Mitochondrial apurinic/apyrimidinic endonuclease 1 enhances mtDNA repair contributing to cell proliferation and mitochondrial integrity in early stages of hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) is the leading cause of primary liver cancers. Surveillance of individuals at specific risk of developing HCC, early diagnostic markers, and new therapeutic approaches are essential to obtain a reduction in disease-related mortality. Apurinic/apyrimidinic endonuclease 1 (APE1) expression levels and its cytoplasmic localization have been reported to correlate with a lower degree of differentiation and shorter survival rate. The aim of this study is to fully investigate, for the first time, the role of the mitochondrial form of APE1 in HCC. Methods: As a study model, we analyzed samples from a cohort of patients diagnosed with HCC who underwent surgical resection. Mitochondrial APE1 content, expression levels of the mitochondrial import protein Mia40, and mtDNA damage of tumor tissue and distal non-tumor liver of each patient were analyzed. In parallel, we generated a stable HeLa clone for inducible silencing of endogenous APE1 and re-expression of the recombinant shRNA resistant mitochondrially targeted APE1 form (MTS-APE1). We evaluated mtDNA damage, cell growth, and mitochondrial respiration. Results: APE1's cytoplasmic positivity in Grades 1 and 2 HCC patients showed a significantly higher expression of mitochondrial APE1, which accounted for lower levels of mtDNA damage observed in the tumor tissue with respect to the distal area. In the contrast, the cytoplasmic positivity in Grade 3 was not associated with APE1's mitochondrial accumulation even when accounting for the higher number of mtDNA lesions measured. Loss of APE1 expression negatively affected mitochondrial respiration, cell viability, and proliferation as well as levels of mtDNA damage. Remarkably, the phenotype was efficiently rescued in MTS-APE1 clone, where APE1 is present only within the mitochondrial matrix. Conclusions: Our study confirms the prominent role of the mitochondrial form of APE1 in the early stages of HCC development and the relevance of the non-nuclear fraction of APE1 in the disease progression. We have also confirmed overexpression of Mia40 and the role of the MIA pathway in the APE1 import process. Based on our data, inhibition of the APE1 transport by blocking the MIA pathway could represent a new therapeutic approach for reducing mitochondrial metabolism by preventing the efficient repair of mtDNA

UV irradiated graphene-based nanocomposites: Change in the mechanical properties by local harmoniX atomic force microscopy detection

Epoxy based coatings are susceptible to ultra violet (UV) damage and their durability can be significantly reduced in outdoor environments. This paper highlights a relevant property of graphene-based nanoparticles: Graphene Nanoplatelets (GNPs) incorporated in an epoxy-based free-standing film determine a strong decrease of the mechanical damages caused by UV irradiation. The effects of UV light on the morphology and mechanical properties of the solidified nanocharged epoxy films are investigated by Atomic Force Microscopy (AFM), in the acquisition mode "HarmoniX." Nanometric-resolved maps of the mechanical properties of the multi-phase material evidence that the incorporation of low percentages, between 0.1% and 1.0% by weight, of graphene nanoplatelets (GNPs) in the polymeric film causes a relevant enhancement in the mechanical stability of the irradiated films. The beneficial effect progressively increases with increasing GNP percentage. The paper also highlights the potentiality of AFM microscopy, in the acquisition mode "HarmoniX" for studying multiphase polymeric systems

Morphological, rheological and electrical properties of composites filled with carbon nanotubes functionalized with 1-pyrenebutyric acid

Non-covalent functionalization of Multi Wall Carbon Nanotubes (MWCNTs) could provide a solution for preserving their electronic structure facilitating the nanocomposite process preparation. Functionalization of MWCNTs by π-stacking interaction between nanofiller and a pyrene derivative has been explored. The rheological properties of filled epoxy resins highlight very interesting benefits from this kind of functionalization. Besides its peculiar capability for preventing agglomeration in the nanofiller dispersion step, it also efficiently contributes to a decrease in the viscosity of the nanocomposites; hence contrasting one of the most relevant drawback related to the manufacturing processes of the nanocomposites at MWCNTs loading rates beyond the Electrical Percolation Threshold (EPT). Because no damage of MWCNTs occurs, sp2 hybridization of carbon atoms is preserved together with the π-electron delocalization typical of polynuclear aromatic rings. Consequently, no deterioration in the electrical properties are detected; the measured EPT values are typical of nanocomposites containing embedded unfunctionalized MWCNTs (lower than 0.28 wt%), whereas for the electrical conductivity beyond the EPT, an enhancement is observed

