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Ricardo J. Bosch (ESPAÑA): Parathyroid hormone-related protein as a renal regulatory factor: from vessels to glomeruli and tubular epithelium

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Session Start: Thu Feb 21 22:00:17 2000
[22:04] (MJesus) gaucho!!!
[22:04] * MJesus present gaucho: he is from Uruguay!.... Dr. Caporale
[22:04] (gerado) Nelson bienvenido
[22:05] (gaucho) hola gerardo
[22:05] (JBover) hello santiago
[22:06] (RJ_Bosch) hola a todos
[22:06] (JBover) ricardo!!!!!!!
[22:06] (santiago) buenas noches a todos
[22:06] (JBover) Ricardo J Bosch is one of the guest speakers tonight
[22:06] (MJesus) ricardo, en español o ingles ?
[22:06] (JBover) May I say something????
[22:06] (RJ_Bosch) ingles
[22:07] (JBover) Great Ricardo
[22:07] (JBover) well...alll of you know that today we should have all invited speakers on renal physiology
[22:08] (JBover) Dr Halperin could not be here with us to talk about Integrative physiology
[22:08] (JBover) He had a conference somewhere else
[22:09] (JBover) On the other hand..Dr Soriano declined the offer of being here online unfortunately. Obviously, most of our invited speakers are famous and have busy schedules
[22:10] (JBover) Dr Charra should be here as well as DR SHaldon
[22:10] (JBover) Unfortunately we made a great mistake and Dr SHaldon was not properly announced
[22:10] (JBover) Dr Shaldon will be with us next Monday the 28th
[22:10] (JBover) Dr Charra is on duty
[22:11] (JBover) HE will be probably be here sometime
[22:11] (Gainza) Dr Soriano told me to excuse his absence
[22:11] (JBover) but obviously, patients go first
[22:11] (JBover) Thanks Dr GAinza
[22:11] (JBover) as I said, most of our invited speakers have very busy schedules
[22:12] (JBover) if you agree....we can start with Dr Bosh
[22:12] (gerado) podemos enterndernos....
[22:12] (JBover) Dr Ricardo Bosch is originally from Mar del Plata Argentina
[22:12] (JBover) He is now Proffesor of Physiology in ALcala de HEnraes
[22:12] (JBover) MAdrid
[22:12] (JBover) Among many contributions in the field of Nephrology....
[22:13] (JBover) (I would like to remark that he is THE FIRST who included A GENE inside the kidney!) ...first step in renal gene therapy....
[22:13] (RJ_Bosch) tal vez sea interesante hacer preguntas en ambos idiomas
[22:14] (gerado) me parece estupendo ricardo
[22:14] * JBover remarks that questions will be addressed in any language
[22:15] (MJesus) el que desee leer esto en español que vaya al canal #media sin salir de este
[22:15] (JBover) However, he wont be talking today about renal gene therapy, but his contributions to PTH related peptide (PTHrp) to Renal physiology
[22:15] (MJesus) aqui se escribe en ingles.. y en #media se vertira al español ok?
[22:15] * JBover remarks whomever wants to read this speech in spanish may go to channel #media
[22:15] (JBover) simultaneous translation
[22:18] (JBover) PARATHYROID HORMONE-RELATED PROTEIN AS A RENAL REGULATORY FACTOR:
[22:18] (JBover) FROM VESSELS TO GLOMERULI AND TUBULAR EPITHELIUM
[22:18] (JBover) Introduction
[22:19] (JBover) Parathyroid hormone (PTH)-related protein (PTHrP) was initially
[22:19] (JBover) isolated from tumors associated with humoral hypercalcemia of malignancy.
[22:19] (JBover) Both PTH and PTHrP share homology in their N terminal region, and bind to the type 1 PTH/PTHrP receptor (PTHR), leading to activation of both adenylate cyclase and phospholipase C/protein quinase C.
