Different patterns of HIV-1 DNA after therapy discontinuation

Background: By persisting in infected cells for a long period of time, proviral HIV-1 DNA can represent an alternative viral marker to RNA viral load during the follow-up of HIV-1 infected individuals. In the present study sequential blood samples of 10 patients under antiretroviral treatment from 1997 with two NRTIs, who refused to continue any antiviral regimen, were analyzed for 16-24 weeks to study the possible relationship between DNA and RNA viral load. Methods: The amount of proviral DNA was quantified by SYBR green real-time PCR in peripheral blood mononuclear cells from a selected group of ten patients with different levels of plasmatic viremia (RNA viral load). Results: Variable levels of proviral DNA were found without any significant correlation between proviral load and plasma HIV-1 RNA levels. Results obtained showed an increase or a rebound in viral DNA in most patients, suggesting that the absence of therapy reflects an increase and/or a persistence of cells containing viral DNA. Conclusion: Even though plasma HIV RNA levels remain the basic parameter to monitor the intensity of viral replication, the results obtained seem to indicate that DNA levels could represent an adjunct prognostic marker in monitoring HIV-1 infected subjects

Modeling Human Muscular Dystrophies in Zebrafish: Mutant Lines, Transgenic Fluorescent Biosensors, and Phenotyping Assays

: Muscular dystrophies (MDs) are a heterogeneous group of myopathies characterized by progressive muscle weakness leading to death from heart or respiratory failure. MDs are caused by mutations in genes involved in both the development and organization of muscle fibers. Several animal models harboring mutations in MD-associated genes have been developed so far. Together with rodents, the zebrafish is one of the most popular animal models used to reproduce MDs because of the high level of sequence homology with the human genome and its genetic manipulability. This review describes the most important zebrafish mutant models of MD and the most advanced tools used to generate and characterize all these valuable transgenic lines. Zebrafish models of MDs have been generated by introducing mutations to muscle-specific genes with different genetic techniques, such as (i) N-ethyl-N-nitrosourea (ENU) treatment, (ii) the injection of specific morpholino, (iii) tol2-based transgenesis, (iv) TALEN, (v) and CRISPR/Cas9 technology. All these models are extensively used either to study muscle development and function or understand the pathogenetic mechanisms of MDs. Several tools have also been developed to characterize these zebrafish models by checking (i) motor behavior, (ii) muscle fiber structure, (iii) oxidative stress, and (iv) mitochondrial function and dynamics. Further, living biosensor models, based on the expression of fluorescent reporter proteins under the control of muscle-specific promoters or responsive elements, have been revealed to be powerful tools to follow molecular dynamics at the level of a single muscle fiber. Thus, zebrafish models of MDs can also be a powerful tool to search for new drugs or gene therapies able to block or slow down disease progression

Growth, longevity, and mortality of the Largemouth Bass Micropterus salmoides (Lacépède, 1802) in a mediterranean lake (Rome, Italy)

The analysis of the structure and dynamics of an invasive population of the Largemouth Bass (Micropterus salmoides Lacépède, 1802) in a Mediterranean wetland was the main goal of this study. Demographic potentialities were described by means of the Von Bertalanffy parameters, the latter obtained using length-at-age data. The mean values of the observed four age classes (recognized by the scale reading) were analysed by the Gulland & Holt analytical method, in order to calculate the curvature parameter (k = 0.49) and the asymptotic length (L∞ = 31). The Von Bertalanffy parameters were not considered to be affected by sexual dimorphism since no between-sexes differences were found in size and population structure. Besides the two previous parameters, we computed longevity of 5 years, with only a limited number of Largemouth Bass reaching this age (due to a high fishing mortality rate). Structure and dynamics of this population seem to reveal its stability and spreading potential, as an expression of the favourable ecological conditions encountered in the study area. Growth parameters reflect the acclimation capability of M. salmoides in a non-native habitat and show the ecological plasticity of this species, capable of colonizing many temperate and warm habitats worldwide, Mediterranean wetlands includedCroissance, longévité et mortalité de l'Achigan à grande bouche Micropterus salmoides (Lacépède, 1802) dans un lac méditerranéen (Rome, Italie).- L'objectif de notre étude est l'analyse de la structure et de la dynamique d'une population invasive de l'Achigan à grande bouche (Micropterus salmoides Lacépède, 1802) dans un lac méditerranéen. La croissance a été décrite par les paramètres de Von Bertalanffy, eux-mêmes obtenus en utilisant les données « taille à l'âge ». Les valeurs moyennes des quatre classes d'âge observées (identifiées par la lecture des écailles) ont été analysées par la méthode analytique de Gulland & Holt, dans le but de calculer le paramètre de courbure (k = 0.49) et la longueur asymptotique (L∞ = 31). Les paramètres de Von Bertalanffy ne semblent pas liés au dimorphisme sexuel car nous n'avons pas trouvé de différences dans la taille ou dans la structure de la population entre les deux sexes. La longévité observée est de cinq ans, mais peu de spécimens atteignent cet âge, du fait d'un taux élevé de mortalité liée à la pêche. La structure et la dynamique de cette population semblent révéler sa stabilité et sa capacité d'expansion, expressions des conditions favorables rencontrées par l'espèce sur le site d'étude. Les paramètres de croissance reflètent les capacités d'acclimatation de Micropterus salmoides dans un habitat qui n'est pas celui d'origine et montrent la plasticité de l'espèce, capable de coloniser de nombreux habitats tempérés et chauds à travers le monde et notamment les zones humides méditerranéennes

