The systemic control of circadian gene expression.

The mammalian circadian timing system consists of a central pacemaker in the brain's suprachiasmatic nucleus (SCN) and subsidiary oscillators in nearly all body cells. The SCN clock, which is adjusted to geophysical time by the photoperiod, synchronizes peripheral clocks through a wide variety of systemic cues. The latter include signals depending on feeding cycles, glucocorticoid hormones, rhythmic blood-borne signals eliciting daily changes in actin dynamics and serum response factor (SRF) activity, and sensors of body temperature rhythms, such as heat shock transcription factors and the cold-inducible RNA-binding protein CIRP. To study these systemic signalling pathways, we designed and engineered a novel, highly photosensitive apparatus, dubbed RT-Biolumicorder. This device enables us to record circadian luciferase reporter gene expression in the liver and other organs of freely moving mice over months in real time. Owing to the multitude of systemic signalling pathway involved in the phase resetting of peripheral clocks the disruption of any particular one has only minor effects on the steady state phase of circadian gene expression in organs such as the liver. Nonetheless, the implication of specific pathways in the synchronization of clock gene expression can readily be assessed by monitoring the phase-shifting kinetics using the RT-Biolumicorder.

A retrospective analysis of early and late outcome of biodegradable stent placement in the management of refractory anastomotic colorectal strictures.

BACKGROUND: Benign colorectal strictures are treated conventionally by endoscopic dilation. Experience using SEMS for benign colonic strictures is limited, and outcomes to date have been disappointing. Refractory colorectal strictures remain challenging to be treated with surgery. Polydioxanone-based stent are biodegradable (BD) stent CE approved for esophageal strictures. This study was designed to investigate retrospectively the safety and the efficacy of these stents for the management of strictures refractory to multiple sessions of dilation. METHODS: Patients with postsurgical benign strictures located within 20 cm from anal verge, refractory to mechanical or pneumatic dilation (at least 3 sessions) were included in this analysis. Clinical success was defined as the absence of occlusive symptoms and the ability to pass through the stricture with a regular size colonoscope. All patients were predilated before stent placement. Stents were released under fluoroscopic control. All patients were under stool softeners for 3 months. Follow-up was scheduled with endoscopic and fluoroscopic controls within 90 days from stent deployment and afterwards by telephone interview and/or ambulatory consultation. RESULTS: Eleven patients (7 males, mean age 62.3 ± 8.5 years) were included. Technical success was achieved in all the patients. Stent migration was observed in four patients within the first 2 weeks after stent placement. Stent migration was followed by recurrence of stricture and obstructive symptoms in all the cases. Among the seven patients who completed the process of stent biodegradation, five of them had complete resolution of the stricture and relief of symptoms. Two of 11 patients required surgical treatment during the follow-up period (mean 19.8 (range 42-15) months). The overall success rate of the BD stent was 45 %. CONCLUSIONS: This retrospective analysis of a limited number of patients demonstrated that nondedicated esophageal BD stents are associated with high risk of migration and clinical success in less than 50 % of patients. Dedicated stents with large diameter and antimigration findings could potentially improve the outcome of patients with refractory benign colorectal strictures.

Safety of cold polypectomy for <10mm polyps at colonoscopy: a prospective multicenter study.

