Arabidopsis thaliana cdd1 mutant uncouples the constitutive activation of salicylic acid signalling from growth defects

Arabidopsis genotypes with a hyperactive salicylic acid-mediated signalling pathway exhibit enhanced disease resistance, which is often coupled with growth and developmental defects, such as dwarfing and spontaneous necrotic lesions on the leaves, resulting in reduced biomass yield. In this article, we report a novel recessive mutant of Arabidopsis, cdd1 (constitutive defence without defect in growth and development1), that exhibits enhanced disease resistance associated with constitutive salicylic acid signalling, but without any observable pleiotropic phenotype. Both NPR1 (NON-EXPRESSOR OF PATHOGENESIS-RELATED GENES1)-dependent and NPR1-independent salicylic acid-regulated defence pathways are hyperactivated in cdd1 mutant plants, conferring enhanced resistance against bacterial pathogens. However, a functional NPR1 allele is required for the cdd1-conferred heightened resistance against the oomycete pathogen Hyaloperonospora arabidopsidis. Salicylic acid accumulates at elevated levels in cdd1 and cdd1 npr1 mutant plants and is necessary for cdd1-mediated PR1 expression and disease resistance phenotypes. In addition, we provide data which indicate that the cdd1 mutation negatively regulates the npr1 mutation-induced hyperactivation of ethylene/jasmonic acid signalling.     

Localization of vitamin D3-responsive alkaline phosphatase in cultured chondrocytes

Alkaline phosphatase activity appears to be altered when chondrocyte cultures are incubated with 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). This study examined whether the hormone-responsive enzyme activity is associated with alkaline phosphatase-enriched extracellular membrane organelles called matrix vesicles. Confluent, third passage cultures of rat costochondral growth cartilage (GC) or resting zone chondrocytes (RC) were incubated with 1,25-(OH)2D3 or 24,25-dihydroxyvitamin D3 (24,25-(OH)2D3) and enzyme specific activity was assayed in the cell layer or in isolated matrix vesicle and plasma membrane fractions. Alkaline phosphatase-specific activity in the matrix vesicles was enriched at least 2-fold over that of the plasma membrane and 10-fold over that of the cell layer. Matrix vesicle alkaline phosphatase was stimulated by 1,25-(OH)2D3 in GC cultures and by 24,25-(OH)2D3 in RC cultures. The cell layer failed to reveal these subtle differences. 1,25-(OH)2D3 increased GC enzyme activity but the effect was one-half that observed in the matrix vesicles alone. No effect of 1,25-(OH)2D3 on enzyme activity of the RC cell layer or of 24,25-(OH)2D3 on either GC or RC cell layers was detected. Thus, response to the metabolites is dependent on chondrocytic differentiation and is site specific: the matrix vesicle fraction is targeted and not the cells per se.     

A Mathematical Model of the Stability Control of Human Thorax and Pelvis Movements During Walking

A mathematical model is developed to study the human thorax and pelvis movements in the frontal plane during normal walking. The model comprises of two-link base-excited inverted pendulums with one-degree of rotational freedom for each link. Since the linear motion of the pelvis has a significant effect on the upper body stability, this effect is included in the model by having a base point moving in the frontal plane in a general way. Furthermore, because the postural stability is the primary requirement of normal human walking, the control law is developed based on Lyapunov's stability theory, which guarantees the stability of the pendulum system around the up-right position. To evaluate the model, the simulation results, including the angular displacement of each link and the torque applied on each link, are compared with those from gait measurements. It is shown that the simulation results match those from gait measurements closely. These results suggest that the proposed model can provide a useful framework for analysis of postural control mechanisms.     

SAP enables T cells to help B cells by a mechanism distinct from Th cell programming or CD40 ligand regulation