[22:19] (JBover) However, while PTH is a regulator of mineral homeostasis, acting mainly on bone and kidney,
[22:19] (JBover) PTHrP is found in many nonmalignant tissues.
[22:19] (JBover) Despite the widespread production of PTHrP in healthy individuals, its
[22:19] (JBover) circulating concentration is below the detectable limit of the majority of current assays.
[22:19] (JBover) Thus, in contrast to the situation of humoral hypercalcemia
[22:19] (JBover) of malignancy in which PTHrP plays the role of a classical "endocrine"
[22:20] (JBover) hormone, under normal circumstances, PTHrP seems to play a paracrine and/or autocrine role.
[22:20] (JBover) These physiological functions appear to include:
[22:20] (JBover) 1) regulation of smooth muscle (vascular, intestinal, uterine, bladder) tone;
[22:20] (JBover) 2) modulation of transepithelial (renal, placental, oviduct, mammary gland) calcium transport;
[22:20] (JBover) and 3) regulation of tissue and organ development, differentiation, and proliferation.
[22:20] (JBover) PTHrP as a vasoactive hormone
[22:20] (JBover) PTH has long been known to have acute hypotensive vasodilatory effects.
[22:20] (JBover) However, the putative physiological role of PTH as a systemic vasoregulatory hormone has been difficult to understand in homeostatic terms,
[22:21] (JBover) since PTH synthesis is confined to the parathyroid gland. On the other hand,
[22:21] (JBover) it is now clear that PTHrP is produced throughout the cardiovascular system, and binds to vascular smooth muscle cells through the PTHR, acting in a paracrine/autocrine, and perhaps even intracrine manner.
[22:21] (JBover) PTHrP has been shown to be a potent smooth muscle relaxant in every tissue examined,
[22:21] (JBover) including vascular smooth muscle cells.
[22:21] (JBover) Interestingly, in these tissues, both PTHrP mRNA and protein are dramatically upregulated in response to mechanical stretch.
[22:21] (JBover) Pirola et al demonstrated that vasoconstrictors such as angiotensin II,
[22:21] (JBover) serotonin, and bradykinin markedly induce PTHrP gene expression in the
[22:21] (JBover) vascular tree, whereas other vasoactive substances such as atrial natriuretic peptide, neurokinin, and substance P are ineffective.
[22:21] (JBover) In addition, PTHrP inhibits angiotensin II-induced smooth muscle cell growth,
[22:21] (JBover) suggesting that local production of PTHrP may serve as a counterbalancing
[22:22] (JBover) modulator of the contractile and/or growth-promoting effects of
[22:22] (JBover) angiotensin II, and possibly of other vasoconstrictors. Thus, PTHrP might
[22:22] (JBover) serve as a local peptide, limiting or antagonizing the biological activity
[22:22] (JBover) of at least some contractile stimuli in the arterial wall.
[22:22] (JBover) PTHrP and its receptors in renal tissue
[22:22] (JBover) Recently, the localization of PTHrP and PTHR mRNA in the developing mouse
[22:22] (JBover) kidney has been examined. High PTHrP mRNA levels were found in the
[22:22] (JBover) collecting duct, urothelium of the pelvis, and immature elements of the
[22:23] (JBover) glomeruli in this model. PTHR mRNA increased, associated with the maturation
[22:23] (JBover) process, in the developing tubules and glomeruli.
[22:23] (JBover) These findings suggest a role for PTHrP in renal development. In the adult
[22:23] (JBover) kidney, PTHrP has been identified by immunohistochemistry or in situ
[22:23] (JBover) hybridization in the glomerular podocytes, and proximal, distal, and
[22:23] (JBover) collecting tubules, as well as in the intrarenal arterial tree, including afferent and efferent arterioles, and in renal macula densa.
[22:23] (JBover) Using immunohistochemistry or various mRNA detection methods, the PTHR
[22:23] (JBover) was detected in convoluted and straight proximal tubules, cortical straight ascending limbs, and distal convoluted tubules, consistent with known sites of PTH action.
