The limit to wheat water-use efficiency in eastern Australia. II. Influence of rainfall patterns

We investigated the influence of rainfall patterns on the water-use efficiency of wheat in a transect between Horsham (36°S) and Emerald (23°S) in eastern Australia. Water-use efficiency was defined in terms of biomass and transpiration, WUEB/T, and grain yield and evapotranspiration, WUEY/ET. Our working hypothesis is that latitudinal trends in WUEY/ET of water-limited crops are the complex result of southward increasing WUEB/T and soil evaporation, and season-dependent trends in harvest index. Our approach included: (a) analysis of long-term records to establish latitudinal gradients of amount, seasonality, and size-structure of rainfall; and (b) modelling wheat development, growth, yield, water budget components, and derived variables including WUEB/T and WUEY/ET. Annual median rainfall declined from around 600 mm in northern locations to 380 mm in the south. Median seasonal rain (from sowing to harvest) doubled between Emerald and Horsham, whereas median off-season rainfall (harvest to sowing) ranged from 460 mm at Emerald to 156 mm at Horsham. The contribution of small events (≤ 5 mm) to seasonal rainfall was negligible at Emerald (median 15 mm) and substantial at Horsham (105 mm). Power law coefficients (τ), i.e. the slopes of the regression between size and number of events in a log-log scale, captured the latitudinal gradient characterised by an increasing dominance of small events from north to south during the growing season. Median modelled WUEB/T increased from 46 kg/ha.mm at Emerald to 73 kg/ha.mm at Horsham, in response to decreasing atmospheric demand. Median modelled soil evaporation during the growing season increased from 70 mm at Emerald to 172 mm at Horsham. This was explained by the size-structure of rainfall characterised with parameter τ, rather than by the total amount of rainfall. Median modelled harvest index ranged from 0.25 to 0.34 across locations, and had a season-dependent latitudinal pattern, i.e. it was greater in northern locations in dry seasons in association with wetter soil profiles at sowing. There was a season-dependent latitudinal pattern in modelled WUEY/ET. In drier seasons, high soil evaporation driven by a very strong dominance of small events, and lower harvest index override the putative advantage of low atmospheric demand and associated higher WUEB/T in southern locations, hence the significant southwards decrease in WUEY/ET. In wetter seasons, when large events contribute a significant proportion of seasonal rain, higher WUEB/T in southern locations may translate into high WUEY/ET. Linear boundary functions (French-Schultz type models) accounting for latitudinal gradients in its parameters, slope, and x-intercept, were fitted to scatter-plots of modelled yield v. evapotranspiration. The x-intercept of the model is re-interpreted in terms of rainfall size structure, and the slope or efficiency multiplier is described in terms of the radiation, temperature, and air humidity properties of the environment. Implications for crop management and breeding are discussed.

Sulfur-associated polioencephalomalacia in cattle grazing plants in the Family Brassicaceae

Polioencephalomalacia was diagnosed histologically in cattle from two herds on the Darling Downs, Queensland, during July-August 2007. In the first incident, 8 of 20 18-month-old Aberdeen Angus steers died while grazing pastures comprising 60% Sisymbrium irio (London rocket) and 40% Capsella bursapastoris (shepherd's purse). In the second incident, 2 of 150 mixed-breed adult cattle died, and another was successfully treated with thiamine, while grazing a pasture comprising almost 100% Raphanus raphanistrum (wild radish). Affected cattle were either found dead or comatose or were seen apparently blind and head-pressing in some cases. For both incidents, plant and water assays were used to calculate the total dietary sulfur content in dry matter as 0.62% and 1.01% respectively, both exceeding the recommended 0.5% for cattle eating more than 40% forage. Blood and tissue assays for lead were negative in both cases. No access to thiaminase, concentrated sodium ion or extrinsic hydrogen sulfide sources were identified in either incident. Below-median late summer and autumn rainfall followed by above-median unseasonal winter rainfall promoted weed growth at the expense of wholesome pasture species before these incidents.

