Expression and characterization of Gag protein of HIV-1（CN） in Pichia pastoris

To express the core protein of HIV-1 of Chinese prevalent strain （HIV-1 （CN）） in Pichia pastoris, the fulllength gag gene was inserted into the secretory expression vector pHILS1. Linearized recombinant plasmid pHILGAG by Sail was electrotransformed into the yeast strain GS115, and the yeast transformants were identified by PCR. To induce the interest protein to be expressed, the PCR positive transformants were inoculated in the medium of BMGY and BMMY, mRNA of the strain was detected by RT-PCR, and the expressed protein was analyzed by SDS-PAGE, Western blotting and thin layer scanning. mRNA （1.3 kb） was amplified by RT-PCR. SDS-PAGE and Western blotting analysis showed that the molecular mass of the expressed protein was 55 kD, which was similar to the expected value, and the expressed protein could react with McAb to HIV-1 p24. Thin layer scanning analysis demonstrated that the whole amount of the expressed protein was approximately 13 % of the soluble protein in the supernatant. The recombinant yeast had good genetic stability. The optimal expression conditions of the engineering yeast were as follows： BMMY medium, 80-90% of dissolved oxygen, 1% methanol, and 3-day-cultivation course. Gag proteins were expressed under the optimal expression condition and purified via gel filtration chromatography. The purity of the interest protein was up to 85 %. After the purified proteins were inoculated into BALB/c mice, the anti-HIV-1 antibodies in the immunized mice could be detected by Western blotting.     

Older peoples' perceptions of oral health: 'it's just not that simple'

Abstract: Objectives: Little is known about older persons’ perceptions of oral health and oral health care. The purpose of this study was to explore the viewpoint of older adults’ regarding their oral health care practices. Methods: A qualitative interpretive methodology was employed comprising three analytic levels: coding of data into concepts, analysis of concepts into themes, followed by an in‐depth analysis of relationships within concepts and between themes. In‐depth individual interviews were conducted with 19 participants aged 65 to 87 years. Results: Older people’s decision to access oral health care involves complex and personally meaningful strategies. A dental visit surfaces hopes and fears based on past and present experiences. Mouth and teeth are not merely objects of dental care; they represent a person’s social and relational self. Age‐related changes challenge the relational self as represented in societal ideal images of youth and perfection (the perfect smile). This study highlights older peoples’ resilience and determination when faced with the dilemmas in accessing oral health care – it costs, personally as well as financially. Contrary to the assumption that older peoples’ oral health status is related to neglect, rather for many, it is the result of the intersection of their history with technological advances. Conclusions: These findings challenge oral health care practitioners to be sensitive to the contexts affecting their older client’s oral health care status. They do not ‘just go’ to the dentist; they bring with them their past dental experiences and their hopes for the future. It matters how one is treated at this vulnerable time.     

Corticospinal excitability is dependent on the parameters of peripheral electric stimulation: a preliminary study

Chipchase LS, Schabrun SM, Hodges PW. Corticospinal excitability is dependent on the parameters of peripheral electric stimulation: a preliminary study.

Objective

To evaluate the effect of 6 electric stimulation paradigms on corticospinal excitability.

Design

Using a same subject pre-post test design, transcranial magnetic stimulation (TMS) was used to measure the responsiveness of corticomotor pathway to biceps and triceps brachii muscles before and after 30 minutes of electric stimulation over the biceps brachii. Six different electric stimulation paradigms were applied in random order, at least 3 days apart.

Setting

Motor control research laboratory.

Participants

Healthy subjects (N=10; 5 women, 5 men; mean age ± SD, 26±3.6y).

Interventions

Six different electric stimulation paradigms with varied stimulus amplitude, frequency, and ramp settings.

Main Outcome Measure

Amplitudes of TMS-induced motor evoked potentials at biceps and triceps brachii normalized to maximal M-wave amplitudes.

Results

Electric stimulation delivered at stimulus amplitude sufficient to evoke a sensory response at both 10Hz and 100Hz, and stimulus amplitude to create a noxious response at 10Hz decreased corticomotor responsiveness (all P<0.01). Stimulation sufficient to induce a motor contraction (30Hz) applied in a ramped pattern to mimic a voluntary activation increased corticomotor responsiveness (P=0.002), whereas constant low- and high-intensity motor stimulation at 10Hz did not. Corticomotor excitability changes were similar for both the stimulated muscle and its antagonist.

Conclusions

Stimulus amplitude (intensity) and the nature (muscle flicker vs contraction) of motor stimulation have a significant impact on changes in corticospinal excitability induced by electric stimulation. Here, we demonstrate that peripheral electric stimulation at stimulus amplitude to create a sensory response reduces corticomotor responsiveness. Conversely, stimulus amplitude to create a motor response increases corticomotor responsiveness, but only the parameters that create a motor response that mimics a voluntary muscle contraction.     

