Assessment of Peri-Implant Soft Tissues Conditions around Short and Ultra-Short Implant-Supported Single Crowns: A 3-Year Retrospective Study on Periodontally Healthy Patients and Patients with a History of Periodontal Disease

The aim of this retrospective study was to evaluate implant survival, marginal bone loss and peri-implant complications in 326 short and ultra-short implants. Implants were placed in the maxillary and mandibular posterior regions of 140 patients with (PP) and without (NPP) a history of periodontal disease. Clinical and radiographic examinations were performed at 3-year recall appointments. The 8.0, 6.0 and 5.0 mm-length implants placed in PP and NPP were respectively 43.75% and 38.46%, 35.10% and 34.19%, 21.15% and 27.35%; 325 implants (one early failure) were rehabilitated with single crowns in 139 patients. Overall implant survival after 3 years of follow-up was 97.55%, 98.08% and 96.61% for PP and NPP (p = 0.46). Crestal bone level variations were not statistically different among PP and NPP; 15.41% of implants presented signs of mucositis, 14.71% and 16.67% in PP and NPP (p = 0.64). Setting the threshold for bone loss at 2 mm after 36 months, peri-implantitis prevalence was 2.2%, 1.96% and 2.63% in PP and NPP (p = 0.7). Overall implant success was 82.39%, 83.33% and 80.7% for PP and NPP (p = 0.55). Short-term outcomes suggest that short and ultra-short locking-taper implants can successfully be restored with single crowns in the posterior jaws both in PP and NPP

Composition and Epigenetic Markers of Heterochromatin in the Aphid Aphis nerii (Hemiptera: Aphididae).

A detailed karyotype analysis of the oleander aphid Aphis nerii focusing on the distribution, molecular composition and epigenetic modifications of heterochromatin was done in order to better understand the structure and evolution of holocentric/holokinetic chromosomes in aphids. The female karyotype (2n = 8) consisted of 3 pairs of autosomes and a pair of X chromosomes that were the longest elements in the karyotype and carried a single, terminally located nucleolar organizer region. Males showed 2n = 7 chromosomes due to the presence of a single X chromosome. Heterochromatin was located in the X chromosomes only and consisted of 4 satellite DNAs that have been identified. A. nerii constitutive heterochromatin was enriched in mono-, di- and tri-methylated H3 histones and HP1 proteins but, interestingly, it lacked DNA methylation that was widespread in euchromatic chromosomal regions. These results suggest that aphid heterochromatin is assembled and condensed without any involvement of DNA methylation

Serum levels of Sex hormone Binding globulin (ShBg) are not predictive of prostate cancer diagnosis and aggressiveness: results from an italian biopsy cohort

Purpose To explore the association between serum levels of Sex Hormone Binding Globulin (SHBG) and the risk of developing prostate cancer (PCa) as well as high grade disease in men undergoing prostate biopsy. Materials and Methods Between 2006 and 2012, we prospectively enrolled 740 patients with no history of PCa undergoing prostate biopsy. Before biopsy general data of the patient DRE, PSA and BMI were recorded. The risk of detecting cancer and high grade cancer was assessed as a function of SHBG using crude and adjusted logistic regressions. Results Serum levels of SHBG were not associated with an increased risk of PCa or high grade disease. Age (OR 1.027 95% CI 1.003-1.052 p = 0.027), DRE (OR 3.391 95% CI 2.258-5.092 p = 0.000) and PSA (OR 1.078 95% CI 1.037-1.120 p = 0.000) were found to be independent predictors of prostate cancer risk. Age (OR 1.051 95% CI 1.009-1.095 p = 0.016), DRE (OR 2.519 95% CI 1.384-4.584 p = 0.000), BMI (OR 1.098 95% CI 1.011-1.193 p = 0.027) and PSA (OR 1.074 95% CI 1.014-1.137 p = 0.015) were found to be independent predictors of high grade disease. Conclusions In our cohort of patients, serum levels of SHBG are not predictive of PCa or high grade disease. According to our experience SHBG should not be considered a biomarker in PCa diagnosis neither a marker for high grade disease

Single-Crown, Short and Ultra-Short Implants, in Association with Simultaneous Internal Sinus Lift in the Atrophic Posterior Maxilla: A Three-Year Retrospective Study

