An orchestrated intron retention program in meiosis controls timely usage of transcripts during germ cell differentiation

Global transcriptome reprogramming during sper-matogenesis ensures timely expression of factors in each phase of male germ cell differentiation. Sper-matocytes and spermatids require particularly exten-sive reprogramming of gene expression to switch from mitosis to meiosis and to support gamete morphogenesis. Here, we uncovered an extensive alternative splicing program during this transmeiotic differentiation. Notably, intron retention was largely the most enriched pattern, with spermatocytes showing generally higher levels of retention compared with spermatids. Retained introns are characterized by weak splice sites and are enriched in genes with strong relevance for gamete func-tion. Meiotic intron-retaining transcripts (IRTs) were exclusively localized in the nucleus. However, differ-ently from other developmentally regulated IRTs, they are stable RNAs, showing longer half-life than properly spliced transcripts. Strikingly, fate-mapping experiments revealed that IRTs are recruited onto polyribosomes days after synthesis. These studies reveal an unexpected function for regulated intron retention in modulation of the timely expression of select transcripts during spermatogenesis

Identification of murine phosphodiesterase 5A isoforms and their functional characterization in HL-1 cardiac cell line

Phosphodiesterase 5A (PDE5A) specifically degrades the ubiquitous second messenger cGMP and experimental and clinical data highlight its important role in cardiac diseases. To address PDE5A role in cardiac physiology, three splice variants of the PDE5A were cloned for the first time from mouse cDNA library (mPde5a1, mPde5a2 and mPde5a3). The predicted amino acidic sequences of the three murine isoforms are different in the N-terminal regulatory domain. mPDE5A isoforms were transfected in HEK293T cells and they showed high affinity for cGMP and similar sensitivity to sildenafil inhibition. RT-PCR analysis showed that mPde5a1, mPde5a2 and mPde5a3 had differential tissue distribution. In the adult heart, mPde5a1 and mPde5a2 were expressed at different levels whereas mPde5a3 was undetectable. Overexpression of mPDE5As induced an increase of HL-1 number cells which progress into cell cycle. mPDE5A1 and mPDE5A3 overexpression increased the number of polyploid and binucleated cells, mPDE5A3 widened HL-1 areas and modulated hypertrophic markers more efficiently respect to the other mPDE5A isoforms. Moreover, mPDE5A isoforms had differential subcellular localization: mPDE5A1 was mainly localized in the cytoplasm, mPDE5A2 and mPDE5A3 were also nuclear localized. These results demonstrate for the first time the existence of three PDE5A isoforms in mouse and highlight their potential role in the induction of hypertrophy. This article is protected by copyright. All rights reserved

SINCONAPP: A Computerized learning tool for CBCT normal anatomy and variants of the nose and paranasal sinuses

1. Purpose To supply an useful learning tool aimed to interactively display on mobile devices normal anatomy and variants of the nose and paranasal sinuses as seen on CBCT images. 2. Methods and Materials Images Images of the nose and paranasal sinuses were derived by a study series acquired by a CBCT device. CBCT studies of the paranasal sinuses were acquired in patients referred for nasal obstruction or sinusitis with the following parameters: 90 kVp, 12.5 mA, 20 s rotation time, FOV 13 x 14.5 cm, 0.25 x 0.25 x 0.25 mm voxel size. Software The application has been developed for iOS based mobile devices through the platform XCode provided by Apple®, and it is developed using the Objective-C programming language. The application has been configured as Master-Detail. This configuration splits the mobile device display in two panels. The left panel displays a list of the interesting items, while the right panel shows the relative details. Touching an item from the menu on the left panel, the textual description is shown on the same side, while the panel on the right will show the relative image. The application allows interactively navigation through normal anatomy and variants of the nose and paranasal sinuses, as represented on CBCT images in axial, sagittal and coronal planes. Cross-reference images to localize the same anatomic structures on different section planes are available. The navigation is intuitive, with multiple shortcuts. Different labels have been proposed in accordance with the specific anatomic lessic of the district and current literature references. High image quality with a zooming tool are available. 4. Conclusion An App for IOs devices was developed, that can represent an useful educational tool for medical students, residents and continuous medical education in radiology and other medical specialties dealing with nose and paranasal sinuses. This interactive atlas based on CBCT images could be also an useful option to be implemented on CBCT software

Exploring the Role of Fusobacterium nucleatum in Preterm Birth: A Narrative Review

