Combining germline, tissue and liquid biopsy analysis by comprehensive genomic profiling to improve the yield of actionable variants in a real-world cancer cohort.

BACKGROUND: Comprehensive next-generation sequencing is widely used for precision oncology and precision prevention approaches. We aimed to determine the yield of actionable gene variants, the capacity to uncover hereditary predisposition and liquid biopsy appropriateness instead of, or in addition to, tumor tissue analysis, in a real-world cohort of cancer patients, who may benefit the most from comprehensive genomic profiling. METHODS: Seventy-eight matched germline/tumor tissue/liquid biopsy DNA and RNA samples were profiled using the Hereditary Cancer Panel (germline) and the TruSight Oncology 500 panel (tumor tissue/cfDNA) from 23 patients consecutively enrolled at our center according to at least one of the following criteria: no available therapeutic options; long responding patients potentially fit for other therapies; rare tumor; suspected hereditary cancer; primary cancer with high metastatic potential; tumor of unknown primary origin. Variants were annotated for OncoKB and AMP/ASCO/CAP classification. RESULTS: The overall yield of actionable somatic and germline variants was 57% (13/23 patients), and 43.5%, excluding variants previously identified by somatic or germline routine testing. The accuracy of tumor/cfDNA germline-focused analysis was demonstrated by overlapping results of germline testing. Five germline variants in BRCA1, VHL, CHEK1, ATM genes would have been missed without extended genomic profiling. A previously undetected BRAF p.V600E mutation was emblematic of the clinical utility of this approach in a patient with a liver undifferentiated embryonal sarcoma responsive to BRAF/MEK inhibition. CONCLUSIONS: Our study confirms the clinical relevance of performing extended parallel tumor DNA and cfDNA testing to broaden therapeutic options, to longitudinally monitor cfDNA during patient treatment, and to uncover possible hereditary predisposition following tumor sequencing in patient care.

Femoral Neck Fractures in HIV-Positive Patients: Analysis of 10 Years Short-Term Post-operative Complications.

INTRODUCTION: Aging and effect of antiretroviral therapy on bone mass could increase the risk of femoral neck fractures (FNF) in HIV patient. The aim of this study was specifically to determine whether intracapsular FNF in HIV-positive patients are more prone to short-term post-operative complications than similar fractures occurring in HIV-negative patients. MATERIALS AND METHODS: A group of 25 HIV-positive patients with intracapsular FNF were enrolled and matched to HIV-negative patient with similar fractures according to gender, age, a modified Charlson Comorbidity Index (CCI), fracture classification, surgical treatment and time interval between fracture event and surgery. For each group, length of stay, surgical time, early clinical outcomes and short-term surgical and medical complications were compared to determine the impact on the early outcome. RESULTS: At the time of the fracture occurrence, 56% of HIV-positive patients were on antiretroviral therapy and 12% started with therapy in the perioperative period. At three months follow-up, there were no statistically significant differences between the two study groups in length of stay, Harris hip score and total number of early complications. However, a statistically significant increase in urinary tract infections and longer surgical time using hip sliding screw fixation were seen in the HIV-positive group. The poorest post-operative result was seen in a patient who failed to adequately adhere to the HIV therapy protocol. CONCLUSIONS: This study failed to show any statistically significant increase in short-term complications or worse clinical outcomes for intracapsular FNF in HIV-positive patients compared to HIV-negative patients to recommend their treatment in dedicated centres.

E-learning course improves knowledge in tobacco dependence, electronic nicotine delivery systems and heat-not-burn products in Medical School students.

