Liofilchem? O.A. Listeria agar and direct CAMP test provided sooner Listeria monocytogenes identification from neonatal bacteremia

Listeria monocytogenes infection in pregnant women and newborns is a cause for serious concern, and invasive disease outcome strongly depends on prompt antibiotic therapy. To provide sooner identification from neonatal bacteremia we performed a CAMP test directly on positive blood aliquots and inoculated the Liofilchem^® O.A. Listeria chromogenic agar as well, thus providing a 24-h turn-around time for response.     

<Emphasis Type="Italic">Streptococcus agalactiae</Emphasis> CAMP factor/protein B does not bind to human IgG

CAMP factor is an extracellular cytolytic protein produced by Streptococcus agalactiae. CAMP factor has been reported to bind the Fc fragments of immunoglobulin G (IgG) and has therefore also been called protein B, in analogy to protein A of Staphylococcus aureus. We attempted to characterize the interaction of protein B with IgG in more detail. In contrast to protein A, CAMP factor does not inhibit the activation of complement by hemolysin antibodies bound to sheep red cell surfaces. IgG also failed to inhibit the co-hemolytic activity of CAMP factor, which is in disagreement with previous findings. After co-incubation, CAMP factor and IgG were cleanly separated by gel filtration, indicating that no binding had occurred. Waseem El-Huneidi and Ryan Mui contributed equally to this work.     

洛伐他汀对糖尿病大鼠肾功能及肾脏组织p38丝裂原激活蛋白激酶表达的影响

目的 探讨洛伐他汀对糖尿病(DM)大鼠肾小球结构、功能及肾组织p38丝裂原活化蛋白激酶(MAPK)及其下游转录因子cAMP反应元件结合蛋白(cREB)表达的影响。方法 18只雄性Wistar大鼠行右肾切除术2周后,随机分为3组:右肾切除对照组、DM组和洛伐他汀治疗组每组6只。DM组和洛伐他汀组腹腔注射链脲佐菌素(STZ,65 mg／kg)诱发DM模型,洛伐他汀组制模后1 d每日给予洛伐他汀20 mg／kg灌胃。分别于注射STZ后4周收集大鼠尿液,测定尿蛋白(Upro)、尿肌酐(UCr);股动脉放血分离血清,测定血糖(Glu)、血肌酐(SCr)、尿素氮(BUN)、胆固醇(CHO)和甘油三酯(TG),并计算肌酐清除率(CCr)。免疫组化检测肾皮质磷酸化p38 MAPK(P-p38 MAPK)及磷酸化CREB(P-CREB)的表达特征,并检测转化生长因子-β1(TGF-β1)、纤维粘连蛋白(FN)及层粘连蛋白(LN)的表达,应用图像分析系统进行定量分析。流式细胞术检测P-p38 MAPK和P-CREB蛋白的表达,Western印迹法检测肾皮质P-p38 MAPK和P-CREB活性的变化。结果 与对照组相比,4周时DM组P-p38 MAPK、P-CREB、TGF-β1、FN及LN表达均明显升高(P均<0.01);而与DM组相比,洛伐他汀组各指标的表达有不同程度的降低(P均<0.01)。结论 洛伐他汀对DM大鼠肾脏结构和功能具有保护作用,可能通过调节p38 MAPK和CREB蛋白的表达     

Interaction Between α and β-Adrenergic Receptor Agonists Modulating the Slow Ca2+-activated K+ Current - IAHP in Hippocampd Neurons