Prophylactic mesh augmentation after laparotomy for elective and emergency surgery: meta-analysis

Background: Incisional hernia is a common short- and long-term complication of laparotomy and can lead to significant morbidity. The aim of this systematic review and meta-analysis is to provide an up-to-date overview of the laparotomy closure method in elective and emergency settings with the prophylactic mesh augmentation technique. Methods: The Scopus, PubMed, and Web of Science databases were screened without time restrictions up to 21 June 2022 using the keywords 'laparotomy closure', 'mesh', 'mesh positioning', and 'prophylactic mesh', and including medical subject headings terms. Only RCTs reporting the incidence of incisional hernia and other wound complications after elective or emergency midline laparotomy, where patients were treated with prophylactic mesh augmentation or without mesh positioning, were included. The primary endpoint was to explore the risk of incisional hernia at different follow-up time points. The secondary endpoint was the risk of wound complications. The risk of bias for individual studies was assessed according to the Revised Cochrane risk-of-bias tools for randomized trials. Results: Eighteen RCTs, including 2659 patients, were retrieved. A reduction in the risk of incisional hernia at every time point was highlighted in the prophylactic mesh augmentation group (1 year, risk ratio 0.31, P = 0.0011; 2 years, risk ratio 0.44, P < 0.0001; 3 years, risk ratio 0.38, P = 0.0026; 4 years, risk ratio 0.38, P = 0.0257). An increased risk of wound complications was highlighted for patients undergoing mesh augmentation, although this was not significant. Conclusions: Midline laparotomy closure with prophylactic mesh augmentation can be considered safe and effective in reducing the incidence of incisional hernia. Further trials are needed to identify the ideal type of mesh and technique for mesh positioning, but surgeons should consider prophylactic mesh augmentation to decrease incisional hernia rate, especially in high-risk patients for fascial dehiscence and even in emergency settings

The Cost and the Use of Serials in Italian Astronomical Libraries

We present the results of a two year study on the nature, cost and use of periodicals in Italian Astronomical Libraries (IALs). In the year 2000, IALs spent two-thirds of the budget (∼440.000 Euros) for serial subscriptions. This fact provided an incentive for the 12 Observatory librarians to investigate the potential for developing a consortium. In so doing, we first analysed characteristics of our individual collections and then evaluated the cost of on-line access as opposed to paper subscriptions. Then we studied the relationship between cost and serial usage

How to identify intersegmental planes in performing sublobar anatomical resections

Pulmonary segmentectomy is a common surgical procedure in thoracic surgery nowadays. Though this technique helps preventing postoperative pulmonary function loss, potential challenges are the management of air leaks and the difficulty of palpating ground-glass components during surgery, as well as how to deal with the intersegmental planes. Several techniques have been proposed for the identification and treatment of the intersegmental planes during sublobar anatomical resections: this review focuses on preoperative planning and workup and intraoperative strategies. Three-dimensional computed tomography bronchography and angiography (3D-CTBA), virtual-assisted mapping (VAL-MAP) using bronchoscopy multi-spot dye marking and three-dimensional computed tomography (3D-CT) are preoperative tools that may facilitate the planning of operation. Inflation-deflation techniques, infrared-fluorescence-enhanced method combined with bronchial and intravenous injection of indocyanine green (ICG) and near-infrared fluorescence (NIF) mapping with ICG have been described as intraoperative strategies to identify the intersegmental plane. The treatment and section of the intersegmental planes is mainly accomplished by stapler and electrocautery or energy devices. The use of staplers reduces postoperative air leaks, bleeding risks and operative time but seems to reduce preserved lung volume, compromising adjacent lung expansion; in addition, higher costs and sometimes non-adequate oncological margins, being a non-anatomical technique have been described. The electrocautery and energy devices allow for a more anatomical and precise dissection maintaining safe oncological margins, with a better lung expansion and so an increased postoperative lung function. Time consuming procedure and frequent requirement of aero-haemostatic tools to treat air and blood leaks are the main drawbacks. In conclusion, there are several methods to identify and treat the intersegmental planes but there are no significant differences between the different tools, therefore the use of one technique rather than another depends overall on surgeon’s preference and the location of the segment