Defining and predicting the 'break of the season' for north-east Queensland grazing areas

For pasture growth in the semi-arid tropics of north-east Australia, where up to 80% of annual rainfall occurs between December and March, the timing and distribution of rainfall events is often more important than the total amount. In particular, the timing of the 'green break of the season' (GBOS) at the end of the dry season, when new pasture growth becomes available as forage and a live-weight gain is measured in cattle, affects several important management decisions that prevent overgrazing and pasture degradation. Currently, beef producers in the region use a GBOS rule based on rainfall (e. g. 40mm of rain over three days by 1 December) to define the event and make their management decisions. A survey of 16 beef producers in north-east Queensland shows three quarters of respondents use a rainfall amount that occurs in only half or less than half of all years at their location. In addition, only half the producers expect the GBOS to occur within two weeks of the median date calculated by the CSIRO plant growth days model GRIM. This result suggests that in the producer rules, either the rainfall quantity or the period of time over which the rain is expected, is unrealistic. Despite only 37% of beef producers indicating that they use a southern oscillation index (SOI) forecast in their decisions, cross validated LEPS (linear error in probability space) analyses showed both the average 3 month July-September SOI and the 2 month August-September SOI have significant forecast skill in predicting the probability of both the amount of wet season rainfall and the timing of the GBOS. The communication and implementation of a rigorous and realistic definition of the GBOS, and the likely impacts of anthropogenic climate change on the region are discussed in the context of the sustainable management of northern Australian rangelands.

Cassowary dispersal of the invasive pond apple in a tropical rainforest: the contribution of subordinate dispersal modes in invasion.

Dispersal is a significant determinant of the pattern and process of invasions; however, weed dispersal distances are rarely described and descriptions of dispersal kernels are completely lacking for vertebrate-dispersed weeds. Here, we describe dispersal kernels generated by a native disperser, the endangered southern cassowary (Casuarius casuarius, L.) for an invasive, tropical rainforest plant, pond apple (Annona glabra, L.). Pond apple is primarily water-dispersed and is managed as such. We consider whether cassowary dispersal, as a numerically subordinate dispersal mode, provides an additional dispersal service that may modify the invasion process. In infested areas, pond apple seed was common in cassowary dung. Gut passage had no effect on the probability of single seed germination but deposition in clumps or as whole fruits reduced the probability of germination below that of single seeds. Gut passage times ranged from 65 to 1675 min. Combined with cassowary movement data, this resulted in estimated dispersal distances of 12.5-5212 m, with a median distance of 387 m (quartile range 112-787 m). Native frugivores can be effective dispersers of weeds in rainforest and even terrestrial dispersers can provide long-distance dispersal. Importantly, though pond apple might be expected to be almost entirely dispersed downstream and along the margins of aquatic and marine habitats, cassowaries provide dispersal upstream and between drainages, leading to novel dispersal outcomes. Even through the provision of small quantities of novel dispersal outcomes, subordinate dispersal modes can play a significant role in determining invasion pattern and influence the ultimate success of control programs by providing dispersal to locations unattainable via the primary mode.

Lähi-itä - mielikuvat, media ja konflikti : Tutkimuskohteena opettajaopiskelijat

This study examines Finnish student teachers' images of The Middle East conflict and its parties and medias' influence on these images. The research problems are 1) how student teachers understand the Middle East as a geographical and a cultural area 2) what kind of conceptions they have of the Middle East conflict and its parties; Israelis, Palestinians and Arabs and societies and religions closely connected to them and 3) how the media has influenced on these conceptions. Theoretical background of this study concerns with different mental images, their construction and meaning, stereotypes, intercultural competence, media and its influence as well as media literacy. The methods used in this study were survey and theme interview. Also headlines of the news were analysed. The survey was directed at student teachers of the University of Helsinki (n = 75). Six of them were interviewed. The survey was conducted in February and interviews were made in the turn of May and June 2003. The headlines analysed dealt with the Middle East conflict in the newspaper "Helsingin Sanomat" in January, April and July 2003. Main results: Images of the Middle East area differed largely from respondent to another. Student teachers didn't know much about the Middle East conflict. Stereotypes occurred in the conceptions connected to the parties of the conflict. Stereotypes appeared especially in the negative conceptions of islam and muslims and one-sided conceptions of Arabs. The influence of the conflict was noticeable in conceptions related to Israelis, Israel and Palestinians. Palestinians were sympathized. Attitudes towards Israelis, Palestinians and Arabs were negative to some extend and their societies were not valued very equal, open or modern. On the basis of the research, student teachers' intercultural competence was not good enough. Media had influence on the conceptions. Also skills in critical use of media varied largely. Some trusted strongly in the objectivity of a picture and the news. It can be argued, on the grounds of the results, that teacher education should concentrate more on themes of intercultural competence and media literacy. These two are interrelated and are both needed in understanding and conceptualising the world.

