Quantum sensing of microRNAs with nitrogen-vacancy centers in diamond.

Label-free detection of nucleic acids such as microRNAs holds great potential for early diagnostics of various types of cancers. Measuring intrinsic biomolecular charge using methods based on field effect has been a promising way to accomplish label-free detection. However, the charges of biomolecules are screened by counter ions in solutions over a short distance (Debye length), thereby limiting the sensitivity of these methods. Here, we measure the intrinsic magnetic noise of paramagnetic counter ions, such as Mn2+, interacting with microRNAs using nitrogen-vacancy (NV) centers in diamond. All-atom molecular dynamics simulations show that microRNA interacts with the diamond surface resulting in excess accumulation of Mn ions and stronger magnetic noise. We confirm this prediction by observing an increase in spin relaxation contrast of the NV centers, indicating higher Mn2+ local concentration. This opens new possibilities for next-generation quantum sensing of charged biomolecules, overcoming limitations due to the Debye screening.

ND0701, A Novel Formulation of Apomorphine for Subcutaneous Infusion, in Comparison to a Commercial Apomorphine Formulation: 28-Day Pharmacokinetic Study in Minipigs and a Phase I Study in Healthy Volunteers to Assess the Safety, Tolerability, Pharmacokinetics and Relative Bioavailability.

BACKGROUND: Subcutaneous apomorphine is used for the treatment of Parkinson's disease (PD); however, infusion site reactions are a common adverse event (AE), which can lead to treatment discontinuation. Apomorphine formulations that are more tolerable and convenient for use are needed. OBJECTIVE: Our aim was to compare the toxicity and bioavailability of ND0701, a new concentrated formulation of apomorphine free base, with one of the commercially available apomorphine HCl formulations (APO-go®, Britannia Pharmaceuticals Ltd). METHODS: (1) Preclinical study: 16 minipigs were randomly assigned to placebo, APO-go®, and ND0701 groups, and treated for 28 days. Pharmacokinetic, clinical, and pathological assessments were performed. (2) Phase I study: 18 healthy volunteers participated in an open-label, two-sequence, randomized, three single-dose, partial crossover study to compare the pharmacokinetics, safety, and tolerability of ND0701 with APO-go® (1%). RESULTS: (1) Preclinical study: No systemic toxicity was observed in apomorphine-treated minipigs, but local skin reactions were observed at the infusion sites. These effects were less frequent and less severe and recovery was more rapid for ND0701 compared with APO-go®. (2) Phase I study: Both formulations were safe and well tolerated under the conditions of the study and no severe or serious treatment-emergent AEs were reported. Infusion site nodules were reported more frequently, with higher severity, and recovered slower at APO-go®-treated sites compared with ND0701-treated sites. Bioavailability of apomorphine was comparable between the two formulations. CONCLUSION: Based on these pilot studies, ND0701 appears to be superior to APO-go® in terms of tolerability and safety, while maintaining comparable bioavailability with APO-go®, and shows promise as a future treatment for PD.

Ninety-day Local Tolerability and Toxicity Study of ND0612, a Novel Formulation of Levodopa/Carbidopa, Administered by Subcutaneous Continuous Infusion in Minipigs.

A 90-day study in Göttingen minipigs was conducted to test the local tolerability and systemic toxicity of ND0612, a novel aqueous solution of carbidopa (CD)/levodopa (LD) intended for the treatment of Parkinson's disease by continuous subcutaneous administration using a discrete infusion pump. To evaluate tissue site reactions, we used a unique study design involving multiple infusion sites to evaluate the effect of dose per site (270/63, 360/45, and 360/84 mg LD/CD), volume of infusion per site (4.5 and 6 ml per site), formulation concentration (60/14 and 60/7.5 mg/ml LD/CD), daily rate of infusion per site (240 µl/hr for16 hr and 80 µl/hr for 8 hr, 320 µl/hr for 16 hr and 100 µl/hr for 8 hr, or 750 µl/hr for 8 hr), frequency (once every 5, 10, 15, or 20 days), and number of infusions (4, 6, or 9) to the same infusion site. No systemic adverse effects were observed. Histopathological changes at infusion sites started with localized minimal necrosis and acute inflammation that progressed to subacute and chronic inflammatory and reparative changes with evidence of progressive recovery following the final infusion. None of the infusion site effects were judged to be adverse, and clinical exposures to ND0612 are not expected to result in adverse responses.

