Prolongation of incubation time improves clinical diagnosis of Mycobacterium xenopi infection and allows susceptibility testing of mycobacterial strains against multiple antibiotics.

Objectives: Mycobacterium xenopi is a nontuberculous mycobacterium (NTM) whose clinical diagnosis and drug susceptibility studies are frequently hampered by poor in vitro growth. Extending the culture incubation time from 42 days (common-standard) to 56 days could improve the likelihood of diagnosis and provide strains for phenotypic drug susceptibility profiling of this poorly studied but clinically relevant mycobacterium. Methods: Time-to-positivity of mycobacterial cultures incubated for 56 days were analysed and compared. Clinical mycobacteriosis was defined by ATS/IDSA criteria. In vitro susceptibility of M. xenopi isolates was tested by broth microdilution. Results: Of 3852 mycobacteria-positive cultures (26 different mycobacterial species),M. xenopi required by far the longest growth time in culture, exceeding the 42 days commonly used in routine diagnostics in 41.2% of cases versus 4.7% for other NTM and 2.0% for Mycobacterium tuberculosis complex (P < 0.001). Prolonging the incubation time to 56 days had a great impact on M. xenopi diagnosis, as 56.3% (27/48) of patients would have not fulfilled the ATS/IDSA criteria at an incubation limited to 42 days. All 40 M. xenopi isolates from patients with clinical mycobacteriosis were fully susceptibility to macrolides and rifamycins in vitro and to moxifloxacin, amikacin and linezolid. Conclusion: These results indicate that a significant percentage (56.3%) of positive culture forM. xenopi would have incorrectly been reported as negative to clinicians without prolonging the incubation time to 56 days. Moreover, 56.3% of patients with M. xenopi disease would have missed the diagnosis along with an appropriate germ-based antimycobacterial treatment, otherwise fully effective

One-stage vs two-stage bilateral THA in Lombardy: a cost-effectiveness analysis

Abstract Background Total hip arthroplasty (THA) is the most common treatment for primary and secondary end-stage hip osteoarthritis (OA). Almost 20% of all patients undergoing primary THA suffer from bilateral hip OA and, consequently, will need a contralateral procedure to be performed in the following years. The aim of this study is to evaluate the cost-effectiveness and the reliability of one-stage bilateral THA (1-BTHA) compared to two-stage bilateral THA (2-BTHA), in low-risk patients, performed with anterior minimally invasive surgery (AMIS). Methods Single patient’s costs were obtained by dividing the annual costs report by the number of hospitalizations, considering the diagnosis related group (DRG) of the two procedures. Then, 16 patients undergoing 1-BTHA and 8 undergoing 2-BTHA were examined. Hemoglobin (Hb) values before surgery and before discharge, transfusion rate and the occurrence of post-operative complications were observed. Results Procedural costs were divided in different subgroups: pre-hospitalization, operating room, hospital stay, post-operative follow-up and other costs. 1-BTHA total costs amount to 5.754,82€, while performing 2-BTHA costs 7.624,32€. However, considering DRG reimbursement, the hospital’s profit margin following 1-BTHA is lower than that following 2-BTHA (6.346,18€ versus 9.261,68€). Surgical time was found not to be significantly different between 1-BTHA and 2-BTHA (141,13 ± 26,1 min vs 164,8 ± 44,3 min; p = 0,111). The two groups showed a statistically significant difference in Hb decrease (4,8 ± 1,3 g/dl vs 3,3 ± 0,9; p = 0,001), despite no variances in transfusion rate. No further complications were observed in either group. Conclusions This study demonstrates how, in carefully selected patients, 1-BTHA performed with AMIS is a cost-effective and safe technique compared to 2-BTHA, resulting in a shorter OR time, LOS and lower overall costs. Level of evidence II

COVID-19 Elderly Patients Treated for Proximal Femoral Fractures during the Second Wave of Pandemic in Italy and Iran: A Comparison between Two Countries

