Postnatal growth and column spacing in cat primary visual cortex

The primary visual cortex (area 17) of cats continues to grow substantially during early postnatal development. To assess the implications of this growth for the organization of visual cortical maps, we analysed both postnatal growth of area 17 and the spacing of ocular dominance columns in the same animals using 2-deoxyglucose autoradiography to label cortical activation patterns. Quantitative analyses of area size and column spacing were performed on flat-mount sections of the cortical hemispheres. Our analysis of the surface area revealed an average increase of the size of area 17 between the third and tenth postnatal weeks by about 51%. About 75% of this increase occurred during the third and sixth postnatal weeks (size increase of 37%). However, the distance between adjacent columns did not exhibit a similar increase but rather remained constant during the same postnatal period. Since cortical growth is not accompanied by an increased spacing of ocular dominance columns, new functional modules must somehow be added during the first postnatal weeks to occupy the enlarging cortical sheet. Possible mechanisms underlying the formation of new modules are discussed. Electronic Publication     

Whole-genome sequence assembly for mammalian genomes: Arachne 2

We previously described the whole-genome assembly program Arachne, presenting assemblies of simulated data for small to mid-sized genomes. Here we describe algorithmic adaptations to the program, allowing for assembly of mammalian-size genomes, and also improving the assembly of smaller genomes. Three principal changes were simultaneously made and applied to the assembly of the mouse genome, during a six-month period of development: (1) Supercontigs (scaffolds) were iteratively broken and rejoined using several criteria, yielding a 64-fold increase in length (N50), and apparent elimination of all global misjoins; (2) gaps between contigs in supercontigs were filled (partially or completely) by insertion of reads, as suggested by pairing within the supercontig, increasing the N50 contig length by 50%; (3) memory usage was reduced fourfold. The outcome of this mouse assembly and its analysis are described in (Mouse Genome Sequencing Consortium 2002).     

ICOS+ Th cells produce distinct cytokines in different mucosal immune responses

T cell activation, differentiation and effector functions depend on signals delivered through the antigen-specific TCR and non-clonal costimulatory receptors on the T cell. Activated T cells express the inducible costimulator (ICOS). We examined the co-expression of ICOS with Th cytokines in mucosal immune responses. ICOS+CD4+ Th cells expressed strikingly different cytokines depending on the type of infection encountered and the cells' anatomical localization. In the Th2-dominated response to Schistosoma mansoni, ICOS expression of CD4+ cells isolated from the liver was strongly associated with the expression of IL-5, IL-10, IL-13, and T1/ST2, but not with the chemokine receptor CXCR5, a pattern consistent with Th2 effector cells. In the secondary lymphatic organs of schistosome-infected mice, ICOS expression was randomly correlated with Th2 effector-cytokines, but positively correlated with CXCR5 expression; a pattern consistent with follicular Th cells. In Th cells isolated from gut or liver of mice infected with Toxoplasma gondii, ICOS expression was positively correlated with IFN-gamma production. Finally, in the severe combined immunodeficiency transfer colitis model, ICOS expression was strongly positively associated with IFN-gamma and IL-2. Thus, ICOS appears to costimulate distinct effector functions in different immune responses, depending on factors such as the nature of the antigen encountered and localization and chronicity of the immune response.     

Increased expression of HMGB-1 in the skin lesions of erythema toxicum

Abstract:  At birth, commensal microbes penetrate into the skin of the human newborn, eliciting an acute rash, erythema toxicumn neonatorum . Histologically, the rash is characterized by an upregulation of proinflammatory activity and a local recruitment of immunocytes, including macrophages. High mobility group box chromosomal protein 1, a nuclear and cytosolic protein, is also a pro-inflammatory cytokine released by macrophages in response to microbial stimulation. Here, we reasoned that macrophages but also keratinocytes might upregulate this protein in response to the first colonization and that high mobility group box chromosomal protein 1 might play a role as a proinflammatory mediator in the development and progression of erythema toxicum . Punch biopsy specimens from 1-day-old healthy infants, seven with and four without erythema toxicum were analyzed with indirect immunohistochemistry and two different antihigh mobility group box chromosomal protein 1 antibodies, immunofluorescence, nuclear counterstaining, confocal and immunoelectron imaging. We found relocation of nuclear high mobility group box chromosomal protein 1 into the cytoplasm in keratinocytes and macrophages in erythema toxicum. Cytoplasmatic high mobility group box chromosomal protein 1 was also found in melanocytes and did neither co-locate with lysosomal-associated membrane proteins nor with melanosomes. We speculate that terrestrial adaptation triggers the induction of the endogenous "danger signal" high mobility group box chromosomal protein 1 in the skin of the newborn infant, perhaps in response to the first commensal colonization and that this signal may contribute to alert the immune system and promote a protective immune response.     

