Expansion of endogenous T cells in CSF of pediatric CNS tumor patients undergoing locoregional delivery of IL13R〿2-targeting CAR T cells: an interim analysis.

Outcomes for pediatric brain tumor patients remain poor, and there is optimism that chimeric antigen receptor (CAR) T cell therapy can improve prognosis. Here, we present interim results from the first six pediatric patients treated on an ongoing phase I clinical trial (NCT04510051) of IL13BBζ-CAR T cells delivered weekly into the lateral cerebral ventricles, identifying clonal expansion of endogenous CAR-negative CD8+ T cells in the cerebrospinal fluid (CSF) over time. Additionally, of the five patients evaluable for disease response, three experienced transient radiographic and/or clinical benefit not meeting protocol criteria for response. The first three patients received CAR T cells alone; later patients received lymphodepletion before the first infusion. There were no dose limiting toxicities (DLTs). Aside from expected cytopenias in patients receiving lymphodepletion, serious adverse events possibly attributed to CAR T cell infusion were limited to one episode of headache and one of liver enzyme elevation. One patient withdrew from treatment during the DLT period due to a Grade 3 catheter-related infection and was not evaluable for disease response, although this was not attributed to CAR T cell infusion. Importantly, scRNA- and scTCR-sequence analyses provided insights into CAR T cell interaction with the endogenous immune system. In particular, clonally expanded endogenous CAR- T cells were recovered from the CSF, but not the peripheral blood, of patients who received intraventricular IL13BBζ-CAR T cell therapy. Additionally, although immune infiltrates in CSF and post-therapy tumor did not generally correlate, a fraction of expanded T cell receptors (TCRs) was seen to overlap between CSF and tumor. This has important implications for what samples are collected on these trials and how they are analyzed. These initial findings provide support for continued investigation into locoregionally-delivered IL13BBζ-CAR T cells for children with brain tumors.

Ferric Carboxymaltose Improves the Quality of Life of Patients with Inflammatory Bowel Disease and Iron Deficiency without Anaemia.

Background: Iron deficiency (ID) without anaemia is a common comorbidity associated with inflammatory bowel disease (IBD) that has a negative impact on health-related quality of life (HRQoL). Methods: This multicentre, prospective, observational study examined the response to, safety of and impact on HRQoL of a single 500 mg dose of intravenous ferric carboxymaltose (FCM) in patients with IBD and ID without anaemia. The diagnostic criteria for ID were low serum ferritin (<30 µg/L in the absence of inflammatory activity or <100 µg/L with inflammation) and transferrin saturation index (TSAT) < 16%. The effect on iron levels and HRQoL, according to the health status questionnaires SF-12v2 and EQ-5D, was evaluated 1 month after FCM infusion in an outpatient setting. Results: Of the 105 patients who received FCM, 98 patients completed the study. After 1 month, a single dose of FCM significantly increased serum ferritin, serum iron and TSAT. Importantly, patients reported fewer ID symptoms and problems on all EQ-5D dimensions. They also had higher EQ-5D visual analogue scale and SF-12v2 scores after treatment. FCM had similar clinical effects on men and women and on patients with Crohn's disease (n = 66) and ulcerative colitis (n = 32). Conclusion: A single dose of FCM rapidly restored iron parameters and significantly improved patients' symptoms and HRQoL at 1 month after treatment.

Underwater hybrid endoscopic submucosal dissection in a rectal polyp: a case report of a new application of "underwater endoscopy"

Background: Hybrid endoscopic submucosal dissection (ESD) has been described as an alternative to traditional ESD. This technique is less time consuming, but the en bloc resection rate is lower than in ESD. Similar to endoscopic mucosal resection, the underwater technique could improve preliminary disadvantages of hybrid ESD. Case report: We attempted a mixture resection technique of Hybrid ESD with underwater endoscopic mucosal resection (EMR). Using this approach, an underwater hybrid ESD was successfully performed without adverse events on a 71 year old woman with a 20 mm adenoma. The histologic analysis identified a tubulovillious adenoma with high grade dysplasia and tumor free margins. Discussion: Underwater hybrid ESD could be an alternative to ESD. Moreover the modification of the "underwater method" provides a suitable way to overcome the technical drawbacks of the hybrid ESD

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Underwater hybrid endoscopic submucosal dissection in a rectal polyp: a case report of a new application of "underwater endoscopy".

