Photochemical internalization (PCI)--a novel technology for release of macromolecules from endocytic vesicles

The utilisation of macromolecules in therapy of cancer and other diseases is becoming increasingly relevant. Recent advances in molecular biology and biotechnology have made it possible to improve targeting and design of cytotoxic agents or DNA complexes for clinical applications. To achieve the expected biological effect of these macromolecules in many cases internalization to the cell cytosol is crucial. A number of different methods for internalization of membrane impermeable molecules has been established, including electroporation, liposome fusion, antibodies/targeting ligands as protein carriers and the utilisation of various types of vectors such as cationic polymers and viruses, for gene therapy. Although new delivery systems have improved the cellular uptake of macromolecules, tissue penetration, cellular uptake and efficient transfer of the molecules into the cytosol of the target cell are still fundamental obstacles. At an intracellular level, the most fundamental obstruction for cytosolic release of the therapeutic molecule is the membrane-barrier of the endocytic vesicles. Photochemical internalization (PCI) is a novel technology for release of endocytosed macromolecules into the cytosol. The technology is based on the use of photosensitizers located in endocytic vesicles that upon activation by light induce a release of macromolecules from their compartmentalization in endocytic vesicles. PCI has been shown to potentiate the biological activity of a large variety of macromolecules and other molecules that do not readily penetrate the plasma membrane, including proteins, peptides, and DNA delivered as a complex with cationic polymers or incorporated in adenovirus. The basis as well as the utilization of this technology will be briefly reviewed in this paper.     

Poliovirus detection in wastewater and stools following an immunization campaign in Havana, Cuba

BACKGROUND: Recent outbreaks of poliomyelitis caused by vaccine-derived virus have raised concerns that vaccine-derived poliovirus may continue to circulate after eradication. In these outbreaks, the virus appears to have replicated for > or =2 years before detection. Early detection is critical for an effective response to these outbreaks. Although acute flaccid paralysis (AFP) surveillance will remain the standard for poliovirus detection, wastewater sampling could be a useful supplement. In this study, we evaluated the sensitivity of wastewater sampling by concurrently collecting stools from children aged < 3 years attending two neighbourhood clinics in Havana, Cuba, and wastewater from the same neighbourhoods. METHODS: Sample collection was begun during the third week after the national immunization campaign, continued weekly through the seventh week, and was repeated during weeks 15 and 19. Virus detection and titration were performed using both cell culture and polymerase chain reaction techniques. RESULTS: Wastewater sampling was found to be at least as sensitive as stool sampling under these conditions. Poliovirus was isolated from children through week 7, suggesting that viral shedding reached undetectable levels between weeks 8 and 14. The last virus-positive wastewater sample was collected during week 15. CONCLUSIONS: Wastewater sampling under the conditions studied can be a sensitive supplement to AFP surveillance. Similar studies under different conditions are needed to determine the role of wastewater sampling in post-eradication surveillance.     

Living with bodily strangeness: women's experiences of their changing and unpredictable body following a stroke

The authors' aim in this phenomenologial and feminist study was to gain a deeper understanding of how female stroke survivors experienced their body after a stroke. They recruited 25 women in a rural area in eastern Norway who had suffered a first-time stroke and interviewed them in depth three times each during the first 1 1/2 to 2 years following the stroke. The data analysis was inspired by phenomenological method. The stroke survivors' experiences of their bodies were characterized by profound, disturbing, and, in part, unintelligible changes during the onset and the process of recovery from the stroke. Their experiences can be summarized under three major themes: The Unpredictable Body, The Demanding Body, and The Extended Body.     

Driving ability after acute and sub-chronic administration of levocetirizine and diphenhydramine: a randomized,double-blind,placebo-controlled trial

Rationale Sedation following antihistamine use poses a danger to ambulant patients involved in daily activities such as driving. Objective To investigate effects of levocetirizine (5 mg), diphenhydramine (50 mg), and placebo on driving ability during normal traffic. Methods Forty-eight healthy volunteers participated in a double-blind, placebo-controlled, randomized clinical trial. Treatments were administrated on days 1, 2, 3 and 4, exactly 1.5 h before the start of the standardized driving test (performed on day 1 and day 4). In the standardized driving test, subjects were instructed to drive with a steady lateral position, while maintaining a constant speed (95 km/h). Primary parameter was the standard deviation of lateral position (SDLP; cm). Statistical analyses were performed separately for day 1 and day 4, using analysis of variance and an equivalence test. Equivalence to placebo was evidenced if the 95% confidence interval lay between −2.6 cm and +2.6 cm. Results SDLP after levocetirizine was equivalent to placebo on both day 1 (−0.66 cm; +1.12 cm) and day 4 (−0.37 cm; +1.28 cm). In contrast, SDLP after diphenhydramine differed significantly from placebo on both day 1 (P<0.0001) and day 4 (P<0.0003). On day 1, the 95% confidence interval of diphenhydramine (+1.85 cm; +3.63 cm) was partially above the upper equivalence limit (+2.6 cm), indicating clinically relevant driving impairment. On day 4, however, the 95% confidence interval of diphenhydramine (+0.74 cm; +2.38 cm) was contained within the acceptance range. Conclusion In contrast to diphenhydramine, driving performance was not significantly affected while using 5 mg levocetirizine once daily.     

