The non-vesicle cell-free DNA (cfDNA) induces cell transformation associated with horizontal DNA transfer.

BACKGROUND: Cell-free DNA (cfDNA) is a source for liquid biopsy used for cancer diagnosis, therapy selection, and disease monitoring due to its non-invasive nature and ease of extraction. However, cfDNA also participates in cancer development and progression by horizontal transfer. In humans, cfDNA circulates complexed with extracellular vesicles (EV) and macromolecular complexes such as nucleosomes, lipids, and serum proteins. The present study aimed to demonstrate whether cfDNA not associated with EV induces cell transformation and tumorigenesis. METHODS: Supernatant of the SW480 human colon cancer cell line was processed by ultracentrifugation to obtain a soluble fraction (SF) and a fraction associated with EV (EVF). Primary murine embryonic fibroblast cells (NIH3T3) underwent passive transfection with these fractions, and cell proliferation, cell cycle, apoptosis, cell transformation, and tumorigenic assays were performed. Next, cfDNA was analyzed by electronic microscopy, and horizontal transfer was assessed by human mutant KRAS in recipient cells via PCR and recipient cell internalization via fluorescence microscopy. RESULTS: The results showed that the SF but not the EVF of cfDNA induced proliferative and antiapoptotic effects, cell transformation, and tumorigenesis in nude mice, which were reduced by digestion with DNAse I and proteinase K. These effects were associated with horizontal DNA transfer and cfDNA internalization into recipient cells. CONCLUSIONS: The results suggest pro-tumorigenic effects of cfDNA in the SF that can be offset by enzyme treatment. Further exploration of the horizontal tumor progression phenomenon mediated by cfDNA is needed to determine whether its manipulation may play a role in cancer therapy.

Crossover from super- to subdiffusive motion and memory effects in crystalline organic semiconductors.

The transport properties at finite temperature of crystalline organic semiconductors are investigated, within the Su-Schrieffer-Heeger model, by combining an exact diagonalization technique, Monte Carlo approaches, and a maximum entropy method. The temperature-dependent mobility data measured in single crystals of rubrene are successfully reproduced: a crossover from super- to subdiffusive motion occurs in the range 150≤T≤200 K, where the mean free path becomes of the order of the lattice parameter and strong memory effects start to appear. We provide an effective model, which can successfully explain features of the absorption spectra at low frequencies. The observed response to slowly varying electric field is interpreted by means of a simple model where the interaction between the charge carrier and lattice polarization modes is simulated by a harmonic interaction between a fictitious particle and an electron embedded in a viscous fluid.

Environmental analysis of sunflower production with different forms of mineral nitrogen fertilizers.

Environmental profiles of mineral nitrogen fertilizers were used to evaluate the environmental disturbances related to their use in cultivation systems in Europe. Since the production of mineral fertilizers requires a large amount of energy, the present study of bioenergy systems is relevant in order to achieve crop yields less dependent on fossil fuels and to reduce the environmental impact due to fertilization. In this study, the suitability of the LCA methodology to analyze the environmental impact of sunflower cultivation systems with different forms of mineral nitrogen fertilizers urea and ammonium nitrate was investigated. Effects on climate change were estimated by the use of Ecoinvent 2.2 database default value for soil N2O emission factor (1%) and local emission data (0.8%) of mineral nitrogen applied to soils. LCA analysis showed a higher impact on environmental categories (human health and ecosystem quality) for the system in which urea was used as a nitrogen source. Use of urea fertilizer showed a higher impact on resource consumption due to fossil fuel consumption. Use of mineral nitrogen fertilizers showed a higher environmental burden than other inputs required for sunflower cultivation systems under study. Urea and ammonium nitrate showed, respectively, a 7.8% and 4.9% reduced impact of N2O as greenhouse gas by using direct field data of soil N2O emission factor compared to the default soil emission factor of 2006 IPCC Guidelines. Use of ammonium nitrate as mineral nitrogen fertilizer in sunflower cultivation would have a lower impact on environmental categories considered. Further environmental analysis of available technologies for fertilizer production might be also evaluated in order to reduce the environmental impacts of each fertilizer.

Nanoscale Dynamics of Streptococcal Adhesion to AGE-Modified Collagen.