Safety and Biodistribution of an Equine Infectious Anemia Virus-Based Gene Therapy, RetinoStat(®), for Age-Related Macular Degeneration

Abstract RetinoStat(?) is an equine infectious anemia virus-based lentiviral gene therapy vector that expresses the angiostatic proteins endostatin and angiostatin that is delivered via a subretinal injection for the treatment of the wet form of age-related macular degeneration. We initiated 6-month safety and biodistribution studies in two species; rhesus macaques and Dutch belted rabbits. After subretinal administration of RetinoStat the level of human endostatin and angiostatin proteins in the vitreous of treated rabbit eyes peaked at ～1 month after dosing and remained elevated for the duration of the study. Regular ocular examinations revealed a mild to moderate transient ocular inflammation that resolved within 1 month of dosing in both species. There were no significant long-term changes in the electroretinograms or intraocular pressure measurements in either rabbits or macaques postdosing compared with the baseline reading in RetinoStat-treated eyes. Histological evaluation did not reveal any structural changes in the eye although there was an infiltration of mononuclear cells in the vitreous, retina, and choroid. No antibodies to any of the RetinoStat vector components or the transgenes could be detected in the serum from either species, and biodistribution analysis demonstrated that the RetinoStat vector was maintained within the ocular compartment. In summary, these studies found RetinoStat to be well tolerated, localized, and capable of persistent expression after subretinal delivery.     

Continuing professional development: the missing link

Attending formal course-based learning forms a major part of continuing professional development for physiotherapists. There is a vast array of formal courses available to physiotherapists each holding the tantalising prospect of improving knowledrge, skills and patient outcomes. However, educational evidence suggests that, while participation in traditionally organised workshops and conferences improves knowledge and practice behaviours of the individual attendee, there is no corollary improvement in patient outcomes. This paper discusses reasons why formal course-based learning has yet to be successful at improving the patient outcomes of those who participate. Suggestions and strategies for reconceptualising this aspect of continuing professional development are provided.     

High-definition imaging of trabeculectomy blebs using spectral domain optical coherence tomography adapted for the anterior segment

Background:  The aim of this work was to image trabeculectomy blebs using spectral domain optical coherence tomography (SDOCT).
Methods:  In this prospective cross-sectional study, patients who had undergone trabeculectomy with at least 3 months of follow up were included. Blebs were imaged using an adapted SDOCT system (Cirrus HD-OCT, Carl Zeiss Meditec Inc., Dublin, CA, USA) and time domain anterior segment optical coherence tomography (ASOCT) (Visante OCT, Carl Zeiss Meditec Inc.). An observer masked to clinical data assessed the utility of SDOCT and ASOCT in visualizing structures in successful and failed blebs.
Results:  Fifty-one eyes were imaged, of which 43 (84.3%) were successful. SDOCT showed wall thickening (93.0% vs. 67.4%, P  = 0.006) and discrete hyporeflective spaces in the wall (88.4% vs. 14.0%, P  < 0.0001) in a greater proportion of successful blebs than ASOCT. SDOCT showed the bleb cavity (23.3% vs. 48.8%, P  = 0.02), scleral flap (34.9% vs. 90.7%, P  < 0.0001), subflap space (20.9% vs . 72.1%, P  < 0.0001) and ostium (9.3% vs. 88.4%, P  < 0.0001) in fewer successful blebs than ASOCT. The internal ostium was not visualized in any failed bleb using SDOCT, whereas ASOCT showed the ostium in 87.5% of failed blebs ( P  = 0.001). SDOCT showed cystic spaces in the bleb wall in a greater proportion of successful blebs than failed blebs (88.4% vs. 37.5%, P  = 0.005).
Conclusions:  SDOCT imaging was able to show fine superficial features in the bleb wall. However, SDOCT had limited clinical utility in that it did not provide useful information about deep features such as flap position, bleb cavity formation or patency of the subflap space and internal ostium.     

沙利度胺联合拓扑特肯与单一拓扑特肯在复发性上皮性卵巢癌的前瞻性随机研究

根据沙利度胺和拓扑特肯的活性机制和具抗血管生成的特性,本研究比较了沙利度胺联合拓扑特肯与拓扑特肯单用在复发的上皮性卵巢、输卵管、腹膜癌的疗效。 此项历时4年的多中心的前瞻性随机2期研究得到明尼苏达大学妇女癌症中心的协助。人选者具有下列条件之一：①在以铂类和紫杉醇为基础的初始化疗后未控。