Retinal photography screening programs to prevent vision loss from diabetic retinopathy in rural and urban Australia: A review

Purpose This review assessed the effectiveness of diabetic retinopathy (DR) screening programs, using retinal photography in Australian urban and rural settings, and considered implications for public health strategy and policy. Methods An electronic search of MEDLINE, PubMed, and Embase for studies published between 1 January 1996 and the 30 June 2013 was undertaken. Key search terms were “diabetic retinopathy,” “screening,” “retinal photography” and “Australia.” Results Twelve peer-reviewed publications were identified. The 14 DR screening programs identified from the 12 publications were successfully undertaken in urban, rural and remote communities across Australia. Locations included a pathology collection center, and Indigenous primary health care and Aboriginal community controlled organizations. Each intervention using retinal photography was highly effective at increasing the number of people who underwent screening for DR. The review identified that prior to commencement of the screening programs a median of 48% (range 16–85%) of those screened had not undergone a retinal examination within the recommended time frame (every year for Indigenous people and every 2 years for non-Indigenous people in Australia). A median of 16% (range 0–45%) of study participants had evidence of DR. Conclusions This review has shown there have been many pilot and demonstration projects in rural and urban Australia that confirm the effectiveness of retinal photography-based screening for DR

Modelling the nitrogen-driven trade-off between nitrogen utilisation efficiency and water use efficiency of wheat in eastern Australia.

The nitrogen-driven trade-off between nitrogen utilisation efficiency (yield per unit nitrogen uptake) and water use efficiency (yield per unit evapotranspiration) is widespread and results from well established, multiple effects of nitrogen availability on the water, carbon and nitrogen economy of crops. Here we used a crop model (APSIM) to simulate the yield, evapotranspiration, soil evaporation and nitrogen uptake of wheat, and analysed yield responses to water, nitrogen and climate using a framework analogous to the rate-duration model of determinate growth. The relationship between modelled grain yield (Y) and evapotranspiration (ET) was fitted to a linear-plateau function to derive three parameters: maximum yield (Ymax), the ET break-point when yield reaches its maximum (ET#), and the rate of yield response in the linear phase ([Delta]Y/[Delta]ET). Against this framework, we tested the hypothesis that nitrogen deficit reduces maximum yield by reducing both the rate ([Delta]Y/[Delta]ET) and the range of yield response to evapotranspiration, i.e. ET# - Es, where Es is modelled median soil evaporation. Modelled data reproduced the nitrogen-driven trade-off between nitrogen utilisation efficiency and water use efficiency in a transect from Horsham (36°S) to Emerald (23°S) in eastern Australia. Increasing nitrogen supply from 50 to 250 kg N ha-1 reduced yield per unit nitrogen uptake from 29 to 12 kg grain kg-1 N and increased yield per unit evapotranspiration from 6 to 15 kg grain ha-1 mm-1 at Emerald. The same increment in nitrogen supply reduced yield per unit nitrogen uptake from 30 to 25 kg grain kg-1 N and increased yield per unit evapotranspiration from 6 to 25 kg grain ha-1 mm-1 at Horsham. Maximum yield ranged from 0.9 to 6.4 t ha-1. Consistent with our working hypothesis, reductions in maximum yield with nitrogen deficit were associated with both reduction in the rate of yield response to ET and compression of the range of yield response to ET. Against the notion of managing crops to maximise water use efficiency in low rainfall environments, we emphasise the trade-off between water use efficiency and nitrogen utilisation efficiency, particularly under conditions of high nitrogen-to-grain price ratio. The rate-range framework to characterise the relationship between yield and evapotranspiration is useful to capture this trade-off as the parameters were responsive to both nitrogen supply and climatic factors.