The adhesion of initial colonizers such as Streptococcus mutans to collagen is critical for dentinal and root caries progression. One of the most described pathological and aging-associated changes in collagen-including dentinal collagen-is the generation of advanced glycation end-products (AGEs) such as methylglyoxal (MGO)-derived AGEs. Despite previous reports suggesting that AGEs alter bacterial adhesion to collagen, the biophysics driving oral streptococcal attachment to MGO-modified collagen remains largely understudied. Thus, the aim of this work was to unravel the dynamics of the initial adhesion of S. mutans to type I collagen in the presence and absence of MGO-derived AGEs by employing bacterial cell force spectroscopy with atomic force microscopy (AFM). Type I collagen gels were treated with 10 mM MGO to induce AGE formation, which was characterized with microscopy and enzyme-linked immunosorbent assay. Subsequently, AFM cantilevers were functionalized with living S. mutans UA 159 or Streptococcus sanguinis SK 36 cells and probed against collagen surfaces to obtain force curves displaying bacterial attachment in real time, from which the adhesion force, number of events, Poisson analysis, and contour and rupture lengths for each individual detachment event were computed. Furthermore, in silico computer simulation docking studies between the relevant S. mutans UA 159 collagen-binding protein SpaP and collagen were computed, in the presence and absence of MGO. Overall, results showed that MGO modification increased both the number and adhesion force of single-unbinding events between S. mutans and collagen, without altering the contour or rupture lengths. Both experimental and in silico simulations suggest that this effect is due to increased specific and nonspecific forces and interactions between S. mutans UA 159 and MGO-modified collagen substrates. In summary, these results suggest that collagen alterations due to aging and glycation may play a role in early bacterial adherence to oral tissues, associated with conditions such as aging or chronic hyperglycemia, among others.

Management of Hypothyroidism in Internal Medicine: Patient Profile and Effects of an Educational Programme in the Cluster-Randomized FADOI TIAMO Study.

Background and aims: There is still limited knowledge regarding the clinical profile and appropriateness of treatment in patients with hypothyroidism hospitalized in Internal Medicine (IM) Departments in Italy. The aim of this study is to evaluate: 1) the characteristics of patients and possible deviations from national and international clinical practice recommendations (CPRs) in evidence-based guidelines (EBGs); 2) the improvement of patient management by means of a standardized educational programme (EP). Methods: A nationwide multicentre study, comprising two replications of a retrospective survey (phases 1 and 3) with an intervening EP (phase 2) in half of the centres and no EP in the other half, was conducted. The EP was based on outreach visits. Centres were assigned to the two arms of the study, labelled the training group (TG) and control group (CG) respectively, by cluster randomization. Four EBGs and 39 CPRs provided the basis on which 22 treatment management indicators were identified (7 referring to the time of hospital admission, 15 to post-admission). Results: The 21 participating centres recruited 587 hospitalized patients with hypothyroidism, 421 of which were females (71.7%, mean age 74.1 + 14.4 yrs): 318 in phase 1 and 269 in phase 3. The cause of hypothyroidism was unknown in 282 patients (48%). Evaluation at the time of admission identified satisfactory adherence to CPRs (>50%) for 63.6% of the indicators. In the phase 3, TG centres showed significant improvement vs CG in 4 of the 15 post-admission indicators, while 1 out of 15 was significantly worse. Conclusions: The EP based on outreach visits significantly improved some indicators in the management of patients with hypothyroidism, with specific reference to appropriateness of TSH dosage and levothyroxine (LT4) treatment modality. Clinical Trial Registration: ClinicalTrials.gov, identifier NCT05314790.

Hereditary diffuse gastric cancer (HDGC). An overview.

It is estimated that up to 10% of gastric carcinomas show familial aggregation. In contrast, around 1-3 % (approximately 33,000 yearly) are genuinely hereditary. Hereditary diffuse gastric cancer (HDGC) is a rare malignancy characterized by autosomal dominant inheritance of pathological variants of the CDH1 and CTNNA1 genes encoding the adhesion molecules E-cadherin and α-catenin, respectively. The multifocal nature of the disease and the difficulty of visualizing precursor lesions by endoscopy underscore the need to be aware of this malignancy as surgical prevention can be fully protective. Here, we provide an overview of the main epidemiological, clinical, genetic, and pathological features of HDGC, as well as updated guidelines for its diagnosis, genetic testing, counseling, surveillance, and management. We conclude that HDGC is a rare, highly penetrant disease that is difficult to diagnose and manage, so it is necessary to correctly identify it to offer patients and their families' adequate management following the recommendations of the IGCL. A critical point is identifying a mutation in HDGC families to determine whether unaffected relatives are at risk for cancer.