BACKGROUND: Hybrid endoscopic submucosal dissection (ESD) has been described as an alternative to traditional ESD. This technique is less time consuming, but the en bloc resection rate is lower than in ESD. Similar to endoscopic mucosal resection, the underwater technique could improve preliminary disadvantages of hybrid ESD. CASE REPORT: We attempted a mixture resection technique of Hybrid ESD with underwater endoscopic mucosal resection (EMR). Using this approach, an underwater hybrid ESD was successfully performed without adverse events on a 71 year old woman with a 20 mm adenoma. The histologic analysis identified a tubulovillious adenoma with high grade dysplasia and tumor free margins. DISCUSSION: Underwater hybrid ESD could be an alternative to ESD. Moreover the modification of the "underwater method" provides a suitable way to overcome the technical drawbacks of the hybrid ESD.

Multiband mucosectomy for the treatment of challenging non-pedunculated colorectal polyps.

BACKGROUND AND STUDY AIMS: Endoscopic mucosal resection is the gold standard treatment for non-pedunculated colorectal polyps; however, some specific situations (location behind folds, scarred or flat morphology) can make this technique challenging. We aimed to assess the efficacy and safety of multiband mucosectomy (MBM) for resection of non-pedunculated colorectal polyps. PATIENTS AND METHODS: This was a retrospective study of patients in whom MBM was performed to resect large non-pedunculated colorectal polyps. All procedures were carried out using the Shooter multiband ligator kit (Cook Medical, Limerick, Ireland). A 3-month follow-up colonoscopy was performed in all patients. RESULTS: 10 patients underwent MBM for resection of 10 large (median 33.5 mm) non-pedunculated polyps. A total of 45 MBM sessions were carried out to resect all of the lesions using on average one rubber band per 1.5 cm2 of resected tissue. Complete resection was possible in 9 out of 10 lesions, although en bloc resection was only feasible in one lesion. Follow-up colonoscopy revealed residual adenoma in just one patient. No major complications were registered. CONCLUSIONS: In this small series of patients, MBM proved to be a safe and effective endoscopic resection technique for challenging non-pedunculated colorectal polyps.

Endoscopic ultrasound in gastric cancer staging before and after neoadjuvant chemotherapy. A comparison with PET-CT in a clinical series.

BACKGROUND: Treatment of gastric cancer is based on accurate staging. Emerging methods, such as PET-CT, are increasingly being used for this purpose. Our aim was to analyze the results of EUS and PET-CT in staging and restaging our patients with gastric cancer, comparing both of them with the histological results. METHODS: Patients with confirmed gastric cancer were prospectively enrolled. Inclusion criteria for the final analysis included only patients who finally received a surgical resection. All patients underwent preoperative TNM staging by means of EUS and PET-CT within 21 days prior to the surgical treatment. RESULTS: A total of 256 patients were included. The overall EUS accuracy for T staging was 78% and 80.2% in restaging. EUS showed its best accuracy when distinguishing T1-T2 tumors vs. T3-T4, with an increased accuracy in restaging. Regarding N staging, the overall accuracy of EUS was 76.2%, and 72.5% for PET-CT (p = 0.02). With regards to restaging, accuracy of EUS and PET-CT for N staging was 88.5% and 69%, respectively, with significant differences (p < 0.0001). CONCLUSIONS: EUS performed better than PET-CT in gastric cancer N staging and restaging. EUS accuracy in this setting is still suboptimal and probably more than one single diagnostic procedure should be used.