Background and objevtive: The worldwide spread of SARS-CoV-2 has affected the various regions of the world differently. Italy and Iran have experienced a different adaptation to coexistence with the pandemic. Above all, fractures of the femur represent a large part of the necessary care for elderly patients. The aim of this study was to compare the treatment in Italy and Iran of COVID-19-positive patients suffering from proximal femur fractures in terms of characteristics, comorbidities, outcomes and complications. Materials and Methods: Medical records of COVID-19-positive patients with proximal femoral fractures treated at IRCCS Istituto Ortopedico Galeazzi in Milan (Italy) and at Salamat Farda and Parsa hospitals in the province of Tehran (Iran), in the time frame from 1 October 2020 to 16 January 2021, were analyzed and compared. Results: Records from 37 Italian patients and 33 Iranian patients were analyzed. The Italian group (mean age: 83.89 &plusmn; 1.60 years) was statistically older than the Iranian group (mean age: 75.18 &plusmn; 1.62 years) (p value = 0.0003). The mean number of transfusions for each patient in Italy was higher than the Iranian mean number (p value = 0.0062). The length of hospital stay in Italy was longer than in Iran (p value &lt; 0.0001). Furthermore, laboratory values were different in the post-operative value of WBC and admission and post-operative values of CRP. Conclusions: The present study shows that differences were found between COVID-19-positive patients with proximal femoral fractures in these two countries. Further studies are required to validate these results and to better explain the reasons behind these differences

Internet Use and Access, Behavior, Cyberbullying and Grooming. Revised questionnaire

The way children use the Internet and mobile technologies has changed in the last few years according to the Digital Agenda for Europe. The growing phenomenon of cyberbullying is among the risks associated with the increasing Internet use. Methods of access and use by teenagers are highly important factors in assessing those risks.In order to fight cyberbullying we expanded our previous study adding to our questionnaire a new cluster of 5 more items, specifically developed to investigate cyberbullying and cyber-grooming among the teenage population. This new cluster, proposed by the “Cyber Expert” program created by the Spanish Policia Nacional, is structured to gauge the students’ awareness and knowledge of what cyberbullying is through multiple choice questions. We submitted this updated survey to students of the Liceo Scientifico “G. Keplero” in Rome, while also inviting them to provide their feedback on the study itself. On the methodological side, since the data we gathered strongly hinted at the need to spread surveys and informational material using language and concepts adolescents can relate to more easily, we verified the need to review and update the questionnaire as often as required by the constant evolution of browsing habits in daily life. Here we present our revised questionnair

Full Mimicking of Coronary Hemodynamics for Ex-Vivo Stimulation of Human Saphenous Veins

After coronary artery bypass grafting, structural modifications of the saphenous vein wall lead to lumen narrowing in response to the altered hemodynamic conditions. Here we present the design of a novel ex vivo culture system conceived for mimicking central coronary artery hemodynamics, and we report the results of biomechanical stimulation experiments using human saphenous vein samples. The novel pulsatile system used an aortic-like pressure for forcing a time-dependent coronary-like resistance to obtain the corresponding coronary-like flow rate. The obtained pulsatile pressures and flow rates (diastolic/systolic: 80/120 mmHg and 200/100 mL/min, respectively) showed a reliable mimicking of the complex coronary hemodynamic environment. Saphenous vein segments from patients undergoing coronary artery bypass grafting (n = 12) were subjected to stimulation in our bioreactor with coronary pulsatile pressure/flow patterns or with venous-like perfusion. After 7-day stimulation, SVs were fixed and stained for morphometric evaluation and immunofluorescence. Results were compared with untreated segments of the same veins. Morphometric and immunofluorescence analysis revealed that 7 days of pulsatile stimulation: (i) did not affect integrity of the vessel wall and lumen perimeter, (ii) significantly decreased both intima and media thickness, (iii) led to partial endothelial denudation, and (iv) induced apoptosis in the vessel wall. These data are consistent with the early vessel remodeling events involved in venous bypass adaptation to arterial flow/pressure patterns. The pulsatile system proved to be a suitable device to identify ex vivo mechanical cues leading to graft adaptation

