Distribution of human colonic dendritic cells and macrophages

To define the phenotype of intestinal dendritic cells and macrophages, resected colonic specimens were used to obtain lamina propria cell suspensions by EDTA treatment, then enzymatic digestion. The phenotype of dendritic cell-enriched suspensions was compared with that of macrophage-enriched populations by immunocytochemistry using the avidin-biotin-peroxidase (ABC) system and immunoelectron microscopy. Dendritic cells expressed HLA-DR (L243) and HLA-DQ-associated (RFD1) antigens and CD68 in a perinuclear distribution. Staining for S100 was weak or absent. Macrophages also expressed HLA markers (L243 and RFD1) and CD68. The 25F9 antigen was expressed strongly, whilst CD14 was absent from cells isolated from non-inflamed tissues. To determine their anatomic distribution, immunohistochemistry was performed using single- and double-labelling techniques (ABC ± alkaline phosphatase anti-alkaline phosphatase method). Mutually exclusive subsets of 25F9⁺ and S100⁺cells were seen: 25F9⁺ macrophages were concentrated in a band immediately beneath the luminal epithelium; S100⁺/HLA-DR⁺ dendritic cells formed a reticular network throughout the lamina propria and beneath the basement membrane of the crypts. This distribution suggests that macrophages may help regulate intestinal responses by acting as the first line of defence against the entry of luminal antigens. A breach of the macrophage ‘barrier’ by invading antigens may necessitate the recruitment of T cell responses by immunostimulatory dendritic cells.     

CD8+ lymphocyte phenotype and cytokine production in long-term non-progressor and in progressor patients with HIV-1 infection

In most HIV-1-infected patients, clinical and immunological progression develops within a few years. Few infected people, termed long-term non-progressors (LTNP), remain healthy and immunologically stable for a long time. The factors governing the maintenance of this condition are not well known, but it is conceivable that CD8⁺ lymphocytes, cells that play a central role in controlling in vitro HIV replication, may have a part in vivo in this process. The aim of this study was to characterize the phenotypic profile and the cytokine production of CD8⁺ cells in a group of LTNP patients who had stable CD4⁺ cell counts (>500/mm³) for at least 7 years. Their CD8⁺ absolute numbers were similar to a control group composed of HIV-1⁺ patients who have a progressive decline of their CD4⁺ cell counts. However, our multiparameter immunofluorescence studies show that a clinical and immunologically stable condition is associated with the presence of a CD28⁺, CD95 strongly positive CD8⁺ population, while disease progression is marked by the CD28⁻CD95⁺CD8⁺ subset. Purified CD8⁺ cells from LTNP retain their ability to produce IL-2, interferon-gamma (IFN-γ) and, to a lesser degree, to produce IL-10 and IL-4. In contrast, CD8⁺ cells from progressors are unable to secrete IL-2 and IL-10. Although CD8⁺ cytokine profile does not fit with the proposed T helper (Th)1/Th2 switch in progressive HIV infection, LTNP CD8⁺ T cells maintain their capacity to produce IL-2 and IL-10 (Th0-like), a pattern very similar to that observed in normal HIV healthy controls. We suggest that CD8⁺ cells expressing CD28, CD95 and having a Th0-like profile may be considered to be associated with long-term survival.     

Viral Persistence in Neurons Alters Synaptic Plasticity and Cognitive Functions without Destruction of Brain Cells

Neurons have a restricted expression of MHC heavy chain molecules which prevents presentation of antigens of infecting viruses. As a result, such infected cells escape immune surveillance and allow the establishment of noncytolytic persistent infection. Here we show that a chronic noncytolytic viral infection bothin vitroandin vivoselectively perturbed the expression of GAP-43, a protein that plays a central role in neuronal plasticity processes accompanying learning and memory. GAP-43 expression was greatly decreased in the hippocampus, an area of heightened viral replication, while synaptic density was preserved. Concurrently, the ability to learn tasks was significantly impaired in these persistently infected mice. Yet, infected neurons remained free from structural injury.     

Who needs what from a national health research system: lessons from reforms to the English Department of Health's R&D system

Health research systems consist of diverse groups who have some role in health research, but the boundaries around such a system are not clear-cut. To explore what various stakeholders need we reviewed the literature including that on the history of English health R&D reforms, and we also applied some relevant conceptual frameworks.     

Relation between IgG antibodies to foods and IgE antibodies to milk, egg, cat, dog and/or mite in a cross-sectional study

BACKGROUND: Because IgG antibodies to foods can be detected before IgE antibodies to inhalants, increased levels of IgG antibodies to foods might be used as a predictor of IgE-mediated allergy in initially nonatopic children. OBJECTIVE: To examine the cross-sectional relation between IgG to foods (i.e. mixture of wheat and rice, mixture of soybean and peanut, egg white, cow's milk, meat, orange and potato) and specific IgE to cat, dog, mite, milk and egg white in 1-year-old children. METHODS: All atopic children (n = 120; 58 with and 62 without eczema) and a random sample of the nonatopic children (n = 144) of the Bokaal study were tested on their IgG response to foods. The IgG results of the food assays were dichotomized high or low using the 66th centile as a cut-off value. RESULTS: Atopic children more often had high IgG levels to foods than nonatopic children. IgG to egg white (OR = 7.50) and mixture of wheat and rice (OR = 4.79) were most strongly associated with positive specific IgE. In a stepwise logistic regression analysis egg white, mixture of wheat and rice, and orange were selected (OR = 3.76, OR = 2.43, and OR = 2.11, respectively). In children without eczema higher levels of IgG to foods were still significantly associated with atopy, which was most prominent for egg white, orange and cow's milk. CONCLUSION: An increased IgG antibody level to foods, especially to egg white, orange, and mixture of wheat and rice, indicates an increased risk of having IgE to cat, dog, mite, egg and/or milk allergens, even in the noneczematous group. Therefore, in another prospective study we are currently investigating the usefulness of IgG in early identification, i.e. before IgE antibodies can be detected, of children with an increased risk of developing allergic diseases in the future.     