An aquarium hobbyist poisoning: Identification of new palytoxins in Palythoa cf. toxica and complete detoxification of the aquarium water by activated carbon.

Palytoxin (PLTX) is a lethal natural toxin often found in Palythoa zoantharians that, together with its congeners, may induce adverse effects in humans after inhalation of toxic aerosols both in open-air and domestic environments, namely in the vicinity of public and private aquaria. In this study, we describe a poisoning of an aquarium hobbyist who was hospitalized after handling a PLTXs-containing zoantharian hexacoral. Furthermore, we provide evidence for water detoxification. The zoantharian was morphologically and genetically identified as Palythoa cf. toxica (Cnidaria: Anthozoa). Palytoxin itself and two new PLTX congeners, a hydroxyPLTX and a deoxyPLTX, were detected and structurally identified by liquid chromatography high resolution multiple stage mass spectrometry (LC-HRMSn, n = 1, 2). Total and individual toxins were quantified by LC-HRMS and sandwich ELISA both in the zoantharian (93.4 and 96.80 µg/g, respectively) and in the transport water (48.3 and 42.56 µg/mL, respectively), with an excellent mean bias of 1.3% between the techniques. Activated carbon adsorbed 99.7% of PLTXs contained in the seawater and this represents a good strategy for preventing aquarium hobbyist poisonings.

Combined antihypertensive and cardioprotective effects of nebivolol and hydrochlorothiazide in spontaneous hypertensive rats.

The antihypertensive and cardioprotective effects of the combination of nebivolol (NEB) and hydrochlorothiazide (HCTZ) in spontaneously hypertensive rats (SHR) were investigated. SHR and age-matched Wistar-Kyoto normotensive rats were randomly assigned to one of six groups: SHR treated with NEB (1 mg/kg/day, intragastric); SHR treated with HCTZ (5 mg/kg/day, intragastric); SHR treated with NEB plus HCTZ (1 + 2.5 mg/kg/day); SHR treated with NEB plus HCTZ (1 + 5 mg/kg/day), along with a SHR and a WKY control group. The effects of NEB and HCTZ on clinical chemistry parameters and on cardiac function and structure were evaluated. NEB and HCTZ in combination had synergistic antihypertensive effects and significantly reduced heart rate and blood pressure compared with monotherapy with either agent. These effects were independent of water intake, urine output and electrolyte excretion. NEB plus HCTZ was associated with reduced oxidative stress in terms of glutathione availability, lower angiotensin I levels as index of plasma renin activity and reduced clearance of urinary sodium compared with HCTZ alone. Cardiac morphometric data and antioxidant parameters indicate that NEB may influence cardiac structure by reducing hypertrophy and by enhancing the availability of endothelial nitric oxide - a unique characteristic of NEB. The authors suggest that a combination of NEB and HCTZ for the treatment of hypertension may be useful for the synergistic characteristics of both drugs.

Factors affecting injury severity among recreational skiers and snowboarders: an epidemiology study.

Different results have been reported for skiing and snowboarding injuries worldwide. Few studies consider the injury severity score (ISS) for the evaluation of differences among injured skiers-snowboarders. The aim of this study is to identify possible risk factors that affect the severity of skiing and snowboarding injuries in three winter seasons (2002-2005) in South Tyrol. For every injured skier or snowboarder referred to our emergency department in three consecutive seasons, the following data were collected: date of birth, gender, self-declared technical skills level, place of residence (local/non-local), as well as the date, time, and place of the accident. Type of injury and ISS were retrospectively assigned. Data concerning the snowfall in the last 24 h, average snow level, and outdoor air temperature values were obtained from four weather stations that were located inside the ski resorts. A multiple linear regression model was used to evaluate the association between ISS and potential determinants. In the analyzed seasons, 2,511 injured skiers and 843 injured snowboarders were evaluated at our emergency department. There was a significant change in the ISS value for subjects with different self-reported skills levels (P < 0.001). Men and non-local residents experienced more severe injuries than women and local residents, respectively (P < 0.013, P < 0.001). The ISS was higher for people aged over 60 (P < 0.001). Snowfalls brought about a decrease in accident severity (P = 0.009). The severity of the injuries increases with age. Prevention and information programs should be targeted to people who are at high risk of severe injury. A 24-h fresh snowfall seems to reduce the severity of injuries. Very little is known about snow conditions and winter sports injury. Further studies are needed to explore this field.

