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Background Bruton tyrosine kinase (BTK) inhibition targets B-cell and other non-T-cell immune cells implicated in the pathophysiology of pemphigus, an autoimmune disease driven by anti-desmoglein autoantibodies. Rilzabrutinib is a new reversible, covalent BTK inhibitor demonstrating preclinical efficacy as monotherapy in canine pemphigus foliaceus. Objectives To evaluate the efficacy and safety of oral rilzabrutinib in patients with pemphigus vulgaris in a multicentre, proof-of-concept, phase II trial. Methods Patients with Pemphigus Disease Area Index severity scores 8-45 received 12 weeks of oral rilzabrutinib 400-600 mg twice daily and 12 weeks of follow-up. Patients initially received between 0 and

Clovers (Trifoliumspecies) are a large and widespread genus of legumes. Recent work supports the Mediterranean origin of the genus in the Early Miocene period, and centres of diversity for clovers occur in the Eastern Mediterranean, East Africa and South America. A number of clovers are of agricultural importance as forage species in grassland agriculture around the world, particularly in temperate areas. White clover (Trifolium repensL.) is the most important legume of grazed pastures, and red clover (T. pratense) is widely cut and conserved as a winter feed. We consider the current state of collected resources in clovers, which have been collated in the most important databases. These are European Internet Search Catalogue (EURISCO), germplasm resources information network and system-wide information network for genetic resources together with the collection held at AgResearch, New Zealand. In total, collections have been made of 204 species with 48 species having more than 100 accessions in these databases. As expected, the majority of accessions are of the agriculturally important species. The geographical origin of collected material is outlined and, for the major species, accessions are broken down according to their status, e.g. wild population, breeders' lines. We then describe some of the ways in which genetic resources of white and red clovers and their relatives have been used in the breeding of these species. These include introgression of stress tolerance traits, targetting improvements in resource use efficiency and increasing seed yield.

We describe an association mapping approach using natural populations of perennial ryegrass (Lolium perenne L.) to identify molecular markers associated with heading date, an important trait affecting seasonal production, tillering, digestibility and grassland management regimes. Twenty-three natural populations originating from throughout Europe, with heading date phenotypes ranging from very early to very late, as well as three synthetic populations (varieties) were used for molecular marker genotyping using AFLP. In total, 589 polymorphic markers were identified. Hierarchical clustering, principal coordinate and other statistical analyses identified four outlying populations forming a clearly distinct sub-group. Removal of those four populations from the subsequent analysis reduced population sub-structure twofold. However, this made relatively little difference to the result of the association analysis. Linear regression identified three markers whose frequency of occurrence correlated with the heading date phenotype. Moreover, these markers were shown to be closely linked to each other within a major QTL on Chromosome 7, explaining 70% of the total variation in heading date. Pairwise linkage disequilibrium among them was also significant. These results suggest that association mapping approaches may be feasible in L. perenne, and that the use of natural populations could provide a useful source of genetic variation in traits of importance in crop improvement.

Festuca species form a polyploid series but only two of the diploid species have been firmly proposed as progenitors of any polyploid. The number and distribution of rDNA sites on the chromosomes of F. scariosa (section Scariosae) and the four diploid species that comprise section Montanae are presented with their relative DNA amounts and key morphological features. Comparisons of the results with those of some polyploid Festuca species from section Bovinae published previously indicate that F. scariosa and F. altissima could be diploid progenitors of the polyploids. It is unlikely that any one of the other three Montanae species is a progenitor of these polyploids.

Molecular genecology is the study of geographical clines in frequencies of molecular markers and their relationship to ecological clines in environmental conditions. This study outlines the principles underlying the selection of populations, focusing on avoiding 'false positives'- noncausal correlations between allele frequency and the environment. The principles are illustrated by identifying a set of populations of Lolium perenne for the study of temperature responses. The selected set of populations encompasses a 20 degrees C range in mean January temperature. Their freezing tolerance shows a linear trend with winter temperature, LT50 decreasing by 0.25 degrees C for each 1 degrees C reduction in mean January temperature.

The Plant Genetic Resources Unit of the Institute of Grassland and Environmental Research, U.K., and the Department of Genetics and Biotechnology, University of Tras-os-Montes and Alto Douro, Portugal, carried out a joint plant collecting expedition in Portugal in May 1995. Seven regions of Portugal were explored from the north to the south. A diverse range of habitats was sampled covering different altitudes, management systems and ecological conditions, where semi-natural grassland and wild unmanaged grassland was found. Vegetative samples of forage grass and legume populations were collected wherever possible. Seed samples were collected in the absence of live plants and from farmers' stores. Detailed collection site data was recorded at every site. 113 populations of Lolium spp., Festuca spp., Agrostis spp., Dactylis spp. and Briza spp. and 53 populations of Trifolium spp., Medicago spp. and Lupinus spp. were collected from 115 sites. It was possible to find traditional agricultural systems throughout Portugal but they are changing in response to European Union farming policies. For example, Lolium multiflorum Lam. is still intercropped with maize, although seed production of this crop on the farm is no longer permissible and, therefore, local land races are being replaced by commercial varieties. These changes to traditional agricultural systems are contributing to a major loss of genetic diversity.