Human Osteochondral Explants as an Ex Vivo Model of Osteoarthritis for the Assessment of a Novel Class of Orthobiologics

Osteoarthritis (OA) is a highly prevalent joint disease still lacking effective treatments. Its multifactorial etiology hampers the development of relevant preclinical models to evaluate innovative therapeutic solutions. In the last decade, the potential of Mesenchymal Stem Cell (MSC) secretome, or conditioned medium (CM), has emerged as an alternative to cell therapy. Here, we investigated the effects of the CM from adipose MSCs (ASCs), accounting for both soluble factors and extracellular vesicles, on human osteochondral explants. Biopsies, isolated from total knee replacement surgery, were cultured without additional treatment or with the CM from 106 ASCs, both in the absence and in the presence of 10 ng/mL TNFα. Tissue viability and several OA-related hallmarks were monitored at 1, 3 and 6 days. Specimen viability was maintained over culture. After 3 days, TNFα induced the enhancement of matrix metalloproteinase activity and glycosaminoglycan release, both efficiently counteracted by CM. The screening of inflammatory lipids, proteases and cytokines outlined interesting modulations, driving the attention to new players in the OA process. Here, we confirmed the promising beneficial action of ASC secretome in the OA context and profiled several bioactive factors involved in its progression, in the perspective of accelerating an answer to its unmet clinical needs

Mycoplasma pneumoniae: IgG and IgM antibody response in presence of different antigens. Evaluation of commercial tests

Background. P1 adhesin protein (170-kDa) is responsible for the interaction between M. pneumoniae and the host. Such protein is the target of specific antibodies produced by the host in response to M. pneumoniae infection. Objectives. The aim of this study was to evaluate the performance of serological tests for Mycoplasma pneumoniae IgG and IgM on chemiluminescence analyzer LIAISON® and to demonstrate how the determination of specific antibodies is the main tool for accurate diagnosis of Mycoplasma infection. Study Design. Eighty-one unselected samples were assayed by ELISA kits Vircell IgG and IgM (bacterial lysate) and LIAISON® Mycoplasma IgG and IgM (recombinant protein P1). The discordant samples were tested with ELISA Savyon Serorecombinant IgG and IgM (recombinant protein P1) and reclassified according to the consensus rule. Results. Fifty-one samples were concordant between ELISA Vircell IgG and LIAISON® Mycoplasma IgG. After resolution of the 30 discordant: twenty-eight samples were classified concordant negative and one positive. Seventy-seven samples were concordant between ELISA Vircell IgM and LIAISON® Mycoplasma IgM. After resolution of 4 discordant samples: three samples were classified as negative. Conclusions. The higher correlation between LIAISON® and Savyon Serorecombinant is determined by the use of recombinant protein P1 in the solid phase and the type of cut-off calculation allows high specificity and sensitivity. The Mycoplasma pneumoniae IgG and IgM kits, performed in full automation on LIAISON® analyzer, show characteristics comparable to other kits and an easy handling of the sample

Regulation of oxytocin receptor gene expression in obsessive-compulsive disorder: a possible role for the microbiota-host epigenetic axis

