HPV16 E5 expression induces switching from FGFR2b to FGFR2c and epithelial-mesenchymal transition

The E5 oncoprotein of the human papillomavirus type 16 (HPV16 E5) deregulates epithelial homeostasis through the modulation of receptor tyrosine kinases and their signaling. Accordingly, the fibroblast growth factor receptor 2b (FGFR2b/KGFR), epithelial splicing transcript variant of the FGFR2, is down-modulated by the viral protein expression, leading to impairment of keratinocyte differentiation. Here, we report that, in cell models of transfected human keratinocytes as well as in cervical epithelial cells containing episomal HPV16, the down-regulation of FGFR2b induced by 16E5 is associated with the aberrant expression of the mesenchymal FGFR2c isoform as a consequence of splicing switch: in fact, quantitative RT-PCR analysis showed that this molecular event is transcriptionally regulated by the epithelial splicing regulatory proteins 1 and 2 (ESRP1 and ESRP2) and is able to produce effects synergistic with those caused by TGFβ treatment. Immunofluorescence analysis revealed that this altered FGFR2 splicing leads to changes in the specificity for the ligands FGFs and in the cellular response, triggering epithelial-mesenchymal transition (EMT). Through 16E5 or FGFR2 silencing as well as inhibition of FGFR2 activity we demonstrated the direct role of the viral protein in the receptor isoform switching and EMT, suggesting that these early molecular events during HPV infection might represent additional mechanisms driving cervical transformation and tumor progression

Correlates of spinal deforming index (SDI) in HIV-positive patients naive and on treatment

Methods HIV-infected subjects naive or on stable HAART were included. Vertebral deformities were identified using SDI (according to semiquantitative method by Genant), calculated by summing the deformity grades of all vertebrae (T4 to L4); pathological deformities are defined as follow: grade 1 between 20–25%, grade 2 between 26–40%, and grade 3 > >40%. According to WHO criteria, osteopenia and osteoporosis were diagnosed in patients having spine BMD calculated as -1 << T-score << -2.5 and T-score ≤≤2.5, respectively. The correlation between SDI and spine BMD was evaluated by univariate and multivariate linear regression. [Other variables considered: gender, age, current CD4 count, CD4 nadir, BMI, lipid parameters, alcohol intake, smoking habit, physical activity, family history for bone fracture, months of ARV exposure, and co-infection with hepatitis viruses; only the variables with p <<0.2 in univariate analyses were included in the final model.

The nucleolar protein nucleophosmin is physiologically secreted by endothelial cells in response to stress exerting proangiogenic activity both in vitro and in vivo

Nucleophosmin (NPM), a nucleolar multifunctional phosphoprotein, acts as a stress sensor in different cell types. NPM can be actively secreted by inflammatory cells, however its biology on endothelium remains unexplored. In this study, we show for the first time that NPM is secreted by human vein endothelial cells (HUVEC) in the early response to serum deprivation and that NPM acts as a pro-inflammatory and angiogenic molecule both in vitro and in vivo. Accordingly, 24 h of serum starvation condition induced NPM relocalization from the nucleus to cytoplasm. Interestingly, NPM was increasingly excreted in HUVEC-derived conditioned media in a time dependent fashion upon stress conditions up to 24 h. The secretion of NPM was unrelated to cell necrosis within 24 h. The treatment with exogenous and recombinant NPM (rNPM) enhanced migration as well as the Intercellular Adhesion Molecule 1 (ICAM-1) but not Vascular cell adhesion protein 1 (VCAM-1) expression and it did not affect cell proliferation. Notably, in vitro tube formation by Matrigel assay was significantly increased in HUVEC treated with rNPM compared to controls. This result was confirmed by the in vivo injection of Matrigel plug assay upon stimulation with rNPM, displaying significant enhanced number of functional capillaries in the plugs. The stimulation with rNPM in HUVEC was also associated to the increased expression of master genes regulating angiogenesis and migration, including Vascular Endothelial Growth Factor-A (VEGF-A), Hepatocyte Growth Factor (HGF), Stromal derived factor-1 (SDF-1), Fibroblast growth factor-2 (FGF-2), Platelet Derived Growth Factor-B (PDGF-B), and Matrix metallopeptidase 9 (MMP9). Our study demonstrates for the first time that NPM is physiologically secreted by somatic cells under stress condition and in the absence of cell necrosis. The analysis of the biological effects induced by NPM mainly related to a pro-angiogenic and inflammatory activity might suggest an important autocrine/paracrine role for NPM in the regulation of both phenomena

