Use of a short hPDE6b promoter for rod gene transfer in a model of severe retinal cystrophy, the Rd10 mouse

Purpose: Gene therapy of severe retinal dystrophies directly affecting photoreceptor is still a challenge in terms of clinical application. One of the main hurdles is to generate high transgene expression specifically in rods or cones. In the present study, we are investigating the possibility to drive hPDE6b expression in the Rd10 mouse retina using a specific sequence of the human PDE6b promoter. Methods: Two 5' flanking fragments of the human PDE6b gene: (-93 to +53 (146 bp) and -297 to +53 (350 bp, see Di Polo and Farber, 1995) were cloned in different plasmids in order to check their expression in vitro and in vivo. These elements drove the activity of either luciferase (pGL3 plasmids) or EGFP (AAV2/8 backbone). Then, an AAV2/8 vector carrying the PDE6b cDNA was tested with subretinal injections at P9 in the Rd10 eyes. Eye fundus, OCT, ERG recordings and histological investigations were performed to assess the efficacy of the gene transfer. Results: The short PDE6b promoter containing 146bp (-93 to +53) showed the highest activity in the Y-79 cells, as described previously (Di Polo and Farber, 1995). Subretinal administrations of AAV2/8-PDE6bpromoter-EGFP allowed a rapid expression specifically in rods and not in cones. The expression is faster than a vector containing the CMV promoter. The AAV2/8-PDE6bpromoter-PDE6b and the control vector were injected at P9 in the Rd10 mouse retina and investigated 5 weeks post-injection. Out of 14 eyes, 6 presented an increased rod sensitivity of about 300 fold, and increased a- and b-wave responses in ERG recordings. Flicker stimulations revealed that cones are also functional. OCT images and histological analyses revealed an increased ONL size in the injected area. The retina treated with the therapeutic vector presented 4-6 rows of photoreceptors with outersegments containing PDE6b. In the control eyes, only 2-4 rows of photoreceptors with almost no OS were observed . Conclusions: The 146 bp promoter sequence (-93 to + 53) is the shortest regulatory element described to date which allows to obtain efficient rod-specific expression in the context of somatic gene transfer. This first result is of great interest for AAV vector design in general allowing more space for the accommodation of transgenes of interest and good expression in rods. Moreover we showed the proof of principle of the efficacy of AAV2/8-PDE6bp-PDE6b vector in the Rd10 mouse model of severe photoreceptor degeneration without using neither AAV mutated capsids, nor self-complementary vectors.

Evaluation of bedbath in critically ill patients: impact of water temperature on the pulse oximetry variation

This is a participant study, quasi-experimental, of a before and after type. A quantitative approach of biophysiological measures was used, represented by the saturation of oxygen measured by pulse oximeter (SpO2), and recorded on three occasions: before, during and after the bedbath in critically ill patients hospitalized at the ICU of a University Hospital in Brazil. Objective: to compare the SpO2 in various stages of the bath, with and without control of water temperature. Data collection was performed between December 2007 and April 2008 on a convenience sample consisting of 30 patients aged over 18 who had classification in TISS-28 from level II. Results show that water temperature control means a lower variation of SpO2 (p<0.05). No marked differences in variation of saturation between men and women or between age groups were established. In conclusion, heated and constant water temperature during the bedbath is able to minimize the fall of SpO2 that occurs while handling patients during procedures.

Ultra-Thin PCC Overlays, January 2005

· Evaluate conventional methods of slab removal and asphalt surface preparation for subsequent overlays of portland cement concrete (PCC) in the “remove and replace” areas. · Evaluate existing asphaltic concrete surface under the “remove and patch” areas of rehabilitation areas and evaluate joint formation in the areas of patching. · Evaluate polypropylene fiber enhanced concrete at the three-inch depth to determine the cost/benefit of its inclusion. · Evaluate the performance of the rehabilitated ultra-thin whitetopping sections and the extended performance of the existing ultra-thin sections with and without patching. · Validate existing ultra-thin whitetopping design procedures of the Portland Cement Association (PCA) and American Concrete Pavement Association (ACPA) for application in Iowa.

Intrauterine growth restriction and absent or reverse end-diastolic blood flow in umbilical artery (Doppler class II or III): A retrospective study of short- and long-term fetal morbidity and mortality.

