Neural retina limits the nonviral gene transfer to retinal pigment epithelium in an in vitro bovine eye model

We investigated the permeation of liposomal and polymeric gene delivery systems through neural retina into retinal pigment epithelium (RPE) and determined the roles of various factors in permeation and subsequent uptake of the delivery systems by RPE. Anterior parts and vitreous of fresh bovine eyes were removed. Retina was left intact or peeled away. Complexes of ethidium monoazide (EMA)-labeled plasmid DNA and cationic carriers (polyethyleneimine, poly-L-lysine, DOTAP liposomes) were pipetted on the retina or RPE. Two hours later the neural retina was removed, if present, and the RPE cells were detached. Contaminants were removed by sucrose centrifugation, and the RPE cells were analyzed for DNA uptake by flow cytometry. Cellular uptake of FITC-dextrans (molecular weight [mw] 20,000, 500,000 and 2,000,000), FITC-poly-L-lysine (mw 20,000), FITC-labeled oligonucleotide (15-mer), and naked EMA-labeled plasmid DNA was determined after pipetting the solutions on the RPE or neural retina. Location of the fluorescent materials in the retina was visualized with fluorescence microscopy. Neural retina decreased the cellular uptake of DNA complexes by an order of magnitude, the uptake of FITC-dextrans slightly, whereas delivery of polycationic FITC-poly-L-lysine to RPE was almost completely inhibited. Neural retina decreased the cellular uptake of FITC-oligonucleotides, while the uptake of uncomplexed plasmid was always negligible. Conclusions from FACS and fluorescence microscopy were similar: delivery of polymeric and liposomal DNA complexes into RPE are limited by the neural retina. This is due to the size and positive charge of the complexes.     

Phenotypic expression of the targeted null-mutation in the dopamine transporter gene varies as a function of the genetic background

The dopamine transporter (DAT) plays a critical role in calibrating the duration and intensity of dopamine (DA) neurotransmission. Mice in which the DAT gene has been genetically deleted exhibit constitutively high levels of extrasynaptic DA and spontaneous hyperactivity. Numerous studies have characterized the adaptive molecular, physiological, and behavioural consequences of abnormal DA neurotransmission in these mice. In order to determine the genetic background contribution to these phenotypes, the DAT mutation was transferred on C57BL/6JOrl (B6) or DBA/2JOrl (D2) inbred backgrounds for more than ten generations of back-crossing to derive three B6-, D2-, and B6xD2(F(1))-DAT strains. We observed that the genetic background dramatically affects phenotypes previously reported on DAT knockout (KO) mice. Depending on the genetic background, it was possible to restore survival, growth rate and ability to lactate. Interactions with the genetic background were found to modulate both quantitative and qualitative patterns of novelty-driven spontaneous hyperactivity. The paradoxical calming effect of cocaine was observed for all DAT-KO mice. However, the genetic background influenced individual threshold responses to both locomotor and rewarding effects of cocaine. These findings reveal the extent of phenotypic variation associated with the DAT mutation. They also provide concrete arguments against the assumption that the normal function of a gene can be inferred directly from its mutant phenotype.     

Low platelet MAO activity associated with high dysfunctional impulsivity and antisocial behavior: evidence from drunk drivers

Rationale Low platelet monoamine oxidase (MAO) activity is associated with problem drinking and other deviant behaviors. Since the majority of alcohol abusers are smokers, and tobacco smoke has a direct inhibitory effect on the enzyme, these associations may not be meaningful.Objective The authors compared platelet MAO activity and impulsivity in police-referred subjects caught driving while intoxicated and in control subjects, controlling for smoking.Methods Platelet MAO activity was measured radioenzymatically and impulsivity scores obtained from questionnaires. Smoking status was self-reported.Results Subjects caught driving while intoxicated had significantly higher dysfunctional impulsivity and lower platelet MAO activity than control subjects. This difference in platelet MAO activity between the two groups was significant in non-smokers and ex-smokers.Conclusions These findings demonstrate that platelet MAO activity is lower in subjects with socially deviant behavior, and the association of low platelet MAO and problem drinking is not an artifact of smoking.     