Nocturnal sleep, daytime sleepiness, and napping among women with significant emotional/behavioral premenstrual symptoms

The objective of this study is to examine daytime sleepiness and alertness and nap characteristics among women with significant emotional/behavioral premenstrual symptoms, and to determine their relationship with nocturnal sleep. Participants spent one night during the follicular phase and two nights during the late-luteal phase, one of which occurred after a 40 min opportunity to nap, sleeping in the laboratory. Subjective measures of sleepiness and alertness were completed during the afternoon of each recording. Setting took place at the sleep laboratory at the University of Ottawa. A total number of participants were 10 women with significant and nine women with minimal emotional/behavioral premenstrual symptoms (mean age 26 years). The results were compared with the follicular phase, both groups of women had less slow wave sleep and more stage 2 sleep at night, as well as a higher daytime and nocturnal mean and maximum temperature during the late-luteal phase. Women with significant symptoms were sleepier and less alert during the late-luteal phase and had a higher overall mean nocturnal temperature compared with women with minimal symptoms. No significant differences were found between the two groups on nap characteristics and nocturnal sleep characteristics. Results show that women with more severe premenstrual symptoms are sleepier during the late-luteal phase than women with minimal symptoms. The increased daytime sleepiness seems to be unrelated to nocturnal sleep or nap characteristics.     

Comparative anticoagulant activity and influence on thrombin generation of dextran derivatives and of a fucoidan fraction

CMDBS compounds are synthetic dextran derivatives with a random distribution of glucosyl units substituted with carboxymethyl, benzylamide, sulfonate, and sulfate groups. Fucoidans are sulfated polysaccharides extracted from brown seaweeds. CMDBS and fucoidans exhibit anticoagulant activity which depends on their chemical composition and molecular weight. Tested with purified proteins, these compounds catalyse thrombin (EC 3.4.21.5) inhibition mainly via heparin cofactor II (HCII). We investigated the mechanism involved in the anticoagulant activity of these polysaccharides relative to that of heparin. Three CMDBS with different chemical compositions were studied to evaluate the effect of sulfate and sulfonate groups on the anticoagulant activity. The fucoidan fraction was extracted from the brown seaweed Ascophylum nodosum. The clotting assays (activated partial thromboplastin time, thrombin time, prothrombin time) were significantly prolonged in the presence of CMDBS and fucoidan, which were less active than heparin. To investigate the action mechanism of these polysaccharides, thrombin generation tests (TGT) were performed on human plasma in the presence of several CMDBS and a fucoidan fraction. The results showed an inhibition of thrombin generation in contact-activated plasma in the presence of both polysaccharides, with a prolonged lag phase preceding the burst of thrombin generation. In thromboplastin-activated plasma, thrombin generation was inhibited by CMDBS and fucoidan, with a prolonged lag phase only in the presence of CMDBS. The data obtained with each polysaccharide, compared to those obtained with heparin (our study) and hirudin (published data), led to hypothesize that fucoidan could act, like heparin, by forming complexes with the inhibitor (although antithrombin (AT) in the case of heparin, and HCII for fucoidan), while CMDBS could act, like hirudin, by forming complexes with thrombin.     

MUSHROOM BODY INFLUENCE ON LOCOMOTOR ACTIVITY AND CIRCADIAN RHYTHMS IN DROSOPHILA MELANOGASTER

Whether or not mechanisms underlying circadian locomotor rhythms and learning are related anatomically through the mushroom bodies (MBs) was investigated by monitoring behavioral rhythmicity in flies with MB lesions induced by chemical ablation and by mutations in five different genes. All flies tested were later examined histologically to assess (1) MB neuroanatomy, and (2) the condition of the putative pacemaker cells -- the ventral Lateral Neurons (LN v s) and their terminals that project to the vicinity of the MB calyces. All groups of flies had normal rhythms except for mushroom body miniature ( mbm ; only in a wild-type Berlin genetic background) and mushroom body defect ( mud ). MB ablation had no effect on the gender-specific differences in the rhythmic activity profile that are typical of wild-type flies. However, ablated males had a slightly longer period than untreated males and were more active under constant dark conditions. LN v s and their arborization patterns appeared normal in MB-ablated and in most mutant flies. Activity defects of mbm flies were attributed to genetic background rather than to the mutation alone. Misrouted LN v projections and ～14% arrhythmia of mud flies were uncorrelated and attributed to pleiotropy rather than to specific effects of MB lesions. Our results imply that MBs are not involved in circadian activity rhythms but that they do have an inhibitory effect on activity levels of male flies.     

Epiphyseal osteoblastoma-like osteosarcoma

Osteoblastoma-like osteosarcoma is a rare variant of osteosarcoma occurring in this instance in a highly unusual location: the lateral femoral condyle of a 13-year-old girl. The radiological features were non-aggressive and, although slightly unusual, were most suggestive of chondroblastoma.Presented at the Closed Meeting of the International Skeletal Society, Geneva, Switzerland, September 2003