BACKGROUND: Cold polypectomy techniques (without electrocautery) by means of biopsy forceps or snare are widely adopted for the removal of subcentimetric polyps. However, few data are available on the safety of this approach. The aim of this study was to assess the safety of cold polypectomy for subcentimetric polyps, as well as the rate of advanced neoplasia in these lesions. PATIENTS AND METHODS: In a prospective multicenter trial, consecutive patients with at least one < 10-mm polyp at colonoscopy were prospectively included. All of the < 10-mm polyps detected within the study period were removed by cold polypectomy. The rates of immediate or delayed bleeding and other complications were assessed at 7 and 30 days after cold polypectomy by telephone calls. The rate of advanced histology was also assessed. Predictive variables of postpolypectomy bleeding or advanced neoplasia were identified by multivariate analysis. RESULTS: A total of 1015 < 10-mm polyps in 823 patients (15.5 % on antiplatelet agents) were removed. Of these, 822 (81 %) were ≤ 5 mm and 193 (19 %) were 6 - 9 mm. Immediate postpolypectomy bleeding occurred in 18 patients, corresponding to a per-patient and per-polyp bleeding rate of 2.2 % (95 % confidence interval [CI] 1.2 % - 3.2 %) and 1.8 % (95 %CI 1 % - 2.6 %), respectively. Therapy with antiplatelet agents (odds ratio [OR] 4; 95 %CI 1.5 - 10.6) and larger polyp size (OR 2; 95 %CI 1.1 - 6.9) were independent predictors of bleeding. Bleeding was successfully treated by endoscopic hemostasis in all cases and required no further medical intervention. Advanced neoplasia prevalence in polyps ≤ 5 mm was as high as 8.7 %. CONCLUSIONS: The results from this study showed the high safety of a cold polypectomy approach for subcentimetric polyps. This was due to the low rate of postpolypectomy bleeding and to the high efficacy of endoscopic hemostasis in its treatment. The high rate of advanced neoplasia in polyps ≤ 5 mm should prompt some caution on the management of these lesions following detection at computed tomography colonography or colon capsule endoscopy.

The environmental chemical tributyltin chloride (TBT) shows both estrogenic and adipogenic activities in mice which might depend on the exposure dose.

Exposure during early development to chemicals with hormonal action may be associated with weight gain during adulthood because of altered body homeostasis. It is known that organotins affect adipose mass when exposure occurs during fetal development, although no knowledge of effects are available for exposures after birth. Here we show that the environmental organotin tributyltin chloride (TBT) exerts adipogenic action when peripubertal and sexually mature mice are exposed to the chemical. The duration and extent of these effects depend on the sex and on the dose of the compound, and the effects are relevant at doses close to the estimated human intake (0.5µg/kg). At higher doses (50-500µg/kg), TBT also activated estrogen receptors (ERs) in adipose cells in vitro and in vivo, based on results from acute and longitudinal studies in ERE/luciferase reporter mice. In 3T3-L1 cells (which have no ERs), transiently transfected with the ERE-dependent reporter plus or minus ERα or ERß, TBT (in a dose range of 1-100nM) directly targets each ER subtype in a receptor-specific manner through a direct mechanism mediated by ERα in undifferentiated preadipocytic cells and by ERß in differentiating adipocytes. The ER antagonist ICI-182,780 inhibits this effect. In summary, the results of this work suggest that TBT is adipogenic at all ages and in both sexes and that it might be an ER activator in fat cells. These findings might help to resolve the apparent paradox of an adipogenic chemical being also an estrogen receptor activator by showing that the two apparently opposite actions are separated by the different doses to which the organism is exposed.

Intra-articular Hymovis injection for managing hip OA in active sportsmen. A 24-month observational retrospective clinical investigation.

BACKGROUND: Hip Osteoarthitis (OA) risk is sport-specific and depends on frequency, intensity, and type of mechanic stress the hip is subjected to. This retrospective observational study aims to investigate the safety and performance of Hymovis (HYADD-4) injection, a hexadecyl (C-16) HA-derivative, when used to manage symptomatic hip OA in active middle-aged sportsmen over a 24-month observation period. METHODS: The retrospective analysis included clinical records of active sportsmen, aged between 40 and 65 years, and suffering from symptomatic Kellgren-Lawrence grade II to III hip OA, treated with two (24 mg/3 ml) Hymovis injections, two weeks apart, every 3-4 months, for two years. When available, data on MRI examination were included in the analysis as well as Heidelberg Sports Activity Score (HAS) and Copenhagen Hip and Groin Outcome Score (HAGOS) questionnaires. RESULTS: Thirty patients (56.4 ± 7.3 years) were included in the study, sixteen cyclists and 14 tennis players. For all patients, HAS and most HAGOS scores improved significantly (p < 0.05) at the first control visit (4 months) and further improved over time. For all other scores an important clinical benefit was experienced by more than 50% of participants. No adverse events were recorded. CONCLUSION: Treatment of hip OA in active sportsmen with Hymovis seems a safe and effective approach for the management of OA symptoms, by potentially protecting cartilage and subchondral bone from further damage.