[22:24] (JBover) Whether this receptor results from alternative splicing of
[22:24] (JBover) the PTHR mRNA, as shown to occur in the human kidney cortex, is unknown.
[22:24] (JBover) Recently, we have found that both PTH and PTHrP counteract the contracting effects of platelet activating factor on human mesangial cells in vitro.
[22:24] (JBover) However, while we could identify the PTHR mRNA in the human kidney cortex by Northern blot analisys and RT-PCR, no PTHR transcript was found in mesangial cells.
[22:24] (JBover) Our results strongly suggest that the PTHR is not responsible for
[22:24] (JBover) the effects of PTH and PTHrP in the human mesangium.
[22:24] (JBover) Glomerular actions of PTHrP
[22:25] (JBover) Several lines of evidence support a direct action of PTH and PTHrP on the
[22:25] (JBover) glomerulus. In a recent study, using a hydronephrotic rat kidney model, local
[22:25] (JBover) administration of PTH and PTHrP induced vasodilatation of all preglomerular
[22:25] (JBover) vascular segments, including the afferent arteriole, and an increase in renal
[22:25] (JBover) blood flow. Moreover, Massfelder et al studied the renal effects of PTHrP
[22:25] (JBover) infused directly into the left renal artery of anaesthetized rats.
[22:25] (JBover) PTHrP increased renal blood flow by 10%, and GFR by 20%, without
[22:25] (JBover) significantly increasing the filtration fraction, and it increased urine
[22:25] (JBover) flow by 57% in the left kidney. Meanwhile, in the right control kidney,
[22:25] (JBover) GFR and diuresis did not change. These findings support the renal
[22:26] (JBover) vasodilatory effect of PTHrP.
[22:26] (JBover) More recently, we found a direct relaxant effect of PTHrP on mesangial cells
[22:26] (JBover) in vitro, which seems to involve cAMP and G-proteins. Our findings suggest a
[22:26] (JBover) modulatory effect of PTHrP on glomerular function by counteracting the
[22:26] (JBover) effects of vasoconstrictor agents on mesangial cells. Moreover, these in
[22:26] (JBover) vitro data support the notion that PTHrP (and PTH as well) has a direct
[22:26] (JBover) relaxant effect on the mesangium, which would tend to increase both Kf and
[22:26] (JBover) GFR. Thus, taken together, the results from these studies suggest that the
[22:26] (JBover) local action of PTHrP would predominantly induce an increase of both Kf and
[22:27] (JBover) GFR.
[22:27] (JBover) Physiology and pathophysiology of PTHrP in the tubular epithelium
[22:27] (JBover) PTHrP is mitogenic for various renal cells, including renal carcinoma cells,
[22:27] (JBover) mesangial cells, distal tubule-like cells MDCK, and subconfluent proximal
[22:27] (JBover) tubule cells. Furthermore, PTHrP mRNA increases, associated with a
[22:27] (JBover) decreased PTHR gene expression, in renal tubular cells during the recovery
[22:27] (JBover) phase after ischemic injury. We recently found a similar response pattern
[22:28] (JBover) for the renal expression of PTHrP and the PTHR in folic acid-injected rats,
[22:28] (JBover) another model of acute renal failure, which is associated with mild kidney
[22:28] (JBover) damage but dramatic tubular hyperplasia. These findings suggest that
[22:28] (JBover) PTHrP is an autocrine factor that might participate in the renal regenerative
[22:28] (JBover) process after acute injury.
[22:28] (JBover) PTHrP and renal disease progression
[22:28] (JBover) Recent data suggest a role of PTHrP in the mechanisms associated to
[22:29] (JBover) progression of renal damage. In this regard, PTHrP mRNA was found to
[22:29] (JBover) increase sequentially in the renal cortex during the development of
[22:29] (JBover) proteinuria in a rat model of tubulointerstitial nephropathy after protein
[22:29] (JBover) overload. PTHrP immunostaining also increased in both proximal and distal
[22:29] (JBover) tubules, and in the glomerulus, where PTHrP positivity was found in both
[22:29] (JBover) mesangial and endothelial cells.
