Functionality and opposite roles of two interleukin 4 haplotypes in immune cells.

Cytokines expression can be influenced by polymorphisms in their respective coding genes. We associated the CTI/TTD haplotype (Hap-1) and TCI/CCI haplotype (Hap-2) in the IL4 gene formed by the -590, +33 and variable number of tandem repeat polymorphisms with the severity of chronic periodontitis in humans. The functionality of these IL4 haplotypes in the response of immune cells to phorbol 12-myristate 13-acetate (PMA) with Ionomycin and IL-1ß (as inflammatory stimuli) was evaluated. Gene expression (quantitative real-time PCR), profile of secreted cytokines (multiplex) and phenotypic polarization of T cells (flow cytometry) were the outcomes assessed. Green fluorescent protein reporter plasmid constructs containing specific IL4 haplotype were transiently transfected into JM cells to assess the influence of the individual haplotypes on promoter activity. In response to inflammatory stimuli the immune cells from Hap-1 haplotype had increased expression of anti-inflammatory IL4; conversely, the Hap-2 haplotype showed higher levels of pro-inflammatory cytokines. The haplotype CTI proved to be the most important for the regulation of IL4 promoter, regardless of the nature of the inflammatory stimulation; whereas the polymorphism in the promoter region had the least functional effect. In conclusion, IL4 haplotypes studied are functional and trigger opposite immune responses: anti-inflammatory (Hap-1) and pro-inflammatory (Hap-2). In addition, we identified the CTI haplotype as the main responsible for the regulation of IL4 transcriptional activity.

A new surgical positioning system for robotic assisted minimally invasive spine surgery and transpedicular approach to the disc.

Minimally Invasive Spine Surgery (MISS) procedures for the treatment of spinal pathologies have experienced exponential growth due to improved techniques and decreased trauma to the patient. Several MISS procedures that require the use of a trans-pedicular cannula as a guiding tool for pedicle screw placement, delivery of biomaterials to the vertebral body or injection of biologics to the disc space have been described. Although these are clear advantages of MISS, the limited dissection and exposure may reduce the accuracy and stability of operation and make spine surgeons rely heavily on intraoperative fluoroscopy, raising concerns over the level of radiation exposure. Robot-assisted minimal invasive surgery has aroused more attention for its high precision and stability, minimizing risks of damage to neurovascular structures and diminishing harmful exposure to ionizing radiation. The aim of this paper is to describe and characterize a new surgical positioning system for for robotic assisted MISS. The system is conceived to be integrated in a surgical platform capable of supporting the surgeon in a new procedure to treat degenerative intervertebral disc disease. For this purpose, it is necessary to orientate a cannula in order to guide the bone drill along a planned route, to access the intervertebral disc through the pedicle and endplate. In particular, we describe a mechanism that percutaneously guides a cannula towards the intervertebral disc based on the acquisition of few fluoroscopic images. The design of the positioning system, with its features and constrains imposed by the presence of instrumentation and medical staff in the operating room, as well as the software for trajectory planning during surgery, are here described.

[Cumulative exposure to polychlorodibenzo-p-dioxins (PCDDS), polychlorodibenzofurans (PCDFS) and polychlorobiphenyls (PCB) in the metallurgic workers and the general population in Brescia province, northern Italy]. / Esposizione cumulativa a policlorodibenzodiossine (PCDD), policlorodibenzofurani (PCDF) e policlorobifenili (PCB) in lavoratori metallurgici e nella popolazione generale della provincia di Brescia.