Endoscopic flexible treatment of Zenker's diverticulum: a modification of the needle-knife technique.

BACKGROUND AND STUDY AIM: Endoscopic treatment of Zenker's diverticulum has been successfully reported over the last 10 years using different approaches. The hook-knife is a new device originally developed for endoscopic submucosal dissection procedures. This study aimed to investigate the safety and efficacy of endoscopic myotomy performed with the hook-knife. PATIENTS AND METHOD: From July 2005, 32 consecutive patients (23-male, mean age 74.8 years) with dysphagia secondary to the presence of Zenker's diverticulum were prospectively enrolled. Myotomy was performed using a straight-end transparent hood to the tip of the scope and the hook-knife for the incision of the bridge between the Zenker's diverticulum and the esophagus. Clinical outcome was evaluated assigning a dysphagia symptom score from 0 (symptoms absent) to 4 (inability to swallow saliva). RESULTS: General anesthesia was used in 4 patients, deep sedation with propofol in 23 patients, while midazolam was used in 5 patients. The mean procedural time was 28 minutes. Complications occurred in 2 patients (6.25 %). At 1 month follow-up, the mean dysphagia score was significantly improved from 2.9 to 0.6 ( P < 0.001) with 87.5 % of patients free of symptoms and 4 patients with dysphagia that was persistent but milder than before the treatment. Three of these 4 patients underwent a successful second endoscopic treatment with complete relief of dysphagia; one was not re-treated because of advanced age (92 years). During the follow-up period (23.87 +/- 9.6 months), 2 patients developed dysphagia recurrence. The overall success rate was 90.6 %. CONCLUSIONS: Diverticulectomy with a flexible scope and the hook-knife may represent a safe and effective alternative treatment for patients with Zenker's diverticulum.

[Utility and safety endoscopic digestive procedure in pediatric age]. / Utilità e sicurezza delle procedure endoscopiche digestive in età pediatrica.

The aim of this study is to verify the utility and safety of endoscopic procedures in the evaluation of children with clinically-significant gastrointestinal symptomatology. We report our experience of 87 pediatric endoscopy procedures including esophagogastroduodenoscopy, colonoscopy and tissue biopsies performed in 85 infants, children and adolescent, 3 months-15 years old, over a two-year period, june 2002-november 2004 after complete history, physical examination and basic investigations. General anesthesia was used in all patients after informed consent obtained from parents. Non significant complications were observed in this series of patients. In 81 cases (92.5%) with clinical symptoms and laboratory indications for gastrointestinal disease, the endoscopy and bioptical samples confirmed the utility and safety of procedure. Coeliac disease (39 cases), gastritis (11 cases), esophagitis (6 cases) were the most common organic cause of upper gastrointestinal disease. Allergic and indeterminate colitis (7 cases) were the most common cause of lower gastrointestinal disease. In 4.7% the procedures appear to be particularly helpful in the diagnosis of inflammatory lesions of the esophagus and stomach. In summary, the data demonstrate that endoscopy techniques show low morbidity, provide important diagnostic informations in pediatric gastrointestinal diseases and can be done safely in patients over 3 months of age.

Evaluation of left ventricular diastolic function by pulmonary venous and mitral flow velocity patterns in endurance veteran athletes.