This chapter discusses the data management aspects of germplasm collections. It describes several developments in genetic resources databases, and regional projects aimed at facilitating conservation and use. It focuses solely on germplasm databases, specifically those that are publicly accessible through the World Wide Web and that are hosted and curated by well-established organizations, although there are, or have been, numerous other such databases. An additional criterion for choosing the databases introduced in this chapter is that they catalogue germplasm (generally conserved ex situ in genebanks) much of which is available free of charge (such as the collections held in the centres of the Consultative Group on International Agricultural Research) or may possibly be for purchase at a modest price.

The Plant Genetic Resources Unit (PGRU) of the Institute of Grassland and Environmental Research, Welsh Plant Breeding Station (IGER, WPBS), UK, and the Grassland Research Station, ZubrŨi (GRS, Z), in 1992 part of OSEVA Research Institute for Fodder Crops, Troubsko (OSEVA, RIFC), Czechoslovakia, carried out a collaborative plant collecting expedition in Czechoslovakia between 10 August and 29 August 1992. 14 geographical sub-regions of Czechoslovakia were covered with centres of collection in Bohemia, Moravia and Slovakia. The expedition concentrated on a range of vegetation from managed semi-natural grassland to wild, un-managed grassland. The IGER team collected mainly vegetative samples and the GRS, Z team collected seed samples of forage grass and legume populations. Detailed collection site data was recorded at every site. Samples were collected from 67 sites. Vegetative collections of Lolium spp. (34 populations) and Trifolium spp. (39 populations) and seed collectio ns of other species (57 populations) were made by the IGER team. Seed collections of Lolium spp. (26 populations), Trifolium spp. (19 populations) and other species (28 populations) were made by the GRS, Z team (Table 2). A diverse range of habitats was sampled covering a range of altitude, management systems and ecological conditions. Wild grassland was easily found but semi-natural grassland was uncommon in the collective farming landscape. Entry into a free-market economy is forcing changes on the collective farming system which dominates Czechoslovakian agriculture. Changes in crops and management practices will lead to a further reduction in the area of semi-natural grassland and genetic erosion of the unique forage grass and legume populations in Czechoslovakia. The collecting expedition took place in August 1992, 5 months before Czechoslovakia was divided into the separate states of the Czech Republic and Slovakia. In the text ‘Czechoslovakia’ is used to reflect the situation at the time of the collecting expedition.

The aim was to establish whether temperature and/or elevated [CO2] (-700 μmol mol−1) affects the cell doubling time (cdt) in the different zones of the shoot apex of two natural populations of Dactylis glomerata originating in Portugal (38° S3′ N) and in Sweden (63° 09′ N). In the Portuguese population at ambient [CO2], only the pith rib meristem (PRM) exhibited a significant shortening of cdts from 10 to 30 °C. Elevated [CO2] resulted in a significant shortening of cdt, particularly in the PRM where cdt was reduced 4-8- and 6-1-fold at 10 and 20 °C, respectively, but only 2-fold at 30 °C. In the Swedish population at ambient [CO2], there were no consistent temperature-dependent alterations to cdt and this population was less responsive to elevated [CO2] than the Portuguese population. Nevertheless, elevated [CO2] resulted in a significant shortening of the cdt for some of the zones; the maximum reduction occurred in the PRM at 30 °C. We concluded that in the shoot apex of the Portuguese population, and most notably in the PRM, 10 and 20 °C were non-optimal temperatures for cell division, whilst the Swedish population was relatively buffered against temperature change. Elevated [CO2] resulted in substantially greater reductions in cdts in the shoot meristem of the Portuguese population than in that of the Swedish population.

