Response to Comment on "Late Upper Paleolithic occupation at Cooper's Ferry, Idaho, USA, ~16,000 years ago".

Manning builds an inappropriate Bayesian age model to assert that the initial occupation at Cooper's Ferry began only ~15,935 ± 75 to 15,130 ± 20 cal yr B.P., suggesting that our estimation of ~16,560 to 15,280 cal yr B.P. is unsupported. However, this analysis both ignores evidence of human occupation from the earliest undated cultural deposits and reflects a misapplication of Bayesian age-modeling techniques. Consequently, his results are unreliable.

Incidencia de enfermedad pulmonar obstructiva crónica en pacientes con el virus de la inmunodeficiencia humana. Análisis post-hoc del estudio viva / Incidence of chronic obstructive pulmonary disease in patients with human immunodeficiency virus. Post-hoc analysis of the viva study

Introducción: desde la aparición de la terapia antiretroviral la supervivencia de los pacientes infectados por el virus de la inmunodeficiencia humana (VIH) ha aumentado considerablemente tomando importancia la aparición de otras patologías crónicas en estos pacientes como puede ser la enfermedad pulmonar obstructiva crónica (EPOC). Nuestro objetivo fue conocer la incidencia de EPOC en una cohorte de pacientes VIH derivados en un programa de detección de hipertensión pulmonar (HTP). Material y Métodos: análisis post-hoc, de un prospectivo, pseudo-experimental de pacientes con infección del VIH a los que se les preguntaba por disnea y en caso afirmativo eran derivados a consultas de neumología para despistaje de HTP. Resultado: desde 2014 hasta 2016, reclutamos un total de 32 pacientes, con un predominio de varones (75%). La disnea según la mMRC (Medical Reserach Council) fue grado 1, 2 y 3 en el 37,5%, 43,8% y 18,8%, respectivamente. La prevalencia de tabaquismo fue del 87,1% (intervalo de confianza [IC] 95%: 71- 96,4%), y 18 pacientes fueron catalogados de EPOC (62%; IC95%: 42,2 - 79,3%). Conclusión: la incidencia de EPOC en nuestra serie fue muy superior a la de la población general. Es necesario plantear estrategias de búsqueda activa de EPOC en estos pacientes para un diagnóstico y tratamiento precoz

Introduction: Since the advent of antiretroviral therapy, the survival of patients infected with the human immunodeficiency virus (HIV) has considerably increased, with the occurrence of other chronic diseases such as chronic obstructive pulmonary disease (COPD) gaining importance in these patients. Our objective was to find out the incidence of COPD in a cohort of HIV patients that were referred to a program to detect pulmonary hypertension (PH). Materials and Methods: Post hoc analysis of a prospective, quasi-experimental study on HIV-infected patients who were asked whether they had dyspnea. If this was the case, they were referred to a pulmonologist for PH screening. Results: From 2014 to 2016, we recruited a total of 32 patients, with a predominance of male recruits (75%). According to the mMRC (Modified Medical Research Council) Dyspnea Scale, 37.5%, 43.8% and 18.8% were classified as Grade 1, 2 and 3, respectively. The prevalence of smoking was 87.1% (95% confidence interval [CI]: 71 - 96.4%), and 18 patients were classified with COPD (62%; 95% CI: 42.2 - 79.3%). Conclusion: The incidence of COPD in our sample was much higher than that of the general population. It is necessary to plan active search strategies for COPD in these patients for early diagnosis and treatment

ABO blood group phenotype frequency estimation using molecular phenotyping in rhesus and cynomolgus macaques.