Background Obsessive-compulsive disorder (OCD) is a prevalent and severe clinical condition. Robust evidence suggests a gene-environment interplay in its etiopathogenesis, yet the underlying molecular clues remain only partially understood. In order to further deepen our understanding of OCD, it is essential to ascertain how genes interact with environmental risk factors, a cross-talk that is thought to be mediated by epigenetic mechanisms. The human microbiota may be a key player, because bacterial metabolites can act as epigenetic modulators. We analyzed, in the blood and saliva of OCD subjects and healthy controls, the transcriptional regulation of the oxytocin receptor gene and, in saliva, also the different levels of major phyla. We also investigated the same molecular mechanisms in specific brain regions of socially isolated rats showing stereotyped behaviors reminiscent of OCD as well as short chain fatty acid levels in the feces of rats. Results Higher levels of oxytocin receptor gene DNA methylation, inversely correlated with gene expression, were observed in the blood as well as saliva of OCD subjects when compared to controls. Moreover, Actinobacteria also resulted higher in OCD and directly correlated with oxytocin receptor gene epigenetic alterations. The same pattern of changes was present in the prefrontal cortex of socially-isolated rats, where also altered levels of fecal butyrate were observed at the beginning of the isolation procedure. Conclusions This is the first demonstration of an interplay between microbiota modulation and epigenetic regulation of gene expression in OCD, opening new avenues for the understanding of disease trajectories and for the development of new therapeutic strategies

Platelet-Derived Growth Factor-D Enables Liver Myofibroblasts to Promote Tumor Lymphangiogenesis in Cholangiocarcinoma

BACKGROUND: /Aims. In cholangiocarcinoma, early metastatic spread via lymphatic vessels often precludes curative therapies. Cholangiocarcinoma invasiveness is fostered by an extensive stromal reaction, enriched in cancer-associated fibroblasts (CAF) and lymphatic endothelial cells (LEC). Cholangiocarcinoma cells recruit and activate CAF by secreting PDGF-D. Here we investigated the role of PDGF-D and liver myofibroblasts in promoting lymphangiogenesis in cholangiocarcinoma. METHODS: Human cholangiocarcinoma specimens were immunostained for podoplanin (LEC marker), \u3b1-SMA (CAF), VEGF-A, VEGF-C, and their cognate receptors (VEGFR2, VEGFR3). VEGF-A and VEGF-C secretion (ELISA) was evaluated in human fibroblasts obtained from primary sclerosing cholangitis explants stimulated by PDGF-D, with/without the PDGFR\u3b2 inhibitor imatinib. Using human LEC incubated with conditioned medium from PDGF-D-stimulated fibroblasts (with/without imatinib), we assessed migration (Boyden chambers), 3-D vascular assembly (AngioTool), trans-endothelial electric resistance and trans-endothelial migration of cholangiocarcinoma cells (EGI-1). In Fischer-344 rats transplanted with a syngeneic cholangiocarcinoma cell line, we studied effects of selective CAF depletion induced by the BH3 mimetic navitoclax on LEC density and lymphnode metastases. RESULTS: In cholangiocarcinoma specimens, CAF and LEC were closely adjacent. CAF expressed VEGF-A and VEGF-C, while LEC expressed VEGFR2 and VEGFR3. Upon PDGF-D stimulation, fibroblasts secreted increased levels of VEGF-C and VEGF-A. Fibroblasts, stimulated by PDGF-D induced LEC recruitment and 3-D assembly, increased LEC monolayer permeability, and promoted trans-endothelial EGI-1 migration. These effects were all suppressed by imatinib. In the rat model of cholangiocarcinoma, navitoclax-induced CAF depletion markedly reduced lymphatic vascularization and lymphnode metastases. CONCLUSION: PDGF-D stimulates VEGF-C and VEGF-A production by fibroblasts, resulting in expansion of the lymphatic vasculature and tumor cell intravasation. This process critical for the early metastatization of cholangiocarcinoma, may be blocked by inducing CAF apoptosis or by inhibiting PDGF-D-induced axis. LAY SUMMARY: Cholangiocarcinoma (CCA) is a highly malignant cancer affecting the biliary tree. In CCA, a rich stromal reaction densely populated by cancer-associated fibroblasts promotes early metastatic spread. Here we show that CCA-derived PDGF-D recruiting fibroblasts, also stimulates them to secrete VEGF-A and VEGF-C. These vascular growth factors kindle both lymphatic vascularization and tumour cell vascular invasion. Thus, targeting fibroblasts or PDGF-D-induced signals may represent an effective tool to block tumor-associated lymphangiogenesis and reduce CCA invasiveness