GDF5 regulates TGFß-dependent angiogenesis in breast carcinoma MCF-7 cells: in vitro and in vivo control by anti-TGFß peptides

BACKGROUND: TGFß overproduction in cancer cells is one of the main characteristics of late tumor progression being implicated in metastasis, tumor growth, angiogenesis and immune response. We investigated the therapeutic efficacy of anti-TGFß peptides in the control of angiogenesis elicited by conditional over-expression of TGFß. METHODS: We have inserted in human MCF7 mammary-cancer cells a mutated TGFß gene in a tetracycline-repressible vector to obtain conditional expression of mature TGFß upon transient transfection, evaluated the signaling pathways involved in TGFß-dependent endothelial cells activation and the efficacy of anti-TGFß peptides in the control of MCF7-TGFß-dependent angiogenesis. RESULTS: TGFß over-expression induced in MCF7 several markers of the epithelial-to-mesenchymal transition. Conditioned-medium of TGFß-transfected MCF7 stimulated angiogenesis in vivo and in vitro by subsequent activation of SMAD2/3 and SMAD1/5 signaling in endothelial cells, as well as SMAD4 nuclear translocation, resulting in over-expression of the pro-angiogenic growth and differentiation factor-5 (GDF5). Inhibition or silencing of GDF5 in TGFß-stimulated EC resulted in impairment of GDF5 expression and of TGFß-dependent urokinase-plasminogen activator receptor (uPAR) overproduction, leading to angiogenesis impairment. Two different TGFß antagonist peptides inhibited all the angiogenesis-related properties elicited in EC by exogenous and conditionally-expressed TGFß in vivo and in vitro, including SMAD1/5 phosphorylation, SMAD4 nuclear translocation, GDF5 and uPAR overexpression. Antagonist peptides and anti-GDF5 antibodies efficiently inhibited in vitro and in vivo angiogenesis. CONCLUSIONS: TGFß produced by breast cancer cells induces in endothelial cells expression of GDF5, which in turn stimulates angiogenesis both in vitro and in vivo. Angiogenesis activation is rapid and the involved mechanism is totally opposed to the old and controversial dogma about the AKL5/ALK1 balance. The GDF-dependent pro-angiogenic effects of TGFß are controlled by anti-TGFß peptides and anti-GDF5 antibodies, providing a basis to develop targeted clinical studies

Production of Composite Zinc Oxide–Polylactic Acid Radiopaque Filaments for Fused Deposition Modeling: First Stage of a Feasibility Study

Three-dimensional printing technologies are becoming increasingly attractive for their versatility; the geometrical customizability and manageability of the final product properties are the key points. This work aims to assess the feasibility of producing radiopaque filaments for fused deposition modeling (FDM), a 3D printing technology, starting with zinc oxide (ZnO) and polylactic acid (PLA) as the raw materials. Indeed, ZnO and PLA are promising materials due to their non-toxic and biocompatible nature. Pellets of PLA and ZnO in the form of nanoparticles were mixed together using ethanol; this homogenous mixture was processed by a commercial extruder, optimizing the process parameters for obtaining mechanically stable samples. Scanning electron microscopy analyses were used to assess, in the extruded samples, the homogenous distribution of the ZnO in the PLA matrix. Moreover, X-ray microtomography revealed a certain homogenous radiopacity; this imaging technique also confirmed the correct distribution of the ZnO in the PLA matrix. Thus, our tests showed that mechanically stable radiopaque filaments, ready for FDM systems, were obtained by homogenously loading the PLA with a maximum ZnO content of 6.5% wt. (nominal). This study produced multiple outcomes. We demonstrated the feasibility of producing radiopaque filaments for additive manufacturing using safe materials. Moreover, each phase of the process is cost-effective and green-oriented; in fact, the homogenous mixture of PLA and ZnO requires only a small amount of ethanol, which evaporates in minutes without any temperature adjustment. Finally, both the extruding and the FDM technologies are the most accessible systems for the additive manufacturing commercial apparatuses