A much larger sample (N = 2369) was used to evaluate a previously reported distribution of the A, AB and B blood group phenotypes in rhesus and cynomolgus macaques from six different regional populations. These samples, acquired from 15 different breeding and research facilities in the United States, were analyzed using a real-time quantitative polymerase chain reaction (qPCR) assay that targets single nucleotide polymorphisms (SNPs) responsible for the macaque A, B and AB phenotypes. The frequency distributions of blood group phenotypes of the two species differ significantly from each other and significant regional differentiation within the geographic ranges of each species was also observed. The B blood group phenotype was prevalent in rhesus macaques, especially those from India, while the frequencies of the A, B and AB phenotypes varied significantly among cynomolgus macaques from different geographic regions. The Mauritian cynomolgus macaques, despite having originated in Indonesia, showed significant (P ≪ .01) divergence from the Indonesian animals at the ABO blood group locus. Most Mauritian animals belonged to the B blood group while the Indonesian animals were mostly A. The close similarity in blood group frequency distributions between the Chinese rhesus and Indochinese cynomolgus macaques demonstrates that the introgression between these two species extends beyond the zone of intergradation in Indochina. This study underscores the importance of ABO blood group phenotyping of the domestic supply of macaques and their biospecimens.

Características antropométricas maternas y del lactante, correlacionado a la concentración proteica del calostro y leche madura: un estudio longitudinal / Anthropometric characteristics of the mother and the infant, correlated to the protein concentration of colostrum and mature milk: a longitudinal study

Objetivo: Determinar las características antropométricas maternas y del lactante correlacionados a la concentración proteica del calostro y la leche madura en el Hospital "El Carmen", Huancayo. Método: Estudio analítico, longitudinal, correlacional; Muestra: 35 (madres y lactantes), quienes cumplían criterios de selección. Instrumento: ficha de recolección de datos consignando las medidas antropométricas maternas, del lactante y resultados de concentración proteica del calostro y leche madura; medición de variables en dos fases (Al parto y cuarto mes post- parto). Se utilizó correlación R de Spearman, Pearson y regresión logística multivariada. Resultados: La mediana de edad de las madres fue de 20 años, el IMC promedio pre- gestacional fue 23,55, mediana pre- parto fue de 26,71, y cuarto mes post- parto fue de 24,42; En los lactantes el sexo predominante fue el femenino 54,29%, la mediana del peso al nacer fue 3.030 kg y a los 4 meses tuvieron un peso promedio de 6,580 kg. El análisis bivariado mostró significancia estadística entre la concentración de proteínas del calostro y peso al cuarto mes (p=0.0119) y la concentración de proteínas de leche madura con respecto a la talla del lactante al cuarto mes (p=0.0041). El análisis multivariado para el peso del recién nacido, tiene relación con el Índice de Masa Corporal Pre- gestacional (p<0.011), R2 :0.481. Conclusión: Las características antropométricas de la madre y del lactante están correlacionados a la concentración proteica del calostro y leche madura Palabras clave: Índice de masa corporal, Recién nacido, Peso al nacer, Leche Materna, Proteínas.

Objective: To determine maternal and infant anthropometric characteristics correlated to protein concentration of colostrum and mature milk in the Hospital "El Carmen", Huancayo Method: analytical, correlational longitudinal study; Shows: 35 (mothers and infants) who met the selection criteria. Instrument: data collection sheet consigning maternal anthropometric measurements, infant and results of protein concentration of colostrum and mature milk; measurement variables in two phases (Al childbirth and fourth month postpartum). Spearman correlation R, Pearson and multivariate logistic regression was used. Results: The median age of the mothers was 20 years, mean BMI was pre- gestational 23.55, median pre- delivery was 26.71, and fourth month postpartum was 24.42; In infants the predominant female sex was 54.29%, the median birth weight was 3,030 kg and at 4 months had an average weight of 6,580 kg. The bivariate analysis showed statistical significance between protein concentration of colostrum and fourth month weight (p = 0.0119) and the protein concentration of mature milk with respect to the size of the infant to four months (p = 0.0041). Multivariate for birth weight, analysis is related to the gestational Body Mass Index Pre (p <0.011), R2: 0.481. Conclusion: The anthropometric characteristics of the mother and infant are correlated to protein concentration of colostrum and mature milk Keywords: Body mass index, Newborn, Birth weight, Breast Milk, Proteins.

Should IFN-γ, IL-17 and IL-2 be considered predictive biomarkers of acute rejection in liver and kidney transplant? Results of a multicentric study.