BACKGROUND: Adequate training in tobacco, nicotine dependence and treatment is lacking in Medical School education. With the rise in popularity of electronic alternatives to cigarettes, future physicians should also be provided with the more recent scientific evidence on these products during their undergraduate studies. We introduced an e-learning course for Medical School students and assessed its effec-tiveness of increasing knowledge on these topics. METHODS: We developed 16 didactic modules divided in 3 courses: tobacco dependence (TDI), treating tobacco dependence (TDII) and electronic products and tobacco control (TDIII). The course was offered to 4th, 5th, and 6th year Medical School students in Italy. To assess learning outcomes, we examined the pre- to post- changes in knowledge scores associated with each course. Paired and independent samples t-tests were performed overall, and among smokers and non-smokers separately. RESULTS: A total of 1318 students completed at least one of the courses; 21% were self-reported smokers. A significant increase in knowledge was observed at the end of TDI (pre-course: 52.1±15.9, post-course: 79.9±13.5, p<0.001), TDII (pre-course: 52.5±13.0, post-course: 66.5±12.0, p<0.001) and TDIII (pre-course: 52.2±15.3, post-course: 76.1±17.7, p<0.001). Smokers showed significantly lower improvements compared to non-smokers. CONCLUSIONS: The e-learning course was effective in increasing knowledge about tobacco dependence, treatments, and electronic ni-cotine products in advanced medical students. Given the fundamental role for healthcare practitioners in encouraging and assisting people in quitting smoking, e-learning may be a useful tool in providing up-to-date and standardized training in the area during Medical School.

Skeletal muscle repair in a rat muscle injury model: the role of growth hormone (GH) injection.

OBJECTIVE: Muscle injury tends to heal with incomplete functional recovery. Among the growth factors released in the physio-pathological response of muscle lesion, the Insulin-like Growth-Factor-1 (IGF-1) results in an engine factor of the reparation program. The therapeutic use of growth factors has been exploited to improve healing. As IGF-1 is a primary mediator of the effects of growth hormone (GH), we exploited its systemic administration to muscle recovery in a rat model of muscle injury. MATERIALS AND METHODS: Monolateral lesion of the longissimus dorsi muscle of rats was performed. Animals were divided into 5 groups: four groups for histological studies and serum hormone dosage, whilst the fifth group represented the uninjured control. Rat GH was intraperitoneally administered after 24h from the surgical lesion at three different concentrations (0.1, 0.2, 0.4 mg/kg). At 3 days from surgery, immunohistochemical and histological analyses evaluated the expression of MyoD and Myogenin, and the presence of neovascularization and inflammation, respectively. After 2 months, we analyzed the presence of muscle regeneration and fibrosis. RESULTS: The treatment with GH resulted in a significant increase in neovascularization and Myogenin expression at 24h from injury in comparison with the control. This suggested speed up biological recovery times. After two-months, a dose-dependent increase of the connective component was observed. CONCLUSIONS: The potential effect of GH on muscle repair and regeneration, through the activation of satellite cells already demonstrated in vitro, was confirmed in this in vivo experimental approach. This study sheds light on the role of growth factors in damage repair mechanisms to find an appropriate biological treatment for muscle injury.

Global Health Professions Student Survey among Healthcare students: a cross sectional study.

BACKGROUND: Healthcare professionals have the potential to influence patients' smoking status; however, their smoking habits and their opinions about tobacco use can hamper an effective counselling about smoking to patients. METHODS: A cross-sectional survey was carried out among healthcare students from the Sapienza University of Rome using a previously validated Italian version of Global Health Professions Student Survey (GHPSS) online questionnaire. RESULTS: Overall, 349 students filled out the online survey: 158 (45%) were current smokers and 191 (55%) were nonsmokers. Nurses students had the highest percentage of smokers (55%). Healthcare students declared they had learned about the health consequences about tobacco use during their academic course (90%), however, only 24% received specific training on smoking cessation. CONCLUSIONS: It is necessary to implement a smoking cessation course in the undergraduate curriculum in order to train healthcare professionals to provide patients an effective counselling against smoking.

Unilateral multifocal choroiditis following EBV-positive mononucleosis responsive to immunosuppression: a case report.

