Survivin in autoimmune diseases.

Survivin is a protein functionally important for cell division, apoptosis, and possibly, for micro-RNA biogenesis. It is an established marker of malignant cell transformation. In non-malignant conditions, the unique properties of survivin make it indispensable for homeostasis of the immune system. Indeed, it is required for the innate and adaptive immune responses, controlling differentiation and maintenance of CD4+ and CD8+ memory T-cells, and in B cell maturation. Recently, survivin has emerged as an important player in the pathogenesis of autoimmune diseases. Under the conditions of unreserved inflammation, survivin enhances antigen presentation, maintains persistence of autoreactive cells, and supports production of autoantibodies. In this context, survivin takes its place as a diagnostic and prognostic marker in rheumatoid arthritis, psoriasis, systemic sclerosis and pulmonary arterial hypertension, neuropathology and multiple sclerosis, inflammatory bowel diseases and oral lichen planus. In this review, we summarise the knowledge about non-malignant properties of survivin and focus on its engagement in cellular and molecular pathology of autoimmune diseases. The review highlights utility of survivin measures for clinical applications. It provides rational for the survivin inhibiting strategies and presents results of recent reports on survivin inhibition in modern therapies of cancers and autoimmune diseases.

Metal artefact reduction in CT imaging of hip prostheses­an evaluation of commercial techniques provided by four vendors.

OBJECTIVE: The aim of this study was to evaluate commercial metal artefact reduction (MAR) techniques in X-ray CT imaging of hip prostheses. METHODS: Monoenergetic reconstructions of dual-energy CT (DECT) data and several different MAR algorithms, combined with single-energy CT or DECT, were evaluated by imaging a bilateral hip prosthesis phantom. The MAR images were compared with uncorrected images based on CT number accuracy and noise in different regions of interest. RESULTS: The three MAR algorithms studied implied a general noise reduction (up to 67%, 74% and 77%) and an improvement in CT number accuracy, both in regions close to the prostheses and between the two prostheses. The application of monoenergetic reconstruction, without any MAR algorithm, did not decrease the noise in the regions close to the prostheses to the same extent as did the MAR algorithms and even increased the noise in the region between the prostheses. CONCLUSION: The MAR algorithms evaluated generally improved CT number accuracy and substantially reduced the noise in the hip prostheses phantom images, both close to the prostheses and between the two prostheses. The study showed that the monoenergetic reconstructions evaluated did not sufficiently reduce the severe metal artefact caused by large orthopaedic implants. ADVANCES IN KNOWLEDGE: This study evaluates several commercially available MAR techniques in CT imaging of large orthopaedic implants.

Down-regulation of survivin alleviates experimental arthritis.

Survivin is a proto-oncogene that regulates cell division and apoptosis. It is a molecular marker of cancer. Recently, survivin has emerged as a feature of RA, associated with severe joint damage and poor treatment response. The present study examined if inhibition of survivin affects experimental arthritis, which was induced in mBSA-immunized mice by an injection of mBSA in the knee joint or developed spontaneously in collagen type II-immunized mice. The inhibition of survivin transcription by a lentivirus shRNA construct alleviated joint inflammation and reduced bone damage. The inhibition of survivin reduced the levels of metalloproteinases, ß-catenin, and vimentin, limiting the invasive capacity of synovia, while no inhibition of osteoclastogenesis could be found. The inhibition of survivin led to a p53-independent reduction of T cell proliferation and favored the transcription and activity of Blimp-1, which limited IL-2 production and facilitated formation of regulatory Foxp3(+)CD4(+) and effector CD8(+) T cells. The study shows that the inhibition of survivin is sufficient to reduce joint inflammation and bone damage in preclinical models of arthritis. Antiarthritic effects of survivin inhibition are related to p53-independent control of lymphocyte proliferation.

Anticipated changes in sexual risk behaviour following vaccination with a low-efficacy HIV vaccine: survey results from a South African township.

We assessed the potential for anticipated changes in sexual risk-taking behaviour following hypothetical administration of a low-efficacy preventive HIV vaccine. We developed a survey and collected self-reported data from 158 HIV-negative volunteers in a cohort undergoing prescreening for Phase I/II HIV vaccine trials in Soweto. Overall, 22% reported they might use condoms less frequently; 9% reported that they might increase their frequency of sex with casual/anonymous partners; and 55% reported their sexual partners might want to use condoms less frequently knowing they were vaccinated. Multivariate analyses revealed that anticipated decrease in condom use was predicted by poor comprehension and by young age. Individuals may increase their risk-taking behaviour knowing that a vaccine would provide only incomplete protection against HIV transmission. In HIV vaccine trials and future vaccination programmes, education and risk-reduction counselling will be needed for vaccinated individuals and their partners, and mass media education campaigns may be necessary.

Sexual risk behaviour of the first cohort undergoing screening for enrollment into Phase I/II HIV vaccine trials in South Africa.

We assessed risk behaviour in a heterosexual cohort undergoing prescreening for the first Phase I/II HIV vaccine trials in Soweto. We developed a survey and collected self-reported data from HIV-negative potential volunteers. Of 488 participants, most were single and approximately half were from households with incomes below the poverty level. Males reported higher rates of heavy alcohol use (P < 0.001), marijuana use (P < 0.001) and other recreational drug use (P < 0.01). Males reported more sex partners than females in the previous six months (P < 0.001), as well as more casual/anonymous partners (P < 0.001) and one-night stands (P < 0.001). Multivariate analyses revealed substance use and male gender predicted higher risk behaviours, including <100% condom use with known/suspected HIV-positive partners, having casual/anonymous partners and having more than two partners. For this population, male volunteers may need increased risk-reduction counselling during Phase I/II trials and additional recruitment methods may be necessary to identify high-risk female volunteers for Phase III efficacy trials.