Acute rejection (AR) remains a major challenge in organ transplantation, and there is a need for predictive biomarkers. In the present multicenter study, we prospectively examined a series of biomarkers in liver and kidney recipients. Intracellular expression of IFN-γ, IL-17 and IL-2 and IL-17 soluble production were evaluated both pre-transplantation and post-transplantation (1st and 2nd week, 1st, 2nd and 3rd month). 142 transplant patients (63 liver/79 kidney) were included in the study. Twenty-eight recipients (14 liver/14 kidney) developed AR. Pre- and post-transplantation intracellular expression of %IFN-γ(+) in CD4(+)CD69(+) and in CD8(+)CD69(+) and soluble IL17 identified liver and kidney transplant patients at high risk of AR. Pre-transplantation, %IL-2(+) in CD8(+)CD69(+) also identified kidney patients at high risk. We constructed pre- and post-transplantation risk prediction models, based on a composite panel of biomarkers, which could provide the basis for future studies and will be a useful tool for the selection and adjustment of immunosuppressive treatments.

Intracellular IFN-γ and IL-2 expression monitoring as surrogate markers of the risk of acute rejection and personal drug response in de novo liver transplant recipients.

Biomarker monitoring is needed in transplantation to reflect individual response to immunosuppressive drugs and graft outcome. We evaluated intracellular expression and soluble production of interferon-(IFN)-γ and interleukin-(IL)-2 as predictive biomarkers of acute rejection (AR) and personal drug response. Pharmacokinetic-pharmacodynamic profiles were determined in 47 de novo liver recipients treated with tacrolimus, mycophenolate mofetil and prednisone. Of the 47 patients, AR occurred in nine. There were no differences in drug concentrations between rejectors and non-rejectors. A pre-transplantation cut-off value of 55.80% for %CD8(+)-IFN-γ(+) identified patients at high risk of AR with a sensitivity of 75% and a specificity of 82%. In the first week post-transplantation, patients with a % inhibition for soluble IFN-γ, %CD8(+)-IFN-γ(+) and %CD8(+)-IL2(+) lower than 40% developed AR, showing low susceptibility to immunosuppressive drugs. Therefore, effector-T-cell response monitoring may help physicians to identify personal response to treatment and patients at high risk of AR.

How long should initiation of calcineurin inhibitors be delayed to protect renal function in liver transplantation?

BACKGROUND AND AIM: Delayed introduction of calcineurin inhibitors (CNI) in liver transplantation (OLT) seeks to protect renal function, although the optimal length of the delay is not well established. The aim of this study was to analyze the effects on renal function of CNI initiation on different days after OLT. METHODS: We reviewed the charts of 260 OLT recipients. Group D1-a (n = 36) underwent the standard initial immunosuppression at our center: namely, CNI introduction on day 1 with further daily administration to achieve target levels of 8 to 15 ng/mL for tacrolimus or 150 to 300 ng/mL for cyclosporine. Due to renal concerns, 126 patients (group D1-b) had CNI introduced on day 1 either not daily or at doses to achieve less than the target on at least two occasions. In 43 patients (group D2), CNI were introduced on day 2 in 23 on day 3 (group D3), in 12 on day 4 (group D4), and at least at day 5 in 20 others (group D5). In periods without CNI treatment, patients received mycophenolate mofetil. Steroids were administered to all patients. The study period included the first 3 months post-OLT. Renal function was estimated as creatinine clearance (CrCl) using the Cockcroft-Gault equation. RESULTS: Changes in CrCl from pre-OLT to month 3 were -19% ± 28% in group D1-a; -27% ± 19% in group D1-b; -29% ± 19% in group D2; -23% ± 26% in group D3; -4% ± 38% in group D4, and +4% ± 33% in group D5 (P < .05 vs groups D1-a, D1-b, D2, and D3). On multivariate analysis, CNI introduction at day ≥ 5 was protective for kidneys when adjusted for other variables that potentially influence renal function. CONCLUSION: CNI should be introduced at day 5 after OLT to protect renal function.