Stratification and monitoring of natalizumab-associated progressive multifocal leukoencephalopathy risk: recommendations from an expert group

The use of natalizumab for highly active relapsing-remitting multiple sclerosis (MS) is influenced by the occurrence of progressive multifocal leukoencephalopathy (PML). Through measurement of the anti-JCV antibody index, and in combination with the presence or absence of other known risk factors, it may be possible to stratify patients with MS according to their risk of developing PML during treatment with natalizumab and detect early suspected PML using MRI including a diffusion-weighted imaging sequence. This paper describes a practical consensus guideline for treating neurologists, based on current evidence, for the introduction into routine clinical practice of anti-JCV antibody index testing of immunosuppressant-naïve patients with MS, either currently being treated with, or initiating, natalizumab, based on their anti-JCV antibody status. Recommendations for the frequency and type of MRI screening in patients with varying index-associated PML risks are also discussed. This consensus paper presents a simple and pragmatic algorithm to support the introduction of anti-JCV antibody index testing and MRI monitoring into standard PML safety protocols, in order to allow some JCV positive patients who wish to begin or continue natalizumab treatment to be managed with a more individualised analysis of their PML risk

Mycobacterium tuberculosis Surgical Site Infection after Cardiac Surgery in the COVID-19 Era: A Case Report

Infection of surgical wounds with acid-fast bacilli, including tubercle bacilli, is rare, and is poorly described in the literature. We present the case of a 74-year-old male who developed a sternal wound infection after cardiac surgery due to Mycobacterium tuberculosis complex, diagnosed post-mortem. SARS-CoV-2 infection contributed to worsened clinical conditions and surgical site infection. A high degree of suspicion to avoid unnecessary treatments and progression to severe disease with dismal prognosis is necessary in these types of infections

Estrogen activation of microglia underlies the sexually dimorphic differences in Nf1 optic glioma-induced retinal pathology

Children with neurofibromatosis type 1 (NF1) develop low-grade brain tumors throughout the optic pathway. Nearly 50% of children with optic pathway gliomas (OPGs) experience visual impairment, and few regain their vision after chemotherapy. Recent studies have revealed that girls with optic nerve gliomas are five times more likely to lose vision and require treatment than boys. To determine the mechanism underlying this sexually dimorphic difference in clinical outcome, we leveraged Nf1 optic glioma (Nf1-OPG) mice. We demonstrate that female Nf1-OPG mice exhibit greater retinal ganglion cell (RGC) loss and only females have retinal nerve fiber layer (RNFL) thinning, despite mice of both sexes harboring tumors of identical volumes and proliferation. Female gonadal sex hormones are responsible for this sexual dimorphism, as ovariectomy, but not castration, of Nf1-OPG mice normalizes RGC survival and RNFL thickness. In addition, female Nf1-OPG mice have threefold more microglia than their male counterparts, and minocycline inhibition of microglia corrects the retinal pathology. Moreover, pharmacologic inhibition of microglial estrogen receptor-β (ERβ) function corrects the retinal abnormalities in female Nf1-OPG mice. Collectively, these studies establish that female gonadal sex hormones underlie the sexual dimorphic differences in Nf1 optic glioma–induced retinal dysfunction by operating at the level of tumor-associated microglial activation

Fractional Microablative CO2 Laser-Related Histological Changes on Vulvar Tissue in Patients With Genitourinary Syndrome of Menopause