Adenovirus 5 vector genetically re-targeted by an Affibody molecule with specificity for tumor antigen HER2/neu.

In order to use adenovirus (Ad) type 5 (Ad5) for cancer gene therapy, Ad needs to be de-targeted from its native receptors and re-targeted to a tumor antigen. A limiting factor for this has been to find a ligand that (i) binds a relevant target, (ii) is able to fold correctly in the reducing environment of the cytoplasm and (iii) when incorporated at an optimal position on the virion results in a virus with a low physical particle to plaque-forming units ratio to diminish the viral load to be administered to a future patient. Here, we present a solution to these problems by producing a genetically re-targeted Ad with a tandem repeat of the HER2/neu reactive Affibody molecule (ZH) in the HI-loop of a Coxsackie B virus and Ad receptor (CAR) binding ablated fiber genetically modified to contain sequences for flexible linkers between the ZH and the knob sequences. ZH is an Affibody molecule specific for the extracellular domain of human epidermal growth factor receptor 2 (HER2/neu) that is overexpressed in inter alia breast and ovarian carcinomas. The virus presented here exhibits near wild-type growth characteristics, infects cells via HER2/neu instead of CAR and represents an important step toward the development of genetically re-targeted adenoviruses with clinical relevance.

Tumor cell targeted gene delivery by adenovirus 5 vectors carrying knobless fibers with antibody-binding domains.

Most human carcinoma cell lines lack the high-affinity receptors for adenovirus serotype 5 (Ad5) at their surface and are nonpermissive to Ad5. We therefore tested the efficiency of retargeting Ad5 to alternative cellular receptors via immunoglobulin (Ig)-binding domains inserted at the extremity of short-shafted, knobless fibers. The two recombinant Ad5's constructed, Ad5/R7-Z(wt)-Z(wt) and Ad5/R7-C2-C2, carried tandem Ig-binding domains from Staphylococcal protein A (abbreviated Z(wt)) and from Streptococcal protein G (C2), respectively. Both viruses bound their specific Ig isotypes with the expected affinity. They transduced human carcinoma cells independently of the CAR pathway, via cell surface receptors targeted by specific monoclonal antibodies, that is, EGF-R on A549, HT29 and SW1116, HER-2/neu on SK-OV-3 and SK-BR-3, CA242 (epitope recognized by the monoclonal antibody C242) antigen on HT29 and SW1116, and PSMA (prostate-specific membrane antigen) expressed on HEK-293 cells, respectively. However, Colo201 and Colo205 cells were neither transduced by targeting CA242 or EGF-R nor were LNCaP cells transduced by targeting PSMA. Our results suggested that one given surface receptor could mediate transduction of certain cells but not others, indicating that factors and steps other than cell surface expression and virus-receptor interaction are additional determinants of Ad5-mediated transduction of tumor cells. Using penton base RGD mutants, we found that one of these limiting steps was virus endocytosis.

The protein content in crystals and packing coefficients in different space groups.

A precise way of estimating the packing coefficient, i.e. the ratio between the protein and unit-cell volume, or solvent content in protein crystals is given. At present, the solvent content is not given for most proteins in the Protein Data Bank and in many cases where it is given the values are dubious. The mean density of proteins in the crystalline form is around 1.22 g cm(-3), not 1.35 g cm(-3) as usually stated. This is equivalent to 19.5 A(3) per non-H atom. A statistical investigation of the average protein content and packing coefficient in different space groups is presented. The packing coefficients are generally higher in the most frequently occurring space groups than in the uncommon space groups. There is also a remarkable difference in frequency distribution for enantiomorphous pairs of space groups.

Phasing proteins at low resolution.

A method for obtaining phases of low-order reflections is presented. It is based on four observations: (1) the electron density inside proteins is smooth and uniform at low resolution. (2) Since all proteins have almost the same density, the total volume of the protein is known if the molecular weight is known. (3) The overall shape of many proteins is fairly spherical. (4) The total scattering from a sphere of uniform density is in phase with a point scatterer at its centre of gravity, up to a well defined cross-over. After the first cross-over the total protein molecule scatters out of phase with its centre. If the centre of the protein can be found, the phases of typically the ten lowest resolution reflections can be very accurately determined. The method works, provided low-order reflections can be measured accurately and the centre of gravity can be well positioned from these data. The correctly phased low-resolution reflections may be used as a starting set for phase extension. By combining the measured amplitudes with these phases we believe that the size and low-resolution shape of an unknown protein, i.e. the envelope of the molecule, can be obtained.

Conceptual problems in primary health care research.

Concepts are basic elements in all scientific work. New fields of investigation, like primary health care, have to adopt new concepts. These may have a, more or less, physical appearance or may be phenomenological by nature. Concepts may be defined by description or, more appropriate, by explication of necessary and sufficient attributes or characteristics, or by criteria to be fulfilled. One model for concept analysis contains steps as: determine the aims of analysis, identify all uses of the concept, determine the defining attributes, construct a model case, and define empirical referents. Another model, developed for analysis of phenomenological concepts, is based on accumulation of hundreds of explications of the meaning of the concept. Those are elaborated by Grouping, Reduction, Elimination, Hypothetical identification, Application, Final identification. Methods for concept analysis need to be further emphasized in doctoral programs. Even experienced scientists may need more training in such methods, when entering new fields. Students need accurate guidance through passages of conceptual work. Concept development should be accepted and encouraged as research projects in its own capacity. It makes quite a contribution to scientific knowledge, to elaborate an important concept in all aspects. In addressing new empirical problems, much work should be spared, if the researcher could use concepts already studied and described in the literature.