[Human papillomavirus (HPV) genotypes in cervix uterine cancer patients in a public hospital and private clinic from Santiago, Chile]. / Genotipos de virus papiloma humano (VPH) en pacientes con cáncer cervico-uterino en un hospital público y una clínica privada de Santiago, Chile.

We compared HPV genotypes among squamous cervical cancer samples from a public hospital (n = 55) and a private clinic (n = 35 cases) of Santiago. Paraffin-embedded specimens were analyzed by PCR followed by an immunoenzimatic assay. Reverse line blotting was used for the identification of 36 HPV genotypes. We found HPVDNAm 94.4% of all cancers. Single mfections: HPV16: 40.0%, (clinic 37.1%, hospital 41.8%) VPH18:7.8% (clinic 2.9%, hospital 10.9%); single+multiple mfections: VPH16: 61.1% (clinic 53.1%, hospital 71.7%), VPH18: 34.4% (clinic 21.9%, hospital 45.2%). HPV16 orHPV18 occurredin 75.6% of cases, higher in the hospital than the clinic (87.3%-95% CI: 84.9-96.3 - and 57. l%-95% CI: 46.6-66 - respectively, p = 0.002). Other genotypes in single mfections: HPV 26, 31, 33, 45, 58, 67; in co-mfections: HPV 35,52,56,59 and 66. HPV16 but specially HPV 18 were significantly more frequent in the public hospital; 75.6% of squamous cervical cancer were associated to the vaccine preventable HPV16/18.

Genotipos de virus papiloma humano (VPH) en pacientes con cáncer cervico-uterino en un hospital público y una clínica privada de Santiago, Chile / Human papillomavirus (HPV) genotypes in cervix uterine cancer patients in a public hospital and private clinic from Santiago, Chile

We compared HPV genotypes among squamous cervical cancer samples from a public hospital (n = 55) and a private clinic (n = 35 cases) of Santiago. Paraffin-embedded specimens were analyzed by PCR followed by an immunoenzimatic assay. Reverse line blotting was used for the identification of 36 HPV genotypes. We found HPVDNAm 94.4 percent of all cancers. Single mfections: HPV16: 40.0 percent, (clinic 37.1 percent, hospital 41.8 percent) VPH18:7.8 percent (clinic 2.9 percent, hospital 10.9 percent); single+multiple mfections: VPH16: 61.1 percent (clinic 53.1 percent, hospital 71.7 percent), VPH18: 34.4 percent (clinic 21.9 percent, hospital 45.2 percent). HPV16 orHPV18 occurredin 75.6 percent of cases, higher inthe hospital than the clinic (87.3 percent-95 percent CI: 84.9-96.3 - and 57. l percent-95 percent CI: 46.6-66 - respectively, p = 0.002). Other genotypes in single mfections: HPV 26, 31, 33, 45, 58, 67; in co-mfections: HPV 35,52,56,59 and 66. HPV16 but specially HPV 18 were significantly more frequent in the public hospital; 75.6 percent of squamous cervical cancer were associated to the vaccine preventable HPV16/18.

Se comparan los genotipos de VPH en casos de cáncer cérvico-uterino escamocelular de una clínica privada (n: 35) y de un hospital público (n: 55) atendidos entre 1996 y 2006 en Santiago, Chile. Se analizaron por RPC y ensayo inmunoenzimático muestras tumorales en bloques de parafina, genotipificándose con reverse Une blotting para 36 genotipos de VPH. Se detectó VPH en 94,4 por ciento de los casos: infecciones únicas por: VPH 16: 40,0 por ciento>, (clínica 37,1 por ciento, hospital 41,8 por ciento) VPH 18: 7,8 por ciento (clínica 2,9 por ciento, hospital 10,9 por ciento); total de infecciones por VPH 16 61,1 por ciento (clínica 53,1 por ciento, hospital 71,7 por ciento), por VPH 18 34,4 por ciento (clínica 21,9 por ciento, hospital 45,2 por ciento). Co-infección: VPH 16/18 75,6 por ciento (clínica 57,1 por ciento; IC95 por ciento = 46,6-66,0 hospital 87,3 por ciento; IC95 por ciento = 84,9-96,3, p = 0,002). Se identificó otros 11 genotipos oncogénicos en infecciones únicas (VPH: 26, 31, 33, 45, 58, 67) o en co-infección con VPH 16/18 (VPH: 35, 52, 56, 59, 66). VPH 16 y VPH 18 fueron significativamente más frecuentes en el hospital público, particularmente VPH18; 75,6 por ciento> de los cánceres se asociaron a los genotipos VPH 16/18, tipos prevenibles por vacuna.