Background and Objectives: Fractional CO2 laser has been proposed as an effective treatment for the genitourinary syndrome of menopause (GSM). However, the effects of laser treatment on vulvar tissue have never been assessed. We aimed to assess histological changes related to fractional CO2 laser in vulvar tissue from GSM patients. Study Design/Materials and Methods: A single-center observational prospective cohort study was performed enrolling all GSM patients from July 2017 to October 2018. Patients underwent three outpatient vulvovaginal applications of fractional CO2 laser and vulvar biopsy before and after treatment. Rates of histological changes in vulvar tissue, the difference in means of Vulva Health Index (VuHI), Vaginal Health Index (VHI), Visual Analogue Scale scores for GSM symptoms, and procedure-related pain, and rate of patient's overall satisfaction with treatment were assessed. Univariate comparisons between continuous variables were performed by using the paired t-test (α error = 0.05). Results: Of 20 enrolled patients, 18 underwent all laser applications, and 15 underwent both vulvar biopsies. 93.3% of patients showed remodeling of vulvar connective tissue; 80% showed improvement in vulvar epithelium trophism and 86.7% showed neovascularization. Differences in means between before and after treatment were significant for VuHI, VHI, and all GSM symptoms. Means ± standard deviation of the degree of pain at each laser application were 4.4 ± 0.9, 3.7 ± 1.6, and 2.9 ± 1.9. The rate of overall satisfaction with the treatment was 72.2%. Conclusions: Fractional CO2 laser leads to a restoration of the normal architecture of vulvar tissue, with significant improvement in GSM-related signs and symptoms, and overall satisfaction with the treatment in most GSM patients. Lasers Surg. Med. © 2020 Wiley Periodicals LLC

Esbl/ampc-producing escherichia coli in wild boar: Epidemiology and risk factors

The complex health problem of antimicrobial resistance (AMR) involves many host species, numerous bacteria and several routes of transmission. Extended-spectrum β-lactamase and AmpC (ESBL/AmpC)-producing Escherichia coli are among the most important strains. Moreover, wildlife hosts are of interest as they are likely antibiotics free and are assumed as environmental indicators of AMR contamination. Particularly, wild boar (Sus scrofa) deserves attention because of its increased population densities, with consequent health risks at the wildlife–domestic–human interface, and the limited data available on AMR. Here, 1504 wild boar fecal samples were microbiologically and molecularly analyzed to investigate ESBL/AmpC-producing E. coli and, through generalized linear models, the effects of host-related factors and of human population density on their spread. A prevalence of 15.96% of ESBL/AmpC-producing E. coli, supported by blaCTX-M (12.3%), blaTEM (6.98%), blaCMY (0.86%) and blaSHV (0.47%) gene detection, emerged. Young animals were more colonized by ESBL/AmpC strains than older subjects, as observed in domestic animals. Increased human population density leads to increased blaTEM prevalence in wild boar, suggesting that spatial overlap may favor this transmission. Our results show a high level of AMR contamination in the study area that should be further investigated. However, a role of wild boar as a maintenance host of AMR strains emerged

Quantifying physiological trait variation with automated hyperspectral imaging in rice

Advancements in hyperspectral imaging (HSI) together with the establishment of dedicated plant phenotyping facilities worldwide have enabled high-throughput collection of plant spectral images with the aim of inferring target phenotypes. Here, we test the utility of HSI-derived canopy data, which were collected as part of an automated plant phenotyping system, to predict physiological traits in cultivated Asian rice (Oryza sativa). We evaluated 23 genetically diverse rice accessions from two subpopulations under two contrasting nitrogen conditions and measured 14 leaf- and canopy-level parameters to serve as ground-reference observations. HSI-derived data were used to (1) classify treatment groups across multiple vegetative stages using support vector machines (≥ 83% accuracy) and (2) predict leaf-level nitrogen content (N, %, n=88) and carbon to nitrogen ratio (C:N, n=88) with Partial Least Squares Regression (PLSR) following RReliefF wavelength selection (validation: R2 = 0.797 and RMSEP = 0.264 for N; R2 = 0.592 and RMSEP = 1.688 for C:N). Results demonstrated that models developed using training data from one rice subpopulation were able to predict N and C:N in the other subpopulation, while models trained on a single treatment group were not able to predict samples from the other treatment. Finally, optimization of PLSR-RReliefF hyperparameters showed that 300-400 wavelengths generally yielded the best model performance with a minimum calibration sample size of 62. Results support the use of canopy-level hyperspectral imaging data to estimate leaf-level N and C:N across diverse rice, and this work highlights the importance of considering calibration set design prior to data collection as well as hyperparameter optimization for model development in future studies