BACKGROUND: Multifocal choroiditis (MFC) is a relatively uncommon bilateral inflammatory chorioretinopathy affecting Caucasian young women with myopia. We present images from a case of completely unilateral multifocal choroiditis following EBV-positive mononucleosis that demonstrated a dramatic clinical response to immunosuppression. CASE PRESENTATION: A 20-year-old woman with bilateral high myopia (-6D) and a documented normal prior retinal examination presented with visual loss in the right eye 2 months following confirmed Epstein-Barr virus (EBV) positive mononucleosis. Ophthalmoscopic examination showed completely unilateral placoid lesions of variable age. The left eye was unaffected. Fluorescein angiography revealed active leakage, especially in the parafovea. Spectral domain optical coherence tomography (SD-OCT) demonstrated sub-retinal pigment epithelial nodular deposits, some of which were confluent with overlying intra-retinal fluid and indistinct margins. Upon treatment with the immunosuppressant azathioprine there was significant resolution of the lesions in her right eye along with improvement in vision. CONCLUSION: This is a rare case of completely unilateral MFC following an episode of EBV positive mononucleosis that showed a dramatic response to immunosuppression.

Gastrointestinal stromal tumors and second primary malignancies: a retrospective monocentric analysis.

In the post-Imatinib era, the median survival of patients diagnosed with GIST has reached almost 5 years. Prolonging GIST-specific survival, GIST patients have an increased incidence of secondary neoplasia. Data on the prognostic impact of second tumors in GIST patients are very poor with few and small retrospective analyses available in the literature. We conducted a retrospective monocentric analysis on 145 patients diagnosed with GIST between April 2001 and October 2018. Kaplan-Meier and Cox hazard methods were used for survival analysis. A total of 154 GIST patients were included and 31 patients of them (21%) were diagnosed with at least one additional malignancy. The most common second tumors associated with GIST were gastrointestinal tumors. GIST patients with additional malignancies showed to have lower size (>5 cm: 35% vs 45%; p=0.75), higher mitotic rate (>5/50 HPFs: 42% vs 29%; p=0.24), higher presence of c-KIT mutation (85% vs 69%), a lower presence of PDGFRα mutation (8% vs 17%; p=0.05) and shorter survival (mOS: 9.6 vs 15.5 years; p=0.30). In conclusion, our study did not find any significant correlation between clinicopathological characteristics and the development of a second tumor in GIST patients. Further analyses and strict follow up protocols are needed in order to early diagnose and promptly treat a second primary tumor in the GIST population.

Effectiveness of a training course on smoking cessation knowledge and behaviour for health profession students: the SISMA project.

INTRODUCTION: University students are at risk of starting smoking or continuing and increasing the consumption of tobacco products. The aim of the study was to assess the impact of the training course, Sisma Project, about smoking in healthcare degree courses, in terms of knowledge, behaviour and to evaluate the course. METHODS: SISMA project was a pre- post study about an intervention delivered to healthcare profession students about smoking and smoking cessation. It had a before-after design and was an online optional course available on the eLearning platform Moodle 2. The course was structured in four lessons of sixty minutes, a debate among experts and a final test of evaluation. The McNemar test was used to measure the effectiveness of Sisma on smoking behaviour of students after the intervention. Students rated the course assigning a score from one to ten, and expressed free comments about point of strength and weakness of Sisma project. RESULTS: The participants were 365 students, 28.5% males and 71.5% females, most were nursing 194 (53.2%) and dental hygienists students 105 (28.8%). Current smokers were 161 (44.1%) before and 142 (38.9%) after the course, there was statistical significant difference in smoking status after attending the course (p < 0.001). Students evaluated the course giving a high score with a mean of 8.13 (SD: 1.1); the main points of strength were the content (33.2%), the structure (15.6%) and knowledge given by the course (12.6%). The main point of weakness were the online structure 62 (37%), problem related to length and time 17 (10%) and the final test 15 (9%). DISCUSSION: Given the central role health professionals play in patient care, students need to be aware and trained in tobacco cessation techniques. Our results indicate that smoking behaviour significantly changed after attending a university course for smoking cessation and students appreciated its contents and structure.