[22:29] (JBover) The mechanisms responsible for the observed PTHrP upregulation in the renal
[22:29] (JBover) tissue during progression of renal injury are yet unknown. Interestingly, an
[22:29] (JBover) increase in angiotensin converting enzyme, an important factor in the
[22:30] (JBover) mechanisms associated with the development of renal damage, and in
[22:30] (JBover) preproendothelin-1 mRNA occurred in the renal cortex of protein-overloaded
[22:30] (JBover) rats. The possibilty that angiotensin II or endothelin-1, which rapidly
[22:30] (JBover) induce PTHrP mRNA in vascular smooth muscle cells, would be responsible for the
[22:30] (JBover) increased PTHrP in the chronically damaged kidney has not yet been tested.
[22:30] (JBover) Renal PTHrP overexpression during chronic renal damage could be part of a
[22:30] (JBover) feedback mechanism to counteract the effects of other vasoactive factors such
[22:30] (JBover) as angiotensin II, considering those of PTHrP on vascular tone and mesangial
[22:31] (JBover) contraction mentioned above. However, other PTHrP effects have opposite
[22:31] (JBover) consequences on glomerular hemodynamics, such as those on renin production
[22:31] (JBover) and mesangial cell proliferation. Thus, PTHrP seems to be a factor
[22:31] (JBover) with complex and partly defined roles in the mechanisms associated with renal
[22:31] (JBover) disease progression.
[22:31] (JBover) Summary
[22:31] (JBover) Current data support the notion that PTHrP can be considered as a renal
[22:31] (JBover) regulatory factor that may limit or antagonize the biological activity of contractile stimuli in the renal arterial wall. Therefore, locally produced PTHrP could participate in the regulation of the glomerular filtration rate.
[22:32] (JBover) Moreover, PTHrP, besides mimicing PTH actions, has important effects on thegrowth of both glomerular and tubular cells. Finally, although much more work needs to be done to further characterize the emerging role of PTHrP as a renal regulating factor, studies of the renal effects of PTHrP may provide new insights for a better understanding of the normal and the injured kidney.
[22:32] (JBover) Pedro Esbrit*, Soledad Santos*, Arantxa Ortega*,
[22:32] (JBover) Teresa Fernández-Agulló, Begonia Gea Contreras, M. Antonia Gutiérrez-Tárres, Jordi Bover**, and Ricardo J. Bosch.
[22:32] (JBover) Bone and Mineral Metabolism Laboratory, Research Unit, Fundación
[22:32] (JBover) Jiménez Díaz*, Madrid; Department of Nephrology, Prínceps
[22:32] (JBover) d´Espanya Hospital, L´Hospitalet de Llobregat**, Barcelona; and
[22:32] (JBover) Department of Physiology, Alcalá School of Medicine, Alcalá de
[22:33] (JBover) Henares, Spain.
[22:33] (RJ_Bosch) thanks
[22:33] (MJesus) I like to ask some question!
[22:34] (MJesus) Dr. Bosch..... are you ready?
[22:34] (JBover) go on MJ
[22:34] (MJesus) I like to know about.... what are the differneces between PTH and PTHrp regarding its renal actions???
[22:35] (RJ_Bosch) I would to mention the contribution in the paper and in on line delivery of Dr Bover
[22:35] * JBover would suggest to write at the end of the questions something like **********
[22:35] (MJesus) yes, thank Dr. Bover.....