In recent years, substantive attention has been drawn to the relationship between oral microbiome homeostatic equilibrium disruption and systemic health, demonstrating the negative impacts of this reciprocal biological interplay. Increasingly, there is a concern over the potential noxious effect of oral microbiome dysbiosis on obstetric poor outcomes, focusing on preterm birth. This epidemiological observation remains unexplained, although biologically plausible mechanism has been proposed. Intrauterine infection has long been associated with adverse pregnancy, when the elicitation of an immune response is determinant. There is evidence that Fusobacterium nucleatum (FN), a Gram-negative anaerobe ubiquitous in the oral cavity, infects the mouse placenta originating in the decidua basalis. Based on the current data in literature, we performed a review to provide resources for the explanation of the potential impact of microbiome dysbiosis on poor obstetric outcomes, focusing on the role of FN

Oral Microbiome and Preterm Birth: Correlation or Coincidence? A Narrative Review

AIM: Physiological changes that occur during pregnancy involve, as a natural consequence, also modifications of oral microbiome. However, the addition with microbial imbalance due to pre-existing periodontal infection might impair a pathological alteration in the phylogenetic community structure and composition in the oral cavity, exacerbating an inflammatory status, and becoming a potential risk factor for preterm birth. From the empirical findings about the relationship between periodontal pathogens and systemic diseases, a clear interest focused on the potential impact of some periodontal pathogens on the preterm birth risk has emerged. In this close emerging link, the potential interdependence existing between dysbiosis of oral microbiome and changes in maternal-fetal barrier in premature rupture of membranes was explored. MATERIALS AND METHODS: In accordance with Preferred Reporting Items for Systematic Reviews and Meta- Analyses guidelines, a Medline search was performed for studies focusing on oral microbioma and its association with pre-term birth, and completed by additional hand searching. Two reviewers independently selected studies and extracted data. The search was restricted to only reports written in English. RESULTS: The electronic search produced 66 items. Six duplicates were found. Among the collected studies, 56 were discarded because they met the exclusion criteria. The articles and reports in our review showed a connection between preterm birth and altered oral microbiome, suggesting a potential key role of Fusobacterium nucleatum, a notable periodontal pathogen involved in several pathological periodontal conditions, in increasing the risk of premature birth. CONCLUSIONS: Since F. nucleatum is frequently associated with preterm birth, it is coherent to hypothesize a potential role for the oral microbiota for preterm birth risk. Further studies should be carried out to determine the changes of the oral microflora in pregnancy and to provide comprehensive knowledge of the diversity of oral bacteria involved in preterm birth

Oral Microbiome and Preterm Birth: Correlation or Coincidence? A Narrative Review

BACKGROUND: Physiological changes that occur during pregnancy involve, as a natural consequence, also modifications of oral microbiome. However, the addition with microbial imbalance due to pre-existing periodontal infection might impair a pathological alteration in the phylogenetic community structure and composition in the oral cavity, exacerbating an inflammatory status, and becoming a potential risk factor for preterm birth. From the empirical findings about the relationship between periodontal pathogens and systemic diseases, a clear interest focused on the potential impact of some periodontal pathogens on the preterm birth risk has emerged. AIM: Exploration of the potential interdependence existing between dysbiosis of oral microbiome and changes in maternal-fetal barrier in premature rupture of membranes. MATERIALS AND METHODS: In accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, a Medline search was performed for studies focusing on oral microbioma and its association with pre-term birth, and completed by additional hand searching. Two reviewers independently selected studies and extracted data. The search was restricted to only reports written in English. RESULTS: The electronic search produced 66 items. Six duplicates were found. Among the collected studies, 56 were discarded because they met the exclusion criteria. The articles and reports in our review showed a connection between preterm birth and altered oral microbiome, suggesting a potential key role of Fusobacterium nucleatum, a notable periodontal pathogen involved in several pathological periodontal conditions, in increasing the risk of premature birth. CONCLUSIONS: Since F. nucleatum is frequently associated with preterm birth, it is coherent to hypothesize a potential role for the oral microbiota for preterm birth risk. Further studies should be carried out to determine the changes of the oral microflora in pregnancy and to provide comprehensive knowledge of the diversity of oral bacteria involved in preterm birth

Targeted delivery of neutralizing anti-C5 antibody to renal endothelium prevents complement- dependent tissue damage

Complement activation is largely implicated in the pathogenesis of several clinical conditions and its therapeutic neutralization has proven effective in preventing tissue and organ damage. A problem that still needs to be solved in the therapeutic control of complement-mediated diseases is how to avoid side effects associated with chronic neutralization of the complement system, in particular, the increased risk of infections. We addressed this issue developing a strategy based on the preferential delivery of a C5 complement inhibitor to the organ involved in the pathologic process. To this end, we generated Ergidina, a neutralizing recombinant anti-C5 human antibody coupled with a cyclic-RGD peptide, with a distinctive homing property for ischemic endothelial cells and effective in controlling tissue damage in a rat model of renal ischemia/reperfusion injury (IRI). As a result of its preferential localization on renal endothelium, the molecule induced complete inhibition of complement activation at tissue level, and local protection from complement-mediated tissue damage without affecting circulating C5. The ex vivo binding of Ergidina to surgically removed kidney exposed to cold ischemia supports its therapeutic use to prevent posttransplant IRI leading to delay of graft function. Moreover, the finding that the ex vivo binding of Ergidina was not restricted to the kidney, but was also seen on ischemic heart, suggests that this RGD-targeted anti-C5 antibody may represent a useful tool to treat organs prior to transplantation. Based on this evidence, we propose preliminary data showing that Ergidina is a novel targeted drug to prevent complement activation on the endothelium of ischemic kidney