Discursos interpretativos y prácticas deliberativas: propuesta metodológica para formulación de políticas públicas sanitarias en Colombia / Interpretative speeches and deliberative practices: a methodological proposal for the formulation of public health policies in Colombia

RESUMEN Objetivo Formular políticas públicas sanitarias en escenarios de gobernanza local mediante la participación comunitaria, requiere reconocer la interacción de todos los actores sociales mediante enfoques argumentativos. Métodos A partir de un enfoque hermenêutico crítico, se utilizó un método retórico argumentativo para la formulación política desde un escenario socio céntrico, a partir de un diálogo de saberes entre la comunidad e instituciones locales, que permitieron identificar y comprender problemáticas sociales relevantes en salud, relacionadas con primera infancia y familia. Resultados Se construyó una política pública integral armonizada con programas sociales en salud existentes, para intervenir aspectos en referencia a la primera infancia, infancia, adolescencia y familia, donde las soluciones y su implementación parten de un núcleo participativo que trasmuta lo existente hacia algo inherente a la comunidad. Conclusiones El enfoque argumentativo en las políticas públicas reconoce el núcleo dialéctico de la política acercando la comunidad a los entes gubernamentales mediante la participación ciudadana activa en procesos de identificación, implementación y puesta en marcha de políticas sociales.(AU)

ABSTRACT Objective To propose public health policies in local governance scenarios through community participation; this requires recognizing the interaction of all social actors through argumentative approaches. Methods Considering a critical hermeneutic approach, an argumentative rhetorical method was used to conduct a political formulation from a socio-centric scenario, based on a dialogue of knowledge between the community and local institutions, which allowed to identify and understand relevant social problems in health, related with early childhood and family. Results A comprehensive public policy was built in accordance with existing social health programs to intervene aspects referring to early childhood, childhood, adolescence and family. Solutions and their implementation arise from a participatory nucleus that reconciles institutional political interests and social needs of the local community. Conclusions The argumentative approach in public policies recognizes the dialectical nucleus of politics by bringing the community closer to government entities through active citizen participation in processes of identification, implementation and execution of social policies.(AU)

Quantitative PCR and Digital PCR for Detection of Ascaris lumbricoides Eggs in Reclaimed Water.

The reuse of reclaimed water from wastewater depuration is a widespread and necessary practice in many areas around the world and must be accompanied by adequate and continuous quality control. Ascaris lumbricoides is one of the soil-transmitted helminths (STH) with risk for humans due to its high infectivity and an important determinant of transmission is the inadequacy of water supplies and sanitation. The World Health Organization (WHO) recommends a limit equal to or lower than one parasitic helminth egg per liter, to reuse reclaimed water for unrestricted irrigation. We present two new protocols of DNA extraction from large volumes of reclaimed water. Quantitative PCR (qPCR) and digital PCR (dPCR) were able to detect low amounts of A. lumbricoides eggs. By using the first extraction protocol, which processes 500 mL of reclaimed water, qPCR can detect DNA concentrations as low as one A. lumbricoides egg equivalent, while dPCR can detect DNA concentrations as low as five A. lumbricoides egg equivalents. By using the second protocol, which processes 10 L of reclaimed water, qPCR was able to detect DNA concentrations equivalent to 20 A. lumbricoides eggs. This fact indicated the importance of developing new methodologies to detect helminth eggs with higher sensitivity and precision avoiding possible human infection risks.

BISAP, RANSON, lactate and others biomarkers in prediction of severe acute pancreatitis in a European cohort.

BACKGROUND AND AIM: The study aims to assess and compare the predicting ability of some scores and biomarkers in acute pancreatitis. METHODS: We prospectively collected data from 269 patients diagnosed of acute pancreatitis, admitted to Virgen de las Nieves University Hospital between June 2010 and June 2012. Blood urea nitrogen (BUN), C-reactive protein, and creatinine were measured on admission and after 48 h, lactate and bedside index for severity acute pancreatitis (BISAP) only on admission and RANSON within the first 48 h. Definitions from 2012 Atlanta Classification were used. Area under the curve (AUC) was calculated for each scoring system for predicting severe acute pancreatitis (SAP), mortality, and intensive care unit (ICU) admission, obtaining optimal cut-off values from the receiver operating characteristic curves. RESULTS: Eight (3%) patients died, 17 (6.3%) were classified as SAP, and 10 (3.7%) were admitted in ICU. BISAP was the best predictor on admission for SAP, mortality, and ICU admission with an AUC of 0.9 (95% CI 0.83-0.97); 0.97 (95% CI 0.95-0.99); and 0.89 (95% CI 0.79-0.99), respectively. After 48 h, BUN 48 h was the best predictor of SAP (AUC = 0.96 CI: 0.92-0.99); BUN 48 h and BISAP were the best predictors for mortality (AUC = 0.97 CI: 0.95-0.99) and creatinine 48 h for ICU admission (AUC = 0.96 CI: 0.92-0.99). Lactate showed an AUC of 0.79 (CI: 0.71-0.88), 0.87 (CI: 0.78-0.96), and 0.77 (CI: 0.67-0.87) for SAP, mortality, and ICU admission, respectively. All parameters were predictors for SAP, mortality, and ICU admission, but C-reactive protein on admission was only a significant predictor of SAP. CONCLUSION: Bedside index for severity acute pancreatitis is a good predictive system for SAP, mortality, and ICU admission, being useful for triaging patients for ICU management. Lactate could be useful for developing new scores.