High temperature metallurgical processes may significantly contribute to the release of polychlorodibenzo-p-dioxins (PCDDs) and polychlorodibenzofurans (PCDFs) into the environment. Aim of this study was to evaluate the exposure to PCDDs, PCDFs, dioxin-like PCBs (DL-PCBs) and non dioxin-like PCBs (NDL-PCBs) in a sample of 300 workers of metallurgical plants operating in the Brescia area and in a control group of 113 men. Serum samples were grouped in pools of 10 and analyzed for their concentration of PCDDs and PCDFs, DL- and NDL-PCBs. Metallurgical workers show serum concentrations of NDL-PCBs similar to those observed in non-occupationally exposed living in Brescia, except in the case of some congeners, such as PCBs 28, 52, 101 which result to be significantly higher in metallurgical workers. Their concentrations range of "dioxins" was of 32.5 - 84.8 pgTEQ g(-1) lb. No significant difference was found between metallurgical workers and the control group on the whole, but the difference became significant when comparing metallurgical workers with the group of the 47 subjects living in remote non industrialised areas. In conclusion, concentrations of total PCBs observed in metallurgy workers are comparable with those observed in the control group, with the exception of a few, more volatile, congeners which resulted to be higher in metallurgy workers. Concentrations of PCDDs and PCDFs showed a range of values similar to that observed in the general population of Brescia not professionally exposed. However, concentrations of PCDDs and PCDFs observed in subjects resident in "remote" areas were significantly lower than those assessed in metallurgy workers and subjects living in the town of Brescia. On the whole, men living in the town of Brescia show levels of PCBs, PCDDs and PCDFs higher than those observed in other groups of the Italian general population, this reflecting the intense industrial activity of the town.

Advances on the immunotoxicity of outdoor particulate matter: A focus on physical and chemical properties and respiratory defence mechanisms.

Particulate matter (PM) is acknowledged to have multiple detrimental effects on human health. In this review, we report literature results on the possible link between outdoor PM and health outcomes with a focus on pulmonary infections and the mechanisms responsible for observed negative effects. PM physical and chemical properties, such as size and chemical composition, as well as major emission sources are described for a more comprehensive view about the role played by atmospheric PM in the observed adverse health effects; to this aim, major processes leading to the deposition of PM in the respiratory tract and how this can pave the way to the onset of pathologies are also presented. From the literature works here reviewed, two ways in which PM can threaten human health promoting respiratory infectious diseases are mostly taken into account. The first pathway is related to an enhanced susceptibility and here we will also report on molecular mechanisms in the lung immune system responsible for the augmented susceptibility to pathogens, such as the damage of mechanical defensive barriers, the alteration of the innate immune response, and the generation of oxidative stress. The second one deals with the relationship between infectious agents and PM; here we recall that viruses and bacteria (BioPM) are themselves part of atmospheric PM and are collected during sampling together with particles of different origin; so, data should be analysed with caution in order to avoid any false cause-effect relation. To face these issues a multidisciplinary approach is mandatory as also evident from the ongoing research about the mechanisms hypothesized for the SARS-CoV-2 airborne spreading, which is still controversial and claims for further investigation. Therefore, we preferred not to include papers dealing with SARS-CoV-2.

Consistent determination of the heating rate of light-absorbing aerosol using wavelength- and time-dependent Aethalometer multiple-scattering correction.

Accurate and temporally consistent measurements of light absorbing aerosol (LAA) heating rate (HR) and of its source apportionment (fossil-fuel, FF; biomass-burning, BB) and speciation (black and brown Carbon; BC, BrC) are needed to evaluate LAA short-term climate forcing. For this purpose, wavelength- and time-dependent accurate LAA absorption coefficients are required. HR was experimentally determined and apportioned (sources/species) in the EMEP/ACTRIS/COLOSSAL-2018 winter campaign in Milan (urban-background site). Two Aethalometers (AE31/AE33) were installed together with a MAAP, CPC, OPC, a low volume sampler (PM2.5) and radiation instruments. AE31/AE33 multiple-scattering correction factors (C) were determined using two reference systems for the absorption coefficient: 1) 5-wavelength PP_UniMI with low time resolution (12 h, applied to PM2.5 samples); 2) timely-resolved MAAP data at a single wavelength. Using wavelength- and time-independent C values for the AE31 and AE33 obtained with the same reference device, the total HR showed a consistency (i.e. reproducibility) with average values comparable at 95% probability. However, if different reference devices/approaches are used, i.e. MAAP is chosen as reference instead of a PP_UniMI, the HR can be overestimated by 23-30% factor (by both AE31/AE33). This became more evident focusing on HR apportionment: AE33 data (corrected by a wavelength- and time-independent C) showed higher HRFF (+24 ± 1%) and higher HRBC (+10 ± 1%) than that of AE31. Conversely, HRBB and HRBrC were -28 ± 1% and -29 ± 1% lower for AE33 compared to AE31. These inconsistencies were overcome by introducing a wavelength-dependent Cλ for both AE31 and AE33, or using multi-wavelength apportionment methods, highlighting the need for further studies on the influence of wavelength corrections for HR determination. Finally, the temporally-resolved determination of C resulted in a diurnal cycle of the HR not statistically different whatever the source- speciation- apportionment used.