Effects of beta-carotene supplementation on free radical mechanism in healthy adult subjects

AIM: The objective of this study was to examine whether increasing doses of beta-carotene supplements have effects on biological markers of lipid peroxidation in healthy volunteers. SUBJECTS AND METHODS: Forty-two healthy subjects were supplemented with 5 mg, 10 mg, 20 mg or 40 mg beta-carotene/day, respectively for five weeks. Plasma beta-carotene and tocopherol levels, malondialdehyde-thiobarbituric reactive substances (MDA-TBARS), and conjugated dienes were determined using high-performance liquid chromatography (HPLC). Concentrations of ascorbic acid, uric acid, and the total antioxidative capacity (TAC) in plasma were measured photometrically. RESULTS: Plasma beta-carotene levels increased significantly according to the intervention dose (p < 0.001), and concentrations of tocopherol equivalents and ascorbic acid were within the physiological range except in the 5 mg intervention group where a significant decrease of vitamin C was assessed (p < 0.05). Uric acid in plasma decreased significantly in all groups (p < 0.05) up to the end of investigation, but was within the normal range. Trolox equivalent antioxidative capacity (TEAC) decreased significantly (p < 0.001) in all groups during supplementation. MDA-TBARS remained unchanged after five weeks except for the 40 mg beta-carotene substitution group, where a significant decrease was observed (p < 0.05). CONCLUSION: Beta-carotene supplementation of healthy subjects significantly increased plasma beta-carotene status without inducing adverse biological effects. Beta-carotene did not especially protect against oxidative stress, except for the 40 mg group. These data suggest that additional effects of beta-carotene supplementation on well-nourished, healthy subjects are limited.     

Time spent by primary care practices on pediatric influenza vaccination visits: implications for universal influenza vaccination

OBJECTIVE: To measure the time currently spent by primary care practice personnel, and the examination room occupancy time for childhood influenza vaccination visits, to assess the practicality of annual influenza vaccination of all preschool children. SETTING: Seven primary care practices serving one fourth of the children living in Rochester, NY. PATIENTS: Ninety-two children seen for influenza vaccination visits in the 2000-2001 vaccination season. METHODS: Using a standardized protocol, practice staff measured the time spent on check-in, nurse or physician examination, and the actual influenza vaccination process. Waiting and "hands-on" times were determined, as well as total visit and room occupancy times. Nonparametric tests and multivariable models were used to analyze the time spent for components of the visits and to compare time spent by different age groups and practice types (suburban or urban). RESULTS: The median duration of the influenza vaccination visit was 14 minutes (25th to 75th percentiles range, 9-25 minutes) across the 7 practices, with visits to urban practices being longer (22 minutes) than visits to suburban practices (9 minutes). Eighty percent of patient time involved waiting, primarily in examination rooms. The major components of influenza vaccination visits included waiting room time (4 minutes in suburban practices vs 8 minutes in urban practices; P<.01), and time in the examination room (5 minutes vs 14 minutes, respectively; P<.001), during which only 1 to 2 minutes (for both suburban and urban practices) were for hands-on vaccinations. Only 5% of visits were examined by a physician or nurse practitioner. Visit times did not vary by age. CONCLUSIONS: Although the personnel time for influenza vaccination visits was short, there was substantial patient waiting and long occupancy of examination rooms. If universal influenza vaccination is to be efficiently managed in primary care practices, it may be necessary to implement "vaccination clinics" or sessions in which large numbers of children are scheduled for influenza vaccinations at times when adequate rooms and dedicated nursing staff are available.     

