Hydrodynamic gene delivery in human skin using a hollow microneedle device.

Microneedle devices have been proposed as a minimally invasive delivery system for the intradermal administration of nucleic acids, both plasmid DNA (pDNA) and siRNA, to treat localised disease or provide vaccination. Different microneedle types and application methods have been investigated in the laboratory, but limited and irreproducible levels of gene expression have proven to be significant challenges to pre-clinical to clinical progression. This study is the first to explore the potential of a hollow microneedle device for the delivery and subsequent expression of pDNA in human skin. The regulatory approved MicronJet600® (MicronJet hereafter) device was used to deliver reporter plasmids (pCMVß and pEGFP-N1) into viable excised human skin. Exogenous gene expression was subsequently detected at multiple locations that were distant from the injection site but within the confines of the bleb created by the intradermal bolus. The observed levels of gene expression in the tissue are at least comparable to that achieved by the most invasive microneedle application methods e.g. lateral application of a microneedle. Gene expression was predominantly located in the epidermis, although also evident in the papillary dermis. Optical coherence tomography permitted real time visualisation of the sub-surface skin architecture and, unlike a conventional intradermal injection, MicronJet administration of a 50µL bolus appears to create multiple superficial microdisruptions in the papillary dermis and epidermis. These were co-localised with expression of the pCMVß reporter plasmid. We have therefore shown, for the first time, that a hollow microneedle device can facilitate efficient and reproducible gene expression of exogenous naked pDNA in human skin using volumes that are considered to be standard for intradermal administration, and postulate a hydrodynamic effect as the mechanism of gene delivery.

Plasmonic mediated interactions between waveguides and nanostructured metal surfaces.

We present the polarization resolved, angular dependent, optical reflectance properties for single TE mode optical waveguides in contact with a nanostructured gold surface. A substantial angle dependent resonant decrease in the TE polarized surface reflectivity is measured which cannot be explained by a simple waveguide coupling due to surface roughness. Rather we show that the resonance is due to the excitation of a coupled waveguide-plasmonic surface mode created by the interaction between the metal nanostructure and the waveguide. A model based on coupled mode theory is introduced in order to explain the experimental data.

Reproducibility of bowel ultrasonography in the evaluation of Crohn's disease.

BACKGROUND: Bowel ultrasonography is increasingly used in the detection and follow-up of patients with Crohn's disease, but a limitation to its further diffusion is the lack of standardisation of ultrasonography parameters. AIMS: This study aimed to standardise the most common bowel ultrasonography parameters in order to develop an unequivocal imaging interpretation and to assess bowel ultrasonography reproducibility. PATIENTS: Twenty patients with Crohn's disease were examined. METHODS: Six ultrasonographers (mean bowel ultrasonography experience=16 years) performed the study. They chose and discussed a common assessment methodology concerning eight ultrasonography parameters: bowel wall thickness, bowel wall pattern, bowel wall blood flow, enlarged mesenteric lymph nodes, mesenteric hypertrophy, abdominal free fluid, and stenosis or fistulae at four preliminary meetings. The day of the study operators were randomised to two rooms where they independently and in turn performed ultrasonography scans. Interobserver agreement was scored by kappa statistics. RESULTS: Excellent k values were observed for bowel wall thickness (0.72-1). k Values were poor for bowel wall pattern (-0.22-0.85) and good for bowel wall blood flow (0.53-0.89). The presence of lymph nodes was reproducible (0.56-0.90) except in one case (0.25). Concordance on free fluid was excellent (0.85-1), whereas that on mesenteric hypertrophy was generally poor (0.14-0.69). Agreement was excellent for stenosis (0.81-1) whereas that for fistula was fair in room abscesses (0.31-0.48) and very good in room B (0.87-1). CONCLUSION: Bowel ultrasonography signs used in Crohn's disease can be standardised as most of them showed a fair to good reproducibility. In particular, bowel wall thickness, the most relevant parameter for Crohn's disease detection, showed an excellent reproducibility.

Clinical outcome of patients examined by capsule endoscopy for suspected small bowel Crohn's disease.