As the atrophic posterior maxilla often presents serious limitations for dental implant procedures, a minimally invasive technique was proposed. The study aimed to retrospectively evaluate the outcomes of short and ultra-short locking-taper implants, placed in combination with a modified osteotome sinus floor elevation procedure (internal sinus lift technique) in the posterior maxilla. A total of 31 patients received 51 locking-taper implants. Clinical and radiographic examinations were performed before treatment, at loading time, and after three years. Seven implants of 8.0 mm, 23 implants of 6.0 mm, and 21 implants 5.0 mm in length were rehabilitated with single-crown restorations. Implant survival at three-year follow-up was 96.08%. Pre-operative residual crestal bone height of 5.2 (1.41) (median (interquartile range)) mm increased to 7.59 (1.97) mm at the 36-month follow-up, with an average intra-sinus bone height gain of 3.17 ± 1.13 (mean ± standard deviation) mm. Mean peri-implant crestal bone loss was 0.29 (0.46) mm and mean first bone-to-implant contact point shifted apically to 0.12 (0.34) mm. It can be suggested with confidence that implants used in the study, placed in conjunction with an internal sinus floor elevation technique, can be restored with single crowns as a predictable treatment for the edentulous regions of the posterior maxilla

Continuous occurrence of intra-individual chromosome rearrangements in the peach potato aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae)

Analysis of the holocentric mitotic chromosomes of the peach-potato aphid, Myzus persicae (Sulzer), from clones labelled 50, 51 and 70 revealed different chromosome numbers, ranging from 12 to 14, even within each embryo, in contrast to the standard karyotype of this species (2n = 12). Chromosome length measurements, combined with fluorescent in situ hybridization experiments, showed that the observed chromosomal mosaicisms are due to recurrent fragmentations of chromosomes X, 1 and 3. Contrary to what has generally been reported in the literature, X chromosomes were frequently involved in recurrent fragmentations, in particular at their telomeric ends opposite to the nucleolar organizer region. Supernumerary B chromosomes have been also observed in clones 50 and 51. The three aphid clones showed recurrent fissions of the same chromosomes in the same regions, thereby suggesting that the M. persicae genome has fragile sites that are at the basis of the observed changes in chromosome number. Experiments to induce males also revealed that M. persicae clones 50, 51 and 70 are obligately parthenogenetic, arguing that the reproduction by apomictic parthenogenesis favoured the stabilization and inheritance of the observed chromosomal fragments

Prevention of left ventricular remodeling after myocardial infarction: efficacy of physical training

Post-myocardial infarction left ventricular remodelling should be considered an important therapeutic target in patients after an acute myocardial infarction, considering the heavy prognostic implication. The therapies used in these patients should reduce the progression of the left ventricular dysfunction to refractory heart failure. In order to prevent post-myocardial infarction cardiac remodelling, different therapies have been tested, and for ACE-inhibitors and betablockers a clear demonstration of efficacy has been obtained. Losartan and valsartan, two widely used angiotensin receptor blockers, demonstrated to be safe and equally useful compared to ACEI. The addition of spironolactone to the standard therapy for heart failure has a clear beneficial effect but the clinical use has been refrained by the risk of iperkaliemia. Aerobic physical training improves the left ventricular ejection fraction in patients with systolic dysfunction, reducing the progressive enlargement after myocardial infarction. The positive effect of aerobic training on cardiac remodelling might be related to the positive effect on neurhormonal assessment, to he improvement of microcirculatory myocardial perfusion and of endothelial function

690. Permanent Epigenetic Silencing of Human Genes With Artificial Transcriptional Repressors

There are several diseases whereby the goal of gene therapy is to silence rather than replace a gene function. Paradigmatic examples are diseases caused by a dominant negative mutation or those in which silencing of a host gene confers resistance to a pathogen or compensates the function of the missing gene. Yet, gene silencing can be used to enhance efficacy of cell therapy and for biotechnological applications. Until now, two technologies have been used to silence gene expression, namely RNA interference with short harping RNAs (shRNA) and gene disruption with Artificial Nucleases (ANs). Although some promising pre-clinical and clinical data have been already obtained, the low efficiency of knock-down with shRNA and of biallelic disruption with ANs may limit efficacy of these treatments, especially when residual gene activity can exert a biological function. To overcome this issue, we have developed a novel modality of gene silencing that exploits endogenous epigenetic mechanisms to convey robust and heritable states of repression at the desired target gene. We have generated Artificial Transcriptional Repressors (ATRs), chimeric proteins containing a custom-made DNA binding domain fused to the effector domain of a chromatinmodifying enzyme involved in silencing of Endogenous RetroViruses (ERVs). By performing iterative rounds of selection in human cell lines and primary cells engineered to report for synergistic activity of candidate effector domains, we identified a combination of 3 domains that, when transiently co-assembled on the promoter of the reporter cassette, fully abrogated transgene expression in up to 90% of treated cells. Importantly, silencing was maintained for more than 250 days in cultured cell lines, was resistant to in vitro differentiation or metabolic activation of primary cells, and was confined to the reporter cassette. Silencing was associated with high levels of de novo DNA methylation at the targeted locus and was dependent on this epigenetic mark for its propagation. Finally, transient transfection of 3 ATRs targeted to the promoter region of the Beta-2-microglobulin (B2M) gene resulted in the loss of surface expression of B2M and, consequently, of the MHC-I molecules in up to 80% of treated cells. This phenotype was associated with a switch in the epigenetic and transcriptional state of the constitutively active B2M gene, which became highly decorated with DNA methylation and deprived of RNA PolII and of its transcript. Of note, silencing was resistant to IFN-γ treatment, a potent B2M inducer. Overall, these data provide the first demonstration of efficient and stable silencing of an endogenous gene upon transient delivery of ATRs. This result was made possible by repurposing the machinery involved in silencing of ERVs, which instructs self-sustaining repressive epigenetic states on the gene of interest. While silencing of B2M might be used to generate universally transplantable allogeneic cells, our hit-and-run strategy provides a powerful new alternative to conventional gene silencing for the treatment of several diseases. (LN & AL co-authorship

