A novel strategy for development of glucocorticoids through non-genomic mechanism

Glucocorticoids (GCs) are routinely believed to take effect through genomic mechanisms, which are also largely responsible for GCs’ side effects. Beneficial non-genomic effects of GCs have been reported as being independent of the genomic pathway. Here, we synthesized a new type of GCs, which took effect mainly via non-genomic mechanisms. Hydrocortisone was conjugated with glycine, lysine and phenylalanine to get a bigger molecular structure, which could hardly go through the cell membrane. Evaluation of the anti-inflammatory efficacy showed that hydrocortisone-conjugated glycine (HG) and lysine could inhibit neutrophil degranulation within 15 min. HG could inhibit IgE-mediated histamine release from mast cells via a non-genomic pathway, and rapidly alleviate allergic reaction. Luciferase reporter assay showed that HG would not activate the glucocorticoid response element within 30 min, which verified the rapid effects independent of the genomic pathway. The work proposes a novel insight into the development of novel GCs, and provides new tools for experimental study on non-genomic mechanisms.     

Inactivation of the proximal NPXY motif impairs early steps in LRP1 biosynthesis

The proximal NPXY and distal NPXYXXL motifs in the intracellular domain of LRP1 play an important role in regulation of the function of the receptor. The impact of single and double inactivating knock-in mutations of these motifs on receptor maturation, cell surface expression, and ligand internalization was analyzed in mutant and control wild-type mice and MEFs. Single inactivation of the proximal NPXY or in combination with inactivation of the distal NPXYXXL motif are both shown to be associated with an impaired maturation and premature proteasomal degradation of full-length LRP1. Therefore, only a small mature LRP1 pool is able to reach the cell surface resulting indirectly in severe impairment of ligand internalization. Single inactivation of the NPXYXXL motif revealed normal maturation, but direct impairment of ligand internalization. In conclusion, the proximal NPXY motif proves to be essential for early steps in the LRP1 biosynthesis, whereas NPXYXXL appears rather relevant for internalization.     

A novel interplay between membrane and nuclear melatonin receptors in human lymphocytes: significance in IL-2 production

Human lymphocyte melatonin, through membrane and nuclear receptors binding, acts as an activator in IL-2 production. Antagonism of membrane melatonin receptors using luzindole exacerbates the drop of the IL-2 production induced by PGE₂ in peripheral blood mononuclear and Jurkat cells. This paper studies the melatonin membrane and nuclear receptors interplay in PGE₂-diminished IL-2 production. The decrease in IL-2 production after PGE₂ and/or luzindole administration correlated with downregulation in the nuclear receptor RORα. We also highlighted a role of cAMP in the pathway, because forskolin mimicked the effects of luzindole and/or PGE₂ in the RORα expression. Finally, a significant RORα downregulation was observed in T cells permanently transfected with inducible MT₁ antisense. In conclusion, we show a novel connection between melatonin membrane receptor signalling and RORα expression, opening a new way to understand melatonin regulation in lymphocyte physiology. Received 23 September 2008; received after revision 19 November 2008; accepted 21 November 2008     

A new aspect of the anti-stress effect of kinetin

Zusammenfassung Die mechanische Schädigung von Blattgeweben (das Abreiben der Blattoberfläche mit Karborundum oder eine rapide Infiltration der Gewebe mit H₂O) führt zu einer Steigerung der Ribonukleaseaktivität. Die Behandlung der Gewebe mit Kinetin, nicht aber die mit anderen Wachstumsregulatoren antagonisiert die Wirkung der mechanischen Verletzung (Stress) auf die Ribonukleaseaktivität.     

Diaryl pyrroles: A new series of anti-inflammatory agents

Zusammenfassung Die antiinflammatorischen Eigenschaften von verschiedenen Diaryl-pyrrol-Derivaten wurden untersucht. Es wurde festgestellt, dass eine repräsentative Verbindung, 2-Methyl-4, 5-bis(p-metoxyphenyl)-pyrrol, eine ausgeprägte Hemmung gegen das akute Ödem und die chronische Arthritis der Ratte zeigt.

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We thank Messrs.K. Wachi, Y. Umemuro andT. Kojima for their technical assistance.     

A comparative study of some anti-inflammatory drugs in wound healing of the rat

Resumen Varias drogas anti-inflamatorias incluyendo una droga antiinflamatoria no esteroidea nueva fueron investigadas en heridas producidas en ratas. AHR-1911 resultó ser la droga menos depresora de la cicatrización de las heridas cuando se comparó con la fenilbutazona y la dexametasona. Un menor contenido de colágeno en las heridas fue observado con el uso de AHR-1911.

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This study was in part presented at IV Latin-american Congress on Pharmacology and Therapeutics, Caracas, Venezuela, July 9–14, 1972.

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AHR-1911 was kindly supplied by ProfessorN. Ercoli. Dexamethasone and phenylbutazone were supplied by Merck Sharp and Dohme and Geigy Laboratories, respectively, Caracas, Venezuela.     

The expression profile of TLR9 mRNA and CpG ODNs immunostimulatory actions in the teleost gilthead seabream points to a major role of lymphocytes

The potential effects of synthetic unmethylated oligodeoxynucleotides (ODN) containing CpG motifs, mimicking bacterial DNA, has never been evaluated on the immune response in the teleost fish gilthead seabream (Sparus aurata), the most important fish species in Mediterranean aquaculture. First, binding and competition studies have demonstrated that binding is saturated and promiscuous, suggesting the participation of several receptors. Moreover, leucocyte cytotoxic (NCC) activity, production of ROIs (reactive oxygen intermediates), and expression of immune-relevant genes was greatly primed by ODNs. Focusing on the mechanism, the TLR9 gene is widely distributed in seabream tissues and differently regulated in vitro by several stimuli. Moreover, and for the first time in fish, TLR9 mRNA has been detected in lymphocytes as the main cell-source. To conclude, ODNs containing GACGTT, GTCGTT (optimal for mouse and human, respectively) or AACGTT motifs are the most potent inducers of seabream immunity, whilst the involvement of TLR9 is under debate.     

A radiometric method for the quantitation of experimental inflammation and anti-inflammatory activity

Zusammenfassung Nach Einspritzung von 5-Hydroxy-tryptamin in den einen Fuss einer Maus und physiologischer Salzlösung in den anderen, wurde die Radioaktivität beider Füsse verglichen. Mit radioaktivem Jod (I¹³¹) behandeltes Serumalbumin wurde dazu benutzt, den Umfang des Plasmaexsudates zu bestimmen. Das Ausmass der Hemmung der durch 5-HT induzierten Entzündung ist von der Dosis der physiologischen NaCl-Lösung abhängig.

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This study was supported in part by a grant from Sunkist Growers, Ontario (California).