[22:35] * JBover would suggest as well to write ****** after the final point of an answer Dr Bosch
[22:35] (MJesus) I like to know about.... what are the differneces between PTH and PTHrp regarding its renal actions??? *
[22:35] (RJ_Bosch) in our studies TPH and PTHrP seems to have the same effects
[22:36] (RJ_Bosch) on mesangial cells at least
[22:37] (JBover) ooooooo
[22:37] (MJesus) ok, thank.....
[22:37] (perico) may I have a question?
[22:37] (MJesus) a moment, please... we have a mail from From: "Kim Solez, M.D." Kim.Solez@ualberta.ca
[22:37] (MJesus) I am in India and Nepal which is why I have not been able to actively
[22:37] (MJesus) participate.
[22:37] (MJesus) Best regards. - Kim
[22:38] (MJesus) it is for us!
[22:38] (RJ_Bosch) it is known tha sistemically PTH stimulates ANG II system but locally produced PTHrP has a vasodilatory effect
[22:38] (Charlie) any role for PTHrP in the management of systemic arterial pressure? *
[22:39] (RJ_Bosch) good question, but the PTHrP concentration in blood is below the RIA
[22:39] (RJ_Bosch) limits
[22:40] (RJ_Bosch) It acts in a autocrine paracrine manner
[22:41] (perico) what does intracrine effect mean? ***
[22:42] (RJ_Bosch) is an interaction in the same cell in a way that can interct to the nucleous
[22:43] (RJ_Bosch) so the peptide can be secreted an in the same time interact into the nucleous
[22:44] (perico) do you think that this have any clinical effect?***
[22:45] (RJ_Bosch) we think that PTHrP pays an importan role in renal physiology and in patological situations
[22:46] (Charlie) if I understood well, PTHrP counteracts the proliferative effect of Angio II at the level of vascular walls, right?*
[22:46] (RJ_Bosch) yes
[22:47] (RJ_Bosch) but in some cells, for instance, mesangial cells it has a proliferative effect
[22:48] (RJ_Bosch) so it depend on what type of cell you are talking about
[22:48] (JBover) any other question for Dr Bosch?
[22:49] (MJesus) why is that interaction with the nucleus for??? feed back???***
[22:49] (Charlie) so it can be one of the factors of progression of renal disease (mesangial proliferation)
[22:49] (RJ_Bosch) it is quite possible
[22:50] (MJesus) ok, thanks ***
[22:50] (RJ_Bosch) for example Dr Esbrit have demostrated the novo production of PTHrP by mesangial cells in a model of glomerular injury
[22:50] (JBover) Dr Bosch.....MJesus asked about the intracrine effect... why is that interaction with the nucleus for??? feed back???
[22:51] (RJ_Bosch) The also demostrated the overexpression of PTHrP in the tubular ephiteliun after an ARF
[22:52] (Charlie) the natural production of PTHrP can be stopped by some known therapeutic substance?
[22:53] (RJ_Bosch) The knowleadge of intracrine effect is limited but Massfelder et al have shown
[22:53] (JBover) any other question????
[22:54] (RJ_Bosch) that when PTHrP is overexpressed by smooth muscle cells may have an proliferative effets
[22:54] * JBover looks at the audience looking for a raised hand
[22:54] (gerado) muchas gracias dr, Boch
[22:54] (RJ_Bosch) No I am not aware of any substance like this yet
[22:55] (RJ_Bosch) thanks again
[22:55] (Charlie) thanks dr.Bosch
[22:55] (JBover) THANK YOU VERY MUCH
[22:55] * JBover wonders if Dr Charra is present in the audience
[22:56] (JBover) You know that Dr Charra requested to advance his online appearance because he had to make a trip
[22:56] (Charlie) good night and see you next time (when: tomorrow?)
[22:57] (JBover) Dr Charra told me he was at the hospital tonight but it may eventually happen that he had some trouble with patients
[22:57] (JBover) we all know how it is
[22:57] (JBover) should we give him 5 more minutes???
[22:57] (JBover) meanwhile....