Is antibiotic prophylaxis necessary in elective laparoscopic surgery for benign gynecologic conditions?

BACKGROUND: The use of prophylactic antibiotics in gynecologic laparoscopic surgery is still popular and many gynecologic surgeons believe that prophylactic antibiotic therapy will decrease the incidence of postoperative infection complications. AIM: The aim of this prospective cohort study was to investigate the opportunity of prophylactic antibiotics to prevent early postoperative infections and febrile morbidity in elective laparoscopic surgery for benign gynecological conditions. METHODS: 300 women who underwent laparoscopic surgery for benign gynecological conditions were included in the study. After the patients were included they were divided in two groups (ratio 1:1). Group A (n = 150) received 2 g of cefazolin (Cefamezin; Pfizer Italia Srl, Italy) 30 min before surgery and group B (n = 150) received no antibiotic prophylaxis. RESULTS: No postoperative infection was diagnosed for a total of 300 study patients. The overall rate of febrile morbidity was 3.3% for a total of 300 patients (1.3% for group A and 2% for group B). No differences in infection complications and febrile morbidity were found between the two groups. CONCLUSION: Postoperative infection complications in laparoscopic gynecologic surgery without antimicrobial prophylaxis are negligible and overlapping to those obtained with cefazolin prophylaxis.

ACE inhibition and protection from doxorubicin-induced cardiotoxicity in the rat.

The angiotensin converting enzyme inhibitor zofenopril has been shown to possess cardioprotective effects toward myocardial damage induced by chronic doxorubicin treatment in the rat. In the present study we have investigated the relationship between cardioprotection exerted by 2 angiotensin converting enzyme inhibitors (zofenopril and lisinopril) and degree of inhibition of cardiac versus serum angiotensin converting enzyme. Both zofenopril and lisinopril produced a dose-dependent inhibition of serum and cardiac angiotensin converting enzyme in rats (0.1, 1 or 10 mg/kg/day in the diet for 1 week). However, zofenopril at 0.1 mg/kg/day showed a significantly (P < 0.05) greater inhibition of angiotensin converting enzyme in the myocardium than in the serum (delta about 20%). Using dose levels (0.1 mg/kg/day and 10 mg/kg/day) which inhibits partially (about 50%) or almost totally (about 80%) serum angiotensin converting enzyme, we evaluated the effects of zofenopril and lisinopril in preventing cardiac alterations (QalphaT prolongation) induced by chronic treatment with doxorubicin (1.5 mg/kg q7dx5 i.v.). Zofenopril, at a dose level (0.1 mg/kg/ day) that did not affect haemodynamics and only partially inhibits serum angiotensin converting enzyme activity, almost totally prevent the QalphaT lengthening induced by doxorubicin, whereas lisinopril was ineffective at this dose level. At the higher dose level (10 mg/kg/day), both angiotensin converting enzyme inhibitors totally prevented the electrocardiographic alteration induced by chronic doxorubicin administration. Cardioprotection exerted by zofenopril at a dose level that partially inhibits serum angiotensin converting enzyme without affecting haemodynamics, suggests that inhibition of cardiac angiotensin converting enzyme and additional cardioprotective mechanism(s) may have a role in its ability to prevent myocardial damages in the rat subjected to chronic anthracycline treatment.

Involvement of nitric oxide in both central and peripheral haemodynamic effect of D/L-nebivolol and its enantiomers in rats.

