Search

Liver resident-memory CD8+ T cells (TRM cells) provide important protection against liver-stage Plasmodium infections. Here, we show that a glycolipid-peptide conjugate vaccine designed to be cleaved in vivo to release the NKT cell agonist a-galactosylceramide (a-GalCer), together with a CD8+ T cell malarial epitope, efficiently induced intrahepatic malaria-specific T cells expressing canonical markers of liver TRM (CD69, CXCR6 and CD101). While sterile protection was achieved in a proportion of mice using a single vaccine dose, a combined prime-boost regimen induced higher numbers of liver TRM cells and more robust protection. Improved liver TRM cell generation and sterile immunity using a single dose of the vaccine were obtained in all mice by extending the C-and N-terminal residues of the minimal epitope, and by modifying the linker to enhance a-GalCer release. Our findings describe an ideal synthetic vaccine platform to generate large numbers of liver TRM cells that is potentially valuable for controlling a variety of hepatotropic infections.

During the past years, evidence accumulated showing that glycine comprises anti-inflammatory activities. These effects occur, at least in part, via the activation of glycine-gated chloride channels (GlyR). Glycine is one of the major structural units of collagen, making up about 30% of the amino acids. This study aims to investigate the anti-inflammatory potential of collagen hydrolysate (CH) using the zymosan-induced ear-skin inflammation mouse model. After oral intake of 12.5, 25 or 50 mg CH the plasma levels of glycine increased in a concentration-dependent manner. CH was able to counteract zymosan-induced ear-skin inflammation locally (ear swelling) as well as systemically (IL-6 production by lipopolysaccharide (LPS)-stimulated whole blood cells). The LPS-stimulated IL-6 production in whole blood correlated positively with the ear swelling response. This correlation was abolished by strychnine (a glycine receptor antagonist), indicating the involvement of GlyR. Collectively, these data show that CH is able to modulate inflammatory responses both locally as well as systemically. This effect might be constituted by inhibiting pro-inflammatory cytokine production via GlyR.

We report a case of a 64 year old woman with a calcified ring at the level of the sinotubular junction. Echocardiography and Computed Tomography showed a supravalvular aortic stenosis, without known associated lesions, except for the existence of an aberrant right subclavian artery. These combination of abnormalities makes it an unique case. Differential diagnosis of sinutubular calcification is added. From the literature a short review of supravalvular aortic stenosis is presented with indications for surgical intervention. Lifelong and regular follow up is necessary.

In recent years, various intervention strategies have reduced malaria morbidity and mortality, but further improvements probably depend upon development of a broadly protective vaccine. To better understand immune requirement for protection, we examined liver-stage immunity after vaccination with irradiated sporozoites, an effective though logistically difficult vaccine. We identified a population of memory CD8

Introduction Crohn's disease (CD) is a chronic inflammatory disease predominantly affecting the gastrointestinal tract. CD usually requires lifelong medication and is accompanied by severe complications, such as fistulae and strictures, resulting in surgery. Infliximab (IFX) is very effective for treating paediatric patients with CD, but is currently only registered for therapy refractory patients—the so-called step-up strategy. We hypothesise that using IFX first-line, that is, top-down, will give more mucosal healing, fewer relapses, less complications, need for surgery and hospitalisation. Methods and analysis This international multicentre open-label randomised controlled trial includes children, aged 3–17 years, with new-onset, untreated CD with moderate-to-severe disease activity (weighted Paediatric Crohn's Disease Activity Index (wPCDAI)>40). Eligible patients will be randomised to top-down or step-up treatment. Top-down treatment consists of 5 IFX infusions combined with azathioprine (AZA). After these 5 infusions, patients will continue AZA. Patients randomised to step-up will receive standard induction treatment, either oral prednisolone or exclusive enteral nutrition, combined with AZA as maintenance treatment. The primary outcome is clinical remission (wPCDAI

This article examines possibilities for linkage between the European Union Emissions Trading System (EU ETS) and Russia, with a view to enhancing cooperation on a broader scale than the project-based approaches that have been tested thus far. Three paths for possible EU-Russia linkage are presented by which the Russian Assigned Amount under the Kyoto Protocol can be greened in order to stimulate emissions trading: 1. Joint implementation—reductions earned via individual projects in Russia; 2. Greened allowances or green investment schemes; and 3. Linked cap-and-trade systems, in which a Russian domestic emissions trading system would link with the European Union Emissions Trading System. The authors conclude that the third option, emissions trading through linked domestic emissions trading systems, offers the best opportunities at the lowest transaction costs. The authors discuss useful innovative instruments like call options and slip level arrangements on government-to-government and business-to-business levels.