Testicular adrenal rest tumors and Leydig and Sertoli cell function in boys with classical congenital adrenal hyperplasia.

CONTEXT: Infertility observed in adult males with congenital adrenal hyperplasia (CAH) has been associated with testicular adrenal rest tumors (TART) that may originate during childhood. OBJECTIVE: Our objective was to describe the prevalence of TART and Sertoli and Leydig cell function in a group of boys aged 2-10 yr with CAH and to compare prevalence with that of a control group. DESIGN: From August 2005 to January 2007, 19 patients with classical CAH (CAH group) were referred from seven endocrinology centers. METHODS: We studied 19 subjects in the CAH group and, as a control group, 13 boys from the community that did not have testicular diseases. A complete physical exam was performed. High-resolution ultrasound was used to determine TART prevalence. Inhibin B and anti-Müllerian hormone were used as Sertoli cell markers. The ratio between basal testosterone levels and testosterone levels 72 h after beta-human chorionic gonadotropin (5000 U/m2) treatment [(T72- T0)/T0] was used to evaluate Leydig cell response. RESULTS: CAH and control groups were comparable in chronological age (5.9 vs. 5.6 yr; P = 0.67) and bone age/chronological age ratio (1.09 vs. 1.03; P = 0.09). TART prevalence was four of 19 (21%) in the CAH group. Lower values for inhibin B (49.2. vs. 65.2 pg/ml; P = 0.018), anti-Müllerian hormone (70.1 vs. 94.2 ng/ml; P = 0.002), and (T72- T0)/T0 (5.6 vs. 13.6; P < 0.01) were observed in the CAH group. CONCLUSION: TART in prepubertal males with classic CAH could be found during childhood. We also report differences in markers of gonadal function in a subgroup of patients, especially in those with inadequate control.

Possible role of alpha-adrenoceptor subtypes in acute migraine therapy.

Even though the underlying mechanisms for the pathophysiology of migraine attacks are not completely understood, little doubt exists that the headache phase is explained by dilatation of cranial, extracerebral blood vessels. In this context, experimental models predictive for anti-migraine activity have shown that both triptans and ergot alkaloids, which abort migraine headache, produce vasoconstriction within the carotid circulation of different species. In contrast to the well-established role of serotonin (5-hydroxytryptamine; 5-HT) 5-HT1B receptors in the common carotid vascular bed, the role of alpha-adrenoceptors and their subtypes has been examined only relatively recently. Using experimental animal models and alpha1- and alpha2-adrenoceptor agonists (phenylephrine and BHT933, respectively) and antagonists (prazosin and rauwolscine, respectively), it was shown that activation of either receptor produces a cranioselective vasoconstriction. Subsequently, investigations employing relatively selective antagonists at alpha1- (alpha1A, alpha1B, alpha1D) and alpha2- (alpha2A, alpha2B, alpha2C) adrenoceptor subtypes revealed that specific receptors mediate the carotid haemodynamic responses in these animals. From these observations, together with the potential limited role of alpha1B- and alpha2C-adrenoceptors in the regulation of systemic haemodynamic responses, it is suggested that selective agonists at these receptors may provide a promising novel avenue for the development of acute anti-migraine drugs.

An introduction to migraine: from ancient treatment to functional pharmacology and antimigraine therapy.

