Cow's milk allergy and the development of tolerance

Aims: To gain insight on the immunological processes behind cow’s milk allergy (CMA) and the development of oral tolerance. To furthermore investigate the associations of HLA II and filaggrin genotypes with humoral responses to early oral antigens. Methods: The study population was from a cohort of 6209 healthy, full-term infants who in a double-blind randomized trial received supplementary feeding at maternity hospitals (mean duration 4 days): cow’s milk (CM) formula, extensively hydrolyzed whey formula or donor breast milk. Infants who developed CM associated symptoms that subsided during elimination diet (n=223) underwent an open oral CM challenge (at mean age 7 months). The challenge was negative in 112, and in 111 it confirmed CMA, which was IgE-mediated in 83. Patients with CMA were followed until recovery, and 94 of them participated in a follow-up study at age 8-9 years. We investigated serum samples at diagnosis (mean age 7 months, n=111), one year later (19 months, n=101) and at follow-up (8.6 years, n=85). At follow-up, also 76 children randomly selected from the original cohort and without CM associated symptoms were included. We measured CM specific IgE levels with UniCAP (Phadia, Uppsala, Sweden), and β-lactoglobulin, α-casein and ovalbumin specific IgA, IgG1, IgG4 and IgG levels with enzyme-linked immunosorbent assay in sera. We applied a microarray based immunoassay to measure the binding of IgE, IgG4 and IgA serum antibodies to sequential epitopes derived from five major CM proteins at the three time points in 11 patients with active IgE-mediated CMA at age 8-9 years and in 12 patients who had recovered from IgE-mediated CMA by age 3 years. We used bioinformatic methods to analyze the microarray data. We studied T cell expression profile in peripheral blood mononuclear cell (PBMC) samples from 57 children aged 5-12 years (median 8.3): 16 with active CMA, 20 who had recovered from CMA by age 3 years, 21 non-atopic control subjects. Following in vitro β-lactoglobulin stimulation, we measured the mRNA expression in PBMCs of 12 T-cell markers (T-bet, GATA-3, IFN-γ, CTLA4, IL-10, IL-16, TGF-β, FOXP3, Nfat-C2, TIM3, TIM4, STIM-1) with quantitative real time polymerase chain reaction, and the protein expression of CD4, CD25, CD127, FoxP3 with flow cytometry. To optimally distinguish the three study groups, we performed artificial neural networks with exhaustive search for all marker combinations. For genetic associations with specific humoral responses, we analyzed 14 HLA class II haplotypes, the PTPN22 1858 SNP (R620W allele) and 5 known filaggrin null mutations from blood samples of 87 patients with CMA and 76 control subjects (age 8.0-9.3 years). Results: High IgG and IgG4 levels to β-lactoglobulin and α-casein were associated with the HLA (DR15)-DQB1*0602 haplotype in patients with CMA, but not in control subjects. Conversely, (DR1/10)-DQB1*0501 was associated with lower IgG and IgG4 levels to these CM antigens, and to ovalbumin, most significantly among control subjects. Infants with IgE-mediated CMA had lower β -lactoglobulin and α-casein specific IgG1, IgG4 and IgG levels (p<0.05) at diagnosis than infants with non-IgE-mediated CMA or control subjects. When CMA persisted beyond age 8 years, CM specific IgE levels were higher at all three time points investigated and IgE epitope binding pattern remained stable (p<0.001) compared with recovery from CMA by age 3 years. Patients with persisting CMA at 8-9 years had lower serum IgA levels to β-lactoglobulin at diagnosis (p=0.01), and lower IgG4 levels to β-lactoglobulin (p=0.04) and α-casein (p=0.05) at follow-up compared with patients who recovered by age 3 years. In early recovery, signal of IgG4 epitope binding increased while that of IgE decreased over time, and binding patterns of IgE and IgG4 overlapped. In T cell expression profile in response to β –lactoglobulin, the combination of markers FoxP3, Nfat-C2, IL-16, GATA-3 distinguished patients with persisting CMA most accurately from patients who had become tolerant and from non-atopic subjects. FoxP3 expression at both RNA and protein level was higher in children with CMA compared with non-atopic children. Conclusions: Genetic factors (the HLA II genotype) are associated with humoral responses to early food allergens. High CM specific IgE levels predict persistence of CMA. Development of tolerance is associated with higher specific IgA and IgG4 levels and lower specific IgE levels, with decreased CM epitope binding by IgE and concurrent increase in corresponding epitope binding by IgG4. Both Th2 and Treg pathways are activated upon CM antigen stimulation in patients with CMA. In the clinical management of CMA, HLA II or filaggrin genotyping are not applicable, whereas the measurement of CM specific antibodies may assist in estimating the prognosis.

