Role of the envelope glycoproteins in the infection cycle of tomato spotted wilt virus

Tomato spotted wilt virus (TSWV) forms the type member of the genus Tospovirus , which today harbors more than twelve different species. TSWV is able to infect an enormous variety of different plants, to which it often causes devastating effects, resulting in severe economical losses. Among the plant viruses, TSWV and the other tospoviruses form a distinct group. Taxonomically, they surprisingly do not belong to a plant virus family, but to a virus family which further consists of animal-infecting viruses, the Bunyaviridae . Consequently, they harbor features that are more common to animal-infecting viruses than to plant viruses. The most eye-catching animal-infecting virus-like feature of the tospoviruses is their envelope, in which two viral surface glycoproteins are embedded, denoted G1 and G2. These surface glycoproteins are designed for interaction with receptors, an important step in the infection of animals, but useless in the infection of plants. The plant-infecting tospoviruses are transmitted by thrips, in which they also replicate, and for the entry and circulation of the virus through this insect the glycoproteins are essential. This ensures their continues presence despite their lack of function during the infection of plants. The structure and function of the TSWV glycoproteins during infection in plants and insects form the subject of this thesis, of which the contents will be summarized in the next paragraphs, and also visually represented in Fig. 1.First develop the tools...Although the TSWV glycoproteins do not have a crucial function in the plant, they do play an essential role in the formation of virus particles. At the onset of these studies, however, no clear view of this morphogenesis process was available, so it was the obvious first goal to unravel this process for TSWV in plants. In the past, virus associated structures observed during the infection of whole plants were reported, but thirty five years of observations had not resulted in a clear model of the particle morphogenesis. Useful antibodies against the separate TSWV proteins had not been available, and most importantly, a system in which a synchronous infection could be investigated lacked, so that interpretation of the chrology of the events in the morphogenesis had been difficult. Chapter 2 of this thesis describes the development of a protoplast infection system for TSWV, which enabled the study of a synchronous TSWV infection in plant cells. Using newly produced antibodies against the viral glycoproteins together with antibodies against the nucleoprotein, it could be shown that a full, synchronous, TSWV infection is achieved, by the PEG-mediated inoculation of freshly isolated Nicotiana rustica protoplasts with freshly (and quickly) isolated TSWV particles. Similar inoculation of Vigna unguiculata protoplasts did not result in a full infection, since the production of enveloped particles was hampered due to low expression of the viral glycoproteins....to reveal the essence of TSWV particle making...Using the system, based on N. rustica protoplasts, the different virus associated structures could be assigned a chronological position in the morphogenesis process. This view of the chronology of the process revealed that the so-called paired parrallel membranes and doubly enveloped particles form essential intermediates in the process, which precede the accumulation of singly enveloped particles in the lumen of ER membranes. Using specific antibodies against the TSWV structural proteins, as well as antibodies against plant cell organels, the model could be completed ( Chapter 3 ). TSWV structural components, nucleocapsids as well as glycoproteins, accumulate at Golgi membranes, which are consequently modified to form the paired parrallel membranes. Doubly enveloped particles are formed by the so-called "wrapping" of these viral glycoprotein containing membranes around nucleocapsid cores, a process unique among plant viruses. The subsequent step is the fusion of these doubly enveloped particles with each other and specifically with ER membranes. This results in the formation of singly enveloped particles that accumulate within the ER....and then look what causes all this...After the complete model of the morphogenesis became available, the next step was to investigate what (molecular) features of especially the glycoproteins regulate the process. An important observation in the morphogenesis process is the apparent accumulation of viral glycoproteins in the Golgi system. This may, analogous to other enveloped viruses, be caused by the specific targeting of the glycoproteins to this organel due to a retention signal. The trafficking and retention behavior of TSWV glycoproteins was investigated in mammalian cells, the results of which are described in Chapter 4 . TSWV G1 and G2 accumulate in the Golgi system when expression together, which indeed implicates that at least one of the proteins must harbor a Golgi retention signal. Separate expression of G1 and G2 revealed that the retention signal is present in G2. G1 on its own is transport incompetent, but this can be rescued by the co-expression with G2, which suggests that G1 is dependent on, and interacts with G2 during transport and retention. These molecular features, identified in mammalian cells, are most probably also functional in plant cells, causing the observed accumulation of glycoproteins in the plant Golgi system during infection. The TSWV glycoproteins thereby show their crucial role in directing the particle morphogenesis process....and how these particles interact with thrips.Once the particles are formed and accumulated inside ER membranes, they await the uptake by the thrips vector to be transferred to another plant. Earlier research has shown that between this uptake and the release of virus there is replication and circulation of TSWV in the thrips. However, nothing was known about the molecular interactions between TSWV proteins and proteins of the thrips during this process. In Chapter 5 an overlay blot technique was used to investigate the possible binding of TSWV structural proteins with thrips proteins, that could be potential receptors involved in entry or circulation of TSWV. A 94 kDa thrips protein was identified, displaying specific binding to TSWV G2 protein. This 94 kDa protein was found in known vectors of TSWV, and also in a non-vector thrips species, albeit it not in the larval stages of the latter. Although a receptor is anticipated in the gut of the vectoring insects, the 94 kDa protein is not found there. It is however present in all other parts of the thrips body, suggesting that it may have a role during the replication and circulation of the virus.Fig. 1Diagram of TSWV infection in a plant cell, indicating the scope of the experimental chapters of this thesis</CENTER