Biomonitoring of the adult population living near the waste incinerator of Turin: Serum concentrations of PCDDs, PCDFs, and PCBs after three years from the plant start-up.

In September 2013 a waste-to-energy (WTE) incinerator located in the Turin area (Piedmont, Northern Italy) started to produce energy by the incineration of municipal solid wastes. The plant, one of the largest WTE incinerator in Europe, burns up to 490,000 tons of waste per year. A health surveillance program was implemented in order to evaluate the potential health effects on the population living near the plant. This program included a biomonitoring study aimed at assessing levels of several environmental contaminants including, among others, PCDDs, PCDFs, and PCBs. Before the WTE incinerator start-up (T0), a group of 85 subjects (41 "exposed" and 44 "not exposed" subjects) was randomly selected for enrollment by the local health units among individuals aged 36-50 years who had been living in the same area for at least five years prior to the study. Subjects were balanced by exposure area, sex and five-year age classes. As from the study design, the same cohort was re-evaluated after three years of incinerator activity (T2). A parallel study was conducted on a group of 12 farmers living and/or working in farms located in an area in the range of 5 km around the incinerator. Results of this study did not evidence any impact of the WTE plant on human exposure to PCDDs, PCDFs, and PCBs. In fact, no significant differences were found in the concentrations of PCDDs + PCDFs, DL-PCBs, and NDL-PCBs measured in the population group residing near the plant after three years of activity (T2) with respect to the control group. A significant decrease of serum concentrations of all the analytes was observed at T2 in both groups compared to T0. Serum concentrations of PCDDs, PCDFs, and PCBs in the group of farmers were higher than those observed in the adult population under study.

Functional significance of eIF5A and its hypusine modification in eukaryotes.

The unusual basic amino acid, hypusine [N(epsilon)-(4-amino-2-hydroxybutyl)-lysine], is a modified lysine with the addition of the 4-aminobutyl moiety from the polyamine spermidine. This naturally occurring amino acid is a product of a unique posttranslational modification that occurs in only one cellular protein, eukaryotic translation initiation factor 5A (eIF5A, eIF-5A). Hypusine is synthesized exclusively in this protein by two sequential enzymatic steps involving deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). The deoxyhypusine/hypusine synthetic pathway has evolved in archaea and eukaryotes, and eIF5A, DHS and DOHH are highly conserved suggesting a vital cellular function of eIF5A. Gene disruption and mutation studies in yeast and higher eukaryotes have provided valuable information on the essential nature of eIF5A and the deoxyhypusine/hypusine modification in cell growth and in protein synthesis. In view of the extraordinary specificity and functional significance of hypusine-containing eIF5A in mammalian cell proliferation, eIF5A and the hypusine biosynthetic enzymes are novel potential targets for intervention in aberrant cell proliferation.

Quorum sensing inhibition activity of garlic extract and p-coumaric acid.

AIMS: The goal of this work was to investigate the influence of DMSO, garlic extract and p-coumaric acid on bacterial quorum sensing (QS). METHODS AND RESULTS: The decreases in the QS responses of QS reporter strains Escherichia coli pSB401 and pSB536, Agrobacterium tumefaciens NTL4, Chromobacterium violaceum 5999 and wt 494, Pseudomonas putida IsoF/gfp and environmental Pseudomonas chlororaphis were quantified in relation to growth inhibitory effects. DMSO showed no significant QS-specific effects on the strains tested even at close-to-lethal concentrations. Garlic extracts antagonized the activity of QS receptors LuxR, AhyR and TraR, but were toxic at higher concentrations. P-coumaric acid fully inhibited QS responses of 5999, NTL4 and P. chlororaphis, with no influence on cell viability. CONCLUSIONS: The quorum sensing inhibition activity of garlic was extended to novel receptors, and p-coumaric acid was found to possess previously undescribed QS antagonist properties. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that p-coumaric acid might act as QS inhibitor. Further studies are required to understand its role in the regulation of QS and investigate structurally related compounds.