Differential regulation of Notch ligands in dendritic cells upon interaction with T helper cells.

The Notch signalling pathway has recently been linked to T helper 1 (Th1)/T helper 2 (Th2) cell polarization via a mechanism involving differential expression of Notch ligands, Delta-like and Jagged, in antigen-presenting cells. However, whether stimuli other than pathogen-derived factors are involved in the regulation of Notch ligand expression in dendritic cells (DCs) remains unknown. Here, we address the effect of T helper cells (Th1 and Th2) on Delta-like 4 and Jagged 2 expression in bone marrow-derived DCs. We demonstrate that both Th1 and Th2 cells induce Delta-like 4 mRNA expression in DCs, in a process that is, in part, mediated by CD40 signalling. In contrast, only Th2 cells induce a significant increase in Jagged 2 mRNA levels in DCs. Additionally, we show that IL-4, a hallmark Th2 cytokine, plays a role in Jagged 2 expression, as evidenced by the fact that cholera toxin, a Th2-promoting stimulus, induces Jagged 2 mRNA expression in DCs only in the presence of IL-4. Finally, we demonstrate that DCs also express Notch 1 and that this expression is downregulated by IL-4. These data suggest that Notch ligands are differentially regulated in DCs: Delta-like 4 is regulated by T helper cells and by pathogen-derived Th1 stimuli, whereas Jagged 2 is regulated by Th2 cells and pathogen-derived Th2-promoting stimuli. Based on our results, we propose that the positive feedback loop that Th2 cells exert on T cell polarization may involve the induction of Jagged 2 expression in DCs.

A simple stochastic lattice gas model for H1N1 pandemic. Application to the Italian epidemiological data.

We construct a very simple epidemic model for influenza spreading in an age-class-distributed population, by coupling a lattice gas model for the population dynamics with a SIR stochastic model for susceptible, infected and removed/immune individuals. We use as a test case the age-distributed Italian epidemiological data for the novel influenza A(H1N1). The most valuable features of this model are its country-independent and virus-independent structure (few demographic, social and virological data are used to fix some parameters), its large statistic due to a very short run-time machine, and its easy generalizability to include mitigation strategies. In spite of its simplicity, the model presented reproduces the epidemiological Italian data, with sensible predictions for the reproduction number and theoretically interesting results for the generation time distribution.

Pharmacokinetics of hydralazine, an antihypertensive and DNA-demethylating agent, using controlled-release formulations designed for use in dosing schedules based on the acetylator phenotype.

PURPOSE: The antihypertensive hydralazine has recently been repositioned as DNA demethylating for the epigenetic therapy of cancer. As the acetylator phenotype is the key determinant of its plasma levels, the dose of hydralazine needs to be adjusted for the acetylation status of patients. METHODS: The pharmacokinetics of orally administered hydralazine was evaluated in 26 healthy volunteers (13 slow and 13 fast acetylators) after a single dose of 182 mg administered as a controlled-release tablet. Plasma levels of hydralazine were analyzed in 85 cancer patients treated with this formulation at a dose of 83 mg/day and 182 mg/day for slow and fast acetylators, respectively. RESULTS: The C(max) and t(max) of hydralazine for fast acetylators were 208.4 ± 56.9 SD ng/ml and 2.8 ± 2.5 h, respectively. The corresponding results for slow acetylators were 470.4 ± 162.8 ng/ml, and 4.4 ± 3.1 h. Healthy volunteers who were fast acetylators had no clinically significant changes in blood pressure and heart rate or any other side-effect, however, slow acetylators had transient episodes of headache, tachycardia and faintness. Among 85 cancer patients that received either 182 mg or 83 mg of hydralazine daily, according to their acetylator status, the mean concentrations of hydralazine in plasma were 239.1 ng/ml and 259.2 ng/ml for fast and slow acetylators, respectively. These differences were not significantly different, p = 0.3868. CONCLUSIONS: The administration of dose-adjusted controlled-release hydralazine according to the acetylation status of cancer patients yields similar levels of hydralazine.