Finger somatotopy is preserved after tetraplegia but deteriorates over time

Previous studies showed reorganised and/or altered activity in the primary sensorimotor cortex after a spinal cord injury (SCI), suggested to reflect abnormal processing. However, little is known about whether somatotopically specific representations can be activated despite reduced or absent afferent hand inputs. In this observational study, we used functional MRI and a (attempted) finger movement task in tetraplegic patients to characterise the somatotopic hand layout in primary somatosensory cortex. We further used structural MRI to assess spared spinal tissue bridges. We found that somatotopic hand representations can be activated through attempted finger movements in the absence of sensory and motor hand functioning, and no spared spinal tissue bridges. Such preserved hand somatotopy could be exploited by rehabilitation approaches that aim to establish new hand-brain functional connections after SCI (e.g. neuroprosthetics). However, over years since SCI the hand representation somatotopy deteriorated, suggesting that somatotopic hand representations are more easily targeted within the first years after SCI

Interaction of the innate immune system with positive-strand RNA virus replication organelles

Molecular basis of virus replication, viral pathogenesis and antiviral strategie

Risk of criminal victimisation in outpatients with common mental health disorders

Crime victimisation is a serious problem in psychiatric patients. However, research has focused on patients with severe mental illness and few studies exist that address victimisation in other outpatient groups, such as patients with depression. Due to large differences in methodology of the studies that address crime victimisation, a comparison of prevalence between psychiatric diagnostic groups is hard to make. Objectives of this study were to determine and compare one-year prevalence of violent and non-violent criminal victimisation among outpatients from different diagnostic psychiatric groups and to examine prevalence differences with the general population.Criminal victimisation prevalence was measured in 300 outpatients living in Amsterdam, The Netherlands. Face-to-face interviews were conducted with outpatients with depressive disorder (n = 102), substance use disorder (SUD, n = 106) and severe mental illness (SMI, n = 92) using a National Crime Victimisation Survey, and compared with a matched general population sample (n = 10865).Of all outpatients, 61% reported experiencing some kind of victimisation over the past year; 33% reported violent victimisation (3.5 times more than the general population) and 36% reported property crimes (1.2 times more than the general population). Outpatients with depression (67%) and SUD (76%) were victimised more often than SMI outpatients (39%). Younger age and hostile behaviour were associated with violent victimisation, while being male and living alone were associated with non-violent victimisation. Moreover, SUD was associated with both violent and non-violent victimisation.Outpatients with depression, SUD, and SMI are at increased risk of victimisation compared to the general population. Furthermore, our results indicate that victimisation of violent and non-violent crimes is more common in outpatients with depression and SUD than in outpatients with SMI living independently in the community

Cost-effectiveness of adherence therapy versus health education for people with schizophrenia: randomised controlled trial in four European countries.

BACKGROUND: Non-adherence to anti-psychotics is common, expensive and affects recovery. We therefore examine the cost-effectiveness of adherence therapy for people with schizophrenia by multi-centre randomised trial in Amsterdam, London, Leipzig and Verona. METHODS: Participants received 8 sessions of adherence therapy or health education. We measured lost productivity and use of health/social care, criminal justice system and informal care at baseline and one year to estimate and compare mean total costs from health/social care and societal perspectives. Outcomes were the Short Form 36 (SF-36) mental component score (MCS) and quality-adjusted life years (QALYs) gained (SF-36 and EuroQoL 5 dimension (EQ5D)). Cost-effectiveness was examined for all cost and outcome combinations using cost-effectiveness acceptability curves (CEACs). RESULTS: 409 participants were recruited. There were no cost or outcome differences between adherence therapy and health education. The probability of adherence therapy being cost-effective compared to health education was between 0.3 and 0.6 for the six cost-outcome combinations at the willingness to pay thresholds we examined. CONCLUSIONS: Adherence therapy appears equivalent to health education. It is unclear whether it would have performed differently against a treatment as usual control, whether such an intervention can impact on quality of life in the short-term, or whether it is likely to be cost-effective in some sites but not others. TRIAL REGISTRATION: Trial registration: Current Controlled Trials ISRCTN01816159.Quality of Life and Management of Living Resources Programme of the European Union (grant number QLG4-CT-2001-01734