Smoking E-CigaRette and HEat-noT-burn products: validation of the SECRHET questionnaire.

BACKGROUND: The nicotine market has rapidly evolved with the emergence of newer forms of smoking device that have been expanded worldwide, such as electronic cigarettes (eCig) which heat a solution (e-liquid) to create vapour and heat-not-burn (HNB) tobacco products, which heats tobacco at a temperature below the point of combustion. Their use is increasing at an alarming rate; it is believed it will surpass the use of traditional cigarettes in next 5 years, mostly among never-smokers and young people. OBJECTIVE: There are not many studies investigating knowledge and behaviour about heat tobacco products (HTP) among teenagers, so the aim of this study is to validate the SECRHET questionnaire (Smoking E-CigaRette and HEat-noT-burn products) on knowledge and behaviour on cigarette and HTP among youth. METHODS: The study was conducted in February and March 2019 inside Sapienza University of Rome and high school Giulio Cesare of Rome. A self-administered anonymous questionnaire was performed to investigate smoking habits and measures knowledge about HTP among Italian teenagers. OUTCOMES: A sample of 60 students took part in the validation of the questionnaire. The overall Cronbach's alpha was 0,635, corresponding to a sufficient reliability. CONCLUSIONS: There is little or no data on consumption or perceptions of HTP products among youth and evidence suggests the usefulness of a standardized and validated questionnaire available to monitoring of awareness, interest in trying and prevalence of use of these novel products among young people.

Knowledge of nicotine dependence and treatment in clinical practice improved after an e-learning course among medical students.

OBJECTIVE: Tobacco smoking is the leading cause of preventable death in developed countries and smokers should be encouraged to quit. Physicians are instrumental in this, but recent reports suggest inadequate training in medical school. We aimed to assess the knowledge of nicotine dependence and its treatment among Italian medical students. STUDY DESIGN: Prospective observational study. METHODS: We developed an online course consisting of 11 Didactic Modules (6 for Tobacco Dependence I, TDI, and 5 for TDII) on nicotine dependence and treatment. The course was administered to 4th and 5th year medical students in Italy in Academic Years 2016-17 (Course A) and 2017-18 (Course B). A validated questionnaire was used before and after each part in order to measure knowledge of smoking epidemiology, health effects and benefits of giving up smoking ("Score 1", TDI), and effectiveness of cessation treatments ("Score 2", TDII). RESULTS: 324 students took both TDI and TDII and completed all questionnaires (Course A, n = 245; Course B, n = 79). 55 students were current smokers (17%). A significant increase in score 1 and 2 was observed at the end of both TDI (pre-course 47.2±13.1, post-course 66.0±12.3, p <0.0001) and TDII (pre-course 55.6±11.5, post-course 68.1±10.9, p <0.0001). The prevalence of students wishing for a smoke-free medical school significantly increased between the beginning of TDI (74.4%, 241/324) and the end of TDII (88%, 285/324; p <0.0001). CONCLUSIONS: This e-learning course has proven to be an effective tool in teaching students on nicotine dependence and treatment.

Epidemiological, clinical and pathological characteristics of gastric neoplasms in the province of Cremona: the experience of the first population-based specialized gastric cancer registry in Italy.

