Production Of Dna Minicircles Less Than 250 Base Pairs Through A Novel Concentrated Dna Circularization Assay Enabling Minicircle Design With Nf-κb Inhibition Activity

Double-stranded DNA minicircles of less than 1000 bp in length have great interest in both fundamental research and therapeutic applications. Although minicircles have shown promising activity in gene therapy thanks to their good biostability and better intracellular trafficking, minicircles down to 250 bp in size have not yet been investigated from the test tube to the cell for lack of an efficient production method. Herein, we report a novel versatile plasmidfree method for the production of DNA minicircles comprising fewer than 250 bp. We designed a linear nicked DNA double-stranded oligonucleotide bluntended substrate for efficient minicircle production in a ligase-mediated and bending protein-assisted circularization reaction at high DNA concentration of 2M. This one pot multi-step reaction based-method yields hundreds of micrograms of minicircle with sequences of any base composition and position and containing or not a variety of site-specifically chemical modifications or physiological supercoiling. Biochemical and cellular studies were then conducted to design a 95 bp minicircle capable of binding in vitro two NF-κB transcription factors per minicircle and to efficiently inhibiting NF-κB-dependent transcriptional activity in human cells. Therefore, our production method could pave the way for the design of minicircles as new decoy nucleic acids. © The Author(s) 2016.45

Structure of the specific combining ability between two species of Eucalyptus. II. A clustering approach and a multiplicative model

International audienceThe Eucalyptus breeding program of URPPI (a partnership between CIRAD-Fore t, Centre National de la Recherche Forestie re du Congo, and Unite dÕAforestation Industrielle du Congo) consists of a re- ciprocal recurrent selection scheme developed in the Congo between the two species Eucalyptus urophylla and Eucalyptus grandis. Two approaches are proposed in order to model and predict the speciÞc combining ability (SCA) between these species. The clustering ap- proach uses a simultaneous clustering procedure of the two species based on SCA and reveals heterotic groups coherent with the geographical origins of E. urophylla genotypes. The second approach uses a multiplicative model to partition the SCA into three multiplicative terms explaining 95% of the interactio

Structure of the specific combining ability between two species of Eucalyptus. I. RAPD data

International audienceWithin the context of the reciprocal recurrent selection scheme developed in 1989 by CIRAD-Fore t on Eucalyptus, RAPD essays were performed to assess the genetic diversity in the two species E. urophylla and E. grandis. The molecular markers were split into two parts: the speciÞc markers (present with di¤erent fre- quencies in the two species) and the common markers (present with similar frequencies in the two species). The study analyses the structure of genetic diversity within and between the two species of Eucalyptus. Dif- ferent genetic distances are worked out for use in pre- diction equations of the individual tree trunk volume of hybrids at 38 months. Each distance is expressed as the sum of the general genetic distance and the speciÞc genetic distance. The general genetic distance based on the double presence plus the double absence of bands seems to be an interesting co-variate to use in a factor regression model. Through this model the distance calculated between species explains the general com- bining ability (GCA) and the speciÞc combining ability (SCA) of the phenotypic character with a global coe¦c- ient of determination of 81.6

Follicular dynamics in Serrana goats

Twenty-two Serrana goats were studied through two successive estrous cycles in order to characterize their follicular dynamics during the breeding season. The ovaries of the goats were scanned daily by realtime ultrasonography and all follicles ≥3mm were measured and classified. The data were classified by the number of follicular waves per goat to test the hypothesis that temporal and morphological differences between the last follicular wave of an ovary, irrespective of ovulation, will affect the selection of the next ovulatory wave. The mean interovulatory interval was 20.7±1.0 days (mean±S.D.). Three to five waves per estrous cycle were observed and 61.3% (19/31) of cycles had fourwaves. In estrous cycles with fourwaves, the day of onset of the first, second, third and fourth wave was 1.4±1.0, 6.9±1.4, 11.6±1.8 and 16.8±1.6, respectively. No differences (P > 0.05) were found between the day of onset of the first and second waves for estrous cycles with three, four or five waves. However, the day of onset of the third and fourth waves occurred later when the number of waves per estrous cycle increased (P < 0.001). The duration of the interwave interval (time between the day of onset of two consecutive waves) was longer when the second wave was ovulatory. The length of the growth phase (2.4±0.9 days) and size (5.9±0.7 mm) of the dominant follicle in the second wave were lower (P < 0.01) than for the first wave (3.3±1.2 days and 6.6±0.9 mm, respectively) and the fifth wave (4.1±1.2 days and 7.5±1.0 mm, respectively). Within pairs of ovaries, the onset of the last wave occurred later (P < 0.05) and was less variable in ovulatory ovaries (day 16.8±1.4, n = 20) than in anovulatory ovaries (day 15.1±3.7, n = 20). The length of the growing phase was longer (P < 0.001) in the last waves of ovulatory ovaries (3.1±0.9 days) than in the last waves of anovulatory ovaries (1.7±0.8 days). These results support the hypothesis that the day of onset of the ovulatory wave is related to or, at least, conditioned by the luteolysis and the decrease in plasma progesterone. In summary, the estrous cycle of Serrana goats is characterized by sequential follicular wave growth with a great variability in their onset and duration, with the exception of the ovulatory wave. The temporal and morphological differences observed in the last wave of estrous cycle provide strong evidence for the role of progesterone in their regulation