More and more older people exercise endurance training. Physical activity regularly exercised has been proven to exert beneficial effects on the cardiovascular system. The aim of the present study was to investigate left ventricular diastolic function by analysis of the pulmonary venous flow velocity pattern (PVFVP) in conjunction with the mitral flow velocity pattern (MFVP) in endurance veteran athletes. The study was performed in 15 trained veteran athletes (mean age 60 +/- 10) and 15 sedentary older subjects (mean age 61 +/- 7). Between the two groups there were no differences of age, body surface area and blood pressure. All subjects were without evidence of cardiovascular diseases. They underwent transthoracal pulsed Doppler echocardiography and the following parameters were measured: early (E) and late (A) peak diastolic filling velocities from mitral flow and E/A ratio; peak forward flow velocities during systole (S) and diastole (D) and peak reverse flow velocity at atrial contraction (Ar) from right upper pulmonary vein. The peak early diastolic filling and E/A ratio resulted significantly increased in the veteran athletes compared with the older sedentary subjects (E 80.0 +/- 13.6 and 62.2 +/- 8.2, respectively, p < 0.01; E/A 1.20 +/- 0.1 and 0.90 +/- 0.1, respectively, p < 0.001), whereas there were no significant differences m the PVFVP between the two groups. Heart rate at rest was significantly lower in the veteran athletes compared with sedentary older subjects (58.3 +/- 8 and 72.8 +/- 7.6, respectively, p < 0.001). These data demonstrate an improvement of left ventricle diastolic function in endurance veteran athletes (E/A ratio > 1 ) in comparison with sedentary older subjects (E/A ratio > 1). Analysis of PVFVP suggests that the left atrial contribution to left ventricular filling increases with aging without any significant differences between the two groups. Therefore, left atrial function, i.e., the main determinant of PVFVP is not likely influenced by training. In conclusion, physical activity was found to attenuate alterations in the cardiovascular system that occur in advanced age by an improvement of left ventricular diastolic filling. These changes at rest may be due to a decreased sympathetic tone, as well as to an improvement of intrinsic properties of myocardium of the endurance veteran athletes.

Importance of training in prevention of the decline of physical performance in elderly sedentary persons and veteran athletes.

Thirty-five male veteran athletes were studied between 55 and 79 years of age, having exercised track-and-field sports in the amateur class for at least 15 years without interruption. These athletes were divided into power and endurance groups. The subjects examined were compared to a male control group of 30 elderly, healthy persons aged between 51 and 77 years, leading a sedentary lifestyle. All the subjects were submitted to a standard protocol using BOSCO ERGOJUMP. These trials permitted us to evaluate in the laboratory the explosive power of the inferior limbs, maximal alactic and lactic capacities. The results obtained suggest interesting correlations between the unwinding of physical activity and a deterioration of explosive and anaerobic power, above all in subjects over 65 years of age. Therefore, a regular training program could attenuate the modifications and the age-associated decline.

[An evaluation of the pulmonary venous flow pattern by transesophageal and transthoracic Doppler echocardiography in a normal subject]. / Valutazione del pattern di flusso venoso polmonare mediante ecocardiografia Doppler transesofagea e transtoracica nel soggetto normale.

Transesophageal echocardiographic studies have permitted a pulmonary venous flow velocity pattern to be identified which is comparable to that recorder using invasive methods. The pattern consists of 4 stages: an anterograde systolic flow with an early (S1) and late (S2) peak velocity, a diastolic anterograde flow (D) and a retrograde flow liked to atrial contraction (Ar). The aim of this study was to evaluate the pattern of pulmonary venous flow velocity using transesophageal and transthoracic colour Doppler echocardiography in normal subjects in an attempt to: 1) determine normal values derived from the pulmonary venous flow pattern which may contribute to future studies; 2) find correlations between the pattern of pulmonary venous flow and a number of physiological, hemodynamic and echocardiographic parameters which take account of the morphological variations of this flow pattern within a normal range; 3) demonstrate the possibility of being able to carry out study using transthoracic colour Doppler echocardiography. The study was carried out in 38 normal subjects aged between 15 and 76 years old (mean 45 +/- 15) who underwent transthoracic and transesophageal colour-Doppler echocardiography. The following parameters were measured: left atrium diameter, diameters and telediastolic and telesystolic volumes of the left ventricle, ejection fraction and systolic percentage shortening of the left ventricle, peak velocity of the mitral flow pattern and the pulmonary venous flow pattern. The results obtained show that: 1) the pattern of pulmonary venous flow alters with ageing causing the prevalence of systolic over diastolic peak velocity; 2) the pulmonary venous flow parameters which appear to be most significant in hemodynamic terms are the peak velocities of the early systolic flow and anterograde diastolic flow and the ratios S1/S2 and S2/D; 3) the echocardiographic parameters most closely correlated with the peak velocity of pulmonary venous flow are the dimensions of the left atrium, telediastolic and telesystolic ventricular volumes, and the pattern of transmitral flow; 4) heart rate does not appear to influence the pulmonary venous flow pattern significantly (at least within the range of 50-100 b/min), except for retrograde flow. Similar results were obtained using transesophageal and transthoracic Doppler echocardiography and therefore if future studies succeed in obtaining useful information, of not only speculative but also practical interest, from the evaluation of pulmonary venous flow, these can be obtained using a simple, widespread and completely non-invasive method such as transthoracic Doppler echocardiography.