Dietary cholesterol supplementation and inhibitory factor 1 serum levels in two dizygotic Smith-Lemli-Opitz syndrome twins: a case report

Smith-Lemli-Opitz syndrome (SLOS) is a rare genetic neurodevelopmental disorder caused by the defect in the 7-dehydrocholesterol reductase. This defect leads to the deficiency of cholesterol biosynthesis with accumulation of 7-dehydrocholesterol. Inhibitory factor 1 (IF1) is a well-known mitochondrial protein. Recently, it has been discovered in the human serum where it is reported to be involved in the HDL-cholesterol intake. Here we report the IF1 presence in the serum of two paediatric SLOS dizygotic twins treated with dietary cholesterol supplementation

PDE5 inhibition counteracts β- adrenergic induction of cardiac hypertrophy

The b-adrenoreceptors play important roles in cardiovascular function regulation mediated by the sympathetic nervous system. It is known that sustained b-adrenergic stimulations promotes cardiac hypertrophy (Oleg et al., 2007). Recently an antihypertrophic role of sildenafil, that acts as a phosphodiesterase 5 (PDE5) inhibitor, has been demonstrated in mice where hypertrophy was mechanically induced (Takimoto et al., 2005). We report the results obtained on a cellular system of cardiac hypertrophy in vitro. By using three-dimensional cultures of mouse ventricular cardiomyocytes (Xiang et al., 2005) and isolated cardiomyocytes we show that: 1) these cells express levels of PDE5 comparable with the ones in normal heart, 2) treatment of the cultures with the b-adrenoreceptors agonist isoproterenol induces cell hypertrophy accompanied by an increment of the level of PDE5 expression and 3) sildenafil prevents the development of such hypertrophy through specific b-adrenoreceptors and signaling pathways 4) the inhibition of other members of PDE family might contribute to the prevention of hypertrophy following b-adrenergic stimulation. In summary, we present a test system that may contribute to clarify intracellular signaling pathways leading to cardiac hypertrophy and to identify molecular targets, like the ones involved in PDE5 activity, on which to steer the development of new drugs and to design new clinical therapies

CBCT in the diagnosis of osteonecrosis of the jaws

Short Summary: ONJ is associated with bisphosphonates, and recently with antiangiogenic drugs. Another cause is radiotherapy. Few papers have previously investigated the value of CBCT in the diagnosis of ONJ. Purpose/Objectives: To describe the CBCT findings in the diagnosis of osteonecrosis of the jaws (ONJ) according to a new staging system of ONJ. Methods and Materials: 9 patients (5 women, 4 men, 54-76 yrs.) affected by ONJ were studied with a CBCT device (90 kV, 12.5 mA, 0.25 mm voxel size) in 2012-2013. 7/9 patients were treated with bisphosphonates (5 for bone metastasis, 2 for osteoporosis), 1/9 patient was treated with an antiangiogenic drug, 1/9 patient received radiotherapy. Patients were divided in 3 stages, according to a new clinical-radiologic classification of ONJ, made by SICMF (Italian Society of Maxillo-Facial Surgery) and SIPMO (Italian Society of Oral Medicine), that gives more prominence to the radiologic findings and includes 3 stages: stage 1 focal ONJ, a) symptomatic, b) asymptomatic; stage 2 diffuse ONJ, a) symptomatic, b) asymptomatic; stage 3 complicated ONJ. Axial, panoramic, cross sectional, multiplanar and 3D reformations were analysed. Results: Radiologic findings of ONJ were found in maxilla (7/9 patients) and mandible (7/9 patients). Patients were staged as follows: 1/9 stage 1b), 1/9 stage 2a), 4/9 stage 2b), and 3/9 stage 3. CBCT findings varied from simple osteosclerosis in stage 1b), to multifragmentary fracture, extraoral fistula, and fistula between oral cavity and maxillary sinus or nasal cavity in stage 3 patients, passing through osteonecrosis and bone sequestrum in 7 patients in stage 2-3. Conclusion: CBCT is a valuable tool in the radiologic assessment of ONJ and proved useful in this new SICMF-SIPMO staging system. Keywords: Staging, jaws, osteonecrosis, CBC