Intraneural colchicine inhibition of adenoviral and adeno-associated viral vector remote spinal cord gene delivery

OBJECTIVE: The mechanism of remote viral gene delivery to the spinal cord is unknown. The present experiment demonstrates that intraneural injection of colchicine is capable of inhibiting remote delivery of both adenoviral and adeno-associated viral (AAV) vectors, implicating axonal transport in this process. METHODS: The right sciatic nerves of adult Sprague-Dawley rats were injected with phosphate-buffered saline (PBS) (n = 5) or 10 (n = 7) or 100 (n = 4) microg colchicine. Two days later, the nerves of all animals were initially injected with 1.2 x 10(7) plaque-forming units of Ad5RSVntLac-Z. Two separate groups were injected concurrently with vector and PBS (n = 5) or 10 microg colchicine (n = 5). In a second experiment, the right sciatic nerves of CD1 mice were preinjected with PBS (n = 6) or 10 microg colchicine (n = 5). Two days later, the nerves were injected with rAAVCAG-EGFPwpre (an adeno-associated vector carrying the green fluorescent protein gene). In both experiments, sciatic nerves and spinal cords were removed and analyzed for gene expression. RESULTS: Sciatic nerve vector injection resulted in expression in both the nerve injection site and neuronal cell bodies located predominantly in the ipsilateral ventral horn. Analysis of variance revealed a significant treatment effect for 10 and 100 microg intraneural colchicine with inhibition of remote adenoviral delivery at 10 microg and blockade of remote delivery at 100 microg (P < 0.001). Colchicine injection concurrent with and before vector injection had similar inhibitory effects. Two-way analysis of variance revealed significant colchicine inhibition of remote delivery in both adenovirus- and AAV-injected animals (P < 0.003) but no dose-by-vector interaction, suggesting that both vectors are equally inhibited by colchicine. CONCLUSION: Colchicine inhibits remote spinal cord delivery of adeno-associated and adenoviral vectors in a dose-dependent manner, suggesting that remote delivery is dependent on retrograde axonal transport.     

Less severe retinopathy of prematurity induced by surfactant replacement therapy

To assess the effect of surfactant replacement therapy (SRT) on the prevalence and severity of retinopathy of prematurity (ROP), we compared data from 160 SRT-treated preterm infants with data from 230 historic controls. The prevalence of ROP was 30.6% in the treatment group and 23.4% in the control group. Severe ROP (stages 3-4) was seen in 6.1% of the infants with ROP in the treatment group and 20.3% of the ROP patients in the control group. Surfactant therapy had no influence on the prevalence of ROP (odds ratio 1.4, 95% confidence interval 0.797-2.459, p = 0.242). However, SRT was associated with a decreased risk for severe ROP, compared to mild ROP (odds ratio 0.226, 95% confidence interval 0.056-0.905, p = 0.036). These data suggest that SRT is associated with a decreased risk for severe ROP.     

What Can Be Expected of Balloon Aortic Valvuloplasty in Adults?

Introduced more than 15 years ago, balloon aortic valvuloplasty (BAV) has clearly delineated indications and limitations. Although, aortic valve replacement is undoubtedly the treatment of choice for healthy patients with aortic stenosis, BAV has been demonstrated as a valuable and safe palliative procedure for patients considered too risky or old for current day surgery. Between January 1991 and December 1999, 858 patients (mean age 76 ± 11 years, 479 [57%] women) underwent BAV at our center. Indications for BAV were contraindication to surgery in 16.2% of the patients, high risk in 40.4%, refusal of surgery in 10.7%, personal decision in 18.5%, and other in 14.2%. The mean gradient decreased from 65 ± 23 to 27 ± 12 mmHg and the valve area increased from 0.56 ± 0.19 cm² to 1.0 ± 0.38 cm². Eleven (1.1%) patients died during the procedure. The other severe complications were complete AV block in 18 (1.8%) patients, stroke in 8 (0.8%), tamponade in 6 (0.6%), and massive aortic insufficiency in 4 (0.4%). In conclusion, BAV does not replace valve replacement, even in the elderly. All those who can be operated on, should be. However, if surgical intervention is not considered reasonable, BAV offers good immediate results with an acceptable complication rate. Furthermore, previous published series have demonstrated that BAV is particularly indicated in specific subgroups, such as very old patients with high operative risk, end-stage heart failure, cardiogenic shock, or general anesthesia for noncardiac surgery.     

Volumetric rendering of MR images

The authors developed new techniques for three-dimensional display of magnetic resonance (MR) images that preserve soft-tissue definition, are fully automatic, and work with routinely used section thicknesses. MR images are segmented, selectively enhanced, and displayed by means of a volumetric rendering algorithm. These techniques were used to illustrate normal anatomy of the brain, knee, and liver. Three-dimensional rendering of balanced spin-echo images shows the ventricles and extracerebral veins and of T1-weighted images, the sulci and gyri. The large hepatic and portal vessels can be seen with these enhancement techniques. Three-dimensional views of the knee reveal articular surfaces of the tibia and clearly depict menisci and posterior and anterior cruciate ligaments. These techniques make it possible to image multiple soft tissues simultaneously while preserving the detail contained in the original images. Three-dimensional presentation of complex, overlapping anatomic regions is helpful in surgical planning and should lead to improved diagnosis.     