Saako terveysuskoja synninpäästön? : Naispappien käsityksiä laihdutus- ja terveyskulttuurista nykyisessä mediayhteiskunnassa

Background and aims. Fatness and dieting have been the object of interest between many fields for a long time. Home economics as a discipline enables a comprehensive inspection of fatness and dieting reviewing different disciplines. In addition to the aspect where the pursuit of dieting and health is seen from the perspective of medical and health science it is also been reviewd as a social and cultural phenomena. This study contemplates the influence of history, religion, medicalization and media on dieting and health culture. The objective is to find out if the modern dieting and health culture has gathered influences from centuries ago and absorbed religious features. The stress deriving from appereance has been discussed in the public and there are many solutions conserning weight issues. The purpose of this study is to find out what personal experiences and thoughts female pastors have conserning these questions. The media – which is one of the most influential systems nowadays – has undeniably a great effect on the consumer. The goal is furthermore to estimate the effect of the media on the changing of dieting and health culture. The three main research questions are: 1. What kind of conseptions do female pastors have of dieting and health culture and of its religious features? 2. What kind of personal experiences and conseptions do female pastors have of dieting and strivines of health? 3. How do female pastors regard the image the media has supplied of dieting and health culture? Material and methods. The qualitative data was gathered in year 2009 using the halfstructured theme interview -method. The data consists of interviews conducted with specialists of spiritual matters, i.e. ten female pastors who are between 35 and 60 years old and live in the metropolitan area. The analytical procedure used is called a theory based context analysis. Results and conclusions. Results of this study show that the idealization of slimness and healthiness is a matter discussed in the public on a daily basis. The problem faced was that the media provided contradictory information regarding fatness and dieting and the standard of slimness in commercials focused on females. The pursuit of dieting and healthiness was believed to include also religious elements. In the Middle Ages and the era after that the fatness, overeating and the pleasure one gets from eating was still seen as a condemnable matter in our culture. One could say this was like a sin. The respondents believed that healthiness, healthy living, optimal eating and good looks were a matter more or less equal than a religion. This was a derivative from the fact that treasuring health has become a life stearing value for many people. In the priest’s profession dieting and the pursuit of health was seen in the light of problems arising from weight issues. In ones profession for example the unhealthy eating in festive situations was seen as a matter that leads to unnecessary weight. Another aspect was the job circumstances that limited the degree of movement. The belief was that the female pastors would in a decreasing fashion confront stress deriving from appearence in their job. Keywords: dieting, fatness, healthiness, slimness, female pastors, religion, medicalization, media

A systematic review of community pharmacist therapeutic knowledge of dietary supplements

Background: Internationally, the use of dietary supplements has been growing rapidly. Patient support for pharmacist sales of nutritional and dietary supplements is also strong. The increase in demand for nutritional and dietary supplements and subsequent advice about these products, however, makes it necessary that pharmacists maintain a contemporary knowledge of the area. Aim of review: This systematic review was conducted to examine the current evidence regarding the level of the nutritional and dietary supplement knowledge of community pharmacists and their understanding of their therapeutic effects. Method: Electronic databases including Medline, Scopus, Embase, CINAHL, Scifinder and the Cochrane Controlled Trials Register were searched. Studies assessing nutritional knowledge of pharmacists in community pharmacies were eligible for inclusion. All languages and study designs were considered. Study results were analysed and pharmacist knowledge scores were given out of 100 . Results: From 5594 studies identified, nine met the inclusion criteria. Each study tested pharmacist knowledge with predetermined questions calculating results as the number of questions answered correctly. These knowledge scores were converted to a percentage score for the purpose of this paper. The median knowledge score across all papers was 64%. A lack of studies assessing community pharmacist's knowledge of commonly sold vitamins and minerals was observed. Conclusions Global community pharmacist knowledge of dietary supplements appears to be poor. Community pharmacists have an professional responsibility to provide accurate health information about dietary supplements as they do for any other therapies they provide to patients. Further research including that which assesses pharmacist's therapeutic knowledge of commonly sold vitamins and minerals is suggested.

To reactivate or not to reactivate : Control of KSHV lytic replication is essential for apoptosis in response to p53 restoration