A digital herbarium for the flora of Karnataka

A herbarium-based database (virtual herbarium) is a referral system for plants that maximizes the usefulness of the collections. The information content of such a database is essentially built on the voucher specimens that the herbarium has in its care. The present article reports on the construction of a `virtual herbarium' for the state-wide collection of flowering plants in the Herbarium JCB housed at the Centre for Ecological Sciences, Indian Institute of Science, Bangalore, that is expected to be launched soon. The taxonomic data on each species include all information presented on the herbarium specimen label, namely species name, author citation, sub-species if any, variety if any, family, subfamily, collection number, locations, date of collection, habitat and the collector's name. The data further comprise `flora' in which the species are described. Additional information includes the nomenclature update according to `The Plant List', a detailed description, phenology, species distribution, threat status and comments on any special features of the taxon. The live images of the species provided in the database form an information synergy on the species. This initiative is the first of its kind for herbaria in peninsular India.

Systematic study of the blue green algae of Ceylon kept at the herbarium of the Royal Botanical Gardens, Peradeniya

The collection of blue green algae kept at the herbarium of the Royal Botanical Gardens, Peradeniya, was collected by Ferguson more than 70 years ago. Since then many changes have taken place in the taxonomy of the blue green algae. West (1902), Lemmermann (1907), Wine (1915), Crow (1923), Bharadwaja (1934) and Holsinger (1935) had described some of the blue green algae of Ceylon. While examining the collections of blue green algae kept at the herbarium, the authors found that most of the identifications were incorrect and required revision. In the present paper 20 blue green algae are described. The classification and key to the species are based on the characters given by Desikachary (1959). Blue green algae are important to fisheries since Chanos larvae feed on them.

NUCLEAR STAINING IN ALGAL HERBARIUM MATERIAL - A REAPPRAISAL OF THE HOLOTYPE OF CALLITHAMNION-DECOMPOSITUM AGARDH,J. (RHODOPHYTA)

The taxonomic importance of uninucleate vs. multinucleate vegetative cells in the Ceramiaceae is emphasized. It has been possible to make visible nuclei in old material, including type specimens, using aceto-carmine and aceto-iron-haematoxylin stains. The holotypes of Ceramium roseum and Callithamnion fasciculatum (currently known as Callithamnion roseum and Pleonosporium borreri var. fasciculatum) have uninucleate cells and belong to Aglaothamnion. In contrast, the holotype of Callithamnion decompositum, a name that has been applied to collections of at least two uninucleate taxa, has multinucleate cells; its morphological and cytological details agree with those of specimens collected in France and Ireland that were previously thought to represent an undescribed species. Female reproductive morphology (described from a thallus with gametangia in addition to tetrasporangia), in conjunction with habit and vegetative features, indicates that this species belongs to Compsothamnion (Compsothamnieae), as a third species, being distinguished from C. thuyoides and C. gracillimum by its sessile, lateral tetrasporangia. The required new combinations under Aglaothamnion and Compsothamnion are made.

Analysis of DNA polymorphism in ancient barley herbarium material: validation of the KASP SNP genotyping platform