This study's central research question is: "How do managers evaluate the desirability of early retirement of their employees, and under what circumstances and for what types of workers are they in favor of delay?" We sought to compare managers' and business students' decision making regarding older workers. We examined the extent to which student samples are appropriate to study organizational behavior. An identical factorial survey was carried out among 26 managers and 25 business school students. The results revealed that business students concentrate on performance-related individual characteristics when making selection decisions, whereas managers also recognize contextual factors (need for downsizing, tight labor market) and older workers' attitudes toward retirement. © 2009 Wiley Periodicals, Inc.

Leptosphaeria biglobosa ‘canadensis’ is reported for the first time in Australia. All 88Leptosphaeria isolates cultured fromBrassicajuncea stubble from northern NSW wereL. biglobosa ‘canadensis’ whilst all 55 isolates cultured from Victorian stubble of the sameB. juncea lines wereL. maculans. BothL. biglobosa ‘canadensis’ andL. maculans formed similar sized lesions onB. juncea cotyledons after 14 days. However,L. biglobosa ‘canadensis’ isolates colonised stems less effectively thanL. maculans and consequently caused less crown cankering.

Extensive study of stresses induced during and after winding of coils has led to significant improvements to the processing of web-like materials for the packaging industries and consumer goods. The current paper presents an inverse solution for the prediction of the winding tension profile required to satisfy a specified residual stress distribution in the coil, for non-linear material properties (in the radial direction), with allowance for large strain and large deformation in the coil. The inverse solution approach has advantage over the forward solution approach by directly determining the winding tension profile, which can be readily controlled in a processing line, once the desired residual stresses in the coil are known. The current paper, together with several recently published ones, completes the study of stresses in the winding of coils using the inverse solution approach.

The intestinal epithelium is the most rapidly self-renewing tissue in adult mammals. It is currently believed that four to six crypt stem cells reside at the +4 position immediately above the Paneth cells in the small intestine; colon stem cells remain undefined. Lgr5 (leucine-rich-repeat-containing G-protein-coupled receptor 5, also known as Gpr49) was selected from a panel of intestinal Wnt target genes for its restricted crypt expression. Here, using two knock-in alleles, we reveal exclusive expression of Lgr5 in cycling columnar cells at the crypt base. In addition, Lgr5 was expressed in rare cells in several other tissues. Using an inducible Cre knock-in allele and the Rosa26-lacZ reporter strain, lineage-tracing experiments were performed in adult mice. The Lgr5-positive crypt base columnar cell generated all epithelial lineages over a 60-day period, suggesting that it represents the stem cell of the small intestine and colon. The expression pattern of Lgr5 suggests that it marks stem cells in multiple adult tissues and cancers.

This paper extends a previous analysis of determining the winding stress in coils, using an inverse solution approach to coils with large deformations of linear orthotropic materials. This approach has the advantage of determining the required winding stress profile explicitly once the desired residual stresses in the coil to avoid post-winding defects are known. Comparisons between the theories for small and large deformations are also made, with coil properties, where the small deformations assumption is inappropriate, are identified.

Transmission blocked by drug resistance Resistance to the antimalarial drug atovaquone might prove to be this parasite's weak spot. Resistance develops rapidly via mutations in the drug's target: the parasite's mitochondrial cytochrome b complex. Goodman et al. have discovered that although resistant Plasmodium berghei parasites persist in mice, in blood-sucking malarial mosquitoes, the mutations disable female parasites too much for them to reproduce. The human-specific Plasmodium falciparum can only be investigated experimentally in mosquitoes, but a similar effect was seen. Thus, atovaquone-resistant parasites cannot be transmitted to another mammal or person. Science , this issue p. 349