Migraine treatment has evolved from the realms of the supernatural into the scientific arena, but it seems still controversial whether migraine is primarily a vascular or a neurological dysfunction. Irrespective of this controversy, the levels of serotonin (5-hydroxytryptamine; 5-HT), a vasoconstrictor and a central neurotransmitter, seem to decrease during migraine (with associated carotid vasodilatation) whereas an i.v. infusion of 5-HT can abort migraine. In fact, 5-HT as well as ergotamine, dihydroergotamine and other antimigraine agents invariably produce vasoconstriction in the external carotid circulation. The last decade has witnessed the advent of sumatriptan and second generation triptans (e.g. zolmitriptan, rizatriptan, naratriptan), which belong to a new class of drugs, now known as 5-HT1B/1D/1F receptor agonists. Compared to sumatriptan, the second-generation triptans have a higher oral bioavailability and longer plasma half-life. In line with the vascular and neurogenic theories of migraine, all triptans produce selective carotid vasoconstriction (via 5-HT1B receptors) and presynaptic inhibition of the trigeminovascular inflammatory responses implicated in migraine (via 5-HT1D/5-ht1F receptors). Moreover, selective agonists at 5-HT1D (PNU-142633) and 5-ht1F (LY344864) receptors inhibit the trigeminovascular system without producing vasoconstriction. Nevertheless, PNU-142633 proved to be ineffective in the acute treatment of migraine, whilst LY344864 did show some efficacy when used in doses which interact with 5-HT1B receptors. Finally, although the triptans are effective antimigraine agents producing selective cranial vasoconstriction, efforts are being made to develop other effective antimigraine alternatives acting via the direct blockade of vasodilator mechanisms (e.g. antagonists at CGRP receptors, antagonists at 5-HT7 receptors, inhibitors of nitric oxide biosynthesis, etc). These alternatives will hopefully lead to fewer side-effects.

Zinc transformations in acidic soil and zinc efficiency on maize by adding six organic zinc complexes.

Experiments under laboratory and greenhouse conditions were conducted to study the response of maize (Zea mays L.) to Zn fertilizer applications (Zn-phenolate, Zn-EDDHA, Zn-EDTA, Zn-lignosulfonate, Zn-polyflavonoid, and Zn-heptagluconate) in an Aquic Haploxeralf soil. The application of Zn complexes significantly increased Zn uptake by the plant compared with that in the control soil. The highest enhancements were obtained in soil treated with Zn-EDTA, Zn-lignosulfonate, and Zn-EDDHA. The highest percentages of Zn taken up by the plants occurred when 20 mg x kg(-1) Zn was applied as Zn-EDTA fertilizer and 10 mg x kg(-1) as Zn-lignosulfonate fertilizer. In the greenhouse experiment, Zn speciation in soil after harvesting showed that almost all Zn was found in the residual fraction followed by metal in the water-soluble plus exchangeable fraction and metal bound to organic matter. The most effective fertilizers maintaining Zn in the most labile fractions were Zn-phenolate, Zn-EDTA, and Zn-lignosulfonate. Conversely, in the incubation experiment, only a small percentage of Zn was found in the water-soluble plus exchangeable fraction and no differences in the Zn distribution were observed between the different fertilizer treatments. The micronutrient content in maize was positively correlated with the water-soluble plus exchangeable Zn as well as with the available Zn determined by the diethylenetriaminepentaacetic acid and Mehlich-3 methods, in the greenhouse experiment. Results of this study showed that the incubation experiment in acidic soil is not a suitable tool to establish the different effectiveness of Zn chelates in plants.

Mobility and leachability of zinc in two soils treated with six organic zinc complexes.

