The effect of acidic polysaccharides and prostaglandin-like substances isolated from Propionibacterium acnes on granulocyte chemotaxis.

Three acidic polysaccharide (AP) fractions and the prostaglandin-like substances (PLS) isolated from P. acnes were investigated regarding their chemotactic activities on polymorphonuclear leukocytes. Both AP's and PLS induced a significant chemotatic response, which suggests their involvement in inflammatory acne vulgaris.     

Evidence that the prostaglandin-like substances fromPropionibacterium acnes are not identical with PGE2

Summary The prostaglandin-like substances (PLS) isolated fromP. acnes were investigated by reversed phase chromatography and gas chromatography-mass spectrometry. These analyses demonstrated that PLS were not identical with PGE₂, which supports a concept of PLS as a potential mediator of the inflammatory process in acne vulgaris.     

Prostaglandin-like substances inPropionibacterium acnes II. Stimulatory effect on ovarian cyclic AMP

Summary The prostaglandin-like substances (PLS) fromPropionibacterium acnes increased the ovarian tissue levels of cyclic AMP (cAMP) approximately 2-fold. The lipid material extracted fromP. acnes thus behaved like PG's of the E-type, and since it is unlikely that other known stimulators of the ovarian cAMP system can be present in the bacterial lipid fraction, these experiments give further evidence in favour of the occurrence of PLS inP. acnes.     

Prostaglandin-like substances inPropionibacterium acnes. IV. Effect on isolated human vessels

Summary The present study demonstrates a powerful vasoconstrictor activity of prostaglandin-like substances (PLS), extracted fromP. acnes, on human blood vessels. PLS is about equipotent to PGE₂ in its effect on human umbilical vessels, but the contractile response pattern is different. PLS therefore seems to have specific and different physiological characteristics.     

Prostaglandin-like substances inPropionibacterium acnes. V. Activity profiles using cascade superfusion bioassay and platelet aggregation

Summary The activity spectrum of prostaglandin-like substances (PLS) fromP. acnes was investigated with cascade superfusion technique and by platelet aggregation assay. The biological activity of PLS resembles that of PGI₂: both relax bovine coronary artery, rabbit mesentric and coeliac arteries; both contract the rat stomach strip as well as both typically inhibit spontaneous movements of isolated guinea pig ileum. Also, similarly to PGI₂, PLS inhibits platelet aggregation regardless the inducer used. However, PLS possesses a specific antiaggregatory pattern on platelet, which indicates that these compounds are not indentical with primary prostaglandins or PGI₂.     

Leukocyte integrins and inflammation

Leukocyte adhesion is of pivotal functional importance. Without adequate adhesion, T lymphocytes and natural killer cells are not cytotoxic, B cells cannot develop into antibody secreting plasma cells, leukocytes do not home into inflamed tissues and myeloid cells are not able to phagocytize or exhibit chemotactic responses. During evolution several leukocyte adhesion molecules have developed belonging to a few molecular families. Among these, the leukocyte-specific integrins (β ₂ integrins, CD11/CD18 molecules) are among the most important. Much progress has taken place during the past few years, and at present we have a considerable knowledge of their structure and function. Inflammation is critically dependent on integrin activity, and its regulation forms the topic of this short review.     

Transduction of the chemotactic cAMP signal across the plasma membrane ofDictyostelium cells