Bouwers die kiezen voor energiezuinig en Nul op de Meter staan voor een aantal uitdagingen. Zowel op sociaal, economisch en technisch gebied is er werk aan de winkel. Er wordt al gepionierd en daar kunnen we van leren. Daarom delen wij, TNO, RIGO, Van Beek en Energiesprong | Platform 31, met plezier de lessen die we hebben getrokken uit de monitoring van een aantal energiezuinige renovatie- en nieuwbouwprojecten. In dit document draait het om de bewoner, een niet te onderschatten aspect dat komt kijken bij (ver)nieuwbouw. Hoe tevreden zijn bewoners met de renovatie en welke aspecten spelen daarbij een rol? We beginnen met de belangrijkste lessen en tips. Verderop in het document zoeken we de verdieping op. Behalve het inzichtelijk maken van onderzoeksresultaten, geven we ook tips. Dit kunnen zowel aanbevelingen voor bouwers als voor opdrachtgevers zijn. Dit document maakt deel uit van een serie van drie documenten, de andere documenten zijn gericht op prestatiegarantie en concepten.

The ascomycete Sclerotinia sclerotiorum is a plant pathogen with a very broad host range. In order to identify and characterize genes involved in S. sclerotiorum infection of Brassica napus (canola), expressed sequence tags (ESTs) were examined from libraries prepared from three tissues: complex appressorium (infection cushions), mycelia grown on agar and lesions formed on leaves of B. napus. A high proportion of genes (68%) had not been previously reported for S. sclerotiorum in public gene or EST databases. The types of novel genes identified in the infection cushion library highlights the functional specificity of these structures and similarities to appressoria in other fungal pathogens. Quantitative real-time PCR was used to analyse tissue specificity and timing of transcription of genes with best matches to MAS3 (appressoria-associated protein from Magnaporthe grisea), cellobiohydrolase I, oxaloacetate acetylhydrolase, metallothionein, pisatin demethylase, and an unknown gene with orthologs in fungal pathogens but not in saprophytic fungi.

The dothideomycete fungus, Leptosphaeria maculans, is poorly described at the genomic level. Two genomic regions have been analysed - one (55 kb) comprises a cluster of 18 genes encoding the biosynthetic enymes for a phytotoxin, sirodesmin, whilst the other (184 kb) is the pericentromeric region of a 2.80 Megabase chromosome. Transcription of all genes in the sirodesmin gene cluster is co-regulated with the production of sirodesmin in culture. Disruption of one of these genes (encoding a two-module non-ribosomal peptide synthetase) is essential for production of sirodesmin. The 184 kb sequence contains 6.980 kb repetitive element named Pholy bordered by two Long Terminal Repeats (LTRs), five Pholy-related sequences, mostly truncated at their 3′ ends; and five solo-LTRs. This element, Pholy, comprises a previously described element, LMR1. Structural features suggest that Pholy corresponds to an ancient copia-like retrotransposon, as it has high sequence similarity to the ELSA retrotransposon of the closely related fungus, Stagonospora nodorum. Comparative analysis of the structure of the Pholy-like sequences in the 184 kb contig and in other parts of the genome suggests that this family of repetitive elements has undergone extensive Repeat Induced Point (RIP) mutation.

The self-renewing epithelium of the small intestine is ordered into stem/progenitor crypt compartments and differentiated villus compartments. Recent evidence indicates that the Wnt cascade is the dominant force in controlling cell fate along the crypt-villus axis. Here we show a rapid, massive conversion of proliferative crypt cells into post-mitotic goblet cells after conditional removal of the common Notch pathway transcription factor CSL/RBP-J. We obtained a similar phenotype by blocking the Notch cascade with a gamma-secretase inhibitor. The inhibitor also induced goblet cell differentiation in adenomas in mice carrying a mutation of the Apc tumour suppressor gene. Thus, maintenance of undifferentiated, proliferative cells in crypts and adenomas requires the concerted activation of the Notch and Wnt cascades. Our data indicate that gamma-secretase inhibitors, developed for Alzheimer's disease, might be of therapeutic benefit in colorectal neoplastic disease.

We report a case of a 39-year-old female with a ventricular septal defect (VSD) and a giant appendiform aneurysm of the membranous septum, illustrated by echocardiography and magnetic resonance imaging. From the literature a short review of the prevalence of spontaneous closure of VSDs together with the possible complications of persisting VSDs is presented. Since patients stay at risk in later years, follow-up at regular intervals is advised.