Clinical and instrumental assessment of herniated discs after nucleoplasty: A preliminary study

Background and Aim: The therapy for low back pain boasts different approaches; one of these is nucleoplasty. We wanted to assess the effectiveness of nucleoplasty both by clinical response both by MR imaging evaluation, including even extrusions larger than one third of the spinal canal. Methods: Fifty-seven patients were treated with nucleoplasty in our hospital, 11 of these patients accepted both clinical and MRI evaluation after six months from treatment. The clinical evaluation was performed with Visual Analogue Scale (VAS) of pain, scored before and after the procedure. MRI evaluation consisted of analysing some imaging parameters of disc protrusions before and after the treatment. Results: In 10 out of 11 (91%) patients, VAS was reduced and only 1 out of 11 (9%) had the same pain after procedure. The mean of decrease of VAS score was 64%. In our population 8/11 (72%) patients had a herniation larger than 1/3 of the sagittal diameter of spinal canal and 100% of them had an improvement with a mean VAS reduction value of 75%. With MRI evaluation, the mean percentage of expulsion before and after treatment was respectively 40% and 34%. The expulsion decreased in 7/13 discs, remained equal in 4/13, and increased in 2/13 discs. Among the 9 larger protrusions, 3 didn\u2019t change, 6 reduced with a decrease mean value of 13%. Other MRI parameters didn\u2019t change significantly. Conclusions: Our preliminary experience supports the success of coblation on pain relief, aiming to show progressively that this treatment is suitable even in case of great extrusions, which are generally treated only with surgical approach. It\u2019s not clear the usefulness of MRI control yet, even if in most of cases we could have found a certain reduction of expulsion degree

Anti-ApoA-1 IgGs in Familial Hypercholesterolemia Display Paradoxical Associations with Lipid Profile and Promote Foam Cell Formation

Anti-Apolipoprotein A-1 autoantibodies (anti-ApoA-1 IgG) promote atherogenesis via innate immune receptors, and may impair cellular cholesterol homeostasis (CH). We explored the presence of anti-ApoA-1 IgG in children (5-15 years old) with or without familial hypercholesterolemia (FH), analyzing their association with lipid profiles, and studied their in vitro effects on foam cell formation, gene regulation, and their functional impact on cholesterol passive diffusion (PD)

The Great Pretender : Rectal Syphilis Mimic a Cancer

Rectal syphilis is a rare expression of the widely recognised sexual transmitted disease, also known as the great imitator for its peculiarity of being confused with mild anorectal diseases because of its vague symptoms or believed rectal malignancy, with the concrete risk of overtreatment. We present the case of a male patient with primary rectal syphilis, firstly diagnosed as rectal cancer; the medical, radiological, and endoscopic features are discussed below

Ultrasound approach as integration of gross anatomy educational path for medical students

For physicians, the human body is the focus of investigation and intervention on a daily basis. It follows that the study of anatomy will continue to be essential to safe medical practice [1]. Anatomical education represents the cultural path that includes the best coexistence of old techniques, and avant-garde. Thus teachers forming future physicians are imposed to find new strategies for the acquisition of adequate professional competences [2]. The gross anatomy course attended by medical students was integrated by ultrasound training. Students were trained either in palpating and recognizing surface body-landmarks, or in the detection of different viscera. Their abilities were then evaluated. For three academic years (since 2009-10 to 2011-12), all the 262 students enrolled in the first year of Medicine and Surgery degree (\u201cSan Paolo\u201d Hospital, Universit\ue0 degli Studi di Milano, Italy) participated. Of them, 16 volunteered in 2009-10, and 17 in each of the next two years, to preliminarily attend ultrasound training that their fellows would attend later. After this preliminary training, volunteers tutored their course fellows as peer tutors. All participants were either models or users. Each training presented three modules: 1) information about ultrasound scanning; 2) musculoskeletal system, major arterial and venous vessels, major nervous trunks, thyroid gland; 3) most thoracic, abdominal and pelvic viscera. Modules 2 and 3 were attended by small groups (6 students, assisted by 2 peer tutors). In module 2, topographical anatomy and subsequent recognition and palpation of surface bodylandmarks were also taught. The study of musculoskeletal system, major vessels and nerve trunks, and thyroid gland was supported by a multi-frequency probe equipped ultrasound machine. Thoracic, abdominal, and pelvic viscera were explored by a new generation pocket-sized ultrasound machine. Acquired skills were verified. The levels of expertise obtained by peer tutors and students were generally satisfactory. Students understood the importance of operative knowledge in human anatomical context. Anatomists found a valid method to consolidate the professionalizing quality of the topic