Rupture of the lateral lobe of the biceps brachii tendon in an Arabian horse.

Rupture of the lateral lobe of the proximal tendon of the biceps brachii muscle was diagnosed in an Arabian horse. To the authors' knowledge, this is the only report of this condition in horses. Although clinical signs were helpful in the identification of the location of disease, ultrasonographic examination was a more definitive and non- invasive means of diagnosing the pathological condition. Bursoscopic examination of the intertubercular bursa was also useful in obtaining confirmation of the diagnosis, and for debridement and lavage of the bursa.

Differential expression of RNA exosome subunits in the amphibian Lithobates catesbeianus during reproductive and non-reproductive periods.

OBJECTIVE: The RNA exosome is an evolutionarily conserved 3'-5' exoribonucleolytic protein complex involved in processing and degradation of different classes of nuclear and cytoplasmic RNAs, and, therefore, important for the posttranscriptional control of gene expression. Despite the extensive in vivo functional studies and the structural data on the RNA exosome, few studies have been performed on the localization and expression of exosome subunits during gametogenesis, process during which gene expression is largely controlled at the posttranscriptional level. RESULTS: We report the identification of exosome subunits in Lithobates catesbeianus and analysis of the differential subcellular localization of RNA exosome core and catalytic subunits in testis cells. In addition, we show seasonal differences in the expression levels of four exosome subunits in different organs. In addition to being part of the RNA exosome complex, its subunits might participate independently of the complex in the control of gene expression during seasonal variation in bullfrog tissues. These results may be relevant for other eukaryotic species.

Lung transplantation for cystic fibrosis in Italy.

Lung transplantation (LT) is the only effective form of therapy for cystic fibrosis (CF) associated with end-stage pulmonary failure. In Italy, the management of CF is regulated by national law, which has instituted regional centers for care and follow-up of all CF patients. LT has been performed since 1992 in only nine LT certified centers. The structured national organization has led to a unified database for LT for CF. As of December 2006, 197 bilateral LT (96 male and 94 female patients; 7 retransplants) have been performed. Of these, four had also liver or heart and liver transplantation, and three are long-term survivors. Overall median survival is 7 years. Mean age at transplantation is 26.5 years, and the mortality on the waiting list is 33.6%. Patients listed for transplant either received a suitable donor within a mean of 10 months or died within a mean of 5.5 months. The most frequent cause of death is bronchiolitis obliterans syndrome (BOS). Our nationwide database indicates the excellent results obtained by LT in FC. Still, mortality on the waiting list remains a challenge and long-term outcome is limited by BOS.

Assessment of light extinction at a European polluted urban area during wintertime: Impact of PM1 composition and sources.

In this paper, results from receptor modelling performed on a well-characterised PM1 dataset were combined to chemical light extinction data (bext) with the aim of assessing the impact of different PM1 components and sources on light extinction and visibility at a European polluted urban area. It is noteworthy that, at the state of the art, there are still very few papers estimating the impact of different emission sources on light extinction as we present here, although being among the major environmental challenges at many polluted areas. Following the concept of the well-known IMPROVE algorithm, here a tailored site-specific approach (recently developed by our group) was applied to assess chemical light extinction due to PM1 components and major sources. PM1 samples collected separately during daytime and nighttime at the urban area of Milan (Italy) were chemically characterised for elements, major ions, elemental and organic carbon, and levoglucosan. Chemical light extinction was estimated and results showed that at the investigated urban site it is heavily impacted by ammonium nitrate and organic matter. Receptor modelling (i.e. Positive Matrix Factorization, EPA-PMF 5.0) was effective to obtain source apportionment; the most reliable solution was found with 7 factors which were tentatively assigned to nitrates, sulphates, wood burning, traffic, industry, fine dust, and a Pb-rich source. The apportionment of aerosol light extinction (bext,aer) according to resolved sources showed that considering all samples together nitrate contributed at most (on average 41.6%), followed by sulphate, traffic, and wood burning accounting for 18.3%, 17.8% and 12.4%, respectively.