Kaposi's sarcoma herpesvirus (KSHV) is an oncogenic human virus and the causative agent of three human malignancies: Kaposi's sarcoma (KS), Multicentric Castleman's Disease (MCD), and primary effusion lymphoma (PEL). In tumors, KSHV establishes latent infection during which it produces no infectious particles. Latently infected cells can enter the lytic replication cycle, and upon provision of appropriate cellular signals, produce progeny virus. PEL, commonly described in patients with AIDS, represents a diffuse large-cell non-Hodgkin's lymphoma, with median survival time less than six months after diagnosis. As tumor suppressor gene TP53 mutations occur rarely in PEL, the aim of this thesis was to investigate whether non-genotoxic activation of the p53 pathway can eradicate malignant PEL cells. This thesis demonstrates that Nutlin-3, a small-molecule inhibitor of the p53-MDM2 interaction, efficiently restored p53 function in PEL cells, leading to cell cycle arrest and massive apoptosis. Furthermore, we found that KSHV infection activated DNA damage signaling, rendering the cells more sensitive to p53-dependent cell death. We also showed in vivo the therapeutic potential of p53 restoration that led to regression of subcutaneous and intraperitoneal PEL tumor xenografts without adversely affecting normal cells. Importantly, we demonstrated that in a small subset of intraperitoneal PEL tumors, spontaneous induction of viral reactivation dramatically impaired Nutlin-3-induced p53-mediated apoptosis. Accordingly, we found that elevated KSHV lytic transcripts correlated with PEL tumor burden in animals and that inhibition of viral reactivation in vitro restored cytotoxic activity of a small-molecule inhibitor of the p53-MDM2 interaction. Latency provides a unique opportunity for KSHV to escape host immune surveillance and to establish persistent infections. However, to maintain viral reservoirs and spread to other hosts, KSHV must be reactivated from latency and enter into the lytic growth phase. We showed that phosphorylation of nucleolar phosphoprotein nucleophosmin (NPM) by viral cyclin-CDK6 is critical for establishment and maintenance of the KSHV latency. In short, this study provides evidence that the switch between latent phase and lytic replication is a critical step that determines the outcome of viral infection and the pathogenesis of KSHV-induced malignancies. Our data may thus contribute to development of novel targeted therapies for intervention and treatment of KSHV-associated cancers.

Technical efficiency of blood component preparation in blood centres of 10 European countries

Various reasons, such as ethical issues in maintaining blood resources, growing costs, and strict requirements for safe blood, have increased the pressure for efficient use of resources in blood banking. The competence of blood establishments can be characterized by their ability to predict the volume of blood collection to be able to provide cellular blood components in a timely manner as dictated by hospital demand. The stochastically varying clinical need for platelets (PLTs) sets a specific challenge for balancing supply with requests. Labour has been proven a primary cost-driver and should be managed efficiently. International comparisons of blood banking could recognize inefficiencies and allow reallocation of resources. Seventeen blood centres from 10 countries in continental Europe, Great Britain, and Scandinavia participated in this study. The centres were national institutes (5), parts of the local Red Cross organisation (5), or integrated into university hospitals (7). This study focused on the departments of blood component preparation of the centres. The data were obtained retrospectively by computerized questionnaires completed via Internet for the years 2000-2002. The data were used in four original articles (numbered I through IV) that form the basis of this thesis. Non-parametric data envelopment analysis (DEA, II-IV) was applied to evaluate and compare the relative efficiency of blood component preparation. Several models were created using different input and output combinations. The focus of comparisons was on the technical efficiency (II-III) and the labour efficiency (I, IV). An empirical cost model was tested to evaluate the cost efficiency (IV). Purchasing power parities (PPP, IV) were used to adjust the costs of the working hours and to make the costs comparable among countries. The total annual number of whole blood (WB) collections varied from 8,880 to 290,352 in the centres (I). Significant variation was also observed in the annual volume of produced red blood cells (RBCs) and PLTs. The annual number of PLTs produced by any method varied from 2,788 to 104,622 units. In 2002, 73% of all PLTs were produced by the buffy coat (BC) method, 23% by aphaeresis and 4% by the platelet-rich plasma (PRP) method. The annual discard rate of PLTs varied from 3.9% to 31%. The mean discard rate (13%) remained in the same range throughout the study period and demonstrated similar levels and variation in 2003-2004 according to a specific follow-up question (14%, range 3.8%-24%). The annual PLT discard rates were, to some extent, associated with production volumes. The mean RBC discard rate was 4.5% (range 0.2%-7.7%). Technical efficiency showed marked variation (median 60%, range 41%-100%) among the centres (II). Compared to the efficient departments, the inefficient departments used excess labour resources (and probably) production equipment to produce RBCs and PLTs. Technical efficiency tended to be higher when the (theoretical) proportion of lost WB collections (total RBC+PLT loss) from all collections was low (III). The labour efficiency varied remarkably, from 25% to 100% (median 47%) when working hours were the only input (IV). Using the estimated total costs as the input (cost efficiency) revealed an even greater variation (13%-100%) and overall lower efficiency level compared to labour only as the input. In cost efficiency only, the savings potential (observed inefficiency) was more than 50% in 10 departments, whereas labour and cost savings potentials were both more than 50% in six departments. The association between department size and efficiency (scale efficiency) could not be verified statistically in the small sample. In conclusion, international evaluation of the technical efficiency in component preparation departments revealed remarkable variation. A suboptimal combination of manpower and production output levels was the major cause of inefficiency, and the efficiency did not directly relate to production volume. Evaluation of the reasons for discarding components may offer a novel approach to study efficiency. DEA was proven applicable in analyses including various factors as inputs and outputs. This study suggests that analytical models can be developed to serve as indicators of technical efficiency and promote improvements in the management of limited resources. The work also demonstrates the importance of integrating efficiency analysis into international comparisons of blood banking.