AggregatingDictyostelium cells secrete cAMP during cell aggregation. cAMP induces two fast responses, the production of more cAMP (relay) and directed cell locomotion (chemotaxis). Extracellular cAMP binds to G-protein-coupled receptors leading to the activation of second messenger pathways, including the activation of adenylyl cyclase, guanylyl cyclase, phospholipase C and the opening of plasma membrane Ca²⁺ channels. Many genes encoding these sensory transduction proteins have been cloned and null mutants of nearly all components have been characterized in detail. Undoubtedly, activation of adenylyl cyclase is the most complex, involving G-proteins, a soluble protein called CRAC and components of the MAP kinase pathway. Null mutants in this pathway do not aggregate, but can exhibit chemotaxis and develop normally when supplied with exogenous cAMP. The pathways leading to the activation of phospholipase C were identified, but unexpectedly, deletion of the phospholipase C gene has no effect on chemotaxis and development, nor on intracellular Ins(1,4,5)P₃ levels; the metabolism of this second messenger will be discussed in some detail. Activation of guanylyl cyclase is G-protein-dependent and essential for chemotaxis. Analysis of a collection of chemotactic mutants reveals that most mutants are defective in either the production or intracellular detection of cGMP, thereby placing this second messenger at the center of chemotactic signal transduction. Analysis of the cAMP-mediated opening of plasma membrane calcium channels in signal transduction mutants suggests that it has two components, one that depends on G-proteins and intracellular cGMP and one that is G-protein-independent.     

Gain of function of TMEM16E/ANO5 scrambling activity caused by a mutation associated with gnathodiaphyseal dysplasia

Mutations in the human TMEM16E (ANO5) gene are associated both with the bone disease gnathodiaphyseal dysplasia (GDD; OMIM: 166260) and muscle dystrophies (OMIM: 611307, 613319). However, the physiological function of TMEM16E has remained unclear. We show here that human TMEM16E, when overexpressed in mammalian cell lines, displayed partial plasma membrane localization and gave rise to phospholipid scrambling (PLS) as well as non-selective ionic currents with slow time-dependent activation at highly depolarized membrane potentials. While the activity of wild-type TMEM16E depended on elevated cytosolic Ca²⁺ levels, a mutant form carrying the GDD-causing T513I substitution showed PLS and large time-dependent ion currents even at low cytosolic Ca²⁺ concentrations. Contrarily, mutation of the homologous position in the Ca²⁺-activated Cl^? channel TMEM16B paralog hardly affected its function. In summary, these data provide the first direct demonstration of Ca²⁺-dependent PLS activity for TMEM16E and suggest a gain-of-function phenotype related to a GDD mutation.     

Prostaglandin-like substances in Propionibacterium acnes II. Stimulatory effect on ovarian cyclic AMP.

The prostaglandin-like substances (PLS) from Propionibacterium acnes increased the ovarian tissue levels of cyclic AMP (cAMP) approximately 2-fold. The lipid material extracted from P. acnes thus behaved like PG's of the E-type, and since it is unlikely that other known stimulators of the ovarian cAMP system can be present in the bacterial lipid fraction, these experiments give further evidence in favour of the occurrence of PLS in P. acnes.     

Kinetics of chemo-attraction of polymorphonuclear leukocytes towards N-formyl peptide studied with a novel polycarbonate (Nucleopore) membrane in the Boyden chamber

The motile responses of human polymorphonuclear leukocytes (PMN) to N-formyl-methionyl-leucyl-phenylalanine (FMLP) in the Boyden chamber using a new sparse-pore polycarbonate membrane (pores 3 micron in diameter and occupying 0.1% of surface area) were compared with those demonstrated by using a standard polycarbonate (Nuclepore) filtration membrane (pores 3 micron in diameter and occupying 5% of surface area). Motility of PMN in gradients of FMLP using the new membrane was not influenced by chemokinetic effects of the factor, and the 'background' migration of the cells was minimal. However, motility of PMN in gradients of FMLP using the standard membrane was found to be influenced by chemokinetic effects of the chemotactic factor, and the 'background' or 'control' migration (in the absence of chemotactic factor) of the cells was substantial. Greater directional migration of PMN according to steepness of the gradient of chemotactic factor was demonstrated with the use of the new membrane. The new membrane may be of considerable value in the further study of the chemotactic responses of PMN.     