A feminist perspective on stroke rehabilitation: the relevance of de Beauvoir's theory

Abstract The dominant view of women has changed radically during the last century. These changes have had an important impact on the way of life of women in general and, undoubtedly, on women as patients. So far, gender differences have received little attention when developing healthcare services. Stroke hits a great number of elderly women. Wyller et al. found that women seemed to be harder hit by stroke than men; they achieved lower scores in tests of motor, cognitive and ADL functions, both in the acute phase and 1 year after stroke. It is reasonable to expect that differences in outcome among male and female sufferers may in part be explained by the fact that rehabilitation services are designed primarily to meet the needs of men. de Beauvoir's feminist theory maintains that one's body is fundamental in creating the person, which is a lifelong process. Traditionally, the female body has been exposed to alienation and oppression through life. This has led women to develop a life in immanence. This we feel can be of significance in connection with rehabilitation after a stroke, particularly for elderly women. In this article we will discuss how de Beauvoir's theory can throw new light on the experiences and rehabilitation of elderly women and point to ways of improving the process of rehabilitation.     

Speech pathologists’ experience of involving people with stroke-induced aphasia in clinical decision making during rehabilitation

Purpose: Although client participation has been part of legislation and clinical guidelines for several years, the evidence of these recommendations being implemented into clinical practice is scarce, especially for people with communication disorders. The aim of this study was to investigate how speech pathologists experienced client participation during the process of goal-setting and clinical decision making for people with aphasia. Methods: Twenty speech pathologists participated in four focus group interviews. A qualitative analysis using Systematic Text Condensation was undertaken. Results: Analysis revealed three different approaches to client participation: (1) client-oriented, (2) next of kin-oriented and (3) professional-oriented participation. Participants perceived client-oriented participation as the gold standard. The three approaches were described as overlapping, with each having individual characteristics incorporating different facilitators and barriers. Conclusions: There is a need for greater emphasis on how to involve people with severe aphasia in goal setting and treatment planning, and frameworks made to enhance collaboration could preferably be used. Participants reported use of next of kin as proxies in goal-setting and clinical decision making for people with moderate-to-severe aphasia, indicating the need for awareness towards maintaining the clients’ autonomy and addressing the goals of next of kin.

• Implications for Rehabilitation

• Speech pathologists, and most likely other professionals, should place greater emphasis on client participation to ensure active involvement of people with severe aphasia.

• To achieve this, existing tools and techniques made to enhance collaborative goal setting and clinical decision making have to be better incorporated into clinical rehabilitation practice.

• To ensure the autonomy of the person with aphasia, as well as to respect next of kin's own goals, professionals need to make ethical considerations when next of kin are used as proxies in collaborative goal setting and clinical decision making.

     

The Norwegian Doloplus-2, a tool for behavioural pain assessment: translation and pilot-validation in nursing home patients with cognitive impairment

BACKGROUND: Pain assessment is challenging in cognitively impaired (CI) patients due to inadequate self-report skills and observational ratings are an alternative. The Doloplus-2 is developed for pain assessment in the CI and rates somatic, psychomotor and psychosocial behaviours as indicators of pain. AIMS: To translate the Doloplus-2 into Norwegian, to test the Doloplus-2 with regard to criterion validity and to obtain the administrators' evaluation of the clinical performance of the Doloplus-2. METHODS: Nurses at three nursing homes, in collaboration with two research assistants, administered the Doloplus-2 to 59 patients with dementia. The results were compared against experienced clinicians' pain ratings. Regression analyses were performed to explore each different item's contribution to the total pain score. The administrators also completed a debriefing questionnaire. RESULTS: The instrument was translated according to international guidelines. Regression analyses demonstrate that the Doloplus-2 score accounts for 62% (R2) of the expert score and that the four most informative items could explain 68% of the expert score. Analyses of the different Doloplus-2 items indicate that facial expressions explain most and social life least of the expert's pain ratings. The administrators reported that Doloplus-2 was helpful and easy to administer, but questioned the validity of the psychosocial domain. CONCLUSIONS: The Norwegian Doloplus-2 demonstrates satisfactory criterion validity and clinical value in this pilot study. However, the content of the instrument needs a general re-evaluation, especially with regard to the psychosocial items.