BACKGROUND: Capsule endoscopy has a greater diagnostic yield than radiology for detecting subtle inflammatory changes of the small bowel mucosa, but the clinical significance of these abnormalities is still uncertain because of the lack of long-term follow-ups. AIM AND METHODS: To verify the accuracy of capsule endoscopy in a cohort of patients with suspected Crohn's disease of the small bowel, taking as 'gold standard' the final diagnosis made after a long follow-up. From April 2002 to March 2005, we enrolled and examined by capsule endoscopy 27 consecutive patients with abdominal pain and diarrhea lasting more than 3 months and at least one of the following: anaemia, weight loss, fever, extra-intestinal manifestation(s) of inflammatory bowel disease. All patients already had an unremarkable pan-endoscopy, serology for celiac disease and intestinal radiology inconclusive for small bowel abnormality. On the basis of capsule endoscopy findings, patients were distributed in three groups; Group A had severe stricturing lesions requiring surgery; Group B, moderate inflammatory lesions further investigated invasively; Group C, minimal inflammatory changes or normal findings, clinically observed every 3 months (median 21 months, range 15-29). RESULTS: Small bowel inflammatory lesions were found in 16 of the 27 patients (diagnostic yield 59%). Three had surgery (Group A) and Crohn's disease was confirmed in two; the remainder had ileal adenocarcinoma in a pathological context of chronic inflammation. Crohn's disease was histologically confirmed in four of the five patients in Group B. Group C comprised 19 patients; Crohn's disease was confirmed in seven out of eight with positive capsule endoscopy, while only one of the patients with normal findings later developed overt ileal Crohn's disease. Sensitivity, specificity, positive and negative likelihood ratio were, respectively, 93%, 84%, 5.8 and 0.08. Assuming a 50% pre-test probability of disease, capsule endoscopy gave a post-test probability of 85%. CONCLUSIONS: In our selected cohort, capsule endoscopy was highly sensitive in detecting small bowel inflammatory changes, enhancing by nearly 35% the pre-test probability of structural small bowel disease. Focal erythema and luminal debris may limit the specificity of capsule endoscopy.

["Mini-rhinoplasty"]. / La mini-rhinoplastie.

OBJECTIVES: To define the interest of the so called "mini-rhinoplasty" in aesthetic nose surgery and to report the surgical technique. METHOD: The experience of the authors, based on more than 500 mini-rhinoplasty surgical procedures is reported. The surgical procedure such as technical tips are reported. RESULTS: Mini-rhinoplasty procedure is indicated in patients with small deformities, particularly in patients with nasal hump or hyper-projected noses, with no deviation. The nasal tip should be normal or slightly drooping. The surgical technique is safe and reproducible. Surgical aesthetic outcomes are excellent. This technique is also indicated in elderly patients willing a facial rejuvenation. CONCLUSION: Mini-rhinoplasty surgical technique is a minimal invasive procedure with no complication in the postoperative period. The postoperative management of patients undergoing this procedure is of main importance.

Vertical-cavity semiconductor devices for fluorescence spectroscopy in biochips and microfluidic platforms.

This paper describes the properties of vertical-cavity semiconductor devices designed to emit light when driven in forward bias mode and detect optical radiation at wavelengths longer than that of emission when driven in reverse bias mode. The study of this type of devices is motivated by the miniaturization and integration into a single unit of the three functions that a microfluorimeter has to perform, optical pumping, optical detection, and optical filtering of weak light sources. The devices produced can generate fluorescence with a low output power since their emission wavelength can be tuned with that of maximum absorption of the fluorescent dye. We demonstrate also that they can detect low power fluorescence generated in a small volume of concentrated solution of a commercial dye. These devices can find useful application in microanalytical systems such as microfluidic devices or optical biochips.

[Urinary hexafluoroisopropanol in the assessment of occupational exposure to sevoflurane: methodologic features and critical points]. / L'esafluoroisopropanolo urinario nella valutazione dell'esposizione professionale a sevofluorano: aspetti metodologici e punti critici.

The increasing use of sevoflurane as anaesthetic leads to the need for finding a biological index to evaluate the occupational exposure in surgical activity. Several studies indicate that Hexafluoroisopropanol (HFIP) is a specific sevoflurane metabolite quickly glucuronidated and excreted as HFIP-glucuronide in the urine (HFIPu). Therefore the HFIP removal kinetics in occupational exposure and the correlation between sevoflurane exposure and HFIPu are poorly understood. We studied no. 86 operating room workers of Novara Hospital to evaluate the correlation between the sevoflurane individual exposure (SE) and the HFIPu at the end of the shift expressed in microgram/L (A-HFIPu) and in microgram/g creat. (C-HFIPu). Therefore, in the same group of subjects we evaluated the HFIPu in the urine sampled at 8.00 a.m. before the work. The correlation coefficient was R2 = 0.782 (p < 0.0001) for SE/A-HFIPu and R2 = 0.862 (p < 0.0001) for SE/C-HFIPu; HFIPu normalized for urinary creatinine (C-HFIPu) is an index more suitable than the A-HFIPu. Furthermore we concluded for the usefulness of pre-shift HFIPu.

Combined treatment with serine protease inhibitor aprotinin and matrix metalloproteinase inhibitor Batimastat (BB-94) does not prevent invasion of human esophageal and ovarian carcinoma cells in vivo.