The performance of poly-epsilon-caprolactone scaffolds in a rabbit femur model with and without autologous stromal cells and BMP4.

The ability of a cellular construct to guide and promote tissue repair strongly relies on three components, namely, cell, scaffold and growth factors. We aimed to investigate the osteopromotive properties of cellular constructs composed of poly-epsilon-caprolactone (PCL) and rabbit bone marrow stromal cells (BMSCs), or BMSCs engineered to express bone morphogenetic protein 4 (BMP4). Highly porous biodegradable PCL scaffolds were obtained via phase inversion/salt leaching technique. BMSCs and transfected BMSCs were seeded within the scaffolds by using an alternate flow perfusion system and implanted into non-critical size defects in New Zealand rabbit femurs. In vivo biocompatibility, osteogenic and angiogenic effects induced by the presence of scaffolds were assessed by histology and histomorphometry of the femurs, retrieved 4 and 8 weeks after surgery. PCL without cells showed scarce bone formation at the scaffold-bone interface (29% bone/implant contact and 62% fibrous tissue/implant contact) and scarce PCL resorption (16%). Conversely, PCL seeded with autologous BMSCs stimulated new tissue formation into the macropores of the implant (20%) and neo-tissue vascularization. Finally, the BMP4-expressing BMSCs strongly favoured osteoinductivity of cellular constructs, as demonstrated by a more extensive bone/scaffold contact.

Is there a role for eIF5A in translation?

The putative translation factor eIF5A is essential for cell viability and is highly conserved from archaebacteria to mammals. This factor is the only cellular protein that undergoes an essential posttranslational modification dependent on the polyamine spermidine, called hypusination. This review focuses on the functional characterization of eIF5A. Although this protein was originally identified as a translation initiation factor, subsequent studies did not support a role for eIF5A in general translation initiation. eIF5A has also been implicated in nuclear export of HIV-1 Rev and mRNA decay, but these findings are controversial in the literature and may reflect secondary effects of eIF-5A function. Next, the involvement of eIF5A and hypusination in the control of the cell cycle and proliferation in various organisms is reviewed. Finally, recent evidence in favor of reconsidering the role of eIF5A as a translation factor is discussed. Future studies may reveal the specific mechanism by which eIF5A affects protein synthesis.

Use of a synthetic lethal screen to identify genes related to TIF51A in Saccharomyces cerevisiae.

The putative eukaryotic translation initiation factor 5A (eIF5A) is an essential protein for cell viability and the only cellular protein known to contain the unusual amino acid residue hypusine. eIF5A has been implicated in translation initiation, cell proliferation, nucleocytoplasmic transport, mRNA decay, and actin polarization, but the precise biological function of this protein is not clear. However, eIF5A was recently shown to be directly involved with the translational machinery. A screen for synthetic lethal mutations was carried out with one of the temperature-sensitive alleles of TIF51A (tif51A-3) to identify factors that functionally interact with eIF5A and revealed the essential gene YPT1. This gene encodes a small GTPase, a member of the rab family involved with secretion, acting in the vesicular trafficking between endoplasmatic reticulum and the Golgi. Thus, the synthetic lethality between TIF51A and YPT1 may reveal the connection between translation and the polarized distribution of membrane components, suggesting that these proteins work together in the cell to guarantee proper protein synthesis and secretion necessary for correct bud formation during G1/S transition. Future studies will investigate the functional interaction between eIF5A and Ypt1 in order to clarify this involvement of eIF5A with vesicular trafficking.

Size-segregated aerosol in a hot-spot pollution urban area: Chemical composition and three-way source apportionment.