The Plant Genetic Resources Unit (PGRU) of the Institute of Grassland and Environmental Research, Welsh Plant Breeding Station (IGER, WPBS), UK, and the Plant Genetic Resources Section of the Plant Breeding and Acclimatization Institute (PBAI), Poland, carried out a joint collecting expedition in Poland between 25 August and 7 September 1990. The expedition was unique in that it was the first time that vegetative sampling had been applied to the perennial forage grass and legume populations of south east Poland. The expedition focused on seminatural vegetation in agriculturally managed situations and detailed collection site data on management systems was obtained from landowners. Samples were collected from 62 sites. Vegetative collections of Lolium spp. (37 populations) and Trifolium spp. (56 populations), and seed collections of Festuca spp. (32 populations) were made by the IGER team (Table 2). The PBAI team made 59 separate seed collections, mainly of Leguminosae. The expedition covered four geographical subregions of south east Poland. (See Fig. 1). These were the Nizina Mazowiecka south of Warszawa, the Wyzyna Malopolska with the town of Kielce at its centre and the Beskidy Zachodnie and Beskidy Wschodnie regions of the Carpathian mountains. A diverse range of habitats was sampled covering a broad range of altitude agricultural management systems and ecological conditions. Polish agriculture is faced with the problems of a rapid orientation to a market economy and it is likely that the diverse range of habitats encoutered will be reduced as agricultural practices change. This will lead to genetic erosion of the unique forage grass and legume populations to be found in Poland.

The drought resistance of 25 accessions of meadow fescue (Festuca pratensis Huds.) from seven countries was investigated in four experiments: two in the glasshouse using pot-grown plants from which water was withheld for various periods, and two in controlled environments under osmotic stress.

Cell suspension colonies from four embryogenic Lolium temulentum lines were selected and plated individually in 25 embryoid maturation treatments which varied in various factors reported to stimulate embryogenesis or improve regeneration. Using a numerical scoring system to compare the cultures against a control, treatments were identified which increased growth, suppressed morphogenesis or encouraged premature shoot formation.

We describe a candidate gene approach for associating SNPs with variation in flowering time and water-soluble carbohydrate (WSC) content and other quality traits in the temperate forage grass species Lolium perenne. Three analysis methods were used, which took the significant population structure into account. First, a linear mixed model was used enabling a structured association analysis to be incorporated with the nine populations identified in the structure analysis as random variables. Second, a within-population analysis of variance was performed. Third, a tree-scanning method was used, in which haplotype trees were associated with phenotypes on the basis of inferred haplotypes. Analysis of variance within populations identified several associations between WSC, nitrogen (N), and dry matter digestibility with allelic variants within an alkaline invertase candidate gene LpcAI. These associations were only detected in material harvested in one of the two years. By contrast, consistent associations between the L. perenne homolog (LpHD1) of the rice photoperiod control gene HD1 and flowering time were identified. One SNP, in the immediate upstream region of the LpHD1 coding sequence (C-4443-A), was significant in the linear mixed model. Within-population analysis of variance and tree-scanning analysis confirmed and extended this result to the 2118 polymorphisms in some of the populations. The merits of the tree-scanning method are compared to the single SNP analysis. The potential usefulness of the 4443 SNP in marker-assisted selection is currently being evaluated in test crosses of genotypes from this work with turf-grass varieties.

Improved winter hardiness is an important breeding objective in the forage grass Lolium perenne. This is a complex trait with several components, including the ability to survive and grow at low temperature, to acclimate to cold, tolerate wind, snow cover and ice encasement. Marker-assisted selection has the potential to increase the efficiency of breeding for improved cold tolerance. Here we describe a genecological approach to identifying molecular markers that are associated with adaptation to low winter temperatures. AFLP was used to assess the genetic diversity in 29 wild populations of ryegrass (Lolium perenne) representing a pan-European temperature cline in terms of their geographical origin. A further 18 populations from a temperature cline in Bulgaria were also analysed. In addition, two varieties and five populations representing parents of mapping families currently in use at IGER were included in the analysis. Principal coordinate (PCoA) and cluster analyses of the molecular marker data showed that the Bulgarian altitude cline populations could be distinguished clearly from the other populations. Two regression analyses were carried out; one to identify AFLP markers that correlated in frequency with low mean January temperature of the geographical origin of the population, and another to identify AFLP markers correlating in frequency with the cold tolerance phenotype of the populations, as determined by LT50 values in freezing tests. In the first analysis six AFLP markers showed significant type II trends with mean January temperature, and in the second analysis 28 bands had a significant univariate relationship with the LT50 value of the accessions. In steps 2 and 3 of the stepwise analysis a further 4 and 5 bands, respectively, improved the fit significantly. The results of the two types of regression analysis are discussed in relation to ecogeography and cold tolerance phenotype of the populations.

Festuca gigantea (L.) Vili. (Giant fescue) is a wild species found in woodland margins and on riverbanks throughout Europe. It has large leaves of high nutritive value and is used in breeding programmes which aim to broaden the adaptability of ryegrasses (Lolium perenne L. and L. multiflorum L.), introducing desirable characteristics from fescue species through intergeneric hybridisation. As well as its agronomic attributes, F. gigantea also has some specific germination character—istics; a high level of seed dormancy, a high temperature threshold for germination (28°C and a relatively slow rate of germination. Such characteristics may limit its use. The germination requirements of populations collected from throughout Europe have been studied. Variation in optimal germination temperature and rate of germination have been identified. A pre-chill of 4 weeks at 4°C prior to germination at 28°C has reduced dormancy and increased the rate of germination of all populations examined. Storage at 4°C for up to 12 months reduced dormancy but subsequent storage for 30 months had little further effect on reducing dormancy. The germination requirements of this species are considered and implications for breeding discussed.

