Cells migrating to sites of tissue damage in response to the danger signal HMGB1 require NF-kappaB activation

Tissue damage is usually followed by healing, as both differentiated and stem cells migrate to replace dead or damaged cells. Mesoangioblasts (vessel-associated stem cells that can repair muscles) and fibroblasts migrate toward soluble factors released by damaged tissue. Two such factors are high mobility group box 1 (HMGB1), a nuclear protein that is released by cells undergoing unscheduled death (necrosis) but not by apoptotic cells, and stromal derived factor (SDF)-1/CXCL12. We find that HMGB1 activates the canonical nuclear factor kappaB (NF-kappaB) pathway via extracellular signal-regulated kinase phosphorylation. NF-kappaB signaling is necessary for chemotaxis toward HMGB1 and SDF-1/CXCL12, but not toward growth factor platelet-derived growth factor, formyl-met-leu-phe (a peptide that mimics bacterial invasion), or the archetypal NF-kappaB-activating signal tumor necrosis factor alpha. In dystrophic mice, mesoangioblasts injected into the general circulation ingress inefficiently into muscles if their NF-kappaB signaling pathway is disabled. These findings suggest that NF-kappaB signaling controls tissue regeneration in addition to early events in inflammation.     

Molecular determinants for the action of general anesthetics at recombinant α2β3γ2γ-aminobutyric acidA receptors

General anesthetics modulate the activity of ligand-gated ion channels including the GABA(A) receptor. Mutational studies mainly on the benzodiazepine-insensitive alpha(2)beta(1(M286W)) and alpha(6)beta(3(N289M))gamma(2) GABA(A) receptors revealed that a serine in transmembrane domain 2 and a methionine in transmembrane domain 3 are essential for the action of most general anesthetics. We investigated whether these residues would similarly be relevant for their action at the benzodiazepine-sensitive GABA receptor subtype, alpha(2)beta(3)gamma(2). We found that not only the N265M but also the M286W mutation nearly abolished the modulatory effect of etomidate. However, the anti-convulsant loreclezole, a structural homologue of etomidate, was inactive on the N265M mutant, but displayed normal modulatory activity on the M286W mutant. Both mutations did not affect the modulatory action of the neurosteroid alphaxalone. The direct action of alphaxalone, however, was dramatically increased in the M286W mutant to about twice the maximal GABA current but not significantly affected in the N265M mutant. These data demonstrate that the structural requirements for modulatory and direct actions of various general anesthetics are distinct. The molecular switches induced by these mutations can be exploited to identify the molecular determinants for the action of general anesthetics.     

Cloning and Gene Mapping of the Chromosome 13q14 Region Deleted in Chronic Lymphocytic Leukemia

Frequent deletions and loss of heterozygosity in a segment of chromosome 13 (13q14) in cases of B-cell chronic lymphocytic leukemia (CLL) have suggested that this malignancy is caused by inactivation of an unknown tumor suppressor gene located in this region. Toward the identification of the putative CLL tumor suppressor, we have constructed a high-resolution physical map of YAC, PAC, and cosmid contigs covering 600 kb of the 13q14 genomic region. In addition to densely positioned genetic markers and STSs, this map was further annotated by localization of 32 transcribed sequences (ESTs) using a combination of exon trapping, direct cDNA selection, sample sequencing of cosmids and PACs, and homology searches. On the basis of these mapping data, allelic loss analyses at 13q14 using CLL tumor samples allowed narrowing of the genomic segment encompassing the putative CLL gene to <300 kb. Twenty-three ESTs located within this minimally deleted region are candidate exons for the CLL-associated tumor suppressor gene.     

Light microscopic autoradiographic visualization of [3H]-arginine vasopressin binding sites in rat brain

Specific [³H]-arginine vasopressin ([³H]-AVP) binding sites were identified in the rat brain by light microscopic autoradiography. Discrete intrahypothalamic nuclei were densely labelled by [³H]-AVP. High specific binding was observed in the paraventricular and supraoptic nuclei. These binding sites may represent specific receptors for AVP, postulated to exist in the mammalian central nervous system.     

Alternative splicing and gene polymorphism of the human TAP3/SEC14L4 gene

Three closely related human SEC14p-like proteins (hTAP1, hTAP2, hTAP3, or SEC14L2, SEC14L3, SEC14L4, respectively) have been described that are related to the Saccharomyces cerevisiae SEC14 protein. These proteins may participate in intracellular lipid transport and influence regulatory lipid-dependent events. Here we report the isolation of an alternatively spliced hTAP3 cDNA and a polymorphism within the coding region of the hTAP3/SEC14L4 gene.     

Soil Patches of Inorganic Nitrogen in Subtropical Brazilian Plant Communities with Araucaria angustifolia

One-year old tubers of two hybrid calla lily (calla) cultivars (Zantedeschia ‘Pot of Gold’ and ‘Majestic Red’) were inoculated with the arbuscular mycorrhizal fungus (AMF), Glomus intraradices, or not, and grown at three different rates of phosphorus (P) supply to asses the effects of AMF-inoculation on plant development (time of shoot emergence and flowering), flowering (number, length and rate of flowering), and tuber biomass and composition over two growing cycles (2002, 2003). Tubers and flowers of calla responded differently to AMF inoculation. Differences in mycorrhizal responsiveness between cultivars was related to differences in P requirements for flower and tuber production, and the influence of P supply on resource allocation to different reproductive strategies. Inoculation increased shoot production and promoted early flowering, particularly in 2003. Inoculated plants also produced larger tubers than non-inoculated plants, but only increased the number of flowers per plant in 2003. High P supply also increased tuber biomass, but decreased the number of flowers per plant in 2002. Plants grown at a moderate P-rate, produced the most flowers in 2003. For ‘Majestic Red’, benefits from AMF were primarily in terms of tuber yield and composition, and AMF effects on marketable flower production could potentially have negative impact on production strategies for growers. Inoculation of ‘Pot of Gold’ primarily influenced flower production and aspects of tuber quality that caused detectable enhancement of tuber yield and flowering in the second growing cycle following inoculation (2003). The results of this study show that the responses of calla to AMF are partially a function of how nutrient supply alters resource allocation to sexual and vegetative reproduction. Whether AMF-induced changes in resource allocation to flowering and tubers significantly alters commercial productivity and quality of calla depends on the crop production goals (e.g. tubers, cut flowers or potted plants). The U.S. Government’s right to retain a non-exclusive, royalty free licence in and to any copyright is acknowledged.