BACKGROUND: The gastric cancer incidence rate differs widely across geographical areas. In Italy, in the province of Cremona the incidence is high, compared to the national situation. For this reason a specialized population-based registry was set up. METHODS: The collection encompasses all gastric cancers diagnosed in the three districts of the province since January 1, 2010. The main data sources were the pathological and Hospital Discharge Records and patient clinical charts. Only diagnoses of primary gastric cancer were considered. For each case the following variables were registered: personal data, medical history and symptoms at diagnosis; imaging assessments performed, details on surgery and other treatments received; genetic background and biomolecular characteristics; social and environmental factors. RESULTS: As of November 2017, 1087 cases were collected; of which 876, diagnosed up to December 2015, were analyzed. Male/female ratio was 1.4. The European Age-standardized Incidence Rate was 41.4 for males and 28.3 for females as compared to a national average of 33.3 and 17.0 respectively. Median age at diagnosis was 73 for male and 78 for female. Helicobacter Pylori infection was present in fewer than 20% of cases. HER-2 gene was amplified in about 25% of cases. Primary tumour location was the gastro-esophageal junction or cardia in 17.5% in males and 8.3% in females. The majority of cases (58.3%) were diagnosed at an advanced stage and overall only 41.2% underwent surgery. Median overall survival was 14.8 months for men and 18.5 for women. Age standardized 5-year relative survival was 31.4% for men and 40.5% for females. Neoadjuvant treatment was performed in fewer than 10% of patients who underwent surgery, and the rate of postoperative therapy adherence was low. DISCUSSION: This study shows a high gastric cancer incidence in the province of Cremona, with a geographical spread across different districts. Moreover, a high percentage of gastric cancers were detected at an advanced stage of disease and a low rate of 5-year relative survival was registered. Based on these findings, effective preventive interventional health strategies and screening procedures need to be implemented to reduce the impact of this pathology in this geographical area.

Secondary breast angiosarcoma: A multicentre retrospective survey by the national Italian association of Breast Surgeons (ANISC).

BACKGROUND: Breast angiosarcoma is a malignant mesenchymal neoplasm, which accounts for approximately 2% of all soft tissue sarcomas. Secondary breast angiosarcoma (SBA) may be related to chronic lymphedema after a mastectomy with lymph node dissection (Stewart Treves syndrome) and previous radiotherapy for complications from breast radiation treatment. It is a very rare condition; therefore, diagnosis and management are still a challenge. METHODS: The ANISC collected SBA data by means of a survey sent to all Italian breast centres in the ANISC. The clinicopathological characteristics and the management of this disease were analysed. RESULTS: Twenty-four centres participated in this survey in which 112 cases of SBA were analysed. The median age of the women with SBA was 68.9 years and it appeared approximately 90 months after the first irradiation for breast cancer. In 92% of cases, a mastectomy was performed without axillary dissection for those patients having a high grade of SBA (74.2%). The prognosis was worse in the high-grade cases (overall survival-OS: 36 months) as compared with the low-grade cases (OS: 48 months). After a follow-up of 5 years, 50.5% of the patients were still alive. Disease-free survival (DFS) was 35 months, and there were no differences between the groups of patients with either high- or low-grade histology. CONCLUSIONS: Secondary breast angiosarcoma is a very aggressive disease associated with a short survival outcome. The surgical approach still remains an important step in the course of treatment; furthermore, an accurate histological examination is helpful in establishing the prognosis of the patient. A mastectomy is mandatory. A longer OS was observed in patients with low-grade angiosarcoma as compared to high-grade angiosarcoma (C.I. 40-57 vs. 31-41 months).

The synergistic effect between Positivity, socio-demographic factors and smoking cessation: results of a cohort study.

OBJECTIVE: To examine the extent to which a effect does exist between Positivity (POS), smoking and socio-demographic factors in determining quitting smoking in subjects participating in a Group Counselling Program (GCP) for smoking cessation. METHODS: 481 subjects were contacted through a telephone call. A logistic regression analysis was carried out. Possible interaction between sociodemographic variables and POS level was tested using the Synergism Index (SI). RESULTS: For individuals with a POS level over or equal to 3.4 the odds of being smoker was significantly higher among females (OR = 1.55), who smoked at home (OR = 2.16) and lower if there had children at home (OR = 0.53). For individuals with a POS level under 3.4, the only significant variable associated with smoking was beinga female (OR = 2.58). As far concerns the synergistic effect between the variables considered does exist between POS levels and having children at home (SI=1.13) and female gender (SI = 2.8). CONCLUSIONS: The synergistic effect between POS and sociodemographic factors adds evidence on the use of POS as possible determinants of individual happiness.