CT in the evaluation of patients on ECMO due to acute respiratory failure

BACKGROUND: In patients with acute severe respiratory failure (ARF) treated with extracorporeal membrane oxygenation (ECMO) the radiological evaluation has until now almost exclusively relied on bedside radiography and US. At St. G?ran/Karolinska ECMO centre CT has become a routine complement to bedside examinations. OBJECTIVE: To review retrospectively the frequency, indications and findings on CT of patients with ARF on ECMO and to evaluate the risk of complications associated with transportation for CT examinations. MATERIALS AND METHODS: One hundred twelve neonates, children and adults were treated with ECMO from May 1994 to January 2001. Forty-six per cent of these patients had CT examinations on one or more occasions during ECMO, giving a total number of 238 examination sites on 104 occasions. All CT examinations were performed in the Paediatric Radiology Department and included a 10-min transport using a mobile ECMO system. RESULTS: CT was more often performed in older patients and in patients with long ECMO runs. The main indications were suspected complications of ECMO and/or the underlying disease or a delay in clinical improvement. In 57% of the CT occasions, significant findings affecting treatment were revealed. There were no complications associated with the examinations or transport. CONCLUSIONS: CT is safe and useful in evaluation of patients with ARF during ECMO.     

Renal transplantation in patients with hemolytic uremic syndrome: high rate of recurrence and increased incidence of acute rejections

BACKGROUND: The reported outcome of renal transplantation in patients with the hemolytic uremic syndrome (HUS) varies greatly, probably related to the diverse causes of HUS. In this single-center retrospective study, we have analyzed the recurrence rate, the incidence of acute rejections, and graft survival in patients suffering from adult-onset and childhood-onset HUS. METHODS: The medical records of 35 patients with end-stage renal disease caused by HUS, who received 50 renal allografts, were reviewed. A definite recurrence of HUS was diagnosed if both clinical and histologic signs of thrombotic microangiopathy (TMA) were present in the absence of any endovasculitis. If there were signs of mild endovasculitis, a probable recurrence was diagnosed. RESULTS: After first renal transplantation, 0 definite and 1 (6%) probable recurrence occurred in 18 patients with childhood-onset HUS, as opposed to 7 (41%) definite and 3 (18%) probable recurrences in 17 adult-onset HUS patients (odds ratio [OR], 13.4; 95% confidence interval [CI], 1.7-105.7). In the latter patients, early use of cyclosporine A increased the risk for recurrence. The incidence of acute rejections was increased compared with matched controls (OR, 1.52; 95% CI, 1.05-2.19 for adult-onset HUS and OR, 1.88; 95% CI, 1.34-2.62 for childhood-onset HUS). One-year graft survival in adult-onset HUS was poor (29%), whereas 1-year graft survival in childhood-onset HUS was comparable to matched controls. CONCLUSIONS: In adult-onset HUS, the recurrence rate and the incidence of acute rejections are high, resulting in a detrimental graft survival. In childhood-onset HUS, the recurrence rate is low, but the posttransplantation course is complicated by an increased incidence of acute rejections.     

Positional vertigo and cochlear implantation.

OBJECTIVE: To identify patients developing positional vertigo after cochlear implantation. STUDY DESIGN: Prospective study on a cohort of patients undergoing cochlear implantation. SETTING: Academic tertiary referral center. PATIENTS: The study included 70 consecutive patients who underwent vestibular evaluation before and after cochlear implantation. INTERVENTION: Medical record review. MAIN OUTCOME MEASURE: Recorded vestibular symptoms after cochlear implantation. Patients with positional vertigo were considered case subjects, whereas those without vestibular symptoms were considered case controls. RESULTS: Benign paroxysmal positional vertigo (BPPV) occurred in 8 patients (on the cochlear implant [CI] side in 7 patients, and in the other ear in 1). One patient had BPPV of the lateral semicircular canal on the implanted side, and 7 patients had BPPV of the posterior semicircular canal (on the same CI side in 6 patients, and on the opposite side in 1), which were detected and presented during the last examination. In 5 patients, the onset of symptoms varied from 7 to 130 days after implant activation; in 2 patients, the onset occurred before activation. CONCLUSION: Three different mechanisms are proposed for the occurrence of BPPV in patients with CI. The first focuses on the fall of bone dust particles into the cochlea during cochleostomy. In the second, the vibration caused by drilling the cochlea would be sufficient to dislodge otoconia into the labyrinth. The third hypothesis suggests dislodging of an otolith because of the electric stimulation. In our patients, conservative approaches have been used with a minimal invasive cochleostomy and without perilymph suction. Thus, the vibratory trauma affecting the cochlea during cochleostomy seems to play a fundamental role in the development of paroxysmal vertigo in patients with implant.