[22:58] * JBover brings some champagne and pastry
[22:58] (MJesus) the programm is in the web: www.uninet.edu/cin2000/confer ences/index.html usuario ---) cin2000 passwd -----) Burgos
[22:58] (JBover) heheehehehehehehehehheheh
[22:59] (MJesus) where is dr. charra ?
[22:59] (MJesus) charli, where are you from ?
[22:59] * JBover fears that Dr Charra is not gonna be available tonight...he might be busy
[23:00] (MJesus) your poster is in the web....... are you seen ?
[23:00] (SCigarran) Aquí estamos los del Super 1 y 2
[23:00] (MJesus) super health center ?
[23:01] (MJesus) alguien no habla español ?
[23:01] (SCigarran) No, Super HD centers
[23:01] (gbarril) no he podido conectar con el servidor hasta ahora
[23:01] (MJesus) haemodializer ??
[23:01] (gbarril) que tal la conferencia?
[23:01] (Gainza) I accept questions about hereditary tubulopathies
[23:01] (JBover) Dear friends
[23:01] (MJesus) ok!!
[23:01] (JBover) GREAT DR GAINZA!!!!!!!!
[23:02] (MJesus) doy you know about nephronoptisis ?
[23:02] * JBover notices that Dr Gainza belongs to Dr Soriano's group
[23:02] (Gainza) Yes I do, but it is not a tubulopathy
[23:02] (JBover) Dr Soriano sent his apologies for not coming tonight
[23:03] (MJesus) nephronoptisis is not a tubulopathy?
[23:03] (JBover) MJEsus if Dr Gainza says it is not......it must be not hahahaahahahahahah
[23:04] (Gainza) It is a hereditary cystic disease
[23:04] * JBover knows that MJesus is a known world-wide about nephronoptisis
[23:04] (MJesus) I think nephronoptisis is a tipycal tubulopatie fron proximal tubule!
[23:04] * JBover erases "a"
[23:04] (MJesus) yes, i'ts hereditary dr. gainza!
[23:05] (JBover) well...I think this is just a matter of clasifications
[23:06] (MJesus) nephronoptisis is lso called hereditary of Fanconi or...medullary cistic disease...
[23:06] (gustl) anyone speak spanish ?
[23:06] (MJesus) yes...
[23:06] (Gainza) well, I can change my clasification
[23:07] (JBover) DOES ANYONE SPEAK ENGLISH NOW????
[23:07] (gustl) soy de argentina, pero estoy viviendo en Peru
[23:07] * JBover quisiera dar una noticia importante a los amigos
[23:07] (gerado) adelante
[23:07] (MJesus) adelante!!
[23:07] (SCigarran) adelante
[23:07] (gustl) adelante
[23:08] (MJesus) boch ??
[23:08] (Mapi) venga
[23:08] (gaucho) adelante
[23:08] (RJ_Bosch) todo :oidos"
[23:08] (JBover) gerardo! EXPLICANOS
[23:08] (MJesus) la mayoria silenciosa ?
[23:08] (gerado) Bugs era el dr. Shaldon
[23:08] (gustl) gaucho, eres de argentina ?
[23:09] (gerado) solo con una N
[23:09] (gaucho) no, soy de Uruguay
[23:09] (gerado) y segun me comunican le ha gustado
[23:10] * JBover se pregunta si todos estais inscritos en el COngreso
[23:10] (gerado) y que vendrá el lunes 28
[23:10] (MJesus) a ver, re-presento a gaucho.....
[23:10] (MJesus) gaucho es el Dr. Nelson Caporale!
[23:11] (JBover) DR CAPORALE!!!!!!!
[23:11] (RJ_Bosch) hola gaucho
[23:11] (MJesus) nuestro mas brillante conferenciante: el 1º en acabar su mesa redonda !
[23:11] (JBover) Dr Caporale-----> RJ Bosch
[23:11] (JBover) RJ Bosch-------> Dr Caporale
[23:12] (gustl) me parece bien.
