CD4 expression decrease by antisense oligonucleotides. Inhibition of rat T CD4+ cell reactivity

In previous studies, we have demonstrated the inhibition of CD4 expression in rat lymphocytes treated with phorbol myristate acetate (PMA) by antisense oligonucleotides (AS-ODNs) directed against the AUG start region of the cd4 gene. The aim of the present study was to inhibit CD4 expression in lymphocytes without promoting CD4 synthesis and to determine the effect of this inhibition on CD4+ T cell function. Four 21-mer ODNs against the rat cd4 gene (AS-CD4-1 to AS-CD4-4) were used. Surface CD4 expression was measured by immunofluorescence staining and flow cytometry, and mRNA CD4 expression was measured by RT-PCR. T CD4+ cell function was determined by specific and unspecific proliferative response of rat-primed lymphocytes. After 24 hours of incubation, AS-CD4-2 and AS-CD4-4 reduced lymphocyte surface CD4 expression by 40%. This effect remained for 72 hours and was not observed on other surface molecules, such as CD3, CD5, or CD8. CD4 mRNA expression was reduced up to 40% at 24 hours with AS-CD4-2 and AS-CD4-4. After 48 hours treatment, CD4 mRNA decreased up to 27% and 29% for AS-CD4-2 and AS-CD4-4, respectively. AS-CD4-2 and AS-CD4-4 inhibited T CD4+ cell proliferative response upon antigen-specific and unspecific stimuli. Therefore, AS-ODNs against CD4 molecules inhibited surface and mRNA CD4 expression, under physiologic turnover and, consequently, modulate T CD4+ cell reactivity.

Acid-sensing ion channel 1a drives AMPA receptor plasticity following ischemia and acidosis in hippocampal CA1 neurons

The CA1 region of the hippocampus is particularly vulnerable to ischemic damage. While NMDA receptors play a major role in excitotoxicity, it is thought to be exacerbated in this region by two forms of post-ischemic AMPA receptor (AMPAR) plasticity - namely, anoxic long-term potentiation (a-LTP), and a delayed increase in the prevalence of Ca2+ -permeable GluA2-lacking AMPARs (CP-AMPARs). The acid-sensing ion channel 1a (ASIC1a) which is expressed in CA1 pyramidal neurons, is also known to contribute to post-ischemic neuronal death and to physiologically induced LTP. This raises the question - does ASIC1a activation drive the post-ischemic forms of AMPAR plasticity in CA1 pyramidal neurons? We have tested this by examining organotypic hippocampal slice cultures (OHSCs) exposed to oxygen glucose deprivation (OGD), and dissociated cultures of hippocampal pyramidal neurons (HPN) exposed to low pH (acidosis). We find that both a-LTP and the delayed increase in the prevalence of CP-AMPARs are dependent on ASIC1a activation during ischemia. Indeed, acidosis alone is sufficient to induce the increase in CP-AMPARs. We also find that inhibition of ASIC1a channels circumvents any potential neuroprotective benefit arising from block of CP-AMPARs. By demonstrating that ASIC1a activation contributes to post-ischemic AMPAR plasticity, our results identify a functional interaction between acidotoxicity and excitotoxicity in hippocampal CA1 cells, and provide insight into the role of ASIC1a and CP-AMPARs as potential drug targets for neuroprotection. We thus propose that ASIC1a activation can drive certain forms of CP-AMPAR plasticity, and that inhibiting ASIC1a affords neuroprotection.

CD4 expression decrease by antisense oligonucleotides. Inhibition of rat T CD4+ cell reactivity