A study of soil columns was conducted to evaluate Zn movement potential in two reconstructed soil profiles. Zn-phenolate, Zn-EDDHA, Zn-EDTA, Zn-lignosulfonate, Zn-polyflavonoid, and Zn-heptagluconate were applied in the upper zone of the column. The different physicochemical properties of the two soils and the micronutrient source may influence Zn leaching, the distribution of Zn among soil fractions, and the Zn available to the plant in the depth of the layers. In Aquic Haploxeralf soil, the application of six fertilizers produced little migration and very small leaching of Zn in the soil profiles. In Calcic Haploxeralf soil, Zn-EDTA migrated and was distributed throughout the soil columns. This Zn chelate produces a loss of Zn by leaching, which was 36% of the added Zn. In the latter soil, Zn leached very little with the other five fertilizer treatments. The same as for these organic Zn complexes, the retention of added Zn indicated the potential of metal accumulation in the A(p) horizons of the two soil profiles. A large portion of applied Zn was available to plants [diethylenetriaminepentaacetic acid (DTPA) and Mehlich-3 extractable Zn] in the depths reached by the different commercial formulations. The relationship between the two methods was highly significant (Mehlich-3-Zn = 1.25 + 1.13 DTPA-Zn, R(2) = 99.19%). When Zn was added as Zn-EDTA, the amounts of the most labile fractions (water-soluble plus exchangeable and organically complexed Zn) increased throughout the entire profile column in comparison with the control columns, although in the B(t) horizon of the Aquic Haploxeralf soil they increased only slightly.

Pharmacological profile of the mechanisms involved in the external carotid vascular effects of the antimigraine agent isometheptene in anaesthetised dogs.

The present study set out to investigate the external carotid vascular effects of isometheptene in vagosympathectomised dogs, anaesthetised with pentobarbital. One-minute intracarotid (intra-arterial; i.a.) infusions of isometheptene (10, 30, 100 and 300 microg/min) produced dose-dependent decreases in external carotid blood flow, without affecting blood pressure or heart rate. The vasoconstrictor responses to 100 microg/min and 300 microg/min of isometheptene were clearly attenuated in animals pretreated with reserpine (5,000 microg/kg). Moreover, after prazosin (an alpha1-adrenoceptor antagonist; 100 microg/kg), the responses to isometheptene remained unaltered in untreated as well as reserpine-pretreated dogs. In contrast, the responses to isometheptene were attenuated by rauwolscine (an alpha2-adrenoceptor antagonist; 300 microg/kg) in untreated animals, and were practically abolished in reserpine-pretreated dogs. Further investigation into the specific alpha2-adrenoceptor subtypes, using selective antagonists, showed that BRL44408 (alpha2A) and MK912 (alpha2C) markedly attenuated this response, while imiloxan (alpha2B) was ineffective. The involvement of 5-HT1B and 5-HT1D receptors seems highly unlikely since antagonists at 5-HT1B (SB224289) and 5-HT1D (BRL15572) receptors (both at 300 microg/kg) were ineffective. On this basis, it is concluded that isometheptene-induced canine external carotid vasoconstriction is mediated by both indirect (a tyramine-like action) and direct (acting at receptors) mechanisms, which mainly involve alphaA- and alpha2C-adrenoceptors, while the involvement of alpha1-adrenoceptors seems rather limited.

Pharmacological identification of the major subtypes of adrenoceptors involved in the canine external carotid vasoconstrictor effects of adrenaline and noradrenaline.

This study investigated the potential effects of adrenaline and noradrenaline on the external carotid blood flow of vagosympathectomised dogs and the receptor mechanisms involved. One minute (1 min) intracarotid infusions of adrenaline and noradrenaline produced dose-dependent decreases in external carotid blood flow without changes in blood pressure or heart rate. These responses, which remained unaffected after saline, were: (i) mimicked by the adrenoceptor agonists, phenylephrine (alpha1) and BHT933 (6-Ethyl-5,6,7,8-tetrahydro-4H-oxazolo [4,5-d] azepin-2-amine dihydrochloride; alpha2); (ii) abolished after phentolamine (2000 microg/kg) unmasking a vasodilator component (subsequently blocked by propranolol; 1000 microg/kg); and (iii) partly blocked by rauwolscine (30 and 100 microg/kg), and subsequently abolished by prazosin (100 microg/kg). Accordingly, rauwolscine (100 and 300 microg/kg) markedly blocked the responses to BHT933 without affecting those to phenylephrine; likewise, prazosin (100 microg/kg) markedly blocked the responses to phenylephrine without affecting those to BHT933. These results show that both alpha1- and alpha2-adrenoceptors mediate vasoconstriction within the canine external carotid circulation. Moreover, after blockade of alpha1/alpha2-adrenoceptors, both adrenaline and noradrenaline exhibit a beta-adrenoceptor-mediated vasodilator component.