[Sub-tenon block for ocular globe anesthesia: a review]. / Anestesia subtenoniana en cirugia del globo ocular: revision.

Sub-Tenon anesthesia is an effective, well-tolerated technique for surgery in the anterior or posterior compartments of the eye. The advantages of this block are comparable to those of peribulbar and retrobulbar anesthesia and complications are minimal. Sub-Tenon anesthesia provides better analgesia than akinesia. Most studies suggest that sub-Tenon anesthesia is a good technique to choose, given that potential adverse effects are fewer than for other regional blocks and analgesia and akinesia are superior.

Headache as main reason for consultation to a hospital Emergency Department in Spain: a prospective study. / La cefalea como motivo principal de consulta a un servicio de urgencia hospitalaria en España: un estudio prospectivo.

INTRODUCTION: Headache is common in the general population and a frequent reason for medical consultation. OBJECTIVES: To describe the characteristics of patients attending the Emergency Department (ED) for headache. METHODS: A descriptive study with prospective collection of 100 consecutive patients over 15 years old who attended our ED due to headache as the main complaint. RESULTS: Headache accounted for 1,4% of ED visits. The most common age range is between 31 and 45 years and the majority of the patients are females (61%). We diagnosed 67 primary and 33 secondary headaches. The most frequent diagnosis was migraine, with 36% of cases. One out of 3 patients had a history of headache and 4 out of 5 consulted by their own decision. Only a small percentage of patients were admitted as inpatients (12%), and 3 out of 5 were referred to Primary Care. Complementary tests were performed on 84% of the patients. One CT scan was performed for every 3 patients. A total of 80% patients was correctly diagnosed by the ED physicians. CONCLUSIONS: Headache is a frequent complaint in the ED, where primary headaches are the most common with migraine being the most frequent reason for consultation. In our setting, there is a good screening and diagnosis of headaches, as well as an adequate use of the available resources in the ED for their diagnosis and management.

Epigenetic therapy and cisplatin chemoradiation in FIGO stage IIIB cervical cancer.

INTRODUCTION: This trial aimed to evaluate the safety and efficacy of epigenetic therapy associated with cisplatin chemoradiation in FIGO Stage IIIB patients. METHODS: Hydralazine containing either 182 mg for rapid-, or 83 mg for slow acetylators and magnesium valproate were administered at 30 mg/kg tid. Both drugs were taken until intracavitary therapy was finished. Pelvic external beam radiation and low-dose rate brachytherapy were administered at a total cumulative dose to point A of at least 85 Gy. Weekly cisplatin at 40 mg/m2 was delivered for six cycles. RESULTS: Twenty-two patients were included and 18 (82%) patients completed treatment. Mean dose to point A was 84.6 + 2.2. Median number of cisplatin cycles was 5.5 (range, 1-6). Brachytherapy was delayed for technical reasons; the mean overall treatment time was 11.8 weeks. Grade 3 anemia, leucopenia, neutropenia, and thrombocytopenia were observed in 9%, 45%, 45%, and 9% of patients, respectively. CONCLUSIONS: Hydralazine and valproate are well-tolerated and safe when administered with cisplatin chemoradiation. Unfortunately, the suboptimal administration of brachytherapy for technical reasons in this study, precluded assessing the efficacy of epigenetic therapy. However, the tolerability of this regimen administered concurrent to radiation needs to be further tested.

Emergence of complex behavior in gelling systems starting from simple behavior of single clusters.

A theoretical and numerically study of dynamical properties in the sol-gel transition is presented. In particular, the complex phenomenology observed experimentally and numerically in gelling systems is reproduced in the framework of percolation theory, under simple assumptions on the relaxation of single clusters. By neglecting the correlation between particles belonging to different clusters, the quantities of interest (such as the self intermediate scattering function, the dynamical susceptibility, the Van-Hove function, and the non-Gaussian parameter) are written as superposition of those due to single clusters. Connection between these behaviors and the critical exponents of percolation are given. The theoretical predictions are checked in a model for permanent gels, where bonds between monomers are described by a finitely extendable nonlinear elastic potential. The data obtained in the numerical simulations are in good agreement with the analytical predictions.