Sirodesmin PL is a phytotoxin produced by the fungus Leptosphaeria maculans, which causes blackleg disease of canola (Brassica napus). This phytotoxin belongs to the epipolythiodioxopiperazine (ETP) class of toxins produced by fungi including mammalian and plant pathogens. We report the cloning of a cluster of genes with predicted roles in the biosynthesis of sirodesmin PL and show via gene disruption that one of these genes (encoding a two-module non-ribosomal peptide synthetase) is essential for sirodesmin PL biosynthesis. Of the nine genes in the cluster tested, all are co-regulated with the production of sirodesmin PL in culture. A similar cluster is present in the genome of the opportunistic human pathogen Aspergillus fumigatus and is most likely responsible for the production of gliotoxin, which is also an ETP. Homologues of the genes in the cluster were also identified in expressed sequence tags of the ETP producing fungus Chaetomium globosum. Two other fungi with publicly available genome sequences, Magnaporthe grisea and Fusarium graminearum, had similar gene clusters. A comparative analysis of all four clusters is presented. This is the first report of the genes responsible for the biosynthesis of an ETP.

In another work, the authors presented an analysis of stresses in wound coils under the assumption that the coil could be modelled as an orthotropic linear material. In this paper the authors extend the theory to the case where the constitutive relations of the coil material are non-linear. The approach adopted for both the linear and non-linear theory differs radically from that usually reported in this field of research. Instead of the traditional ‘forward solution’ of calculating stresses for a given winding policy, an ‘inverse solution’ is obtained in which the winding stresses required to achieve specified stress distributions in a coil are calculated. Stress distributions required to avoid coiling problems can often be specified. This method can therefore offer considerable advantages in industrial applications, since winding stresses required to achieve a specified stress distribution in a coil can be calculated directly. The non-linear solutions are compared to those for the linear case fo...

In another work, the authors presented an analysis of stresses in wound coils under the assumption that the coil could be modelled as an orthotropic linear material. In this paper the authors extend the theory to the case where the constitutive relations of the coil material are non-linear. The approach adopted for both the linear and non-linear theory differs radically from that usually reported in this field of research. Instead of the traditional ‘forward solution’ of calculating stresses for a given winding policy, an ‘inverse solution’ is obtained in which the winding stresses required to achieve specified stress distributions in a coil are calculated. Stress distributions required to avoid coiling problems can often be specified. This method can therefore offer considerable advantages in industrial applications, since winding stresses required to achieve a specified stress distribution in a coil can be calculated directly. The non-linear solutions are compared to those for the linear case for different winding stresses and coil properties. An example of the effect of modifying the stress distribution in a coil on the winding stress is given to illustrate the advantage of this method.

Several physical techniques were used to study the extent of spoilage in apple juice deliberately inoculated with yeast (concentration of Saccharomyces cerevisiae ranged from 25 cells mL(-1) to 2.5 x 10(6) cells mL(-1), respectively) and their performance compared in terms of detection limit achieved. The optical methods used in this investigation rely on the measurement of either absorption [as is the case for classical spectrophotometry (SP) and the so called optothermal window (OW), a variant of a photothermal method], or scattering [examples are turbidimetry (TB), laser scattering (SC), and laser speckle fluctuation (SF)]. It is shown that the presence of yeast increases both optical absorption and scattering. The most favorable detection limit (25 cells mL(-1)) and a highest (nearly 10(4)) dynamic range, combined with a good linearity, were obtained with the experimental set-up for SC. In addition, the extent of correlation between different methods was determined using two markedly different reference substances, i.e., (i) the mixture of apple and blackcurrant juices, representing a strongly absorbing sample, and (ii) diluted (dilution factor of 103) milk as a strong scatterer. Finally, one has monitored the progress of a spontaneous spoilage process in the inoculated juices stored at 5degreesC under aerobic conditions.

Detecting early ageing stages of seeds in storage is of great concern to vegetable seed companies. Despite their reliability conventional germination tests are destructive and time consuming. One alternative towards assessing the quality of seed is to monitor the concentration of certain volatiles (acting as biological markers) evolved from the seeds; examples are ethanol (EtOH), acetaldehyde, ethane, methanol, etc. Most of the presently used methods have only moderate insensitivity and are therefore not suitable for the on-line measurements. In this work CO laser photoacoustic spectroscopy (LPAS) was used to investigate the on-line evolution of EtOH and acetaldehyde from the imbibed nonaged and aged seeds of cabbage. The overall performance of LPAS was superior to that of Fourier transform infrared and of gas chromatography.