Noradrenaline inhibits the Ca²⁺-activated K⁺ current I_AHP, which underlies the slow afterhyperpolarization and spike frequency adaptation in hippocampal and neocortical neurons. The resulting increase in excitability probably contributes to the state control of the forebrain during arousal and attention. The modulation of I_AHP by noradrenaline has previously been shown to be mediated by β₁ receptors, cyclic AMP and protein kinase A, but not by α receptors. We have now tested the possibility that a receptors also contribute to I_AHP modulation through interaction with β receptors, by the use of whole-cell recordings in CA1 pyramidal cells of rat hippocampal slices. The α-receptor agonist 6-fluoro-noradrenaline strongly potentiated the effect of isoproterenol on I_AHP. The synergistic effect of 6-fluoro-noradrenaline and soproterenol was blocked by the β-receptor antagonist timolol, but the receptor type mediating the effect of 6-fluoro-noradrenaline could not be unequivocally identified by using α-receptor antagonists. The effect of high concentrations of noradrenaline on I_AHP was only partly blocked by the β-receptor antagonist timolol, and was further reduced by blocking α receptors, again suggesting a contribution from α receptors. In contrast, the effect of low concentrations of noradrenaline seemed to be potentiated by the α-receptor antagonist phentolamine in 57% of the cells, suggesting concentration dependent antagonistic interaction between α and β receptors. Further tests indicated that the cross-talk between 6-fluoro-noradrenaline and isoproterenol occurs upstream from cyclic AMP production, and that protein kinase A serves as a final common path for the modulation of I_AHP by noradrenaline, and by the combination of 6-fluoro-noradrenaline and isoproterenol.     

Homologous desensitization of human corticotropin-releasing factor, receptor in stable transfected mouse fibroblast cells

Previous radioligand binding and second messenger studies have shown that corticotropin-releasing factor (CRF) modulates its receptor following both in vivo and in vitro treatment. In the present study, we determined the sequence of events leading to CRF-induced downregulation and desensitization of cloned CRF receptors in murine fibroblast cells (Ltk⁻) stably transfected with CRF, DNA (from human pituitary). Treatment of cells with rat/human CRF produced a dose- and time-dependent decrease in [¹²⁵I]Tyr^o-ovine CRF ([¹²⁵I]oCRF) binding and a concomitant decrease in CRF-stimulated adenylate cyclase activity. Significant decreases in [¹²⁵I]oCRF binding and agonist-stimulated cAMP production were evident minutes after CRF treatment with maximal (60–80%) reductions seen following 1 h of CRF treatment. Scatchard analysis revealed that the decrease in [¹²⁵I]oCRF binding was due to the downregulation of the receptor with no significant alteration seen in the affinity of the ligand. Since the transfected cell line is engineered using an artificial promoter, we did not detect any significant changes in CRF₁ receptor mRNA levels following CRF treatment for up to 24 h.     

Effects of dibutyryl cyclic AMP and forskolin on phospholipid biosynthesis in thrombin-stimulated human platelets

The influence of forskolin, an adenylate cyclase activator, and of dibutyryl cyclic AMP (Bt2cAMP) on [3H]glycerol incorporation into glycerolipids was investigated in human platelets. It was found that preincubation with 2.5 mM Bt2cAMP produced a 2-4-fold increase in thrombin-induced incorporation into phospholipids compared to platelets activated by thrombin alone. Pretreatment with forskolin, which increased cellular cAMP content, also resulted in an increase in thrombin-stimulated [3H]glycerol incorporation into phospholipids. These findings demonstrate that a rise in platelet cAMP can accentuate thrombin-induced de novo synthesis of phospholipids from [3H]glycerol. Since the content of cellular cAMP was correlated with its ability to inhibit platelet activation monitored by serotonin release, it seems likely that glycerolipid, in particular phospholipid biosynthesis, is involved in controlling platelet activation by thrombin.     

Early effects of hCG on human testicular cyclic AMP content, protein kinase activity, in-vitro progesterone conversion and the serum concentrations of testosterone and oestradiol

Ten infertile men underwent testicular biopsy. Cyclic AMP concentration and cAMP-dependent protein kinase activity were determined in biopsies obtained before, and at 3, 10, 20 and 30 min after an intravenous injection of hCG (1500-5000 IU). The in-vitro conversion of progesterone by testicular tissue, and the serum concentrations of testosterone and oestradiol were then studied before and at 30 min after hCG injection. Intravenous injection of hCG induced a rapid increase in cAMP concentration and in the activity of cAMP-dependent protein kinase. The kinetics of this response indicated that cAMP and cAMP-dependent protein kinase mediate hCG effects on the human testis, presumably via effects on the Leydig cells. No stimulatory effect on steroid conversion in vitro or on the serum concentrations of testosterone and oestradiol were seen after 30 min.