Cell migration to the chemokine CXCL8: Paxillin is activated and regulates adhesion and cell motility

The chemokine CXCL8 is a powerful inducer of directional cell motility, primarily during inflammation. In this study, we found that CXCL8 stimulation led to paxillin phosphorylation in normal neutrophils, and that both CXCL8 receptors (CXCR1 and CXCR2) mediated CXCL8-induced paxillin phosphorylation. In CXCR2-transfected cells, the process depended on G_αi and G_αs coupling to CXCR2. Dominant negative (DN) paxillin increased CXCL8-induced adhesion and migration, indicating that endogenous paxillin keeps migration at submaximal levels. Furthermore, using activating antibodies to β1 integrins, analyses with focal adhesion kinase (FAK) DN variant (FRNK) and co-immunoprecipitations of FAK and paxillin, we found that β1 integrin ligation cooperates with CXCL8-induced stimulation, leading to FAK activation and thereafter to FAK-mediated paxillin phosphorylation. Our findings indicate that paxillin keeps directional motility at a restrained magnitude, and suggest that perturbations in its activation may lead to chemotactic imbalance and to pathological conditions associated with excessive or reduced leukocyte migration. R. Mintz, T. Meshel: These authors contributed equally to this work. Received 31 July 2008; received after revision 14 December 2008; accepted 16 December 2008     

Prostaglandin-like substances in Propionibacterium acnes. III. Differential contractile effects on smooth muscle layers of the human utero-tubal junction.

The biological activity of a lipid fraction extracted from P. acnes was tested on isolated smooth muscle strips from the human utero-tubal junction. The bioassay experiments support the concept that prostaglandin-like substances (PLS) occur in P. acnes. However, in the bioassay system used, the effect of PLS was different from that of PGF2 alpha and PGI2 but similar, although not identical, to that of arachidonic acid and PGE2.     

Bioassay for hamster macrophage chemotaxis: application to study particle-lung interactions

Attraction of lung macrophages to particle deposition sites has been demonstrated in different animal species. We reported a threefold increase of the number of macrophages to occur within 40 min after polystyrene particle deposition in hamster airways [Geiser et al. (1994) Am. J. Respir. Cell Mol. Biol. 160: 594–603]. Complement-derived chemotactic activity is one of the mechanisms postulated for macrophage recruitment. It was the aim of this study to test whether complement-derived chemotactic activity is involved in the rapid recruitment of macrophages to the site of deposited polystyrene particles in hamster airways. We first developed an in vitro cell migration assay for hamster macrophages to assess complement-derived chemotaxis. Second, the bronchoalveolar lavage fluids (BALF) of four hamsters that had inhaled aerosols of polystyrene microspheres were tested for chemotactic activity by this bioassay and compared with BALF of four sham-exposed hamsters. Chemotactic response of macrophages was found toward complement-activated hamster serum, whereas macrophage migration was not increased toward BALF of particle and sham-exposed hamsters. In contrast, macrophage migration to BALF of both groups was reduced by 1.6-fold. Thus, the stimulus for macrophage recruitment to the site of deposited polystyrene particles in hamster airways could not be demonstrated using this bioassay. Received 10 September 1997; received after revision 24 November 1997; accepted 10 December 1997     

The effect of mouse erythrocyte rosette forming lymphocytes on lymphokine production in T-cell cultures

An enhancement effect of mouse erythrocyte rosette forming (MERF) cells on the production of migration inhibitory factor, chemotactic factor for neutrophils and skin reactive factor in T-lymphocyte cultures stimulated with the purified protein derivative of tuberculin was observed. We consider it likely that the MERF cells, possessing the appropriate cell surface constituents to construct an immunogenic moiety, present antigen on their surfaces to elicit lymphokine production.     