In previous studies, we have demonstrated the inhibition of CD4 expression in rat lymphocytes treated with phorbol myristate acetate (PMA) by antisense oligonucleotides (AS-ODNs) directed against the AUG start region of the cd4 gene. The aim of the present study was to inhibit CD4 expression in lymphocytes without promoting CD4 synthesis and to determine the effect of this inhibition on CD4+ T cell function. Four 21-mer ODNs against the rat cd4 gene (AS-CD4-1 to AS-CD4-4) were used. Surface CD4 expression was measured by immunofluorescence staining and flow cytometry, and mRNA CD4 expression was measured by RT-PCR. T CD4+ cell function was determined by specific and unspecific proliferative response of rat-primed lymphocytes. After 24 hours of incubation, AS-CD4-2 and AS-CD4-4 reduced lymphocyte surface CD4 expression by 40%. This effect remained for 72 hours and was not observed on other surface molecules, such as CD3, CD5, or CD8. CD4 mRNA expression was reduced up to 40% at 24 hours with AS-CD4-2 and AS-CD4-4. After 48 hours treatment, CD4 mRNA decreased up to 27% and 29% for AS-CD4-2 and AS-CD4-4, respectively. AS-CD4-2 and AS-CD4-4 inhibited T CD4+ cell proliferative response upon antigen-specific and unspecific stimuli. Therefore, AS-ODNs against CD4 molecules inhibited surface and mRNA CD4 expression, under physiologic turnover and, consequently, modulate T CD4+ cell reactivity.

The paracaspase MALT1: biological function and potential for therapeutic inhibition.

The paracaspase MALT1 has a central role in the activation of lymphocytes and other immune cells including myeloid cells, mast cells and NK cells. MALT1 activity is required not only for the immune response, but also for the development of natural Treg cells that keep the immune response in check. Exaggerated MALT1 activity has been associated with the development of lymphoid malignancies, and recently developed MALT1 inhibitors show promising anti-tumor effects in xenograft models of diffuse large B cell lymphoma. In this review, we provide an overview of the present understanding of MALT1's function, and discuss possibilities for its therapeutic targeting based on recently developed inhibitors and animal models.

Valors típics de pH i DP del suport de tela en la pintura de cavallet

Ja fa varies decades que els conservadors-restauradors de paper van comenc;ar a tenir en compteel pH de les obres que tractaven, ja que es va veure que I'acidesa incidia molt directament en comde rapid I'objecte es debilitava mecimicament. De fet, trobem tant aviat com el 1936, una patenten registrada per OJ. Schierholtz per desacidificar el paper d'empaperar parets (Porck, 1996).La tela d'un quadre, essent un teixit fet a base de fibres vegetals compastes majoritariament perceHulosa, com les fibres del paper, és lógicament també facilment degradable si I'ambient ñes acid(te un baix pH).Sorprenentment, pero, I'acidesa de la tela deis quadres, tot i que en ocasions puntuals ha estatanalitzada (Bajocchi, 2009; Young, 1999), encara no és un parametre que es miri de forma rutinariai per tant, fins recentment, no se sabia quins eren els valors de pH més típics que pot tenir unquadre.Aquest estudi és una primera recopilació sistematica d'aquest t ipus d' in formació per a partir d'aquícrear un banc de dades que vagi recollint aquest t ipus d'informació sobre el majar nombre possiblede quadres.Si I'acidesa present en la tela ens indica com de rapid es degradara aquesta en el futur, la mesura delgrau de polimerització de la tela (DP), ens indica si la tela té una al ta o baixa resistencia mecanica enel moment actual. Així doncs, aquests dos parametres ens aporten informació molt úti l sobre I'estatde conserva ció del suportoAquesta recerca pretén respondre a les preguntes de quins són els valors de pH i DP que típicamentpodem trabar en la tela deis quadres, així com analitzar la relació entre aquests dos parametres i també la relació entre la data de producció deis quadres i els valors de pH i DP.

Valors típics de pH i DP del suport de tela en la pintura de cavallet

Ja fa varies decades que els conservadors-restauradors de paper van comenc;ar a tenir en compteel pH de les obres que tractaven, ja que es va veure que I'acidesa incidia molt directament en comde rapid I'objecte es debilitava mecimicament. De fet, trobem tant aviat com el 1936, una patenten registrada per OJ. Schierholtz per desacidificar el paper d'empaperar parets (Porck, 1996).La tela d'un quadre, essent un teixit fet a base de fibres vegetals compastes majoritariament perceHulosa, com les fibres del paper, és lógicament també facilment degradable si I'ambient ñes acid(te un baix pH).Sorprenentment, pero, I'acidesa de la tela deis quadres, tot i que en ocasions puntuals ha estatanalitzada (Bajocchi, 2009; Young, 1999), encara no és un parametre que es miri de forma rutinariai per tant, fins recentment, no se sabia quins eren els valors de pH més típics que pot tenir unquadre.Aquest estudi és una primera recopilació sistematica d'aquest t ipus d' in formació per a partir d'aquícrear un banc de dades que vagi recollint aquest t ipus d'informació sobre el majar nombre possiblede quadres.Si I'acidesa present en la tela ens indica com de rapid es degradara aquesta en el futur, la mesura delgrau de polimerització de la tela (DP), ens indica si la tela té una al ta o baixa resistencia mecanica enel moment actual. Així doncs, aquests dos parametres ens aporten informació molt úti l sobre I'estatde conserva ció del suportoAquesta recerca pretén respondre a les preguntes de quins són els valors de pH i DP que típicamentpodem trabar en la tela deis quadres, així com analitzar la relació entre aquests dos parametres i també la relació entre la data de producció deis quadres i els valors de pH i DP.