Genetic mutations disrupting the function of signaling lymphocytic activation molecule-associated protein (SAP) lead to T cell intrinsic defects in T cell-dependent Ab responses. To better understand how SAP enables Th cells to help B cells, we first assessed whether molecules important for B cell help are dysregulated in SAP-deficient (SAP knockout (KO)) mice. CD40 ligand (CD40L) expression was enhanced on unpolarized SAP KO T cells; however, Th2 polarization returned their CD40L expression to wild-type levels without rescuing their ability to help B cells. CD40L also localized normally to the site of contact between SAP KO T cells and Ag-bearing B cells. Finally, CD40L-deficient Th cells and SAP KO Th cells differed in their abilities to help B cells in vitro. These data argue that Ab defects caused by SAP deficiency do not result from a loss of CD40L regulation or CD40L function on CD4 T cells. SAP KO Th cells additionally displayed normal patterns of migration and expression of ICOS and CXCR5. Global gene expression was remarkably similar in activated SAP KO vs wild-type T cells, prompting us to investigate whether SAP is necessary for "programming" T cells to become B cell helpers. By restricting SAP expression during differentiation, we determined that SAP is not required during the first 5 days of T cell activation/differentiation to generate Th cells capable of helping B cells. Instead, SAP is necessary for very late stages of differentiation or, most likely, for allowing Th cells to communicate during cognate T:B interactions.     

A novel colonic repressor element regulates intestinal gene expression by interacting with Cux/CDP

Intestinal gene regulation involves mechanisms that direct temporal expression along the vertical and horizontal axes of the alimentary tract. Sucrase-isomaltase (SI), the product of an enterocyte-specific gene, exhibits a complex pattern of expression. Generation of transgenic mice with a mutated SI transgene showed involvement of an overlapping CDP (CCAAT displacement protein)-GATA element in colonic repression of SI throughout postnatal intestinal development. We define this element as CRESIP (colon-repressive element of the SI promoter). Cux/CDP interacts with SI and represses SI promoter activity in a CRESIP-dependent manner. Cux/CDP homozygous mutant mice displayed increased expression of SI mRNA during early postnatal development. Our results demonstrate that an intestinal gene can be repressed in the distal gut and identify Cux/CDP as a regulator of this repression during development.     

Expression of gibberellin mutations in fruits of Pisum sativum L.

The gibberellin (GA) economy of young pea (Pisum sativum L.) fruits was investigated using a range of mutants with altered GA biosynthesis or deactivation. The synthesis mutation lh-2 substantially reduced the content of both GA₄ and GA₁ in young seeds. Among the other synthesis mutations, ls-1, le-1 and le-3, the largest reduction in seed GA₁ content was only 1.7-fold (le-1), while GA₄ was not reduced in these mutants, and in fact accumulated in some experiments (compared with the wild type). Mutation sln appeared to block the step GA₂₀ to GA₂₉ in young pods and seeds, but not as strongly as in older seeds. Mutations ls-1, le-1 and le-3 markedly reduced pod GA₁ levels, but pod elongation was not affected. After feeds of [¹³C,³H]GA₂₀ to leaves, the pods contained ¹³C,³H-labelled GA₂₀, GA₁, GA₂₉ and GA₈₁, and the seeds, [¹³C,³H]GA₂₀ and [¹³C,³H]GA₂₉. These findings are discussed in relation to recent suggestions regarding the role and origin of GA₁ in pea fruits. Received: 6 June 1997 / Accepted: 15 July 1997     

Lymphokine-mediated activation of a T cell-dependent IgA antipolysaccharide response

Immune responses to bacterial polysaccharides are important to host immunity at mucosal surfaces. We previously showed that BALB/c mice produce substantial T cell-dependent IgA responses to alpha (1,3) glucan determinants on the bacterial capsular polysaccharide dextran B1355. The data in this study demonstrate that the requirement for T cells for the activation of the IgA anti-alpha (1,3) dextran B1355 response can be replaced by T cell-derived nonantigen specific helper factors that appear to act during the late stages of B cell differentiation. Supernatants from the activated T cell lines cr-15 and (DL)C.C3.11.75, which contain interferon and late-acting T cell replacing factor activity, supported terminal differentiation of dextran-stimulated B cells to IgA anti-alpha (1,3) glucan antibody-forming cells and substantially increased IgM anti-alpha (1,3) glucan responses in culture. Although supernatants with interleukin 2 activity did not support optimal antigen-driven plaque-forming cell responses, they synergized with supernatants having interferon and T cell replacing factor activity in the production of IgA and IgM anti-alpha (1,3) glucan responses and IgM anti-SRBC responses. Supernatants from the T cell lines B6.11 and (DL)A.4 contained B cell growth factor activity but did not support activation of IgA anti-alpha (1,3) glucan PFC. These studies suggest that interferon and/or T cell replacing factor play an important role in the antigen-driven differentiation of B cells of the IgA and IgM isotypes to antibody-forming cells.