OBJECTIVE: Absent or reverse end-diastolic flow (Doppler II/III) in umbilical artery is correlated with poor perinatal outcome, particularly in intrauterine growth restricted (IUGR) fetuses. The optimal timing of delivery is still controversial. We studied the short- and long-term morbidity and mortality among these children associated with our defined management. STUDY DESIGN: Sixty-nine IUGR fetuses with umbilical Doppler II/III were divided into three groups; Group 1, severe early IUGR, no therapeutic intervention (n = 7); Group 2, fetuses with pathological biophysical profile, immediate delivery (n = 35); Group 3, fetuses for which expectant management had been decided (n = 27). RESULTS: In Group 1, stillbirth was observed after a mean delay of 6.3 days. Group 2 delivered at an average of 31.6 weeks and two died in the neonatal period (6%). In Group 3 after a mean delay of 8 days, average gestational age at delivery was 31.7 weeks; two intra uterine and four perinatal deaths were observed (22%). Long-term follow-up revealed no sequelae in 25/31 (81%) and 15/18 (83%), and major handicap occurred in 1 (3%) and 2 patients (11%), respectively, for Groups 2 and 3. CONCLUSION: Fetal mortality was observed in 22% of this high risk group. After a mean period of follow-up of 5 years, 82% of infants showed no sequelae. According to our management, IUGR associated with umbilical Doppler II or III does not show any benefit from an expectant management in term of long-term morbidity.

Short term impact of Tribulus terrestris intake on doping control analysis of endogenous steroids.

Tribulus terrestris is a nutritional supplement highly debated regarding its physiological and actual effects on the organism. The main claimed effect is an increase of testosterone anabolic and androgenic action through the activation of endogenous testosterone production. Even if this biological pathway is not entirely proven, T. terrestris is regularly used by athletes. Recently, the analysis of two female urine samples by GC/C/IRMS (gas chromatography/combustion/isotope-ratio-mass-spectrometry) conclusively revealed the administration of exogenous testosterone or its precursors, even if the testosterone glucuronide/epitestosterone glucuronide (T/E) ratio and steroid marker concentrations were below the cut-off values defined by World Anti-Doping Agency (WADA). To argue against this adverse analytical finding, the athletes recognized having used T. terrestris in their diet. In order to test this hypothesis, two female volunteers ingested 500 mg of T. terrestris, three times a day and for two consecutive days. All spot urines were collected during 48 h after the first intake. The (13)C/(12)C ratio of ketosteroids was determined by GC/C/IRMS, the T/E ratio and DHEA concentrations were measured by GC/MS and LH concentrations by radioimmunoassay. None of these parameters revealed a significant variation or increased above the WADA cut-off limits. Hence, the short-term treatment with T. terrestris showed no impact on the endogenous testosterone metabolism of the two subjects.

Alliance patterns over the course of short-term dynamic psychotherapy: The shape of productive relationships.

The shape of alliance processes over the course of psychotherapy has already been studied in several process-outcome studies on very brief psychotherapy. The present study applies the shape-of-change methodology to short-term dynamic psychotherapies and complements this method with hierarchical linear modeling. A total of 50 psychotherapies of up to 40 sessions were included. Alliance was measured at the end of each session. The results indicate that a linear progression model is most adequate. Three main patterns were found: stable, linear, and quadratic growth. The linear growth pattern, along with the slope parameter, was related to treatment outcome. This study sheds additional light on alliance process research, underscores the importance of linear alliance progression for outcome, and also fosters a better understanding of its limitations.

Functional MRI evaluation of liver tumour response after radiofrequency: short- and mid-term evolution of diffusion parameters

Purpose: To evaluate the short- and mid-term evolutions of the apparent diffusion coefficient of lesions treated with RF, in order to determine if the ADC can be used as a marker of tumour response. Methods and Materials: Twenty patients were treated for a liver malignancy with RF and were examined on a 1.5 T/3.0 T machine with T2, gadolinium-enhanced T1 and diffusion sequences: before treatment (< 1 month), just after treatment (< 1 month) and midterm (3-6 months). The ADC was measured in the whole lesion and in the area with the most restricted diffusion (MRDA). The ROI size was also measured on the diffusion map. The Pearson/ANOVA tests were used. Results: All patients were successfully treated with complete disappearance of CE. The lesional size on T2 showed a negative evolution in time (p < 0.002). The ADC in the whole lesion showed a bell-shaped evolution (increasing just after RF, then decreasing, p = 0.02). The ROI size on the diffusion map followed a similar course (p = 0.01). For the MRDA, such evolutions were also found, but they were not significant. There was a negative correlation between CE and the ADC (p < 0.02) and between the lesional size on T2 and ADC (p = 0.03) in the whole lesion. There were also positive correlations between the ROI size and ADC (p = 0.0008) and between CE and the size on T2 (p = 0.0002). The ADC in MRDA showed some non-significant correlations with other variables. Conclusion: The lesions successfully treated with RF have a clear and predictable evolution in terms of T2 size, CE and ADC.