The availability of crop specimens archived in herbaria and old seed collections represent valuable resources for the analysis of plant genetic diversity and crop domestication. The ability to extract ancient DNA (aDNA) from such samples has recently allowed molecular genetic investigations to be undertaken in ancient materials. While analyses of aDNA initially focused on the use of markers which occur in multiple copies such as the internal transcribed spacer region (ITS) within ribosomal DNA and those requiring amplification of short DNA regions of variable length such as simple sequence repeats (SSRs), emphasis is now moving towards the genotyping of single nucleotide polymorphisms (SNPs), traditionally undertaken in aDNA by Sanger sequencing. Here, using a panel of barley aDNA samples previously surveyed by Sanger sequencing for putative causative SNPs within the flowering-time gene PPD-H1, we assess the utility of the Kompetitive Allele Specific PCR (KASP) genotyping platform for aDNA analysis. We find KASP to out-perform Sanger sequencing in the genotyping of aDNA samples (78% versus 61% success, respectively), as well as being robust to contamination. The small template size (≥46 bp) and one-step, closed-tube amplification/genotyping process make this platform ideally suited to the genotypic analysis of aDNA, a process which is often hampered by template DNA degradation and sample cross-contamination. Such attributes, as well as its flexibility of use and relatively low cost, make KASP particularly relevant to the genetic analysis of aDNA samples. Furthermore, KASP provides a common platform for the genotyping and analysis of corresponding SNPs in ancient, landrace and modern plant materials. The extended haplotype analysis of PPD-H1 undertaken here (allelic variation at which is thought to be important for the spread of domestication and local adaptation) provides further resolution to the previously identified geographic cline of flowering-time allele distribution, illustrating how KASP can be used to aid genetic analyses of aDNA from plant species. We further demonstrate the utility of KASP by genotyping ten additional genetic markers diagnostic for morphological traits in barley, shedding light on the phenotypic traits, alleles and allele combinations present in these unviable ancient specimens, as well as their geographic distributions.

Asthma management and asthma control in Sao Paulo, Brazil and Uppsala, Sweden: a questionnaire-based comparison

The Global Initiative Against Asthma (GINA) was developed to meet the global challenge of asthma. GINA has been adopted in most countries and comparison of asthma management in different parts of the world may be of help when assessing the global dissemination of the guideline. The overall goals in GINA include that asthma patients should be free of symptoms, acute asthma attacks and activity limitations. The aim of the present study was to compare asthma management and asthma control in Sao Paulo, Brazil and Uppsala, Sweden. Information was collected from asthmatics in Sao Paulo and Uppsala with a questionnaire. The questionnaire dealt with the following issues: symptoms, smoking, self-management, hospital visits, effect on school/work and medication. The Sao Paulo patients were more likely to have uncontrolled asthma (36% vs 13%, P < 0.001), having made emergency room visits (57% vs 29%, P < 0.001) and having lost days at school or work because of their asthma (46% vs 28%, P = 0.03) than the asthmatics from Uppsala. There were no difference in the use of inhaled corticosteroids, but the Brazilian patients were more likely to be using theophylline (18% vs 1%, P = 0.001) and less likely to be using long-acting beta-2 agonists (18% vs 37%, P < 0.001). We conclude that the level of asthma control was lower among the patients from Sao Paulo than Uppsala. Few of the patients in either city reached the goals set up by GINA. Improved asthma management may therefore lead to health-economic benefits in both locations. Please cite this paper as: Skorup P, Rizzo LV, Machado-Boman L and Janson C. Asthma management and asthma control in Sao Paulo, Brazil and Uppsala, Sweden: a questionnaire-based comparison. The Clinical Respiratory Journal 2009; 3: 22-28.

Fackskolan för huslig ekonomi. En möjlighet till utbild­ning och kvinnoemancipation för de ogifta borgarflick­orna. : En social rekryteringsstudie av lärarinnekandidater till fackskolan för huslig ekonomi i Upp­sala, 1895-1910.

Jag har med denna uppsats velat undersöka betydelsen av Fackskolan för huslig ekonomi i Uppsala,(FHE), för de ogifta borgarflickornas emancipation vad beträffar utbildningsmöjlig­heter. Detta har jag gjort genom att undersöka den sociala re­kryteringen, där de rekryterade flickornas fäders yr­ken, samt en geografisk fördelning har upptecknats, från tiden då skolan grundades, 1895 och fram till 1910. Det undersökta materi­alet, som är matriklar för rekryterade hushållslärarinnor på FHE, är arkivmaterial som lagras på Landsarkivet i Uppsala, där FHE är arkivbildare.   Undersökningen visar att ca 65 % av eleverna var av medelklassfamilj, vilket vid jämförelse överrensstämmer med andra liknande skolor. Blanda annat visar min jämförande undersökning med Folkskollärarinneseminariet i Falun, (FFS), detta. Det som utmärker FHE är dock det höga antalet elever som kom från akademiker-och överklasshem, ca.30 % och det låga antalet med arbetarbakgrund,(2-4%), vilket visar på att skolan hade en övervägande borgerlig profil.