Omgaan met combinatieklassen en dan óók nog recht doen aan de onderwijsbehoeften rekenen is een hele toer. Eward Cozijnsen, leerkracht in groep 7/8 en Henk Rietdijk, opleidingsdocent reken-wiskunde, doen verslag van de werkwijze die zij ontwikkelden in een Pedagogische Proefpraktijk waarin mentoren, studenten en opleidingsdocenten samenwerken. Vanuit Handelingsgerichtwerken en inzichten uit de rekendidactiek hebben de auteurs een aanpak ontwikkeld voor combinatieklassen.

To follow the fate of CD8+ T cells responsive to Plasmodium berghei ANKA (PbA) infection, we generated an MHC I-restricted TCR transgenic mouse line against this pathogen. T cells from this line, termed PbT-I T cells, were able to respond to blood-stage infection by PbA and two other rodent malaria species, P. yoelii XNL and P. chabaudi AS. These PbT-I T cells were also able to respond to sporozoites and to protect mice from liver-stage infection. Examination of the requirements for priming after intravenous administration of irradiated sporozoites, an effective vaccination approach, showed that the spleen rather than the liver was the main site of priming and that responses depended on CD8α+ dendritic cells. Importantly, sequential exposure to irradiated sporozoites followed two days later by blood-stage infection led to augmented PbT-I T cell expansion. These findings indicate that PbT-I T cells are a highly versatile tool for studying multiple stages and species of rodent malaria and suggest that cross-stage reactive CD8+ T cells may be utilized in liver-stage vaccine design to enable boosting by blood-stage infections., Author Summary Malaria is a disease caused by Plasmodium species, which have a highly complex life cycle involving both liver and blood stages of mammalian infection. To prevent disease, one strategy has been to induce CD8+ T cells against liver-stage parasites, usually by immunization with stage-specific antigens. Here we describe a T cell receptor specificity that recognizes an antigen expressed in both the liver and blood stages of several rodent Plasmodium species. We generated a T cell receptor transgenic mouse with this specificity and showed that T cells from this line could protect against liver-stage infection. We used this novel tool to identify the site and cell-type involved in priming to a recently developed intravenous attenuated sporozoite vaccine shown to have efficacy in humans. We showed that CD8+ T cells with this specificity could protect against liver-stage infection while causing pathology to the blood stage. Finally, we provided evidence that T cells with cross-stage specificity can be primed and boosted on alternative stages, raising the possibility that antigens expressed in multiple stages might be ideal vaccine candidates for generating strong immunity to liver-stage parasites.

The aging congenital heart disease (CHD) population is prone to develop a variety of sequelae, including pulmonary arterial hypertension (PAH). Previous prevalence reports have a high degree of variability, with an overrepresentation of tertiary centers. We aimed to report robust prevalence

Objectives:A general population job-exposure matrix (GPJEM) including physical and psychosocial demands as well as psychosocial resources applicable to older and retired workers was developed. Its validity was evaluated by examining associations of physical demands and iso-strain (combination of high psychosocial demands and low resources) with health.Methods:Physical and psychosocial work exposures reported by 55-64 year olds were derived from the Netherlands Working Conditions Survey and linked to the Netherlands Standard Classification of Occupations 1992. A GPJEM with low, moderate, and high probability of exposure to demands and resources was developed. To examine associations with health, two groups of the Longitudinal Aging Study Amsterdam were selected: current (i.e. at the time of the interview, 55-64 years) and former workers (55-84 years). Linear and logistic regression models were applied.Results:Use of force and work in uncomfortable positions were significantly associated with functional limitations and self-perceived health (SPH), but not hip or knee osteoarthritis (OA), in current and former workers. A moderate probability of repetitive movements was associated with functional limitations in former workers. A high probability of repetitive movements was associated with functional limitations in current and former workers as well as with SPH and hip and knee OA in former workers. Respondents formerly exposed to iso-strain had significantly higher diastolic blood pressure and more often hypertension. No such associations were found in current workers. No association was found with cardiovascular disease.Conclusions:The results suggest that our GPJEM accurately classifies jobs according to physical demands and, although less clearly, iso-strain. © The Author 2013.

