Global microRNA profiling identified miR-10b-5p as a regulator of neurofibromatosis 1 (NF1)-glioma migration.

AIMS: Neurofibromatosis 1 (NF1) is an autosomal-dominant cancer predisposition syndrome caused by loss of function alterations involving the NF1 locus on chromosome 17. The most common brain tumours encountered in affected patients are low-grade gliomas (pilocytic astrocytomas), although high-grade gliomas are also observed at increased frequency. While bi-allelic NF1 loss characterizes these tumours, previous studies have suggested noncoding RNA molecules (microRNA, miR) may have important roles in dictating glioma biology. METHODS: To explore the contributions of miRs in NF1-associated gliomas, we analysed five high-grade gliomas (NF1-HGG) and five PAs (NF1-PA) using global microRNA profiling with NanoString-based microarrays followed by functional experiments with glioma cell lines. RESULTS: miR-10b-5p, miR-135b-5p, miR-196a-5p, miR-196b-5p, miR-1247-5p and miR-320a (adjusted P < 0.05) were increased> 3-fold in NF1-HGG relative to NF1-PA tumours. In addition, miR-378b and miR-1305 were decreased 6.8- and 6-fold, respectively, whereas miR-451a was increased 2.7-fold (adjusted P < 0.05) in NF1-PAs compared to non-neoplastic NF1 patient brain specimens (n = 2). As miR-10b-5p was the microRNA overexpressed the most in NF1-high-grade glioma compared to NF1-low-grade glioma (5.76 fold), we examined its levels in glioma cell lines. miR-10b-5p levels were highest in adult glioma cell lines and lowest in paediatric low-grade glioma lines (P = 0.02). miR-10b-5p knockdown resulted in decreased invasion in NF1-deficient LN229 high-grade glioma line, whereas its overexpression in the NF1-PA derived line (JHH-NF1-PA1) led to increased invasion. There was no change in cell growth (viability and proliferation). CONCLUSIONS: These proof-of-concept experiments support a role for microRNA regulation in NF1-glioma biology.

Neutrophil to lymphocyte ratio (NLR) improves the risk assessment of ISS staging in newly diagnosed MM patients treated upfront with novel agents.

Recent reports identify the ratio between absolute neutrophil count (ANC) and absolute lymphocyte count (ALC), called neutrophil to lymphocyte ratio (NLR), as a predictor of progression-free survival (PFS) and overall survival (OS) in various malignancies. We retrospectively examined the NLR in a cohort of 309 newly diagnosed multiple myeloma (MM) patients treated upfront with novel agents. NLR was calculated using data obtained from the complete blood count (CBC) at diagnosis and subsequently correlated with PFS and OS. The median NLR was 1.9 (range 0.4-15.9). Higher NLR was independent of international staging system (ISS) stage, plasma cell infiltration or cytogenetics. The 5-year PFS and OS estimates were, respectively, 18.2 and 36.4 % for patients with NLR ≥ 2 versus 25.5 and 66.6 % in patients with NLR < 2. Among younger patients (age <65 years, N = 179), NLR ≥ 2 had a negative prognostic impact on both PFS and OS, in all ISS stages. By combining ISS stage and NLR in a model limited to young patients, we found that 19 % of the patients were classified as very low risk, 70 % standard risk and 11 % very high risk. The 5-year estimates were 39.3, 19.4 and 10.9 % for PFS and 95.8, 50.9 and 23.6 % for OS for very low, standard-risk and very high-risk groups. We found NLR to be a predictor of PFS and OS in MM patients treated upfront with novel agents. NLR can be combined with ISS staging system to identify patients with dismal outcome. However, larger cohorts and prospective studies are needed to use NLR as additional parameter to personalise MM therapy in the era of novel agents.

Androgen-induced programs for prostate epithelial growth and invasion arise in embryogenesis and are reactivated in cancer.