Addressing the ethical issues associated with fieldwork education in occupational therapy: Results of an empirical study conducted in Quebec

Occupational therapists who contribute to fieldwork education are exposed to ethical issues when supervising trainees. Both the ethical issues and the solutions to address these ethical issues are undocumented in the literature. A qualitative study was conducted to document these issues and their solutions. Twenty-three occupational therapists with supervising experience participated in this study. All the participants reported experiencing ethical issues while supervising trainees. This article aims to present the solutions proposed by the participants in order to address the ethical issues of fieldwork education. Intrinsic solutions are linked to supervisors’ ethical, pedagogical or occupational therapy competences. The extrinsic solutions deal with the appropriate measures which can and should be implemented so as to better support the supervisors’ work and better recognize the important contribution of occupational therapists who train the next generation of occupational therapists in clinical settings. This study is likely to have implications on clinical practice, teaching, research and governance

Management of aneurysms involving branches of the celiac and superior mesenteric arteries: A comparison of surgical and endovascular therapy

ObjectiveAneurysms involving branches of the superior mesenteric and celiac arteries are uncommon and require proper management to prevent rupture and death. This study compares surgical and endovascular treatment of these aneurysms and analyzes outcome.MethodsPatients at the Mount Sinai Medical Center in New York who were treated for aneurysms in the branches of the celiac artery and superior mesenteric artery were identified through a search of the institution’s medical records and endovascular database. Patient demographics, history, clinical presentation, aneurysm characteristics, treatments, and follow-up outcome were retrospectively recorded. Significant differences between patients treated by surgical or endovascular therapy were determined by using Student’s t test and χ 2 analysis.ResultsBetween January 1, 1991, and July 1, 2005, 59 patients with 61 aneurysms were treated at a single institution. Twenty-four patients had surgical repair, and 35 underwent endovascular treatment, which included coil embolization and stent-graft therapy. Splenic (28) and hepatic (22) artery aneurysms predominated. Eighty-nine percent of splenic artery aneurysms were true aneurysms and were treated by endovascular and surgical procedures in near equal numbers (14 and 11, respectively). Pseudoaneurysms were significantly more likely to be treated by endovascular means (P < .01). The technical success rate of endovascular treatment for aneurysms was 89%, and failures were successfully treated by repeat coil embolization in all patients who presented for retreatment. Patients treated by endovascular techniques had a significantly higher incidence of malignancy than patients treated with open surgical techniques (P = .03). Furthermore, patients treated by endovascular means had a shorter in-hospital length of stay (2.4 vs 6.6 days, P < .001).ConclusionEndovascular management of visceral aneurysms is an effective means of treating aneurysms involving branches of the celiac and superior mesenteric arteries and is particularly useful in patients with comorbidities, including cancer. It is associated with a decreased length of stay in the elective setting, and failure of primary treatment can often be successfully managed percutaneously

In vivo imaging of ram spermatozoa in the ewe genital tract using fibered confocal microscopy

National audienceSperm transit in the female tract is one of the key factors in the success of fertilization after artificial insemination in sheep species. However, its study is limited by the absence of in vivo imaging methods. The imaging of ram sperm in the female genital tract was made possible using the confocal fibered microscopy and fluorescent stains adapted to spermatozoa. Our results show the active role of the uterotubal junction in the selection of sperm during their transit

Visual fixation in the intensive care unit: a strong predictor of post-traumatic amnesia and long-term recovery after moderate-to-severe traumatic brain injury

Objective: We examined whether visual fixation at 24h of intensive care unit (ICU) admission is superior to the initial Glasgow Coma Scale (GCS) score to predict PTA duration and long-term TBI recovery. Design: Two-phase cohort study. Setting: Level I trauma ICU. Patients: Moderate-to-severe TBI discharged alive between 2010-2013. Interventions: None. Measurements and Main Results: Presence/absence of visual fixation at 24h of ICU-admission was determined through standard behavioral assessments in 181 TBI patients and compared to the GCS score to predict PTA duration during hospitalization (Phase 1) and performance on the Glasgow Outcome Scale-Extended (GOS-E) 10-40 months after (n=144; Phase 2a). A subgroup also completed a visual attention task (n=35; Phase 2b) and brain magnetic resonance imaging post-TBI (n=23; Phase 2c). Presence/absence of visual fixation at 24h of ICU-admission showed a sensitivity of 84%, a specificity of 82% and an AUC of 0.87 for the prediction of PTA duration. Visual fixation (AUC=0.85) was also found as performant as PTA (AUC=0.81; difference-between-AUC=0.04; 95%CI:-0.03-0.116; p=0.28) for the prediction of GOS-E scores. Conversely, the GCS score was a poor predictor of both PTA and GOS-E. Even when controlling for age/medication/CT scan findings, fixation remained a significant predictor of GOS-E scores (=-0.29, p<0.05). Poorer attention performance and increased regional brain volume deficits were also observed in participants who could not fixate 24h following ICU-admission versus those who could. Conclusions: Visual fixation within 24h of ICU-admission could be as performant as PTA for predicting TBI recovery, introducing a new variable of interest in TBI outcome research