Multidimensional Evaluation of Endogenous and Health Factors Affecting Food Preferences, Taste and Smell Perception.

OBJECTIVE: This study, by taking a holistic approach, investigates the relationships between taste, smell sensitivity and food preference with prognostic (endogenous and health) factors including age, gender, genetic taste markers, body mass, cigarette smoking, and number of drugs used. DESIGN: Cross sectional study. SETTING: Northern Italy. PARTICIPANTS: 203 healthy subjects (160 women/43 men; mean age: 58.2±19.8 years) were examined. MEASUREMENTS: Individual taste sensitivity was determined by saccharose, sodium chloride, acetic acid and caffeine solutions and by 6-n-propylthiouracil (PROP) responsiveness test. Olfactory sensitivity has been assessed by «Sniffin' Sticks¼. Four tag Single nucleotide polymorphisms (SNPs) in regions of interest were genotyped. Factor analysis and multivariate regression were performed for scaling food preferences and screening prognostic factors, respectively. RESULTS: Increasing age is associated with decreased responsiveness to NaCl (P=0.001), sweet solutions (P=0.044), and smell perception (P<0.001). Concerning the food preferences, elderly like the "vegetables" and "fruits" but dislike "spicy" more than younger. Regarding number of drugs taken, there is a significant negative effect on smell perception (P<0.001). In addition, drugs reduce both the "vegetables foods" score (P=0.002) and the "milk-product foods" score (P=0.027). With respect to Body Mass Index (BMI), only a significant effect was shown, on sweet perception (P=0.006). Variation in taste receptor genes can give rise to differential perception of sweet, acid and bitter tastes. No effect of gender and smoking was observed. CONCLUSIONS: Our study suggested that age, genetic markers, BMI and drugs use are the factors which affect taste and smell perception and food preferences.

Smoking cessation practice among family doctors in Lithuania: a Survey.

OBJECTIVES: Relatively low proportion of ex-smokers and high rates of smoking-related diseases are estimated in Lithuania. The aim of this survey was to assess the extent of smoking cessation assistance among family doctors in Lithuania, and to identify factors associated with cessation interventions. MATERIALS AND METHODS: Data were collected using a questionnaire addressing family doctors' cessation assistance practice; knowledge of smoking epidemiology and health effects; knowledge of cessation assistance guidelines; smoking status; and attendance on Courses/Conferences on Smoking and smoking cessation Assistance (CCSA). The study included 718 family doctors in Lithuania in 2014. RESULTS: The results show that 23% of family doctors always ask their patients about smoking habits. Although 59% of family doctors always advise smoking patients to quit smoking, only 26% recommend a follow-up appointment, 22% refer for a specialized smoking cessation treatment, 15% prescribe nicotine replacement therapy, and 6% prescribe Varenicline or Bupropion. The study demonstrated limited knowledge of family doctors in relation to epidemiological aspects of tobacco use and effective cessation assistance methods. Cessation assistance is influenced by family doctors' age, smoking status, knowledge of cessation assistance guidelines and CCSA attendance. CONCLUSIONS: Effective training of family doctors' in smoking epidemiology, health hazards and in smoking cessation assistance is warranted in Lithuania. In addition, coordinated efforts in primary health care institutions are required to improve help for smokers who intend to quit.

Effects of ecologically relevant concentrations of cadmium in a freshwater fish.

