanding substrate colonization. CONCLUSIONS: In conclusion, the NAG concentration was strongly affected by the sample size and sampling procedure. The strategy of fungal colonization on the substrates was well characterized through SEM analysis. The colonization strategy has direct influence on the kinetic parameters of the logistic model. Myceliophthora thermophila has a well-defined dynamic of enzyme secretion to degrade the substrate, although the kinetics of enzyme secretion has shown not adequate to characterize the kinetics of fungal growth. SIGNIFICANCE AND IMPACT OF THE STUDY: The paper provides reliable growth kinetic parameters in the SSC of the cellulase producer fungus M. thermophila M.7·7, as well as a robust analysis on three indirect methods (NAG, enzymes and SEM) for estimation of fungal development.

Visible DNA microarray and loop-mediated isothermal amplification (LAMP) for the identification of Cryptococcus species recovered from culture medium and cerebrospinal fluid of patients with meningitis.

Cryptococcal meningitis affects normal hosts and immunocompromised patients exhibiting high mortality rates. The objective of this study was to design two molecular assays, visible microarray platforms and loop-mediated isothermal amplification (LAMP), to identify Cryptococcus spp. and the species neoformans and gattii from the cerebral spinal fluid (CSF). To identify Cryptococcus and the two species, we designed two microarrays DNA platforms based on the internal transcribed spacer (ITS) region and CAP59 gene and LAMP assays specific for Cryptococcus species. The assays were tested using CSF from patients with cryptococcal meningitis. CSF from patients with cryptococcal meningitis was cultured in Sabouraud culture medium, and the Cryptococcus spp. grown in the culture medium were also tested for LAMP and microarray platforms. The results were compared to DNA sequencing of the same genetic regions. A total of 133 CSF samples were studied. Eleven CSFs were positive for Cryptococcus (9 C. neoformans and 2 C. gattii), 15 were positive for bacteria, and 107 were negative. The CAP59 platform correctly identified 73% of the CSF samples, while the ITS platform identified 45.5%. CAP59 platform correctly identified 100% of the Cryptococcus isolates, and ITS platform identified 70%. The two sets of LAMP primers correctly identified 100% of the Cryptococcus isolates. However, for CSF samples, the amplification occurred only in 55.5% of C. neoformans. The methodologies were reliable in the identification of Cryptococcus species, mainly for isolates from culture medium, and they might be applied as adjunctive tests to identify Cryptococcus species.

E-Coaching: the DianaWeb study to prevent breast cancer recurrences.

INTRODUCTION: Breast cancer is the most common cancer in women worldwide, for which the survival rate is increasing over time. Growing evidence are showing that the effect of lifestyle could have the same weight of the effect of the usual clinical-pathological risk factors on survival rate. The DianaWeb study responds to the pressing request of patients diagnosed with breast cancer to know the most advanced point of scientific research on the prevention of recurrences, to have a virtual space to meet, where to receive advice and practical information for the daily management the lifestyle change. DianaWeb is a community-based participatory research, dedicated to breast cancer patients, aimed to monitor lifestyle, provide them tips to encourage sustainable lifestyle changes, and to analyze clinical outcomes. In order to achieve these aims, DianaWeb uses a specific interactive website (http://www.dianaweb.org/). METHODS: The web architecture has been designed essential and light, with a rigorous implantation that brings the figure of the woman to the center. In order to humanized the project the acronym used to identify the study was Diana (Diet and Androgens), a classical female name, who was illustrated as a female avatar, aimed to symbolize all women. The graphical interface was developed using seven pastel tones colors, which become a fundamental elements of the layout, such as frame, navigation menu, and separation interspaces. RESULTS: The project started in 2015, and in 4 days the web site was visited by more than 1000 people. A total of 2823 persons enrolled in the study, but 2182 did not send the full documentations, 61 persons abandoned the study, 641 timely answered to the questionnaires. CONCLUSIONS: The high number of participants' interaction within the web page, confirmed the high usability of the web page and the great interest of patients. Prevention of breast cancer recurrences with low cost technologies, easily available to everybody, is a priority for both public health and public finances.

Visible DNA microarray and loop-mediated isothermal amplification (LAMP) for the identification of Cryptococcus species recovered from culture medium and cerebrospinal fluid of patients with meningitis

Cryptococcal meningitis affects normal hosts and immunocompromised patients exhibiting high mortality rates. The objective of this study was to design two molecular assays, visible microarray platforms and loop-mediated isothermal amplification (LAMP), to identify Cryptococcus spp. and the species neoformans and gattii from the cerebral spinal fluid (CSF). To identify Cryptococcus and the two species, we designed two microarrays DNA platforms based on the internal transcribed spacer (ITS) region and CAP59 gene and LAMP assays specific for Cryptococcus species. The assays were tested using CSF from patients with cryptococcal meningitis. CSF from patients with cryptococcal meningitis was cultured in Sabouraud culture medium, and the Cryptococcus spp. grown in the culture medium were also tested for LAMP and microarray platforms. The results were compared to DNA sequencing of the same genetic regions. A total of 133 CSF samples were studied. Eleven CSFs were positive for Cryptococcus (9 C. neoformans and 2 C. gattii), 15 were positive for bacteria, and 107 were negative. The CAP59 platform correctly identified 73% of the CSF samples, while the ITS platform identified 45.5%. CAP59 platform correctly identified 100% of the Cryptococcus isolates, and ITS platform identified 70%. The two sets of LAMP primers correctly identified 100% of the Cryptococcus isolates. However, for CSF samples, the amplification occurred only in 55.5% of C. neoformans. The methodologies were reliable in the identification of Cryptococcus species, mainly for isolates from culture medium, and they might be applied as adjunctive tests to identify Cryptococcus species.

Modelling environmental impacts of treated municipal wastewater reuse for tree crops irrigation in the Mediterranean coastal region.

Wastewater reuse provides valuable solutions to solve the societal challenges of decreasing availability and limiting access to secure water resources. The present study quantifies the environmental performance of nectarine orchards irrigation using treated municipal wastewater (TMW) and surface water using a unique dataset based on field experimental data. Climate change, toxicity (for human and freshwater), eutrophication (marine and freshwater) and acidification impacts were analysed using the impact assessment method suggested by the International Reference Life Cycle Data System (ILCD). The water footprint associated to the life cycles of each system has been estimated using the Available WAter REmaining (AWARE) method. Monte Carlo simulation was used to assess data uncertainty. The irrigation of nectarine orchards using TMW performs better than the irrigation using surface water for eutrophication impact categories. Compared with surface water resources, the potential impacts of TMW reuse in agriculture on climate change and toxicity are affected by the wastewater treatment phase (WWT). Only eutrophication and acidification burdens are generated by in-field substitution of surface water with TMW. Considering human and ecosystem water demand, the irrigation with TMW increases water consumption of 19.12 m3 per kg of nectarine produced. Whereas, it shows a positive contribution to water stress (-0.19 m3) if only human water demand is considered. This study provides important results that allow for a better understanding of the potential environmental consequences of TMW reuse in agriculture. It suggests that embracing the type of WWTs, the replacement of fertilizers, the effects on water scarcity and ecosystem quality might be useful to redefine water reuse regulations and increase public acceptance for the reuse of TMW in agriculture. Moreover, this study reveals the need for developing consensus and standardized guidance for life cycle analysis of water reuse applications.