Tracking aid flows for development assistance for health.

BACKGROUND AND OBJECTIVES: The global architecture for providing development assistance for health (DAH) has become increasing complex in the last decade, with many new funding agencies entering the health sector. This study presents a detailed picture of European Union (EU) and EU member state originating DAH between 2006 and 2009; with a specific focus on assessing the extent of complementarity of development assistance sourced from the EU. DESIGN: We use a combination of internal EU reporting systems, OECD-DAC creditor reporting system data and other data sources to estimate DAH flows. Our method uses a line by line project assessment in order to identify and categorise DAH flows. RESULTS AND CONCLUSIONS: Our findings show a complex picture of DAH flows--from source, to channel of assistance, to channel of implementation--that is hard to track at the global level, and rarely comprehensively and regularly tracked at the country level. While the majority of EU DAH is focused on low and lower middle income countries there also remains much disparity between countries; and further analysis is required to better understand whether these imbalances are fair and efficient; or result in overlap. We also recommend investment in quality control of DAH tracking internally within donor agencies, and investment in the development of country based systems in order to enable countries and development partners better harmonise DAH flows.

Hepatic differentiation of murine disease-specific induced pluripotent stem cells allows disease modelling in vitro.

Direct reprogramming of somatic cells into pluripotent cells by retrovirus-mediated expression of OCT4, SOX2, KLF4, and C-MYC is a promising approach to derive disease-specific induced pluripotent stem cells (iPSCs). In this study, we focused on three murine models for metabolic liver disorders: the copper storage disorder Wilson's disease (toxic-milk mice), tyrosinemia type 1 (fumarylacetoacetate-hydrolase deficiency, FAH(-/-) mice), and alpha1-antitrypsin deficiency (PiZ mice). Colonies of iPSCs emerged 2-3 weeks after transduction of fibroblasts, prepared from each mouse strain, and were maintained as individual iPSC lines. RT-PCR and immunofluorescence analyses demonstrated the expression of endogenous pluripotency markers. Hepatic precursor cells could be derived from these disease-specific iPSCs applying an in vitro differentiation protocol and could be visualized after transduction of a lentiviral albumin-GFP reporter construct. Functional characterization of these cells allowed the recapitulation of the disease phenotype for further studies of underlying molecular mechanisms of the respective disease.

Hepatic differentiation of pluripotent stem cells.

In regenerative medicine pluripotent stem cells are considered to be a valuable self-renewing source for therapeutic cell transplantations, given that a functional organ-specific phenotype can be acquired by in vitro differentiation protocols. Furthermore, derivatives of pluripotent stem cells that mimic fetal progenitor stages could serve as an important tool to analyze organ development with in vitro approaches. Because of ethical issues regarding the generation of human embryonic stem (ES) cells, other sources for pluripotent stem cells are intensively studied. Like in less developed vertebrates, pluripotent stem cells can be generated from the female germline even in mammals, via parthenogenetic activation of oocytes. Recently, testis-derived pluripotent stem cells were derived from the male germline. Therefore, we compared two different hepatic differentiation approaches and analyzed the generation of definitive endoderm progenitor cells and their further maturation into a hepatic phenotype using murine parthenogenetic ES cells, germline-derived pluripotent stem cells, and ES cells. Applying quantitative RT-PCR, both germline-derived pluripotent cell lines show similar differentiation capabilities as normal murine ES cells and can be considered an alternative source for pluripotent stem cells in regenerative medicine.

Production and proteomic characterization of pharmaceutical-grade Dermatophagoides pteronyssinus and Dermatophagoides farinae extracts for allergy vaccines.