Kinetics of chemo-attraction of polymorphonuclear leukocytes towards N-formyl peptide studied with a novel polycarbonate (Nuclepore) membrane in the Boyden chamber

Summary The motile responses of human polymorphonuclear leukocytes (PMN) to N-formyl-methionyl-leucyl-phenylalanine (FMLP) in the Boyden chamber using a new sparse-pore polycarbonate membrane (pores 3 m in diameter and occupying 0.1% of surface area) were compared with those demonstrated by using a standard polycarbonate (Nuclepore) filtration membrane (pores 3 m in diameter and occupying 5% of surface area). Motility of PMN in gradients of FMLP using the new membrane was not influenced by chemokinetic effects of the factor, and the background migration of the cells was minimal. However, motility of PMN in gradients of FMLP using the standard membrane was found to be influenced by chemokinetic effects of the chemotactic factor, and the background or control migration (in the absence of chemotactic factor) of the cells was substantial. Greater directional migration of PMN according to steepness of the gradient of chemotactic factor was demonstrated with the use of the new membrane. The new membrane may be of considerable value in the further study of the chemotactic responses of PMN.     

The effect of mouse erythrocyte rosette forming lymphocytes on lymphokine production in T-cell cultures

Summary An enhancement effect of mouse erythrocyte rosette forming (MERF) cells on the production of migration inhibitory factor, chemotactic factor for neutrophils and skin reactive factor in T-lymphocyte cultures stimulated with the purified protein derivative of tuberculin was observed. We consider it likely that the MERF cells, possessing the appropriate cell surface constituents to construct an immunogenic moiety, present antigen on their surfaces to elicit lymphokine production.This work was supported by the Scientific Research Council, Ministry of Health, Hungary, Code No. 421030401 1/s     

SV40 transformed fibroblasts recognize the same 140 kD fibronectin chemotactic fragment as non-transformed cells

Summary SV40 virus-transformed human embryonal fibroblasts show an enhanced chemotactic response to the glycoprotein fibronectin. However, they recognize the same chemotactic active region as non-transformed fibroblasts. The result suggests that an enhancement of chemotaxis by fibroblasts which have been transformed with Simian Virus 40 is due not to the utilization of further chemotactic domains in the molecule, but to an increased sensitivity of the cells to the chemoattractant.     

SV40 transformed fibroblasts recognize the same 140 kD fibronectin chemotactic fragment as non-transformed cells

SV40-virus-transformed human embryonal fibroblasts show an enhanced chemotactic response to the glycoprotein fibronectin. However, they recognize the same chemotactic active region as non-transformed fibroblasts. The result suggests that an enhancement of chemotaxis by fibroblasts which have been transformed with Simian Virus 40 is due not to the utilization of further chemotactic domains in the molecule, but to an increased sensitivity of the cells to the chemoattractant.     

Radioprotective and haemopoietic effects of some lipopolysaccharides from Rhodospirillaceae species in mice

Summary Lipopolysaccharides (LPS) from variousRhodopseudomonas andRhodospirillum species were tested for their radioprotective efficiency against X-irradiation and for their influence on the growth of spleen colony forming units (CFU-s) in mice. The LPS fromRhodopseudomonas gelatinosa Dr₂ gave a high survival rate. It also favoured CFU-s formation and erythroid differentiation.     

基于可见/近红外光谱的土壤有机质含量预测

应用可见/近红外光谱技术对土壤有机质含量进行了定量分析和预测,为土壤肥力快速测定和评价提供依据.利用ASD FieldSpec 3 Hi-Res光谱仪对116份不同有机质含量的土壤样本进行光谱测量,系统分析了土壤有机质含量与350～2500 nm波段范围光谱反射率之间的关系.利用PLS和小波-BP神经网络对350～2500 nm整个波段范围和剔除水波段的光谱数据进行分析.两种建模方法的结果均表明剔除水波段的预测效果较好,其中,PLS模型预测的相关系数R为0.8416,均方根误差RMSEP为0.2848,相对分析误差RPD为1.7768,WT-BP神经网络模型预测的R为0.9167,RMSEP为0.2196,RPD为2.3043.预测结果表明,PLS模型可以对土壤有机质含量进行粗略估测,而BP神经网络可实现较精确的预测.