729 inheritable silencing of endogenous gene by hit and run targeted epigenetic editing

Gene silencing holds great promise for the treatment of several diseases and can be exploited to investigate gene function and activity of the regulatory genome. Here, we develop a novel modality of gene silencing that exploits epigenetics to achieve stable and highly efficient repression of target genes. To this end, we generated Artificial Transcriptional Repressors (ATRs), chimeric proteins containing a custom-made DNA binding domain fused to the effector domain of chromatin-modifying enzymes involved in silencing process of Endogenous RetroViruses (ERVs). By performing iterative rounds of selection in cells engineered to report for synergistic activity of candidate effector domains, we identified a combination of 3 domains (namely KRAB, DNMT3A and DNMT3L) that, when transiently co-assembled on the promoter of the reporter cassette, recreate a powerful embryonic-specific repressive complex capable of inducing full and long-term (>150 days) silencing of transgene expression in up to 90% of the cells. The ATR-induced silencing was cell type and locus independent, and resistant to metabolic activation of the cells. Importantly, these findings were holding true also for endogenous genes embedded in their natural chromatin context, as shown for the highly and ubiquitously expressed B2M gene. Here, transient co-delivery of TALE-based ATRs resulted in loss of surface expression of B2M and, consequently, of the MHC-I molecules in up to 80% of the cells. This phenotype was associated with a drastic switch in the epigenetic and transcriptional state of the constitutively active B2M promoter, which become highly decorated with de novo DNA methylation and deprived of RNAP II. Importantly, silencing was sharply confined to the targeted gene and resistant to INF-γ, a potent natural activator of B2M. We further extended these studies by showing that our silencing approach is portable to the CRISPR/dCas9 DNA binding technology. In this setting, comparable levels of B2M silencing (up to 80%) were achieved using either pools or even individual sgRNAs coupled to dCas9-based ATRs. Yet, adoption of this technology allowed performing simultaneous, highly efficient multiplex gene silencing within the same cell, as shown for B2M, IFNAR1 and VEGFA. Finally, we assessed resistance of the silenced gene to activity of potent artificial transcription activators and chromatin remodelers, and found that only targeted DNA demethylation was able to reawaken the silent gene. This allowed performing iterative cycles of silencing and reactivation of the same gene in the same cell population. Overall, these data provide the first demonstration of efficient and stable epigenetic silencing of endogenous genes upon transient delivery of ATRs. This was accomplished by repurposing the ERVs silencing machinery, which instructs self-sustaining repressive epigenetic states to the target gene. While silencing of B2M might be used to generate universally transplantable allogeneic cells, our hit-and-run strategy provides a powerful new alternative to conventional gene silencing for both basic and translational research

Cover crops for managing weeds, soil chemical fertility and nutritional status of organically grown orange orchard in Sicily

Cover crops can offer significant advantages in the agronomic management of citrus orchards in Mediterranean environments. Therefore, a three-year research was conducted in eastern Sicily aimed at studying the effects of four cover crop sequences (Sinapis arvensis-Trigonella foenum-graecum-T. foenum-graecum; Medicago scutellata-Avena sativa-Lolium perenne; Vicia faba minor-A. sativa-A. sativa; A. sativa-V. faba. minor-L. perenne) on weeds, major soil chemical properties and nutritional status of an organically grown orange orchard. The results highlighted that, among the studied cover crop sequences, Vicia faba-Avena-Avena was the most beneficial for weeds control within the orchard (92%, of cover crop cover, and 586 and 89 g DW m–2 of cover crop aboveground biomass and weeds aboveground biomass, respectively). Overall, the chemical fertility of the soil was positively influenced. In particular, it was observed an increase of the content of total nitrogen and available phosphorus in the soil by both Sinapis-Trigonella-Trigonella (0.75 g kg–1 and 59.0 mg kg–1, respectively) and Vicia faba-Avena-Avena (0.70 g kg–1 and 56.0 mg kg–1, respectively) cover crop sequences. Medicago-Avena-Lolium sequence seemed to be the most useful to ensure a better nutritional status of the orange orchard