Ultra-Thin Portland Cement Concrete Overlay Extended Evaluation, January 2005

In this day of the mature highway systems, a new set of problems is facing the highway engineer. The existing infrastructure has aged to or past the design life of the original pavement design. In many cases, increased commercial traffic is creating the need for additional load carrying capacity, causing state highway engineers to consider new alternatives for rehabilitation of existing surfaces. Alternative surface materials, thicknesses, and methods of installation must be identified to meet the needs of individual pavements and budgets. With overlays being one of the most frequently used rehabilitation alternatives, it is important to learn more about the limitations and potential performance of thin bonded portland cement overlays and subsequent rehabilitation. The Iowa ultra-thin project demonstrated the application of thin portland cement concrete overlays as a rehabilitation technique. It combined the variables of base preparation, overlay thickness, slab size, and fiber enhancement into a series of test sections over a 7.2-mile length. This report identifies the performance of the overlays in terms of deflection reduction, reduced cracking, and improved bonding between the portland cement concrete (PCC) and asphalt cement concrete (ACC) base layers. The original research project was designed to evaluate the variables over a 5-year period of time. A second project provided the opportunity to test overlay rehabilitation techniques and continue measurement of the original overlay performance for 5 additional years. All performance indicators identified exceptional performance over the 10-year evaluation period for each of the variable combinations considered. The report summarizes the research methods, results, and identifies future research ideas to aid the pavement overlay designer in the successful implementation of ultra-thin portland cement concrete overlays as an lternative pavement rehabilitation technique.

The short arm of the law: judicial institutions and local governance in Brazil

This paper estimates the effect of judicial institutions on governance at the local level in Brazil. Our estimation strategy exploits a unique institutional feature of state judiciary branches which assigns prosecutors and judges to the most populous among contiguous counties forming a judiciary district. As a result of this assignment mechanism there are counties with nearly identical populations, some with and some without local judicial presence, which we exploit to impute counterfactual outcomes. Conditional on observable county characteristics, offenses per civil servant are about 35% lower in counties that have a local seat of the state judiciary. The lower incidence of infractions stems mostly from fewer violations of financial management regulations by local administrators, fewer instances of problems in project execution and project managment, fewer cases of non-existent or ineffective civil society oversight and fewer cases of improper handling of remittances to local residents.

Structural development of the Tso Morari ultra-high pressure nappe of the Ladakh Himalaya

A continental subduction-related and multistage exhumation process for the Tso Morari ultra-high pressure nappe is proposed. The model is constrained by published thermo-barometry and age data, combined with new geological and tectonic maps. Additionally, observations on the structural and metamorphic evolution of the Tso Morari area and the North Himalayan nappes are presented. The northern margin of the Indian continental crust was subducted to a depth of >90 km below Asia after continental collision some 55 Ma ago. The underthrusting was accompanied by the detachment and accretion of Late Proterozoic to Early Eocene sediments, creating the North Himalayan accretionary wedge, in front of the active Asian margin and the 103-50 Ma Ladakh arc batholith. The basic dikes in the Ordovician Tso Morari granite were transformed to eclogites with crystallization of coesite, some 53 Ma ago at a depth of >90 kin (>27 kbar) and temperatures of 500 to 600 degrees C. The detachment and extrusion of the low density Tso Morari nappe, composed of 70% of the Tso Morari granite and 30% of graywackes with some eclogitic dikes, occurred by ductile pure and simple shear deformation. It was pushed by buoyancy forces and by squeezing between the underthrusted Indian lithosphere and the Asian mantle wedge. The extruding Tso Morari nappe reached a depth of 35 km at the base of the North Himalayan accretionary wedge some 48 Ma ago. There the whole nappe stack recrystallized under amphibolite facies conditions of a Barrovian regional metamorphism with a metamorphic field gradient of 20 degrees C/km. An intense schistosity with a W-E oriented stretching lineation L, and top-to-the E shear criteria and crystallization of oriented sillimanite needles after kyanite, testify to the Tso Morari nappe extrusion and pressure drop. The whole nappe stack, comprising from the base to top the Tso Morari, Tetraogal, Karzok and Mata-Nyimaling-Tsarap nappes, was overprinted by new schistosities with a first N-directed and a second NE-directed stretching lineation L-2 and L-3 reaching the base of the North Himalayan accretionary wedge. They are characterized by top-to-the S and SW shear criteria. This structural overprint was related to an early N- and a younger NE-directed underthrusting of the Indian plate below Asia that was accompanied by anticlockwise rotation of India. The warping of the Tso Morari dome started already some 48 Ma ago with the formation of an extruding nappe at depth. The Tso Morari dome reached a depth of 15 km about 40 Ma ago in the eastern Kiagar La region and 30 Ma ago in the western Nuruchan region. The extrusion rate was of about 3 cm/yr between 53 and 48 Ma, followed by an uplift rate of 1.2 mm/yr between 48 and 30 Ma and of only 0.5 mm/yr after 30 Ma. Geomorphology observations show that the Tso Morari dome is still affected by faults, open regional dome, and basin and pull-apart structures, in a zone of active dextral transpression parallel to the Indus Suture zone.

Brevissima et facillima in omnes D. Pauli epistolas scholia : ultra priores editiones, ex antiquissimis graecorum authoribus, abunde locupletata ; itidem in septem canonicas epistolas et D. Ioannis Apocalypsin, brevissima scholia recens edita ([Reprod.]) / authore Ioanne Gagnaio,...

Collection : French books before 1601 ; 66.2