Anesthesia for children in Sub-Saharan Africa – a description of settings, common presenting conditions, techniques and outcomes

Anesthesia in developing countries deserves special attention. The most common technique is general anesthesia (with spontaneous or manually assisted ventilation). Nonmedical anesthetists with limited training and supervision and lacking the most common drugs and anesthetic equipment administer anesthesia, usually for emergency surgery. There are important safety issues, especially for pediatric anesthesia. Regarding pediatric surgery, the major workload is due to abdominal emergencies, mainly neonatal bowel obstruction or peritonitis due to typhoid perforation. The morbidity and mortality rate for these conditions is high.     

Cell tracking technologies for acute ischemic brain injury

Stem cell therapy has showed considerable potential in the treatment of stroke over the last decade. In order that these therapies may be optimized, the relative benefits of growth factor release, immunomodulation, and direct tissue replacement by therapeutic stem cells are widely under investigation. Fundamental to the progress of this research are effective imaging techniques that enable cell tracking in vivo. Direct analysis of the benefit of cell therapy includes the study of cell migration, localization, division and/or differentiation, and survival. This review explores the various imaging tools currently used in clinics and laboratories, addressing image resolution, long-term cell monitoring, imaging agents/isotopes, as well as safety and costs associated with each technique. Finally, burgeoning tracking techniques are discussed, with emphasis on multimodal imaging.     

VJ REARRANGEMENTS OF THE TCRγ LOCUS IN PERIPHERAL T-CELL LYMPHOMAS: ANALYSIS BY POLYMERASE CHAIN REACTION AND DENATURING GRADIENT GEL ELECTROPHORESIS

Using Southern blotting for the diagnosis of clonality in peripheral T-cell lymphomas (PTCLs), analysis of the T-cell receptor (TCR) γ gene rearrangement was shown to be more informative than that of the TCR β gene rearrangement. In order to amplify every VJγ rearrangement, a polymerase chain reaction (PCR) procedure using newly designed GC-clamp primers has been developed. All primers can be mixed in a single multiplex PCR. PCR products are analysed by denaturing gradient gel electrophoresis (DGGE), providing tumour-specific imprints inasmuch as the procedure characterizes N sequence polymorphism at the VJ junctions. In a series of 30 PTCL cases, the PCR procedure demonstrated 27 cases to be clonally rearranged and failed in three cases. PCR was more accurate than Southern blotting, showing 47 rearranged γ alleles, four of which were undetectable on the Southern blot. When lymphomas were studied at different sites and at relapse, the DGGE pattern remained unchanged. In PTCL, the proposed PCR is helpful for the diagnosis and staging of the disease and should improve the follow-up monitoring. The undetectability of clonal rearrangements in a few cases is discussed in the light of concepts of lymphomagenesis and T-cell differentiation.     

Nosocomial respiratory syncytial virus infection in Canadian pediatric hospitals: a Pediatric Investigators Collaborative Network on Infections in Canada Study

OBJECTIVE: To determine nosocomial transmission of respiratory syncytial virus (RSV) in Canadian pediatric hospitals, outcomes associated with nosocomial disease, and infection control practices. DESIGN: A prospective cohort study in the 1992 to 1994 winter respiratory seasons. SETTING: Nine Canadian pediatric university-affiliated hospitals. PARTICIPANTS: Hospitalized children with symptoms of lower respiratory tract infection (at least one of cough, wheezing, dyspnea, tachypnea, and apnea) and RSV antigen identified in a nasopharyngeal aspirate. RESULTS: Of 1516 children, 91 (6%) had nosocomial RSV (NRSV), defined as symptoms of lower respiratory tract infection and RSV antigen beginning >72 hours after admission. The nosocomial ratio (NRSV/[com-munity-acquired RSV {CARSV})] + NRSV) varied by site from 2.8% to 13%. The median length of stay attributable to RSV for community-acquired illness was 5 days, but 10 days for nosocomial illness. Four children with NRSV (4. 4%) died within 2 weeks of infection, compared with 6 (0.42%) with CARSV (relative risk = 10.4, 95% confidence interval: 3.0, 36.4). All sites isolated RSV-positive patients in single rooms or cohorted them. In a multivariate model, no particular isolation policy was associated with decreased nosocomial ratio, but gowning to enter the room was associated with increased risk of RSV transmission (incidence rate ratio 2.81; confidence interval: 1.65, 4.77). CONCLUSIONS: RSV transmission risk in Canadian pediatric hospitals is generally low. Although use of barrier methods varies, all sites cohort or isolate RSV-positive patients in single rooms. Children with risk factors for severe disease who acquire infection nosocomially have prolonged stays and excess mortality.