In this work, a comprehensive characterisation and source apportionment of size-segregated aerosol collected using a multistage cascade impactor was performed. The samples were collected during wintertime in Milan (Italy), which is located in the Po Valley, one of the main pollution hot-spot areas in Europe. For every sampling, size-segregated mass concentration, elemental and ionic composition, and levoglucosan concentration were determined. Size-segregated data were inverted using the program MICRON to identify and quantify modal contributions of all the measured components. The detailed chemical characterisation allowed the application of a three-way (3-D) receptor model (implemented using Multilinear Engine) for size-segregated source apportionment and chemical profiles identification. It is noteworthy that - as far as we know - this is the first time that three-way source apportionment is attempted using data of aerosol collected by traditional cascade impactors. Seven factors were identified: wood burning, industry, resuspended dust, regional aerosol, construction works, traffic 1, and traffic 2. Further insights into size-segregated factor profiles suggested that the traffic 1 factor can be associated to diesel vehicles and traffic 2 to gasoline vehicles. The regional aerosol factor resulted to be the main contributor (nearly 50%) to the droplet mode (accumulation sub-mode with modal diameter in the range 0.5-1 µm), whereas the overall contribution from the two factors related to traffic was the most important one in the other size modes (34-41%). The results showed that applying a 3-D receptor model to size-segregated samples allows identifying factors of local and regional origin while receptor modelling on integrated PM fractions usually singles out factors characterised by primary (e.g. industry, traffic, soil dust) and secondary (e.g. ammonium sulphate and nitrate) origin. Furthermore, the results suggested that the information on size-segregated chemical composition in different size classes was exploited by the model to relate primary emissions to rapidly-formed secondary compounds.

Occupational exposure to PCDDs, PCDFs, and DL-PCBs in metallurgical plants of the Brescia (Lombardy Region, northern Italy) area.

The concentration values of polychlorodibenzodioxins (PCDDs), polychlorodibenzofurans (PCDFs), and dioxin-like polychlorobiphenyls (DL-PCBs) in blood serum samples (pools) of metallurgical workers in the area of the city of Brescia (northern Italy) were statistically processed. As to workers' exposure characteristics, pools were divided into 34 professionally exposed (PE) and 11 non-professionally exposed (NPE). A further subdivision of PE pools was according to workplaces in which ferrous (N = 24) and non-ferrous (N = 10) materials were handled. To evaluate the aforesaid differences we applied the age-adjusted Generalized Linear Models. We identified significant (P ≤ 0.05) exposure models of the classification groups. The first subdivision was confirmed by the concentrations of 1,2,3,4,6,7,8-H7CDF, DL-PCB 105, and DL-PCB 189; the second was confirmed by the concentrations of PCDF TEQ97, PCDD + PCDF + DL-PCB (TEQTOT) TEQ97, 2,3,4,7,8-P5CDF, 1,2,3,6,7,8-H6CDD, 1,2,3,4,6,7,8-H7CDD, and PCB 189. Based on the literature, all mentioned congeners have been found in stack gas and fly ash samples of metallurgical plants: therefore, these indicators indicate the exposure to such work environments. Specifically, the concentrations measured in the workers' blood serum appear to depend on the type of material processed during work.

A theoretical framework for studying the electromagnetic stimulation of nervous tissue.

In this paper we present a model for calculating the electric field, and its spatial derivatives, produced by arbitrarily shaped, oriented and placed coils carrying time-varying currents. The model has been validated by comparing its results with those obtained using FEM simulations. The model provides a simple and fast computation framework to investigate the electromagnetic stimulation of neural tissues. Some example applications are also provided.

Evaluation of mean echogenicity of tendons and ligaments of the metacarpal region in neonatal foals: A preliminary study.

The aims of this research were to evaluate mean echogenicity (ME) of the deep and superficial digital flexor tendons (DDFT and SDFT), the interosseous muscle (IM), and the accessory ligament of the deep digital flexor tendon (ALDDFT) of the metacarpal region in neonatal foals, and determine the effect of sex, side and body weight on this quantitative ultrasonographic evaluation. Thirteen orthopedically sound neonatal foals were examined. Four areas of study (1A, 1B, 2A, 2B) were identified. Transverse scans of the DDFT, SDFT, IM and ALDDFT were obtained, recorded, and analyzed. The most echogenic structures were the ALDDFT and DDFT, while the SDFT was significantly less echogenic than all other structures (P<0.05). No influence of sex, forelimb, or body weight was observed. The echogenicity of the tenodesmic structures of foals partially overlapped that reported in the metacarpal region in adult horses, except for IM.