BACKGROUND: House dust mites (HDM) such as Dermatophagoides pteronyssinus and Dermatophagoides farinae represent a major cause of type 1 allergies worldwide. Hence large quantities of well-characterized HDM extracts are needed to prepare pharmaceutical-grade allergy vaccines. To this aim, the present study was undertaken to define optimal conditions for large-scale cultures. METHODS: D. pteronyssinus and D. farinae were grown on different media combining various proportions of wheat germ, yeast and synthetic amino acids (the latter resembling the composition of the human stratum corneum). Extracts thus obtained were analyzed for their total allergenic activity, as well as major allergen and protein contents, using immunosorbent assays, HPLC, immunoblotting, two-dimensional electrophoresis and peptide mass fingerprinting. RESULTS: An optimal culture medium (Stalmite APF) based on wheat germ, yeast and amino acids in defined proportion (42, 42 and 15% w/w, respectively) was selected to grow various HDM species with high yields. A detailed proteomic analysis revealed that D. pteronyssinus extracts generated under such conditions did not contain allergens originating from culture medium components and that major prevalent HDM allergens (i.e. groups 1, 2, 7, 10, 13 and 20) are found among the most abundant proteins in the D. pteronyssinus extract. Semiquantitative dot-blot assays confirmed the presence of Der p 3-10 as well as Der p 13 and 14 allergens within the extracts. CONCLUSIONS: We developed a well-defined medium allowing to grow various HDM species at an industrial scale in a highly reproducible manner. Extracts from mites produced under such pharmaceutical conditions contain all the relevant allergens for desensitization purposes and in vivo diagnosis.

Mutation spectrum and functional analysis of epidermis-type lipoxygenases in patients with autosomal recessive congenital ichthyosis.

Autosomal-recessive congenital ichthyosis (ARCI) is a clinically and genetically heterogeneous group of severe hereditary keratinization disorders characterized by intense scaling of the whole integument, and differences in color and shape. It is often associated with erythema. To date, six loci for ARCI have been mapped. Mutations in ALOXE3 and ALOX12B on chromosome 17p13, which code for two different epidermal lipoxygenases, were recently found in patients with ichthyosiform erythroderma from Turkey, France, and North Africa. Here we describe molecular and clinical findings in 17 families with ARCI originating from Central Europe, Turkey, and the Indian subcontinent, with mutations in ALOXE3 or ALOX12B. We identified 11 novel point mutations in ALOX12B (one nonsense mutation and 10 missense mutations) and four different inactivating mutations in ALOXE3. The gene products of ALOX12B and ALOXE3, the epidermal lipoxygenases 12R-LOX and eLOX3, respectively, are preferentially synthesized in the skin. They act in sequence to convert arachidonic acid via 12(R)-HPETE to the corresponding epoxyalcohol, 8(R)-hydroxy-11(R),12(R)-epoxyeicosatrienoic acid. To assess the impairment of enzyme activity, we expressed the mutated genes in vitro and determined the activity of the recombinant proteins toward their genuine substrates. All but one of the recombinant mutants were enzymatically inactive. The characterization of disease-causing mutations in ALOXE3 and ALOX12B and the resulting ARCI phenotypes did not result in clear diagnostic criteria; however, we found a first correlation between the genetic findings and the clinical presentation of ichthyosis.

[Acquired angioneurotic edema induced by hormonal contraception]. / Oedème angio-neurotique acquis induit par la contraception hormonale.

OBJECTIVE: Although frequently pointed out in the aetiology of chronic angioedema or chronic abdominal pain, food allergy is frequently a diagnosis for lack of anything better, as exemplified by disappointing results of eviction regimens. This has resulted in the search for other aetiologies, such as an iatrogenic effect of oral contraceptives. METHODS: Detailed medical history was obtained on 38 young women, aged a mean of 26 years, experiencing chronic angioedema initially ascribed to food hypersensitivity, but in whom a deficit in C1 inhibitor was demonstrated. We investigated the effects of oral contraception on their level of C1-INH, functional C1-INH assay and their clinical symptoms. RESULTS: An interruption of oral contraception induced an increase of the C1-INH level from 0.201 to 0.224 g/L (p < 0.002) and of the C1-INH functional assay fom 0.396 to 0.702 U (p < 0.0001), associated with a recovery or a marked improvement of the clinical symptoms formerly ascribed to food allergy. The replacement of the initial contraception containing ethinyloestradiol by a progestogen maintained or even improved their clinical and biological state. CONCLUSION: Exogenous estrogen such as those contained in most oral contraceptive may play an iatrogenic role in the aetiology of chronic angioedema.