Cancer cells differentiate along specific lineages that largely determine their clinical and biologic behavior. Distinct cancer phenotypes from different cells and organs likely result from unique gene expression repertoires established in the embryo and maintained after malignant transformation. We used comprehensive gene expression analysis to examine this concept in the prostate, an organ with a tractable developmental program and a high propensity for cancer. We focused on gene expression in the murine prostate rudiment at three time points during the first 48 h of exposure to androgen, which initiates proliferation and invasion of prostate epithelial buds into surrounding urogenital sinus mesenchyme. Here, we show that androgen exposure regulates genes previously implicated in prostate carcinogenesis comprising pathways for the phosphatase and tensin homolog (PTEN), fibroblast growth factor (FGF)/mitogen-activated protein kinase (MAPK), and Wnt signaling along with cellular programs regulating such 'hallmarks' of cancer as angiogenesis, apoptosis, migration and proliferation. We found statistically significant evidence for novel androgen-induced gene regulation events that establish and/or maintain prostate cell fate. These include modulation of gene expression through microRNAs, expression of specific transcription factors, and regulation of their predicted targets. By querying public gene expression databases from other tissues, we found that rather than generally characterizing androgen exposure or epithelial budding, the early prostate development program more closely resembles the program for human prostate cancer. Most importantly, early androgen-regulated genes and functional themes associated with prostate development were highly enriched in contrasts between increasingly lethal forms of prostate cancer, confirming a 'reactivation' of embryonic pathways for proliferation and invasion in prostate cancer progression. Among the genes with the most significant links to the development and cancer, we highlight coordinate induction of the transcription factor Sox9 and suppression of the proapoptotic phospholipid-binding protein Annexin A1 that link early prostate development to early prostate carcinogenesis. These results credential early prostate development as a reliable and valid model system for the investigation of genes and pathways that drive prostate cancer.

Discovery of 342 putative new genes from the analysis of 5'-end-sequenced full-length-enriched cDNA human transcripts.

In this work we describe the process that, starting with the production of human full-length-enriched cDNA libraries using the CAP-Trapper method, led us to the discovery of 342 putative new human genes. Twenty-three thousand full-length-enriched clones, obtained from various cell lines and tissues in different developmental stages, were 5'-end sequenced, allowing the identification of a pool of 5300 unique cDNAs. By comparing these sequences to various human and vertebrate nucleotide databases we found that about 40% of our clones extended previously annotated 5' ends, 662 clones were likely to represent splice variants of known genes, and finally 342 clones remained unknown, with no or poor functional annotation. cDNA-microarray gene expression analysis showed that 260 of 342 unknown clones are expressed in at least one cell line and/or tissue. Further analysis of their sequences and the corresponding genomic locations allowed us to conclude that most of them represent potential novel genes, with only a small fraction having protein-coding potential.

Retrograde nasotracheal intubation with a new tracheal tube: a feasibility study.

We have assessed the feasibility of retrograde nasotracheal intubation using a flexometallic tracheal tube with a detachable pilot balloon and connector in a study of 20 consecutive adult patients undergoing oropharyngeal surgery. The technique consisted of: (1) laryngoscope-guided orotracheal intubation; (2) insertion of an 18-gauge Foley catheter through the nose and retraction into the mouth; (3) detachment of the anaesthesia circuit, pilot balloon and connector; (4) insertion of the Foley catheter tip into the proximal end of the tracheal tube and inflation of the Foley catheter cuff; (5) withdrawal of the Foley catheter and attached tracheal tube back through the nose; (6) deflation of the Foley catheter cuff; and (7) re-attachment of the pilot balloon, connector and anaesthesia circuit. The technique was successful at the first attempt in all patients. Mean time taken to insert the Foley catheter and retract it into the mouth was 19 (range 12-30) s. Mean time taken from disconnection to reconnection of the anaesthesia circuit was 8 (6-10) s. Heart rate increased after intubation, but there were no significant changes in arterial pressure. Nasal bleeding, airway problems and hypoxic events did not occur. No anatomical abnormalities or nasal trauma were detected at rhinoscopy. We conclude that retrograde nasotracheal intubation is feasible using a flexometallic tracheal tube with a detachable pilot balloon and connector.