Hamster-to-rat bone marrow xenotransplantation and humoral graft vs. host disease.

Bone marrow transplantation (BMT) may induce tolerance across xenogeneic barriers. We have established a xenogeneic BMT model where hamster BM is transplanted into splenectomized LEW rat recipients resulting in high levels of engraftment. Unfortunately, graft vs. host disease (GVHD) with severe dermatitis developed in all rat recipients. We were successful in treating or preventing the dermatitis of this xenogeneic GVHD by the use of the T-cell suppressant tacrolimus. However, this compound did not prevent the development of a fatal liver injury in the rat recipients. This study was designed to elucidate the pathogenesis of this liver injury appearing in T-cell suppressed rat recipients of hamster BM. Splenectomized and irradiated (10 Gy) LEW rats received 300 x 106 unfractionated hamster BM cells. These BMT recipients were divided in 3 groups: Group I recipients (n = 8) did not receive further immunosuppression. Group II animals (n = 10) received tacrolimus 1 mg/kg/d for 7 d. Group III recipients (n = 6) were given the same daily dose of tacrolimus on a long-term basis. Chimerism was detected by flow cytometry. Cytotoxicity of recipient's sera against rat and hamster lymph node cells was measured by complement-dependent cytotoxicity (CDC) test. Immunofluorescence was used to detect hamster antirat antibodies on several recipient organs. In Group I, 2 out of 8 animals engrafted (25%) and survived for a median of 21 d showing the severe dermatitis characteristic of GVHD. In group II (n = 10), 9/10 rat recipients engrafted (90%) and survival was increased to a median of 53.7 days. However, these surviving recipients developed fatal GVHD not different from that observed in Group I recipients. All animals in Group III (n = 6) engrafted and did not show the characteristic dermatitis of GVHD. Their survival, however, was shortened to a median of 30.3 d by a severe liver injury. This injury was characterized by hepatocyte necrosis in zones 1 and 2 with polymorphonuclear (PMN) cell infiltration. Deposits of hamster immunoglobulins were present around the necrotic areas and in the portal veins. Moreover, antirat antibodies appeared in the circulation. These antibodies were sensitive to dithiothreitol (DTT) treatment indicating that they were of the IgM class. This study shows that xenogeneic GVHD may have a dual presentation in the hamster-to-rat model: a classical cellular GVHD not distinct to the allogeneic one and a humoral GVHD affecting solely the recipient liver. The degree of humoral injury is potentiated by T-cell suppression.

The role of several alpha(1)- and alpha(2)-adrenoceptor subtypes mediating vasoconstriction in the canine external carotid circulation.

1. It has recently been shown that both alpha(1)- and alpha(2)-adrenoceptors mediate vasoconstriction in the canine external carotid circulation. The present study set out to identify the specific subtypes (alpha(1A), alpha(1B) and alpha(1D) as well as alpha(2A), alpha(2B) and alpha(2C)) mediating the above response. 2. Consecutive 1 min intracarotid infusions of phenylephrine (alpha(1)-adrenoceptor agonist) and BHT933 (alpha(2)-adrenoceptor agonist) produced dose-dependent decreases in external carotid blood flow, without affecting mean arterial blood pressure or heart rate. 3. The responses to phenylephrine were selectively antagonized by the antagonists, 5-methylurapidil (alpha(1A)) or BMY7378 (alpha(1D)), but not by L-765,314 (alpha(1B)), BRL44408 (alpha(2A)), imiloxan (alpha(2B)) or MK912 (alpha(2C)). In contrast, only BRL44408 or MK912 affected the responses to BHT933. 4. The above results support our contention that mainly the alpha(1A), alpha(1D), alpha(2A) and alpha(2C)-adrenoceptor subtypes mediate vasoconstriction in the canine external carotid circulation.