Doxorubicin upregulates CXCR4 via miR-200c/ZEB1-dependent mechanism in human cardiac mesenchymal progenitor cells.

Doxorubicin (DOXO) treatment is limited by its cardiotoxicity, since it causes cardiac-progenitor-cell depletion. Although the cardioprotective role of the stromal cell-derived factor-1/C-X-C chemokine receptor type 4 (SDF1/CXCR4) axis is well established, its involvement during DOXO-induced cardiotoxicity has never been investigated. We showed that in a mouse model of DOXO-induced cardiomyopathy, CXCR4 &lt;sup&gt;+&lt;/sup&gt; cells were increased in response to DOXO, mainly in human cardiac mesenchymal progenitor cells (CmPC), a subpopulation with regenerative potential. Our in vitro results showed a CXCR4 induction after 24 h of DOXO exposure in CmPC. SDF1 administration protected from DOXO-induced cell death and promoted CmPC migration. CXCR4 promoter analysis revealed zinc finger E-box binding homeobox 1 (ZEB1) binding sites. Upon DOXO treatment, ZEB1 binding decreased and RNA-polymerase-II increased, suggesting a DOXO-mediated transcriptional increase in CXCR4. Indeed, DOXO induced the upregulation of miR-200c, that directly targets ZEB1. SDF1 administration in DOXO-treated mice partially reverted the adverse remodeling, decreasing left ventricular (LV) end diastolic volume, LV ejection fraction and LV anterior wall thickness in diastole, recovering LV end systolic pressure and reducing±dP/dt. Moreover, in vivo administration of SDF1 partially reverted DOXO-induced miR-200c and p53 protein upregulation in mouse hearts. In addition, downmodulation of ZEB1 mRNA and protein by DOXO was significantly increased by SDF1. In keeping, p21 mRNA, that is induced by p53 and inhibited by ZEB1, is induced by DOXO treatment and is decreased by SDF1 administration. This study showed new players of the DOXO-induced cardiotoxicity, that can be exploited to ameliorate DOXO-associated cardiomyopathy

Virtual dissection by ultrasound: probe handling in the first year of medical education

Objectives The aim of the present study was to assess the educational plan of first-year students of medicine by analyzing their scores in ultrasound body scanning. Methods Since 2009, the San Paolo Medical School (Milan, Italy) has vertically integrated the study of anatomy with ultrasound-assisted virtual body dissection. Three modules were supplied: musculoskeletal system, heart and abdomen pelvis. 653 first-year students were trained. The students alternated as mutual model and operator. A skillfulness score was assigned to each student. The scores were consequently listed. Nonparametric exact multiple contrast tests were employed to determine relative group effects. Results Statistical analysis showed that: no gender-related differences were found (0:49; p = 0.769); peer learners performed less well than peer tutors (0.677; p = 0); between modules, scores in the musculoskeletal system (pMS = 0.726) tend to be higher (p < 0.001) than those obtained in the heart and abdomen pelvis (pH = 0.398; pAP = 0.375 p = 0.270); significant differences were found compared to the beginning of the project\u2019s academic year. Conclusion The students considered this didactic course an engaging and exciting approach. Acceptance of peer teaching was extraordinarily high. Autonomous exercitation allowed the students to improve self-criticism and enhance their own skills. The level of expertise obtained by peer tutors and by peer learners can be considered satisfactory. The main objective of training future physicians on personal stethoechoscope with the necessary competence seems to have been successfully started