The cardiovascular profile of the racemate D/L-nebivolol and its enantiomers administered by intravenous (i.v.) or by intracerebroventricular (i.c.v.) route was investigated in anaesthetized normotensive rats. D/L-Nebivolol (0.1-0.5 mg/kg) induced a dose-related reduction in blood pressure when administered by i.c.v. route. These hypotensive effects were more marked as compared to those achieved by peripheral administration of D/L-nebivolol (0.1-1 mg/kg i.v.). Both enantiomers contributed to the hypotensive effect of D/L-nebivolol by i.c.v. route, while the effects of the drug on blood pressure by i.v. route were due to the d-enantiomer. The bradycardic effect of the racemic form given i.v. was dose-related and, at the highest dose (1 mg/kg), was more pronounced as compared to i.c.v. route. D-Nebivolol was responsible for chronotropic effects by both the i.v. and i.c.v. route, although by i.c.v. route L-nebivolol also induced a reduction in heart rate. The nitric oxide synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) administered at 5 mg/kg i.v. bolus + 0.1 mg/kg/min infusion or at 2.5 mg/kg i.c.v. counteracted the effects of D/L-nebivolol (either 1 mg/kg i.v. or 0.5 mg/kg i.c.v.) on blood pressure, while it did not inhibit the cardiovascular changes induced by isoprenaline (300 ng/kg i.v.) or calcitonin gene-related peptide (CGRP; 400 ng/kg i.v.). In addition, i.c.v. effects of D/L-nebivolol on blood pressure and heart rate were not affected by pre-treatment with atropine (2 mg/kg i.v.). The present findings demonstrate that D/L-nebivolol produced haemodynamic changes following both peripheral and central administration; these latter findings are mainly due to its L-enantiomer and these effects involve the L-arginine/nitric oxide pathway.

Role of hysteroscopy with endometrial biopsy to rule out endometrial cancer in postmenopausal women with abnormal uterine bleeding.

OBJECTIVE: To compare the diagnostic accuracy of ultrasonographic endometrial thickness and outpatient hysteroscopy, to establish the most appropriate exam for the diagnosis of endometrial cancer in postmenopausal women with abnormal uterine bleeding (AUB). The secondary aim was to develop a multivariable approach considering clinical history as an added value for these diagnostic procedures. METHODS: This prospective study was conducted on 220 consecutive postmenopausal patients with AUB, who underwent ultrasonographic evaluation of endometrial thickness, outpatient hysteroscopy and endometrial biopsy. Evaluation of sensitivity, specificity, positive and negative predictive value was performed. Receiver operator characteristic curve (ROC) was calculated to assess the global performance of ultrasonographic measurement of endometrial thickness and diagnostic hysteroscopy as tests for detecting endometrial cancer and atrophy. RESULTS: Histological findings for <4 mm level revealed that atrophy was present in 48 (65%) and in 2 cases (2.7%) endometrial cancer was found; for > or = 4 mm values polyps and myomas were present in 86 (59%) and there were 11 (7.5%) endometrial cancer. Sensibility and specificity for trans-vaginal ultrasound, with a cut-off value > or = 4 mm, was 55.6% and 49.7% while positive predictive value was 83.3% and negative predictive value 98.1% (ROC curve 0.597). Hysteroscopy revealed sensitivity 100%, specificity 49.6%, positive predictive value 81.3% and negative predictive value 100% (ROC curve 0.993). CONCLUSIONS: In conclusion, endometrial thickness <4 mm can miss malignancies but trans-vaginal ultrasound remains the first line diagnostic procedure in postmenopausal women without AUB, because it is not invasive and has high sensitivity for detecting endometrial cancer and other endometrial disease; according to our experience, outpatient hysteroscopy with biopsy is mandatory in all postmenopausal women with AUB.

Hysteroscopic metroplasty under laparoscopic guidance in infertile women with septate uteri: follow-up of reproductive outcome.