The intubating laryngeal mask for maxillo-facial trauma.

We report the successful use of the intubating laryngeal mask airway in a patient with maxillo-facial trauma for whom the facemask and laryngoscope were relatively contraindicated and the fibreoptic scope potentially difficult to use.

Campus Bio-Medico technique for nasolaryngeal ventilation with reinforced laryngeal mask in dental surgery: a patient report.

The authors report the usefulness of a prototype nasal laryngeal mask airway (LMA) used successfully in a disabled 20-year-old woman with severe psychomotor retardation and a compromised airway with predictable indexes of impossible tracheal intubation in direct laryngoscopy. A 16-ch Foley catheter was inserted through the patient's left nostril and guided through her mouth. A size-3 reinforced LMA was positioned and connected to the distal end of the catheter. The LMA-reinforced tube was removed in a retrograde fashion by pulling the catheter up with the patient breathing spontaneously. The duration of the entire operation was 3 hours 20 minutes, and the patient was able to breathe spontaneously and at a 98% saturation average. Nasal reinforced LMA seems to be an interesting solution in patients undergoing 1-day dental or maxillofacial surgery, but is especially appropriate when nasotracheal intubation is too invasive or technically impossible.

The intubating laryngeal mask. Clinical appraisal of ventilation and blind tracheal intubation in 110 patients.

This study assesses the efficacy of the intubating laryngeal mask as a ventilation device and blind intubation guide. Following induction of anaesthesia with propofol, the device was successfully inserted at the first attempt in 110/110 (100%) patients. Placement took less than 10 s in all patients. Size selection was based on nose-chin distance. Adequate ventilation was achieved in 104/110 (95%) patients. Blind tracheal intubation using an 8-mm internal diameter straight silicone cuffed tracheal tube was attempted 3 min after the administration of vecuronium. Passage of a lighted stylet through the intubating laryngeal mask was used to determine the position of the intubating laryngeal mask cuff before blind intubation. If resistance was felt during intubation, a sequence of adjusting manoeuvres was used, based on the depth at which resistance occurred. Tracheal intubation was possible in 104/104 (100%) patients. In 42 (40%) patients, no resistance was encountered and the trachea was intubated at the first attempt. Sixty-two (60%) patients required one adjusting manoeuvre. The mean (range) time taken to successful intubation, i.e. the time from disconnection of the intubating laryngeal mask from the breathing system to successful tracheal intubation, was 79 (12-315) s. Six patients with potential or known intubation problems were included in the study. The tracheas of all six patients were successfully intubated. We conclude that the intubating laryngeal mask is an effective ventilation device and intubation guide with potential for use in patients who may present difficulty in tracheal intubation.

[Intra-cavernous injections of vasoactive drugs. A 10-year experience at an andrologic center]. / Le iniezioni intracavernose con farmaci vasoattivi. Esperienza decennale di un centro di andrologia.

The clinical experience of an andrological center in the use of the intra cavernous injections of vaso-active drugs is revisited. Clinical data concerning 1513 ICIs performed, in the last ten years, in 1038 patients have been considered. Various vasoactive drugs have been utilized. Mainly papaverine and prostaglandin E1 are usually employed. The experience and the establishment of the modalities of use both as diagnostic tool and as therapeutic procedure of this particular type of stimulation of the intracavernous vascular system has greatly reduced the possibility of complications and collateral side effects.

Nocturnal and real-time Rigiscan monitoring in patients with venous incompetence.