Dynamical heterogeneity in a model for permanent gels: different behavior of dynamical susceptibilities.

We present a systematic study of dynamical heterogeneity in a model for permanent gels upon approaching the gelation threshold. We find that the fluctuations of the self-intermediate scattering function are increasing functions of time, reaching a plateau whose value, at large length scales, coincides with the mean cluster size and diverges at the percolation threshold. Another measure of dynamical heterogeneities-i.e., the fluctuations of the self-overlap-displays instead a peak and decays to zero at long times. The peak, however, also scales as the mean cluster size. Arguments are given for this difference in the long-time behavior. We also find that the non-Gaussian parameter reaches a plateau in the long-time limit. The value of the plateau of the non-Gaussian parameter, which is connected to the fluctuations of diffusivity of clusters, increases with the volume fraction and remains finite at the percolation threshold.

Metabolic changes following conversion from an anticalcineurin-based therapy to an everolimus-based one: a single-center experience.

Everolimus (EVL), an antagonist of mammalian target of rapamycin, has been recently introduced into solid organ transplantation either associated with low dose of anticalcineurins (CNI) or replacing them in an attempt to avoid nephrotoxicity and chronic allograft nephropathy. Due to the molecular similarities with sirolimus, it has been expected that there would be the same incidence of metabolic changes and adverse events. We retrospectively studied kidney allograft recipients converted from CNI to EVL during a 12-month period. Patients received a standard dose of EVL starting at 1.5 mg/d and thereafter titrating to achieve trough levels in the range of 3 to 5 ng/mL. Patients achieved mean EVL trough levels of 5.2, 4.0 and 4.5 ng/mL at 1, 6, and 12 months, respectively. One year following conversion, the calculated creatinine clearance increased from 57 to 63 mL/min and proteinuria did not change. Fasting blood glucose levels decreased significantly following conversion to EVL. During the same time, no significant changes were observed in body weight, body mass index, albumin, cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, lipid-lowering medication requirements, blood magnesium, and uric acid. We concluded that EVL did not negatively influence various nutritional parameters.

Alemtuzumab induction in kidney transplantation: clinical results and impact on T-regulatory cells.

Alemtuzumab (ALT), a humanized monoclonal anti-CD52 antibody, was introduced in solid organ transplantation as an induction agent. ALT associated with anticalcineurins has provided a low incidence of acute rejection episodes (ARE) and potential tolerogenic properties. We analyzed the clinical outcomes and effects on peripheral Treg of renal transplant recipients treated with ALT. Six-month data on kidney alone or kidney combined with pancreas or liver patients treated with ALT and tacrolimus (TAC) in standard doses were compared with those on renal transplant recipients of similar demography who were not treated with ALT. We evaluated patient and graft survivals, ARE incidence, hematological parameters, renal function, adverse events, and CD4+CD25+FoxP3+ T cells in peripheral blood. Demographics of recipients, donors, and transplants were similar in both groups. Mean HLA mismatch was slightly greater among ALT-treated patients (3.5 vs 2.5). No combined transplantation was performed in the ALT-untreated group. Patient and graft survivals were 100% without rejection or serious infections in both groups. ALT-treated recipients showed anemia and leukopenia in 3 patients as well as severe lymphopenia in 5 recipients, who partially recovered on day 90. Final mean plasma creatinine was 1.4 mg/dL, while calculated creatinine clearance was approximately 65 mL/min in both groups. Mean Treg cell percentage was higher among ALT-treated recipients than the comparative group or healthy controls (P < .05). In conclusion, renal transplantation results obtained using ALT with rigorous immunosuppressive therapy were excellent; serious adverse events and acute rejection were absent. The effect of the increased proportion of Treg cells must be evaluated with longer observation.

Conversion from a calcineurin inhibitor-based immunosuppressive regimen to everolimus in renal transplant recipients: effect on renal function and proteinuria.