[23:12] (RJ_Bosch) bien
[23:15] (JBover) MENSAJE DE DR CHARRA!!!!!!!
[23:15] (MJesus) carlinho, olá, boas noites
[23:15] (gaucho) carlinho es portugués o brasilero?
[23:16] (carlinho) portugues
[23:16] (carlinho) e nao vos encontrava a algum tempo
[23:16] (JBover) 21-2-2000
[23:17] (JBover) Sorry Jordi,
[23:17] (JBover) But I could not get anything else than a chat in neuropsicologia!
[23:17] (JBover) No way to enter in the chat site, it works until I enter my nickname...but
[23:17] (JBover) then nothing happens and the web does not send me into any room, called
[23:17] (JBover) cin2000 or not.
[23:17] (JBover) Too bad,
[23:17] (JBover) Bernard
[23:18] (MJesus) PARATHYROID HORMONE-RELATED PROTEIN AS A RENAL REGULATORY FACTOR:
[23:18] (MJesus) FROM VESSELS TO GLOMERULI AND TUBULAR EPITHELIUM
[23:20] (gaucho) esperemos que el Dr. Charra pueda entrar,dado que el tema va a dar para mucha discusión
[23:20] (JBover) CHARRA ESTA INTENTANDO ENTRAR
[23:20] (JBover) esperad!
[23:21] (gaucho) esperamos!
[23:21] (gerado) somo todos ojos
[23:22] (SCigarran) Bueno, le damos 5 minutitos,......
[23:24] (MJesus) dr. caporale, cuando le toca esa mesa redonda ??
[23:25] (gaucho) el jueves a las 18 de mi pais,y el 6 de marzo la ponencia sobre fracasos de la DP
[23:26] (gaucho) ENTRE EL 26 Y EL 2 DE MARZO ESTARÉ EN sAN fRANCISCO,ALGUIEN VA POR ALLÁ?
[23:26] * JBover esperaba con ansia la presencia de Charra...realmente el tema es MUY IMPORTANTE
[23:26] (Burgos) Mjesus ¿de donde eres
[23:26] (MJesus) quiza se conecte desde donde haya un firewall ?
[23:27] (carlinho) aguem de portugal
[23:27] (gaucho) le pregunté a Carlinho de que ciudad es
[23:27] (Burgos) ¿en que unidad?
[23:28] (carlinho) lisboa
[23:28] (MJesus) la del hospital general yagüe
[23:29] (MJesus) bueno, me da la impresion de que charra no puede entrar .... quiza sea que tenga alguna dificultad en su linea
[23:29] (gaucho) suerte que no dijo de iberoamérica
[23:29] (MJesus) como que no exageremos ??
[23:29] (gaucho) perdonen pero los uruguayos no nos consideramos extranjeros en españa ni en portugal
[23:29] (MJesus) para que ??
[23:30] (Burgos) para presumir
[23:30] (gerardo) por supuesto gaucho esta es tu casa
[23:30] (MJesus) ejem... muchos creen que el nomnbre completo del hgy es hospital general yagüe premio
[23:30] * JBover agradece a la audiencia por su paciencia
[23:30] * MJesus es uruguaya .... de corazon
[23:30] (gaucho) muchas gracias y ni decirlo que mi país es de ustedes, aun independiente
[23:30] (Burgos) De acuerdo animo y a triunfar
[23:34] (JBover) Mensaje de Dr Charra
[23:34] (JBover) I am sorry too Jordi,
[23:34] (JBover) I followed your instructions but keep going in the psicologia on-line! I am
[23:34] (JBover) going to need their help for sure!
[23:35] (JBover) Unfortunately I am leaving very soon for the States, so we may try again in some weeks?
[23:35] (JBover) I give it a last trial.
[23:35] (JBover) Thanks for your kind help anyway
[23:35] (carlinho) adeus a todos