OBJECTIVES: In cases of failure of the vascular occlusive system, testing with both nocturnal and real-time monitoring will offer the possibility of making an accurate diagnosis of venous occlusive dysfunction. METHODS/RESULTS: The study was conducted in 124 cases suspected as having venous occlusive dysfunction (VOD). All patients were submitted to cavernometry, which confirmed VOD in 41 cases; however, the existence of a venous disorder could not be established in 83 cases. Nocturnal monitoring with the Rigiscan System was performed in 68 cases; no organic disorder was found in 6 cases. CONCLUSIONS: Although the results of nocturnal monitoring with the Rigiscan is not always consistent, in our experience it has been useful in both nocturnal and real-time monitoring of the erectile responses during many diagnostic tests such as VSS or intracavernosal injection of vasoactive agents.

Is it useful to operate on adolescent patients affected by left varicocele?

In order to consider the possibility of avoiding seminal disorders induced by varicocele at puberty, we evaluated 200 patients who were operated on for left varicocele at adolescence. Seventy-five of our 200 adolescent patients had evaluations of seminal fluid prior to their varicocelectomy. Of these patients, 75 were re-evaluated at the age of 21. In this latter group of patients, we observed 24 cases of hydrocele (32%) and 9 cases of varicocele recurrence (12%). These data were compared to 75 patients first presenting with a varicocele at 21-25 years of age. If we exclude the cases with azoospermia, we observed the presence of altered seminal fluid in 35 (46.7%) of the operated at adolescence patients and in 42 (56%) of the adult patients with varicocele. These data support the hypothesis that operating on patients in adolescence may result in maintenance of fertility potential.

Papillary cystadenoma of the epididymis as a cause of obstructive azoospermia.

We report a case of infertility due to obstructive azoospermia determined by papillary cystadenoma of the epididymis. The 28-year-old patient had other signs of von Hippel-Lindau syndrome and died from neurological complications. The possibility of finding an epididymal neoplasm should be kept in mind in any case of azoospermia with epididymal enlargement.

[Life habits and impotence]. / Abitudini di vita ed impotenza.

The authors propose their experience of 595 men complaining of erectile dysfunction. Relationships among erectile dysfunction, taking specific medicaments (hypotensive agents) and habits (smoking) are considered. A collaboration among specialists in the management of impotent patients has been proposed.

[Arterial pressure and heart rate: postural variations in the elderly]. / Pressione arteriosa e frequenza cardiaca: variazioni posturali nell'anziano.

The behaviour of arterial blood pressure and heart rate after an active change from the supine to the upright posture was studied in the aged. A significant decrease in the average values for systolic and diastolic blood pressure and a significant increase in the average values for heart rate in the first minute of the upright posture were observed. The systolic and diastolic pressures are reduced in a higher percentage of the cases in the 80-94 year age class. In the same class the PAS is more frequently reduced in the male and the PAD in the female. The heart rate, in the upright posture, is higher in 80-94 year-old males without a significant difference between the sexes.

Viability and function of platelets frozen at 2 to 3 C per minute with 4 or 5 per cent DMSO and stored at -80 C for 8 months.

Each of 15 healthy male volunteers was treated with 650 mg of aspirin 24 hours before the autologous transfusion of one unit of freeze-preserved platelets. Freeze-thaw-wash recovery values in vitro, viability and function in vivo, and the bleeding time and platelet aggregation response were measured. The platelets were frozen with 4 or 5 per cent dimethylsulfoxide (DMSO) at an overall rate of 2 to 3 C per minute and were stored at -80 C in a mechanical freezer for up to eight months. They were washed by dilution/centrifugation. The mean recovery in vitro of platelets frozen with 4 per cent DMSO was 76+/-16%; the value was 64+/-16% for platelets frozen with 5% DMSO. The mean in vivo 51Cr recovery of autologous platelets frozen with 4% DMSO was 34+/-6%, and for platelets frozen with 5% DMSO it was 33+/-7%. In both groups the platelet lifespan was normal. There was a significant reduction in bleeding time after the transfusion of a single unit of autologous platelets preserved with either 4 or 5% DMSO, but no improvement in the aspirin-induced platelet aggregation pattern.