A genetic and physical map has been developed for Leptosphaeria maculans, which causes blackleg of oilseed Brassicas. The genetic map was constructed from 58 F1 progeny and comprises 154 Amplified Fragment Length Polymorphic (AFLP) markers, 3 Random Amplified Polymorphic DNA (RAPD) markers, a gene encoding a retinal-binding protein, the mating type locus and a host specificity locus. Pulsed field gel electrophoresis experiments showed that the parental isolates each had 16 chromosomes and a genome size of about 33.5 Mb. Many markers bound to every chromosome indicating that L. maculans has a high level of dispersed repetitive DNA sequences. This fungus displays chromosomal length polymorphisms, but in the cross examined, the linkage and physical maps were congruent and there was no evidence of translocations. The host specificity locus is flanked by two AFLP markers 19 and 27 cM distant and is located on a chromosome sized 1.85 Mb in the virulent parent. The mating type locus is on a chromosome sized 2.6 Mb and co-incident on an AFLP marker amplified from the virulent parent.

A genetic and physical map has been developed for the loculoascomycete Leptosphaeria maculans, a pathogen of oilseed Brassicas. The genetic map was constructed from 58 F(1) progeny and comprises 155 amplified fragment length polymorphic (AFLP) markers, three random amplified polymorphic DNA (RAPD) markers, the mating type locus and a host specificity locus conferring the ability to form lesions on Brassica juncea. Twenty-one linkage groups, 5 pairs, and 18 unlinked markers were assigned, and the genome size was 1520 cM. Pulsed field gel electrophoresis experiments showed that the parental isolates each had 16 chromosomes and a genome size of about 33.5 Mb. Attempts to anchor a large number of markers to chromosomes were hampered by difficulties in converting AFLPs into RFLP markers, and because many markers bound to every chromosome, indicating that L. maculans has a high level of dispersed repetitive sequences. This fungus displays chromosomal length polymorphisms, but in the cross examined, the linkage and physical maps were essentially congruent and there was no evidence of translocations. The host specificity locus is 18 cM from the nearest AFLP marker and is located on a chromosome sized 1.85 Mb in the virulent parent. The mating type locus is on a chromosome sized 2.6 Mb and coincident on an AFLP marker amplified from the virulent parent. The derived amino acid sequence of part of this marker has some conserved amino acids present in the High Mobility Group DNA binding domain of MAT-2 mating type genes of other ascomycetes.

Continued innovation of products, services, technology and the organization itself, is one way to keep a business on its feet during these turbulent times. The importance of innovation – the process during which leap‐changes are effected – is generally recognized. However, in practice, the successful conclusion of a total innovation project is by no means matter‐of‐course. Many innovations end in failure. Research in the USA raises the questions of how successful Dutch companies and their managers are in terms of innovation. How many innovation projects succeed, how many fail? This study searches for an answer to this question. The logical next question then is, what are the differences between innovation projects that succeed and those that fail? In short, what are the factors that lead to success, and which factors lead to failure in the projects examined?

Amplified Fragment Length Polymorphism (AFLP) analysis has been used to analyse 100 Australian, European and North American isolates of Leptosphaeria maculans. All isolates had distinct AFLP profiles. They could be classified into five types, which had very few AFLP bands in common and corresponded to classifications made previously on the basis of ability to cause stem cankers on Brassica napus (A group), or inability to do so (B group), and on host range. Four isolates had AFLP profiles completely dissimilar to these groups and to each other. Genetic diversity and geographic differentiation were analysed separately within AFLP types 1 and 2. UPGMA analysis of the 66 AFLP type 1 (A group) isolates using 50 polymorphic bands did not provide evidence for clustering according to geographic origin. Non-metric multidimensional scaling (NMDS) analysis suggested that the Australian and European populations were separate adjacent clusters, while the North American population partially overlapped both the others. This geographic differentiation was supported by Wright's fixation index (Fst) analysis. Three measures of genetic variability between isolates within regions (effective number of alleles, gene diversity, and Shannon index) showed that the North American A group isolates were less diverse than those from Australia and Europe. The 21 AFLP type 2 (B group; NA1 sub-group) isolates did not cluster based on geographic region, which was confirmed by NMDS and Fst analysis. There was a similar degree of genetic diversity within A group and the NA1 sub-group of B group isolates. Unlike other techniques, AFLP analysis can readily discriminate between group B isolates of the L. maculans complex that were previously difficult to classify and also provides individual fingerprints for isolates. Isolates of the A group and of the NA1 sub-group of B group could be also distinguished readily by electrophoretic karyotyping, as the latter isolates had more bands smaller than 1.4 Mb than the A group isolates.