Many studies have highlighted the role played by matrix metalloproteinases MMP-2 and -9, by serine proteases uPA and plasmin in tumor cell invasion. This study investigates the impact of the MMP-inhibitor Batimastat and/or the serine protease inhibitor Aprotinin on the in vitro proteolytic activity and in vivo invasive behavior the of esophageal (OC1) and ovarian (OVCAR-3) carcinoma cells. In presence and absence of inhibitors, proteolytic activity of the tumor cells was determined by caseinolytic and collagenolytic in vitro assays and tumor cell invasion by intraperitoneal inoculation of the tumor cells into nude mice. In vitro, Aprotinin, tested alone or in combination with Batimastat, efficiently inhibited degradation of collagen IV and casein by the tumor cells. Batimastat alone had no effect on caseinolytic activities and only partially blocked collagen-type-IV-degradation by the tumor cells. In vivo, Aprotinin tested alone or in combination with Batimastat did not prevent tumor cell invasion. Treatment of tumor bearing mice with Batimastat significantly inhibited tumor growth but promoted tumor cell invasion into the liver. Our findings demonstrate that the inhibition pattern of cellular proteolytic activity achieved in vitro by a serine protease and an MMP inhibitor may lead to predictions that are not necessarily verified in vivo and may even have adverse effects.

Analysis of the fast recovery dynamics of a semiconductor laser with feedback in the low-frequency fluctuation regime.

We experimentally investigate the temporal evolution of the power of an external cavity semiconductor laser in the low-frequency fluctuation regime with subnanosecond resolution. We show, for the first time to our knowledge, that generally the laser power drops to a value significantly different from the solitary laser power. We demonstrate the analogy between the recovery of the laser intensity and the turn-on transient of a semiconductor laser.

Carpal tunnel syndrome in amyotrophic lateral sclerosis and late onset cerebellar ataxia.

We report on clinical and electrophysiological findings and management in nine patients who developed carpal tunnel syndrome during the course of amyotrophic lateral sclerosis and late onset cerebellar ataxia, two neurodegenerative diseases. The patients were treated with surgical decompression (five cases) and local steroid injections (four cases). Only one showed lasting relief of symptoms and significantly improved distal conduction in the median nerve at follow-up after 2 to 3 months. The symptoms and conduction data remained unchanged in three patients who could be followed for more than 1 year. We think that axonal neuropathy plays an important role in the development of carpal tunnel syndrome in these patients and accounts for the failure of the standard treatments.

Electrophysiological findings in peripheral fibres of subjects with and without post-herpetic neuralgia.

The peripheral nervous system was studied using classical electrophysiological methods in 23 subjects with post-herpetic neuralgia (PHN), and compared with the same parameters in 64 herpes zoster (HZ) patients without PHN. The findings indicated sensory axonopathy, the severity of which varied in different patients. Ten percent of all cases showed segmental paresis corresponding to dermatomes affected by HZ. In another 17% of patients axonal motor damage was only detectable by EMG as denervation. No statistically significant difference was found between the two groups in the mean percentage differences of the electrophysiological data for peripheral sensory fibres with respect to mean control values, or between sides affected by HZ and healthy sides. Hence HZ is associated with sensory axonopathy, the severity of which is similar, on the whole, in the groups with and without PHN and stable in time. This suggests that damage to peripheral large-diameter sensory fibres is not the cause of PHN.

Effects of acyclovir on sensory axonal neuropathy, segmental motor paresis and postherpetic neuralgia in herpes zoster patients.

The effect of oral treatment with acyclovir (ACV) on sensory axonal neuropathy, segmental motor paresis and postherpetic neuralgia (PHN) was studied in 105 patients with herpes zoster. Forty-seven patients were treated with ACV at a dose of 4 g/day in 5 doses for at least a week; the others did not undergo any kind of treatment. Electrodiagnostic examination of the nerves and muscles corresponding to the dermatomeric lesions was performed, including sensory and motor nerve conduction studies, blink reflex and electromyography (EMG). The patients treated with ACV showed a significant reduction in the number of cases in which there was electrophysiological evidence of axonal damage in afferent fibres of nerves arising from roots corresponding to affected dermatomes. The treated group also showed a smaller incidence of segmental motor neuritis, clinically evident or only detectable by EMG as denervation. However, there was no significant difference between groups as far as the incidence of PHN was concerned. Oral treatment with ACV therefore reduces peripheral sensory axonopathy due to ganglion damage and prevents the possibility of spread to anterior roots and spinal motoneurones. In this way it reduces the incidence of segmental motor neuritis, but does not reduce the incidence of PHN.

Electrophysiological evidence of "nerve entrapment syndromes" and subclinical peripheral neuropathy in progressive systemic sclerosis (scleroderma).