Genomic inflation factors under polygenic inheritance

Population structure, including population stratification and cryptic relatedness, can cause spurious associations in genome-wide association studies (GWAS). Usually, the scaled median or mean test statistic for association calculated from multiple single-nucleotide-polymorphisms across the genome is used to assess such effects, and 'genomic control' can be applied subsequently to adjust test statistics at individual loci by a genomic inflation factor. Published GWAS have clearly shown that there are many loci underlying genetic variation for a wide range of complex diseases and traits, implying that a substantial proportion of the genome should show inflation of the test statistic. Here, we show by theory, simulation and analysis of data that in the absence of population structure and other technical artefacts, but in the presence of polygenic inheritance, substantial genomic inflation is expected. Its magnitude depends on sample size, heritability, linkage disequilibrium structure and the number of causal variants. Our predictions are consistent with empirical observations on height in independent samples of ~4000 and ~133,000 individuals.

Modern diagnosis of Epstein-Barr virus infections and post-transplant lymphoproliferative disease

Infection by Epstein-Barr virus (EBV) occurs in approximately 95% of the world s population. EBV was the first human virus implicated in oncogenesis. Characteristic for EBV primary infection are detectable IgM and IgG antibodies against viral capsid antigen (VCA). During convalescence the VCA IgM disappears while the VCA IgG persists for life. Reactivations of EBV occur both among immunocompromised and immunocompetent individuals. In serological diagnosis, measurement of avidity of VCA IgG separates primary from secondary infections. However, in serodiagnosis of mononucleosis it is quite common to encounter, paradoxically, VCA IgM together with high-avidity VCA IgG, indicating past immunity. We determined the etiology of this phenomenon and found that, among patients with cytomegalovirus (CMV) primary infection a large proportion (23%) showed antibody profiles of EBV reactivation. In contrast, EBV primary infection did not appear to induce immunoreactivation of CMV. EBV-associated post-transplant lymphoproliferative disease (PTLD) is a life threatening complication of allogeneic stem cell or solid organ transplantation. PTLD may present with a diverse spectrum of clinical symptoms and signs. Due to rapidity of PTLD progression especially after stem cell transplantation, the diagnosis must be obtained quickly. Pending timely detection, the evolution of the fatal disease may be halted by reduction of immunosuppression. A promising new PTLD treatment (also in Finland) is based on anti-CD-20 monoclonal antibodies. Diagnosis of PTLD has been demanding because of immunosuppression, blood transfusions and the latent nature of the virus. We set up in 1999 to our knowledge first in Finland for any microbial pathogen a real-time quantitative PCR (qPCR) for detection of EBV DNA in blood serum/plasma. In addition, we set up an in situ hybridisation assay for EBV RNA in tissue sections. In collaboration with a group of haematologists at Helsinki University Central Hospital we retrospectively determined the incidence of PTLD among 257 allogenic stem cell transplantations (SCT) performed during 1994-1999. Post-mortem analysis revealed 18 cases of PTLD. From a subset of PTLD cases (12/18) and a series of corresponding controls (36), consecutive samples of serum were studied by the new EBV-qPCR. All the PTLD patients were positive for EBV-DNA with progressively rising copy numbers. In most PTLD patients EBV DNA became detectable within 70 days of SCT. Of note, the appearance of EBV DNA preceded the PTLD symptoms (fever, lymphadenopathy, atypical lymphocytes). Among the SCT controls, EBV DNA occurred only sporadically, and the EBV-DNA levels remained relatively low. We concluded that EBV qPCR is a highly sensitive (100%) and specific (96%) new diagnostic approach. We also looked for and found risk factors for the development of PTLD. Together with a liver transplantation group at the Transplantation and Liver Surgery Clinic we wanted to clarify how often and how severely do EBV infections occur after liver transplantation. We studied by the EBV qPCR 1284 plasma samples obtained from 105 adult liver transplant recipients. EBV DNA was detected in 14 patients (13%) during the first 12 months. The peak viral loads of 13 asymptomatic patients were relatively low (<6600/ml), and EBV DNA subsided quickly from circulation. Fatal PTLD was diagnosed in one patient. Finally, we wanted to determine the number and clinical significance of EBV infections of various types occurring among a large, retrospective, nonselected cohort of allogenic SCT recipients. We analysed by EBV qPCR 5479 serum samples of 406 SCT recipients obtained during 1988-1999. EBV DNA was seen in 57 (14%) patients, of whom 22 (5%) showed progressively rising and ultimately high levels of EBV DNA (median 54 million /ml). Among the SCT survivors, EBV DNA was transiently detectable in 19 (5%) asymptomatic patients. Thereby, low-level EBV-DNA positivity in serum occurs relatively often after SCT and may subside without specific treatment. However, high molecular copy numbers (>50 000) are diagnostic for life-threatening EBV infection. We furthermore developed a mathematical algorithm for the prediction of development of life-threatening EBV infection.