[Longitudinal echocardiographic study of HIV positive patients]. / Studio ecocardiografico longitudinale in pazienti HIV positivi.

BACKGROUND: From May 1992 to June 1996 the authors have studied a group of 39 subjects with positive anti-HIV, with echo 2D color Doppler examination, to evaluate with semi-annual controls, the wide variety of cardiac complications in the various phases of clinical evolution of the illness. METHODS: At the moment of recruiting, all the subjects with HIV infection were asymptomatic A1 (HIV + As). The patients whose average age was 29 +/- 5, were composed of 60% drug addicts, 17% homosexuals, 8% haemophiliacs and for the 15% heterosexual. RESULTS: The most frequent cardiac complications are represented by hypokinesia of the left ventricle (h-aLV) and by pericardial effusion (PE); more rarely of endocardial vegetations (EV), dilatation of the left ventricle (dLV) and tricuspid insufficiency (TI). The entity of damage and the number of cases observed, are correlated with the grade of clinical severity of the illness. CONCLUSIONS: In accordance with the literature data, cardiac pathologies, particularly in the first phases of the illness, are asymptomatic or paucisymptomatic, making the clinical-instrumental observation of the patient useful also in cardiology.

Balanced Propofol Sedation in Patients Undergoing EUS-FNA: A Pilot Study to Assess Feasibility and Safety.

Introduction and aims. Balanced propofol sedation (BPS) administered by gastroenterologists has gained popularity in endoscopic procedures. Few studies exist about the safety of this approach during endosonography with fine needle aspiration (EUS-FNA). We assessed the safety of BPS in EUS-FNA. Materials and methods. 112 consecutive patients, referred to our unit to perform EUS-FNA, from February 2008 to December 2009, were sedated with BPS. A second gastroenterologist administered the drugs and monitorized the patient. Results. All the 112 patients (62 males, mean age 58.35) completed the examination. The mean dose of midazolam and propofol was, respectively, of 2.1 mg (range 1-4 mg) and 350 mg (range 180-400). All patients received oxygen with a mean flux of 4 liter/minute (range 2-6 liters/minute). The mean recovery time after procedure was 25 minutes (range 18-45 minutes). No major complications related to sedation were registered during all procedures. The oxygen saturation of all patients never reduced to less than 85%. Blood systolic pressure during and after the procedure never reduced to less than 100 mmHg. Conclusions. In our experience BPS administered by non-anaesthesiologists provided safe and successful sedation in patients undergoing EUS-FNA.

Clinical experience with a new endoscopic over-the-scope clip system for use in the GI tract.

BACKGROUND: The newly designed over-the-scope clip (OTSC) seems to overcome several limitations of current clipping system, such as size and opening-closing force, allowing better control of gastric or colonic bleeding and/or deep wall defect or perforation. AIMS: The aim of this retrospective analysis was to describe the new endoscopic device and evaluate our first clinical experience. PATIENTS AND METHODS: We treated with the OTSC system 9 patients (range, 58-85 years; 6 men, 3 women) with bleeding and/or deep wall lesions of the GI tract. The OTSC system is composed of an application cap, which is mounted onto the distal tip of the endoscope and a connected releasing mechanism, installed on the handle of the scope. The rotation of the handle allows the release of the clip by a two tube sliding mechanism. RESULTS: All applications resulted successful, i.e. haemostasis was achieved, and/or wall defects could be closed. No complication was observed that could be ascribed to the clip itself or to the technique. CONCLUSIONS: The OTSC system is a new endoscopic tool for compression of large tissue areas and its indications are nonvaricose bleedings difficult to control and lesions or perforations of the GI tract. The initial clinical use of this clipping device proved to be efficient and effective.