Gelatin prepared from tuna skin: a risk factor for fish allergy or sensitization?

BACKGROUND: Although fish gelatin may represent a useful alternative to bovine or porcine gelatin, the clearly recognized high prevalence of fish allergy could increase the risk of anaphylaxis to gelatin. The rationale for investigating tuna gelatin rather than gelatin from more allergenic fishes is the availability of an industrial gelatin under development. The infrequent occurrence of tuna allergy should influence the safety of a derived product. The present study investigated IgE antibodies to tuna-skin-derived gelatin in adults and children with documented fish allergy or sensitization. METHODS: Serum samples were taken from 100 consecutive patients with fish allergy or sensitization and tested for IgE antibodies against hydrolyzed or nonhydrolyzed gelatin extracted from tuna skin as compared to extracts from tuna flesh, tuna skin as well as bovine or porcine gelatin. Patients with tuna allergies or sensitization were sensitive to the same tuna species (yellowfin) as that from which the gelatin was obtained. IgE antibodies to these various extracts were analyzed using SDS-PAGE and immunoblotting. RESULTS: Only 3 of the 100 serum samples tested gave evidence of reactivity to gelatin extracted from tuna skin. Cross-reactivity between bovine/porcine and fish gelatin was not observed. CONCLUSION: The risk of adverse reactions to tuna skin gelatin seems to be significantly lower than the risk of fish allergy. Fish gelatin may represent a valuable alternative to bovine or porcine gelatin.

Exogenous oestrogen as an alternative to food allergy in the aetiology of angioneurotic oedema.

Although frequently reported as an aetiology for chronic angioneurotic oedema or urticaria, food allergy is often a diagnosis proposed in the absence of more convincing evidence, as illustrated by the disappointing results of eviction regimens. We report a series of women with an initial diagnosis of food allergy, but in whom the role of oral contraceptives was subsequently demonstrated. Detailed medical history was obtained from 26 young women presenting with chronic angioneurotic oedema or urticaria initially attributed to food allergy, but in whom C1-esterase inhibitor (C1 INH) deficiency was demonstrated. We investigated the effects of oral contraception on C1 INH levels, C1 INH activity and clinical symptoms of these patients. Discontinuation of oral contraception induced an increase in C1 INH levels and C1 INH activity, associated with recovery or marked improvement of the clinical symptoms formerly attributed to food allergy. The relatively high frequency of women taking cyproterone acetate in this population appeared to be a remarkable finding. Replacement of the initial contraception containing ethinylestradiol by a progestogen maintained or even accentuated these good therapeutic results. Exogenous oestrogens, such as those contained in most oral contraceptives, may play an iatrogenic role in the aetiology of chronic angioneurotic oedema or urticaria.

Butyrate strongly inhibits in vitro stimulated release of cytokines in blood.

We studied the in vitro effects of butyrate on the stimulated release of proinflammatory cytokines and cytokines involved in the Th-1/Th-2 balance using the whole-blood model in nine healthy humans. Cytokines were measured without (control) and after stimulation by bacterial lipopolysaccharides (LPS) and phytohemagglutinin (PHA) alone or with addition of butyrate at six different concentrations (0.0625, 0.125, 0.25, 0.5, 1, and 2 mM). We found that butyrate at the six tested concentrations induced a significant decrease (P < 0.001) in the stimulated release of TNF-alpha, IFN-gamma, and IL-12, whereas IL-6 release was not altered. At concentrations > or = 0.25 mM, butyrate significantly reduced the stimulated release of IL-5, IL-10, and IL-13; the stimulated release of IFN-gamma, IL-12, IL-5, and IL-13 was nearly abolished when compared to the unstimulated control sample. We concluded that butyrate has a wide spectrum of inhibitory activity on cytokine release stimulated by LPS + PHA in a whole-blood model. Confirmation of these results on colonic samples would show that butyrate is a factor by which the luminal contents may modulate the immune system in order to maintain the colonic mucosa in a noninflammatory state.