Exploring Brain Inhibition and Facilitation by Transcranial Magnetic Stimulation

Background and Question Paired-pulse TMS (Transcranial Magnetic Stimulation) paradigms allow explore motor cortex physiology. The Triple Stimulation Technique (TST) improves conventional TMS in quantifying cortico-spinal conduction. The objective of our study was to compare both methods in paired-pulse paradigms of inhibition and of facilitation. Method We investigated paired pulse paradigms of 2 ms (short intra-cortical inhibition) and of 10 ms intervals (intra cortical facilitation) in a randomized order in 22 healthy subjects applying conventional TMS and the TST protocol. Results Paired-pulse paradigms by both TMS and the TST yielded comparable results of short intra- cortical inhibition and intra cortical facilitation. However, the coefficient of variation was significantly smaller for SICI paradigm using TST. Conclusion These results suggest no greater sensitivity of the TST for quantifying inhibition and facilitation. The utility of TST to better quantify the individual amount of inhibition in SICI paradigms and its clinical utility need further studies.

Estudio de laboratorio del efecto de las concentraciones de calcio y caseína, el pH y la temperatura sobre la incorporación de proteínas lácteas al coágulo obtenido por acción enzimática

The effect of casein concentration, Ca2+ concentration, temperature and pH on the amount and size of protein aggregates (fines) in the whey produced by enzimic coagulation of nonfat milk was studied in laboratory conditions. Casein concentrations about 0.3 g/L showed a minimal amount of caseins in the whey, with presence of small aggregates of casein micellles. Ca2+ concentrations higher than 5 mM were neccesary to reduce the whey protein to a minimum constituted by protein particles smaller than casein micelles. The coagulation temperature, in the 35 - 45oC range, produced almost no variations in the whey proteins. The obtention of a minimum amount of whey proteins was possible only in a narrow pH range around 6.4. These results pointed to casein concentration and pH as important variables to be controlled in connection with the process yield.

Inhibition of nociceptive responses after systemic administration of amidated kyotorphin

BACKGROUND AND PURPOSE Kyotorphin (KTP; L-Tyr-L-Arg), an endogenous neuropeptide, is potently analgesic when delivered directly to the central nervous system. Its weak analgesic effects after systemic administration have been explained by inability to cross the blood-brain barrier (BBB) and detract from the possible clinical use of KTP as an analgesic. In this study, we aimed to increase the lipophilicity of KTP by amidation and to evaluate the analgesic efficacy of a new KTP derivative (KTP-amide - KTP-NH 2). EXPERIMENTAL APPROACH We synthesized KTP-NH 2. This peptide was given systemically to assess its ability to cross the BBB. A wide range of pain models, including acute, sustained and chronic inflammatory and neuropathic pain, were used to characterize analgesic efficacies of KTP-NH 2. Binding to opioid receptors and toxicity were also measured. KEY RESULTS KTP-NH 2, unlike its precursor KTP, was lipophilic and highly analgesic following systemic administration in several acute and chronic pain models, without inducing toxic effects or affecting motor responses and blood pressure. Binding to opioid receptors was minimal. KTP-NH 2 inhibited nociceptive responses of spinal neurons. Its analgesic effects were prevented by intrathecal or i.p. administration of naloxone. CONCLUSIONS AND IMPLICATIONS Amidation allowed KTP to show good analgesic ability after systemic delivery in acute and chronic pain models. The indirect opioid-mediated actions of KTP-NH 2 may explain why this compound retained its analgesic effects although the usual side effects of opioids were absent, which is a desired feature in next-generation pain medications