Формування світогляду О. Кониського

Crime victimisation is a serious problem in psychiatric patients. However, research has focused on patients with severe mental illness and few studies exist that address victimisation in other outpatient groups, such as patients with depression. Due to large differences in methodology of the studies that address crime victimisation, a comparison of prevalence between psychiatric diagnostic groups is hard to make. Objectives of this study were to determine and compare one-year prevalence of violent and non-violent criminal victimisation among outpatients from different diagnostic psychiatric groups and to examine prevalence differences with the general population.Criminal victimisation prevalence was measured in 300 outpatients living in Amsterdam, The Netherlands. Face-to-face interviews were conducted with outpatients with depressive disorder (n = 102), substance use disorder (SUD, n = 106) and severe mental illness (SMI, n = 92) using a National Crime Victimisation Survey, and compared with a matched general population sample (n = 10865).Of all outpatients, 61% reported experiencing some kind of victimisation over the past year; 33% reported violent victimisation (3.5 times more than the general population) and 36% reported property crimes (1.2 times more than the general population). Outpatients with depression (67%) and SUD (76%) were victimised more often than SMI outpatients (39%). Younger age and hostile behaviour were associated with violent victimisation, while being male and living alone were associated with non-violent victimisation. Moreover, SUD was associated with both violent and non-violent victimisation.Outpatients with depression, SUD, and SMI are at increased risk of victimisation compared to the general population. Furthermore, our results indicate that victimisation of violent and non-violent crimes is more common in outpatients with depression and SUD than in outpatients with SMI living independently in the community

Role of Interleukin 17 in arthritis chronicity through survival of synoviocytes via regulation of synoviolin expression

Background: The use of TNF inhibitors has been a major progress in the treatment of chronic inflammation. However, not all patients respond. In addition, response will be often lost when treatment is stopped. These clinical aspects indicate that other cytokines might be involved and we focus here on the role of IL-17. In addition, the chronic nature of joint inflammation may contribute to reduced response and enhanced chronicity. Therefore we studied the capacity of IL-17 to regulate synoviolin, an E3 ubiquitin ligase implicated in synovial hyperplasia in human rheumatoid arthritis (RA) FLS and in chronic reactivated streptococcal cell wall (SCW)-induced arthritis.&lt;p&gt;&lt;/p&gt; Methodology/Principal Findings: Chronic reactivated SCW-induced arthritis was examined in IL-17R deficient and wild-type mice. Synoviolin expression was analysed by real-time RT-PCR, Western Blot or immunostaining in RA FLS and tissue, and p53 assessed by Western Blot. Apoptosis was detected by annexin V/propidium iodide staining, SS DNA apoptosis ELISA kit or TUNEL staining and proliferation by PCNA staining. IL-17 receptor A (IL-17RA), IL-17 receptor C (IL-17-RC) or synoviolin inhibition were achieved by small interfering RNA (siRNA) or neutralizing antibodies. IL-17 induced sustained synoviolin expression in RA FLS. Sodium nitroprusside (SNP)-induced RA FLS apoptosis was associated with reduced synoviolin expression and was rescued by IL-17 treatment with a corresponding increase in synoviolin expression. IL-17RC or IL-17RA RNA interference increased SNP-induced apoptosis, and decreased IL-17-induced synoviolin. IL-17 rescued RA FLS from apoptosis induced by synoviolin knockdown. IL-17 and TNF had additive effects on synoviolin expression and protection against apoptosis induced by synoviolin knowndown. In IL-17R deficient mice, a decrease in arthritis severity was characterized by increased synovial apoptosis, reduced proliferation and a marked reduction in synoviolin expression. A distinct absence of synoviolin expressing germinal centres in IL-17R deficient mice contrasted with synoviolin positive B cells and Th17 cells in synovial germinal centre-like structures.&lt;p&gt;&lt;/p&gt; Conclusion/Significance: IL-17 induction of synoviolin may contribute at least in part to RA chronicity by prolonging the survival of RA FLS and immune cells in germinal centre reactions. These results extend the role of IL-17 to synovial hyperplasia.&lt;p&gt;&lt;/p&gt