The collections of Lauraceae in the Herbarium of Henri Van Heurck (AWH)

Background and aims - The collections of Lauraceae in the Van Heurck Herbarium (AWH), which is currently on permanent loan to BR, have remained unknown by most of the specialists and by the general public up until the present. The taxonomic status of its one hundred and forty nine specimens of Lauraceae is here presented.Methods - Standard herbarium taxonomy practices were used in conjunction with the literature available. Specimens from fifty eight different herbaria were studied personally, or by checking their holdings available in the internet, or from digital images received from the curators.Key results - The specimens of Lauraceae from AWH are distributed in twenty four genera and seventy eight species with taxonomic status resolved, excepting for three specimens pertaining to species name of status uncertain (one), or to unresolved determination (two). From them, fifty three specimens are nomenclatural types of any sort, which corroborate the relevance of exsiccates acquired by Van Heurck, even for this relatively small set of specimens for the family. Fifteen lectotypifications and one neotypification of species names are proposed here.

Establishment of Regional Herbarium Leads to More Than 200 New Flora Atlas Records for New York State

Loss of small herbaria is an unfortunate global trend, and initiation of new collections at small academic institutions is an increasingly rare occurrence. In 2006, a new herbarium was established at the State University of New York College at Plattsburgh. The PLAT herbarium has since grown to more than 7,000 specimens, many of them representative of the flora of northeastern New York (especially Clinton County). Previous to 2006, this region was without a recognized herbarium, the nearest in-state collections being more than 150 miles away. Although botanists have previously worked in the region, relatively few plant species were recorded for Clinton County by the New York Flora Atlas – a resource providing species distribution records based on specimens accessioned in herbarium collections. Given the dearth of available distribution data for Clinton County (including the eastern Adirondack Mountains and the western Lake Champlain valley), this project sought to provide records of previously unreported species by comparing NY Flora Atlas maps with current holdings. 203 species will now be added to the NY Flora Atlas for Clinton County, roughly half of those considered exotic. This exercise has amplified the importance of supporting and maintaining small regional herbaria as repositories of valuable biodiversity information. Likewise, this project also highlights the enduring value of training in floristics and taxonomy.

Incidence of severe reproductive tract complications associated with diagnosed genital chlamydial infection: the Uppsala Women's Cohort Study

OBJECTIVE: To estimate the cumulative incidence of severe complications associated with genital chlamydia infection in the general female population. METHODS: The Uppsala Women's Cohort Study was a retrospective population based cohort study in Sweden, linking laboratory, hospital, and population registers. We estimated the cumulative incidence of hospital diagnosed pelvic inflammatory disease, ectopic pregnancy, and infertility, and used multivariable regression models to estimate hazard ratios according to screening status. RESULTS: We analysed complete data from 43 715 women in Uppsala aged 15-24 years between January 1985 and December 1989. Follow up until the end of 1999 included 709 000 woman years and 3025 events. The cumulative incidence of pelvic inflammatory disease by age 35 years was 3.9% (95% CI 3.7% to 4.0%) overall: 5.6% (4.7% to 6.7%) in women who ever tested positive for chlamydia, 4.0% (3.7% to 4.4%) in those with negative tests, and 2.9% (2.7% to 3.2%) in those who were never screened. The corresponding figures were: for ectopic pregnancy, 2.3% (2.2% to 2.5%) overall, 2.7% (2.1% to 3.5%), 2.0% (1.8% to 2.3%), and 1.9% (1.7% to 2.1%); and for infertility, 4.1% (3.9% to 4.3%) overall, 6.7% (5.7% to 7.9%), 4.7% (4.4% to 5.1%), and 3.1% (2.8% to 3.3%). Low educational attainment was strongly associated with the development of all outcomes. CONCLUSIONS: The incidence of severe chlamydia associated complications estimated from ours, and other population based studies, was lower than expected. Studies that incorporate data about pelvic inflammatory disease diagnosed in primary care and behavioural risk factors would further improve our understanding of the natural history of chlamydia. Our results provide reassurance for patients, but mean that the benefits of chlamydia screening programmes might have been overestimated.