OBJECTIVE: To evaluate the effectiveness of hysteroscopic metroplasty under laparoscopic guidance in the treatment of infertile women with a diagnosis of septate uterus and the impact of this surgical procedure on reproductive outcome. STUDY DESIGN: From January 1996 to December 2000, 36 women referred to our endoscopy center underwent hysteroscopic metroplasty. Follow-up covered the incidence of pregnancy, term pregnancies and mode of delivery. RESULTS: The mean age of the patients was 31.9 +/- 3.9 years (range, 23-39). One patient (3%) dropped out during follow-up. There were no intraoperative or postoperative complications. The mean interval from metroplasty to conception was 11.3 +/- 9.2 months (range, 2.0-35.5). Twenty of 35 (57%) women had a history of > or = 1 spontaneous abortions, and 18 of the 20 (90%) achieved pregnancy, with 15 of 18 (83%) term deliveries (10 vaginal deliveries and 5 cesarean sections). Fifteen of 35 patients (43%) were nulliparous, and 8 of 15 (53.3%) had term deliveries (1 cesarean section). CONCLUSION: Hysteroscopic metroplasty under laparoscopic guidance is safe, allows spontaneous delivery and short-term pregnancy planning, is particularly successful in infertile women with a history of > or = 1 spontaneous abortions.

Skiing and snowboarding injuries and their impact on the emergency care system in South Tyrol: a restrospective analysis for the winter season 2001--2002.

OBJECTIVES: To evaluate the incidence and the pattern of skiing and snowboarding injuries in South Tyrol and their impact on the emergency medical system in the winter season 2001--2002 in an attempt to rationalize and improve the emergency care and assist in prevention strategies. METHODS: All medical records of patients referred to our emergency department (ED) that sustained a skiing or snow-boarding injury during the study period were retrospectively reviewed. Age, sex, local or non-local residency, type of injury, data and time of accident, type of transport to the hospital, hospital admission or ED discharge, Injury Severity Score, outcome (including mortality) were evaluated. On site mortality data were obtained from the emergency call-center registry. Ski resorts utilization was estimated from the data published by the Regional Office of Cable Transport. RESULTS: For the period analyzed approximately 2,500,000 skier and snowboarder days were recorded in the whole region of which about 500,000 were attributed to the four nearby ski resorts that refer to our hospital. Of the 1087 patients, 794 were skiers and 294 were snowboarders. Snow-boarders were younger than skiers (mean age 20 and 36 respectively, p = 0.001). Females were equally represented in the two groups. Male patients, children, senior skiers and non-local residents suffered from more severe injuries than their corresponding classes (p < 0.01, p = 0.002, p = 0.02, p = 0.000 respectively). Critical injuries (ISS > or = 25) were homogeneously spread in the groups, with the exception of the non-local resident patients that showed a higher incidence (p < 0.02). No difference in severity was found between skiers and snowboarders. The incidence was 2.05 per 1,000 skier-days. Mortality rate was 1.6 per 1,000,000 skier-days. The pattern of injury was different: snowboarders showed more forearm and wrist trauma and skiers more lower extremity injuries. 208 patients were hospitalized and the mean length of stay was 4.5 days. Head trauma and fractures were the most common diagnosis of admission. The lack of field triage led to 12% of unjustified helicopter transfer and 9.6% of avoidable ambulance transport. CONCLUSIONS: Incidence, pattern of injuries and mortality from skiing and snowboarding accidents in South Tyrol resemble those reported in other part of the world. Nevertheless, strategies for prevention are needed. The routine use of helmets should be enforced by law. Dangerous behaviors should be prosecuted. Skiers and snowboarders should be made aware that skiing beyond their technical ability can be life-threatening.

Carbon dioxide versus normal saline in outpatient hysteroscopy.

BACKGROUND: The aim of this prospective randomized study was to measure patients' discomfort after hysteroscopy with CO(2) or normal saline. METHODS: A total of 415 patients was randomized to two groups according to distension medium (CO(2), n = 201; normal saline, n = 214). The nature of randomization was to alternate distension media on a weekly basis. After hysteroscopy, women were asked to rate the pain experienced on a 100-mm visual analogue scale (0 = no pain; 100 = worst imaginable pain). Pain scores were expressed as mean +/- SD (0-40 = minimal; 41-70 = moderate; 71-100 = severe). Data were analysed using Student's t-test. RESULTS: Irrespective of the distension medium used, pelvic discomfort was worse in nulliparous women (pain score 39.0 +/- 26.5) than in multiparous women (30.4 +/- 25.9) (P < 0.05), especially if they were premenopausal. For all patients and both distension media, pelvic discomfort was generally minimal but higher in patients who had undergone hysteroscopy with normal saline (P < 0.05). CONCLUSIONS: CO(2) and normal saline were comparable with regard to patient discomfort, but in terms of the high frequency of abnormal uterine bleeding, normal saline may be the most appropriate distension medium for outpatient hysteroscopy.