Interaction of Adenosine 5‘-Triphosphate (ATP) with Histamine Release Induced by Compound 48/80

The histamine releasing action of compound 48/80 disappeared after pretreatment of rat mast cells with ATP in the absence of divalent cations. The inhibitory action was evident already with 2 μM of ATP, provided the cells were preincubated with ATP for short periods of time prior to the addition of 48/80. When mast cells were exposed to 48/80 combined with ATP more than 10 μM of ATP was needed to induce inhibition of histamine release. This inhibitory effect was found to be specific for ATP compared with various organic phosphorous compounds tested and was not mimicked by EDTA. With concentrations of ATP less than 10 μM the sensitivity to the action of 48/80 was spontaneously restored after prolonged preincubation of the cells with ATP. The experimental findings suggest that ATP, in the absence of divalent cations, induced configurational changes of the plasma membrane of the mast cells so that 48/80 could no longer exert its degranulating and histamine releasing action.     

Postischaemic changes in protein synthesis in the rat brain: effects of hypothermia

Protein synthesis, measured as [¹⁴C]-leucine incorporation into proteins, was studied in the normothermic rat brain following 15 min of transient cerebral ischaemia and 1 h, 24 h and 48 h of recirculation, and in the hypothermic (33°C) brain following 1 h and 48 h of recirculation. Ischaemia was induced by bilateral common carotid occlusion combined with hypotension. Following normothermic ischaemia, incorporation of [¹⁴C]-leucine was depressed by 40–80% at 1 h of recirculation in all brain regions studied. At 48 h postischaemia, incorporation returned to normal or above normal levels in the inner layers of neocortex, the CA3 region, the striatum and the dentate gyrus, while in the outer layers of neocortex and in the hippocampal CA1 region the incorporation was persistently decreased by 26% and 40% respectively. At 24 and 48 h postischaemia, protein synthesis in the CA1 region and the striatum could be attributed to proliferating microglia. Intra-ischaemic hypothermia ameliorated the persistent depression of protein synthesis in the CA1 region at 48 h postischaemia, and a two-fold increase compared to the normothermic group was observed both in the CA1 region and the striatum. In the cortex, eucaryotic initiation factor 2 activity transiently decreased at 30 min postischaemia. In animals subjected to intra-ischaemic hypothermia, the eucaryotic initiation factor 2 activity was reduced by 50% of control at 30 min of recirculation compared with 77% in normothermic animals. We conclude that the postischaemic depression of protein synthesis is in part caused by a decrease in eucaryotic initiation factor 2 activity. The early postischaemic depression may reflect a reaction of the tissue to stress, while the late persistent depression, which is normalised by intra-ischaemic hypothermia, may be related to the mechanism of cell death.     

Measurement of glucose and metabolites in subcutaneous adipose tissue during hyperglycemia with microdialysis at various perfusion flow rates

BACKGROUND: Microdialysis-based glucose sensors have recently been introduced for monitoring glucose levels in diabetic patients. The flow rate by which the fluid sample is pumped through the microdialysis catheter varies in different studies. AIM: To study the effects of various flow rates on glucose and its metabolites sampled by microdialysis during an oral glucose tolerance test. MATERIAL, METHODS: Glucose, lactate, pyruvate and glycerol were measured with microdialysis in interstitial fluid of subcutaneous adipose tissue in twelve healthy young subjects before and during an oral glucose tolerance test using four different flow rates (0.3, 1, 2 and 5 microL/min) and a 30 mm dialysis membrane. RESULTS: At the basal fasting state the dialysate glucose obtained by 0.3 microL/min was equal to capillary glucose concentration. A decrease in dialysate glucose levels during the basal state was observed for higher flow rates but not for 0.3 microL/min, which indicates a depleting effect. The relative increase after OGTT was similar for capillary glucose and flow rate 0.3 microL/min but not for higher flow rates. CONCLUSION: The low microdialysis flow rate (0.3 microL/min) facilitates the capture of true interstitial glucose concentrations during glucose fluctuations. Thus this low flow rate is preferred in studies of local tissue metabolism.     

Actions of Competence in Occupational Therapy Practice: A phenomenological study of practice in narrative form

This study examined the phenomenon ?what are occupational therapists doing when they feel competent?. Data were provided by eleven occupational therapists who narrated clinical cases in which they had felt themselves to be competent. The empirical phenomenological psychological (EPP) method was used to analyse and interpret the data. The result revealed that on a general level the experience of feeling competent as an occupational therapist derived from achieving results in the rehabilitation project that were satisfying for both participants (the therapist and the client). The strategies for accomplishing this were related to the empathic competence of the therapists. This competence involved interpreting clinical situations as well as understanding the relationship between motive, meaning, decision and time. Further it involved bringing objects, in the form of adaptations, technical aids, structures, simplifications or compensations, into the clinical situation. These abilities together had a great impact on the therapeutic outcome by shaping the clients' lifeworld to make it richer and more active.