The position of the 18S-5.8S-26S and 5S rRNA genes have been physically mapped on the chromosomes of seven Lolium taxa. 18S-5.8S-26S sites were seen on two pairs of chromosomes in the inbreeding taxa. In the outbreeding taxa six sites were found in the L. multiflorum, seven in L. perenne and nine in L. rigidum var. rigidum. Two 5S sites were found in each of the taxa. In the inbreeders, the 5S sites were found adjacent to the 18S-5.8S-26S sites on chromosome 2. In L. multifiorum and L.perenne the 5S sites were on the short arm of chromosome 3. However, in L. rigidum var. rigidum the 5S rDNA site was found in either of the two positions.

The clinical diagnosis in cases of suspected hyperthyroidism is very valuable. Laboratory diagnosis rests largely on measurements of circulating thyroid hormones, with radio-iodine studies playing only a minor part. No one test is infallible and some understanding of basic physiological considerations is essential if serious errors are to be avoided. The choice of treatment plan is logically-based, but should take into account the quality of available services.

The thyroidal I131 uptake rate was measured in 1,116 Australian subjects whose thyroidal status was established at the same time. The logarithm of the uptake rate factor for various functional groups was found to be normally distributed. Diagnostic borderlines are described for premenopausal and postmenopausal females with and without goiter, and for males with and without goiter. The I131 uptake rate was found to decrease progressively with age in male subjects. In females it remained constant until the age of the menopause, following which a progressive decrease was seen. In 76 per cent of subjects unaffected by prior medication a clear diagnosis was established by a single thyroidal I131 uptake test.

The thyroidal I131 uptake rate, one hour after injection of I131, was measured in 450 subjects before and after the administration of ι-thyroxine for about three weeks. In 57 thyrotoxic patients without prior medication, the uptake rate showed a small, but significant mean rise after ingestion of thyroxine; in 51 thyrotoxic patients previously treated with antithyroid drugs there was a small, insignificant mean rise after ingestion of thyroxine. When submitted to the suppression test, most of the 345 euthyroid subjects showed a marked fall in the initially elevated thyroidal I131 uptake rate. There was, however, a small region of overlap between the two types of patients, amounting to about 0.7 per cent of the total euthyroid group and 7.3 per cent of the total hyperthyroid group. The assessment of patients in these overlapping regions is discussed.

In 74 patients uninfluenced by thyroid-affecting medication, the fraction of an intravenously injected tracer dose of I131 present in the thyroid region ten minutes after injection (N5–15) was calculated and plotted against the one-hour uptake rate. The curve thus obtained was used as a calibration curve in subsequent studies. Forty-one patients were studied under two or more of the following regimens: (i) untreated; (ii) receiving a full course of antithyroid drug treatment; (iii) receiving a thyroid hormone preparation in “suppressive” dosage; (iv) receiving a combination of (ii) and (iii). Multiplying the ten-minute thyroidal I131 uptake rate in a patient receiving antithyroid drugs by a factor of 1.22 yields the basic untreated one-hour uptake rate. The ten-minute uptake rate is suppressible by a thyroid hormone preparation in nonthyrotoxic patients but is not suppressible in thyrotoxic patients. INTRODUCTION MOST tests used clinically for the determination of thyroid function by measurement ...

Serial observations of residual radioactivity in the region of the thyroid gland and of urinary excretion of radioiodine were made during the early period after giving a therapeutic dose of I131. Blocking the thyroid with potassium pcrchlorate increased the urinary excretion of I131 and increased the rate at which thyroidal radioactivity diminished. Assuming dynamic equilibrium between the several iodide compartments of the body, the thyroidal iodine uptake rate may be derived from such observations.

Measured thyroidal radiosensitivity factors were found to depend on goiter size but not on sex, thyroidal status (hyperthyroid or euthyroid), type of goiter (diffuse or nodular) or value of uptake rate. A method of radioiodine therapy for thyrotoxicosis is outlined which is predicted to yield a myxedema rate of about 2 per cent, with 60 per cent of the patients requiring only one dose, 30 per cent two doses, and 10 per cent three or more doses. The average time required to achieve a permanently euthyroid uptake rate is about ten weeks. Later than three years after completion of therapy there seems to be a slight fall in the uptake rate beyond that expected on account of the patient's aging.