We report the electrophysiological findings and the management of 5 subjects with progressive systemic sclerosis (PSS) and clinical evidence of nerve entrapment. Three had carpal tunnel syndrome (CTS), 1 bilateral CTS and right tarsal tunnel syndrome (TTS) and 1 Guyon's canal syndrome. Only 1 patient (with CTS) showed significant clinical improvement after surgical decompression; the other 4 demonstrated a slight recovery of conduction without lasting clinical relief after conventional treatment. To explain these failures we hypothesized that these entrapment syndromes were the clinical expression of underlying diffuse damage to the peripheral nervous system (PNS). The conduction values of nerves unaffected by entrapment syndromes were within normal limits, but almost all distal velocities were below the mean of controls. Such subclinical distal peripheral neuropathy was also verified in a selected sample of 17 patients with PSS, without clinical symptoms or signs of PNS involvement. In these 17 cases the mean distal sensory and motor conduction findings of the median, ulnar, sural and tibial nerves were significantly lower than those of a control group, while no significant differences were found in the more proximal tracts of the same nerves. Furthermore, 3 of the 17 patients showed classical electrophysiological evidence of TCS and TTS without any clinical symptoms. We conclude that the subjects with PSS had subclinical polyneuropathy which may become plain polyneuropathy or nerve entrapment syndromes perhaps induced by other risk factors.

[Audiometric results of 81 ossiculoplasties after tympanoplasty with closed technique in chronic cholesteatomatous otitis]. / Résultats audiométriques de 81 ossiculoplasties après tympanoplastie en technique fermée pour otites chroniques cholestéatomateuses.

We report our functional results about 81 ossiculoplastys during tympanoplasty with closed technique in chronic cholesteatomatous otitis. We discuss our functional results and the post-operative assessment of the audiometric gain. Patients presented either cholesteatomatous otitis sequelea or any preliminary status (retraction pockets) with attical impairment.

[The attic. Anatomoclinical correlation]. / Attique. Corrélation anatomo-chirurgicale.

Among the hidden parts of the middle ear, the posterior part of the atrium undoubtedly raises the most interest of otologists. The attic is a frequent localization of cholesteatoma and retraction pouches. Its anatomy is not well known, especially the anterior and internal walls. This anatomic study, based on the dissection of 41 petrus bones was designed to determine the anatomic limits and describe the "noble structures" of this region in order to "guide" future surgical procedures.

Axonal motor and sensory neuropathy in myotonic dystrophy.

We report the neurophysiological findings from 24 subjects with myotonic dystrophy of Steinert and the histological findings in two of them. The conduction data is compared with that of a group of subjects with Landouzy-Déjérine muscular dystrophy. In 46% of cases, the electrophysiological data revealed slight and generalized axonal neuropathy. Histological results of sural nerve confirmed axonal damage of sensory fibres. The neuropathy was not correlated with age of patients, duration or onset of the disease, nor with the state of the deep reflexes; it did not show signs of progressing and is probably one of the multisystemic manifestations of gene pleiotropism.

L-2-oxothiazolidine-4-carboxylic acid, a cysteine prodrug: pharmacokinetics and effects on thiols in plasma and lymphocytes in human.

L-2-oxothiazolidine-4-carboxylic acid (OTC), a prodrug of cysteine, was administered at a dose of 0.15 and 0.45 mmol/kg to healthy volunteers. The plasma concentration of OTC reached a peak between 45 and 60 min after p.o. administration. The p.o. clearance of 0.15 mmol/kg of OTC was 0.57 +/- 0.20 liters/(hr.kg) S.D.. The peak increase in plasma OTC was followed by an increase in the plasma concentration of cysteine which rose by 18 to 75 microM over the average basal concentration of 17 microM. The plasma concentrations of free glutathione and total glutathione (i.e., glutathione in small molecular and protein mixed disulfides) did not change significantly during 8 hr after the ingestion of OTC. In contrast, there was a significant increase in the average concentration of cysteine from 0.37 to 0.99 nmol/mg of protein and of glutathione from 8.7 to 15.6 nmol/mg of protein in lymphocytes 2 to 3 hr after ingestion of OTC. The present data shows that OTC is a prodrug of cysteine after p.o. administration in humans and that OTC can raise the circulating concentration of cysteine and the intracellular concentration of cysteine and glutathione in lymphocytes.

Ferdinando Galiani on population and economic growth.

PIP: The author discusses the views of the eighteenth-century scholar Ferdinando Galiani on population and growth, as published in the Dialogues in 1770. It is claimed that Galiani anticipated features of Classical and Ricardian economic theory. (SUMMARY IN FRE)^ieng