Growth differentiation factor 9 signalling in the ovary

In the ovary, two new members of the large TGF-beta superfamily of growth factors were discovered in the 1990s. The oocyte was shown to express two closely related growth factors that were named growth differentiation factor 9 (GDF-9) and growth differentiation factor 9B (GDF-9B). Both of these proteins are required for normal ovarian follicle development although their individual significance varies between species. GDF-9 and GDF-9B mRNAs are expressed in the human oocytes from the primary follicle stage onwards. This thesis project was aimed to define the signalling mechanisms utilized by the oocyte secreted GDF-9. We used primary cultures of human granulosa luteal cells (hGL) as our cell model, and recombinant adenovirus-mediated gene transfer in manipulating the TGF-b family signalling cascade molecules in these cells. Overexpression of the constitutively active forms of the seven type I receptors, the activin receptor-like kinases 1-7 (ALK1-7), using recombinant adenoviruses caused a specific activation of either the Smad1 or Smad2 pathway proteins depending on the ALK used. Activation of both Smad1 and Smad2 proteins also stimulated the expression of dimeric inhibin B protein in hGL cells. Treatment with recombinant GDF-9 protein induced the specific activation of the Smad2 pathway and stimulated the expression of inhibin betaB subunit mRNA as well as inhibin B protein secretion in our cell model. Recombinant GDF-9 also activated the Smad3-responsive CAGA-luciferase reported construct, and the GDF-9 response in hGL cells was markedly potentiated upon the overexpression of Alk5 by adenoviral gene transduction. Alk5 overexpression also enhanced the GDF-9 induced inhibin B secretion by these cells. Similarly, in a mouse teratocarcinoma cell line P19, GDF-9 could activate the Smad2/3 pathway, and overexpression of ALK5 in COS7 cells rendered them responsive to GDF-9. Furthermore, transfection of rat granulosa cells with small interfering RNA for ALK5 or overexpression of the inhibitory Smad7 resulted in dose-dependent suppression of GDF-9 effects. In conclusion, this thesis shows that both Smad1 and Smad2 pathways are involved in controlling the regulation of inhibin B secretion. Therefore, in addition to endocrine control of inhibin production by the pituitary gonadotropins, also local paracrine factors within in the ovary, like the oocyte-derived growth factors, may contribute to controlling inhibin secretion. This thesis shows as well that like other TGF-beta family ligands, also GDF-9 signalling is mediated by the canonical type I and type II receptors with serine/threonine kinase activity, and the intracellular transcription factors, the Smads. Although GDF-9 binds to the BMP type II receptor, its downstream actions are specifically mediated by the type I receptor, ALK5, and the Smad2 and Smad3 proteins.