New immunosuppressive agents are being actively researched to avoid complications of chronic allograft nephropathy (CAN), calcineurin inhibitor (CNI) nephrotoxicity, and posttransplantation cancer. The family of mTOR inhibitors offers a unique immunosuppressive opportunity to avoid CNI toxicity and reduce the incidence of malignancy. Nevertheless, increasing data have demonstrated that sirolimus (SRL), the first mTOR introduced in the treatment of solid organ transplant recipients, induces proteinuria, an adverse event that could produce deterioration of long-term renal function. In this short-term study of patients followed for 1 to 16 months, we examined changes in renal function and proteinuria among renal transplant recipients converted from a CNI-based regimen to an everolimus (EVL)-based one, a recently introduced mTOR inhibitor. Our data showed that renal function can be optimized after conversion to EVL by up to 42% in recipients showing CAN grade 1 or 2, or CNI nephrotoxicity. Importantly, patients who improved their creatinine clearance did not show increased proteinuria measured in a voided specimen as the ratio of urinary protein and creatinine concentration (P/C). These results, if confirmed with long-term follow-up and a larger number of patients, would allow us to consider EVL as a promising agent for maintenance immunosuppressive regimens in kidney transplantation.

[A new reality in the field of penitentiary medicine: the Complex Protected Medicine Structure of the ASL Rome B. Initial report of our working sanitary care]. / Una nuova realtà nell'ambito della medicina penitenziaria: la struttura Complessa di Medicina Protetta della ASL Roma B. Resoconto iniziale dell'attività assistenziale svolta.

BACKGROUND: The continuous demand for sanitary care from the imprisoned patients to the general public hospitals has carried to the creation of dedicated structures like the Complex Protected Medicine Structure (SCMP) at the Hospital Sandro Pertini of Rome. Aim of this study was to present the preliminary epidemiologic-clinical data gathered from July 26th 2006 to March 1st, 2006 in our SCMP. MATERIAL AND METHODS: This study is based on an observational retrospective protocol. Patients characteristics included sex, age, alcohol, smoke and drug abuse, serology for viral hepatitis and AIDS, and treatment with benzodiazepine. RESULTS: The records of 200 consecutive patients (18 F, 182 M; median age 46 years; range: 20-84 years), attending our Unit were analyzed retrospectively. Out of these patients, 40 (20%) were foreigners, 43 (21.5%) alcohol abusers, and 141 (70.5%) smokers. Forty-eight patients (24%) had a recent or last history of drug addiction (heroin and/or cocaine). The HIV antibodies have been assayed only on 28 patients, turning out positive in 10 (35.71%). Twenty-five patients were positive for HBsAg (12.5%) and 57 (28.5%) for HCV. A consumption of benzodiazepine (BZD) was present in 71 (35.5%) of the patients. CONCLUSIONS: The essential elements of medical attendance of our structure are described in the article. We present the initial report of our working experience. The present data are in keeping with previous studies of the literature. We hope that our experience coul be extended to all italian Countries.

Diffuse abdominal hydatidosis: role of magnetic resonance imaging.

Hydatid cyst disease still is a serious public-health problem in endemic areas. It is a tissue infestation caused by the larval stage of a parasite, Echinococcus granulosus. Although liver and lung are the most commonly involved organs, hydatid disease can occur in all viscera and soft tissues. In 10% of cases, hydatid disease arises in the viscera: mainly in the spleen (0.9-8%), but also in kidney, bone, heart and peritoneal cavity (0.5-5%). Other rare locations such as muscles have been described in less than 1% of cases of hydatid disease. We report magnetic resonance imaging findings of a case of diffuse abdominal hydatidosis.

Comparative life cycle assessment study on environmental impact of oil production from micro-algae and terrestrial oilseed crops.

In this study the LCA methodology is applied in order to satisfy two goals: i) to evaluate the hot spots in site-specific production chain of biodiesel from terrestrial and micro-algae feedstock; ii) to compare quantitatively, utilizing primary data, the impacts of the first generation in respect to the third generation bio-fuels. Results show that micro-algae are neither competitive yet with traditional oil crops nor with fossil fuel. The use of renewable technologies as photovoltaics and biogas self production might increase the competitiveness of micro-algae oil. Further investigations are however necessary to optimize their production chain and to increase the added value of co-products.