Estrogen receptor-mediated transcriptional activity of genistein in the mouse testis.

Here we show that genistein, through an estrogen receptor-mediated action, modulates gene expression in the mouse testis throughout development. Genistein passed from the lactating mother to the suckling offspring at levels sufficient to activate gene expression in the testis of the pups. Testis are already responsive to genistein as well as to estradiol at day 14.5 of fetal development. Activation of luciferase correlates with an activation of cell proliferation. In conclusion, our results show that genistein affects reproductive organs of male mice at all developmental ages.

Multimodality imaging: novel pharmacological applications of reporter systems.

The development of novel drugs is a lengthy process requiring years of preclinical research and many steps indispensable to ensure that the molecule of interest can be administered to humans with a minimal risk of toxic effects. Even a minimal reduction in the initial stages of drug development would result in a tremendous saving in time; therefore, pharmaceutical companies are eager to apply novel methodologies that shorten the time required for pharmacodynamic, pharmacokinetic and toxicological studies to be carried out in vitro and in animal systems. Currently, quantitative analysis of molecular events in living organisms is done with the combined application of imaging and genetic engineering technologies. In vivo imaging provides surrogate endpoints that can improve the identification of new drug candidates and speed up their research at preclinical stages. The integration of reporter systems in animal models of human diseases represents a reachable frontier that will dramatically advance drug development in terms of costs, time and efficacy. The present review outlines the applicability of imaging technologies for drug development and presents a panorama on the state of the art of currently available imaging technologies suitable for preclinical studies, with particular focus on bioluminescence and fluorescence as the methodologies of election.

Major anastomotic dehiscence after repair of esophageal atresia: conservative management or reoperation?

SUMMARY. The authors report a case of recurrent anastomotic dehiscence following surgical repair of type C esophageal atresia according to the Gross classification. Surgical repair was followed by a recurrence, which was successfully managed with conservative treatment. Esophageal atresia with fistulization of the lower pouch in a male newborn with the VACTER association was repaired with a high-tension single-layer anastomosis. On the fifth postoperative day, major anastomotic dehiscence (> 4 mm) was diagnosed. The breach was re-sutured and the anastomosis reinforced with fibrin glue, but dehiscence recurred again 4 days later. Surgery was deferred and the infant was treated conservatively with continued chest-tube drainage and total parenteral nutrition. After 43 days, complete closure of the anastomosis was documented. Even major anastomotic dehiscence can be successfully managed with conservative treatment (chest-tube drainage, suspension of oral feedings, total parenteral nutrition). If the patient is otherwise stable, we feel that this approach should be attempted even when major leakage is present.

Fibromyxoid sarcoma in a 4-year-old boy: case report and review of the literature.

The authors report on the case of a low-grade fibromyxoid sarcoma (LGFMS) with giant rosettes in a 4-year-old boy. The tumor arose in the paravertebral region and had infiltrated the vertebral canal between L2 and S1. A review of the literature indicates that this is one of the youngest patients diagnosed with a tumor of this type, and spinal involvement had never been reported with LGFMS.

Malignant retroperitoneal paraganglioma: case report and review of the literature.

Paraganglioma is a rare neuroendocrine neoplasm observed in patients of all ages but it has not been characterized in children. The authors describe a retroperitoneal paraganglioma diagnosed by chance in an 11-year-old boy. Many aspects of retroperitoneal paraganglioma are still under investigation. The treatment of choice is radical resection. Surgery may be possible following chemotherapeutic debulking with cyclophosphamide, vincristine, and dacarbazine. 131I-MIBG radiotherapy has proved increasingly useful in reducing the pain associated with disseminated disease and also in facilitating surgical resection in cases that appear inoperable. Conventional radiotherapy is purely palliative and used to reduce the pain of bone metastases.