Sub-chronic effects of ecologically relevant concentrations of cadmium (Cd) were evaluated in the catfish Rhamdia quelen. The fish were exposed to Cd (0, 0.1, 1, 10 and 100µgL(-1)) for 15 days. Bioconcentration was observed in the liver of fish exposed to 10 and 100µgL(-1) of cadmium. The liver glutathione S-transferase activity decreased at 0.1 and 1µgL(-1) and increased at 100µgL(-1) and lipoperoxidation increased in all tested concentrations. Fish exposed to 0.1, 1 and 100µgL(-1) Cd presented increase in hepatic lesion index. In the kidney, the catalase activity and LPO reduced in all exposed groups. The gluthatione peroxidase, etoxiresorufin-O-deethylase activities and metallothionein increased at the highest concentration of Cd, but the level of reduced glutathione decreased. The genotoxicity was observed at 0.1 and 100µgL(-1). Neurotoxicity was not observed. The results showed that low concentrations (range of µgL(-1)) of Cd caused hepato-, nephro- and hematological alterations in this freshwater fish species.

Bilateral Chronic Subdural Nontraumatic Hematoma in a Woman Affected by AL Amyloidosis.

Chronic subdural hematoma (CSDH) is a condition frequently seen in neurosurgical practice, especially among the elderly. It is often preceded by head injury, even a trivial trauma. Light chain (AL) amyloidosis is a disorder involving extracellular tissue deposition of misfolded native proteins called amyloids. The several types of amyloidosis differ by source of proteins, organ involvement, treatment, and prognosis. We describe the case of a 59-year-old woman affected by AL amyloidosis, harboring surgically treated bilateral chronic nontraumatic subdural hematoma, and we suggest a possible correlation between the clinical entities of CSDH and AL amyloidosis.

Development of a simple, biocompatible and cost-effective Inulin-Diethylenetriamine based siRNA delivery system.

Small interfering RNAs (siRNAs) have the potential to be of therapeutic value for many human diseases. So far, however, a serious obstacle to their therapeutic use is represented by the absence of appropriate delivery systems able to protect them from degradation and to allow an efficient cellular uptake. In this work we developed a siRNA delivery system based on inulin (Inu), an abundant and natural polysaccharide. Inu was functionalized via the conjugation with diethylenetriamine (DETA) residues to form the complex Inu-DETA. We studied the size, surface charge and the shape of the Inu-DETA/siRNA complexes; additionally, the cytotoxicity, the silencing efficacy and the cell uptake-mechanisms were studied in the human bronchial epithelial cells (16HBE) and in the hepatocellular carcinoma derived cells (JHH6). The results presented here indicate that Inu-DETA copolymers can effectively bind siRNAs, are highly cytocompatible and, in JHH6, can effectively deliver functional siRNAs. Optimal delivery is observed using a weight ratio Inu-DETA/siRNA of 4 that corresponds to polyplexes with an average size of 600nm and a slightly negative surface charge. Moreover, the uptake and trafficking mechanisms, mainly based on micropinocytosis and clatrin mediated endocytosis, allow the homogeneous diffusion of siRNA within the cytoplasm of JHH6. Notably, in 16 HBE where the trafficking mechanism (caveolae mediated endocytosis) does not allow an even distribution of siRNA within the cell cytoplasm, no significant siRNA activity is observed. In conclusion, we developed a novel inulin-based siRNA delivery system able to efficiently release siRNA in JHH6 with negligible cytotoxicity thus opening the way for further testing in more complex in vivo models.

Lateral ankle and hindfoot instability: a new clinical based classification.

Ankle sprains are one of the most common soft tissue injuries accounting for nearly 40% of sports injuries. There are large number of procedures for its treatment reported in the literature with largely good results. The ankle forms a functional unit with the subtalar joint. We present a new classification for peritalar lateral instability. There are two intents of this classification. Firstly, the classification demonstrates an assessment and treatment guideline for the many causes of peritalar lateral instability. The second use of the classification is for research purposes so that cohorts of patients can be accurately described and the efficacy of different operations in different groups can be properly assessed.