Chronic cardiotoxicity of anticancer anthracyclines in the rat: role of secondary metabolites and reduced toxicity by a novel anthracycline with impaired metabolite formation and reactivity.

(1) The anticancer anthracycline doxorubicin (DOX) causes cardiomyopathy upon chronic administration. There is controversy about whether DOX acts directly or after conversion to its secondary alcohol metabolite DOXol. Here, the role of secondary alcohol metabolites was evaluated by treating rats with cumulative doses of DOX or analogues--like epirubicin (EPI) and the novel disaccharide anthracycline MEN 10755--which were previously shown to form less alcohol metabolites than DOX when assessed in vitro. (2) DOX induced electrocardiographic and haemodynamic alterations, like elongation of QalphaT or SalphaT intervals and suppression of isoprenaline-induced dP/dt increases, which developed in a time-dependent manner and were accompanied by cardiomegaly, histologic lesions and mortality. EPI caused less progressive or severe effects, whereas MEN 10755 caused essentially no effect. (3) DOX and EPI exhibited comparable levels of cardiac uptake, but EPI formed approximately 60% lower amounts of its alcohol metabolite EPIol at 4 and 13 weeks after treatment suspension (P<0.001 vs DOX). MEN 10755 exhibited the lowest levels of cardiac uptake; hence, it converted to its alcohol metabolite MEN 10755ol approximately 40% less efficiently than did EPI to EPIol at either 4 or 13 weeks. Cardiotoxicity did not correlate with myocardial levels of DOX or EPI or MEN 10755, but correlated with those of DOXol or EPIol or MEN 10755ol (P=0.008, 0.029 and 0.017, respectively). (4) DOX and EPI inactivated cytoplasmic aconitase, an enzyme containing an Fe-S cluster liable to disassembly induced by anthracycline secondary alcohol metabolites. DOX caused greater inactivation of aconitase than EPI, a finding consistent with the higher formation of DOXol vs EPIol. MEN 10755 did not inactivate aconitase, which was because of both reduced formation and impaired reactivity of MEN 10755ol toward the Fe-S cluster. Aconitase inactivation correlated (P<0.01) with the different levels of cardiotoxicity induced by DOX or EPI or MEN 10755. (5) These results show that (i) secondary alcohol metabolites are important determinants of anthracycline-induced cardiotoxicity, and (ii) MEN 10755 is less cardiotoxic than DOX or EPI, a behaviour attributable to impaired formation and reactivity of its alcohol metabolite.

A new technique of hysteroscopic myomectomy with enucleation in toto.

STUDY OBJECTIVE: To determine the safety and effectiveness of a new technique for hysteroscopic resection of uterine submucous myomas with high intramural involvement (G2 type, European Society of Hysteroscopy classification). DESIGN: Prospective study (Canadian Task Force classification II-1). SETTING: University hospital. PATIENTS: Forty-four women. INTERVENTION: Hysteroscopic myoma enucleation in toto. MEASUREMENTS AND MAIN RESULTS: With a hysteroresectoscope and Collins electrode, an elliptic incision of endometrial mucosa that covers the myoma is made at the level of its reflection on the uterine wall until the cleavage zone of the myoma is reached. Connecting bridges between myoma and surrounding muscle fibers are resected. This allows nearly complete protrusion of the myoma into the uterine cavity, facilitating complete myomectomy by slicing. The procedure was performed in 41 (93.1%) of 44 women. Of these, 38 (92.6%) had myomas between 2 and 4 cm in diameter and 3 (7.4%) had myomas exceeding 4 cm. Mean operating time was 27 minutes (range 10-45 min). CONCLUSION: This technique is efficient and allows complete resection of submucous myomas with large intramural component by favoring intracavitary protrusion of that part.