Extracción endoscópica de cuerpos extraños múltiples, grandes y cortantes. Transitando los límites de la endoscopia / Endoscopic retrieval of multiple large sharp foreign bodies from the stomach. Testing the limits of endoscopy

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[Interpretative speeches and deliberative practices: a methodological proposal for the formulation of public health policies in Colombia]. / Discursos interpretativos y prácticas deliberativas: Propuesta metodológica para formulación de políticas públicas sanitarias en Colombia.

OBJECTIVE: To propose public health policies in local governance scenarios through community participation; this requires recognizing the interaction of all social actors through argumentative approaches. METHODS: Considering a critical hermeneutic approach, an argumentative rhetorical method was used to conduct a political formulation from a socio-centric scenario, based on a dialogue of knowledge between the community and local institutions, which allowed to identify and understand relevant social problems in health, related with early childhood and family. RESULTS: A comprehensive public policy was built in accordance with existing social health programs to intervene aspects referring to early childhood, childhood, adolescence and family. Solutions and their implementation arise from a participatory nucleus that reconciles institutional political interests and social needs of the local community. CONCLUSIONS: The argumentative approach in public policies recognizes the dialectical nucleus of politics by bringing the community closer to government entities through active citizen participation in processes of identification, implementation and execution of social policies.

OBJETIVO: Formular políticas públicas sanitarias en escenarios de gobernanza local mediante la participación comunitaria, requiere reconocer la interacción de todos los actores sociales mediante enfoques argumentativos. MÉTODOS: A partir de un enfoque hermenêutico crítico, se utilizó un método retórico argumentativo para la formulación política desde un escenario socio céntrico, a partir de un diálogo de saberes entre la comunidad e instituciones locales, que permitieron identificar y comprender problemáticas sociales relevantes en salud, relacionadas con primera infancia y familia. RESULTADOS: Se construyó una política pública integral armonizada con programas sociales en salud existentes, para intervenir aspectos en referencia a la primera infancia, infancia, adolescencia y familia, donde las soluciones y su implementación parten de un núcleo participativo que trasmuta lo existente hacia algo inherente a la comunidad. CONCLUSIONES: El enfoque argumentativo en las políticas públicas reconoce el núcleo dialéctico de la política acercando la comunidad a los entes gubernamentales mediante la participación ciudadana activa en procesos de identificación, implementación y puesta en marcha de políticas sociales.

Comparison of AIMS65, Glasgow-Blatchford score, and Rockall score in a European series of patients with upper gastrointestinal bleeding: performance when predicting in-hospital and delayed mortality.

OBJECTIVE: AIMS65 is a score designed to predict in-hospital mortality, length of stay, and costs of gastrointestinal bleeding. Our aims were to revalidate AIMS65 as predictor of inpatient mortality and to compare AIMS65's performance with that of Glasgow-Blatchford (GBS) and Rockall scores (RS) with regard to mortality, and the secondary outcomes of a composite endpoint of severity, transfusion requirements, rebleeding, delayed (6-month) mortality, and length of stay. METHODS: The study included 309 patients. Clinical and biochemical data, transfusion requirements, endoscopic, surgical, or radiological treatments, and outcomes for 6 months after admission were collected. Clinical outcomes were in-hospital mortality, delayed mortality, rebleeding, composite endpoint, blood transfusions, and length of stay. RESULTS: In receiver-operating characteristic curve analyses, AIMS65, GBS, and RS were similar when predicting inpatient mortality (0.76 vs. 0.78 vs. 0.78). Regarding endoscopic intervention, AIMS65 and GBS were identical (0.62 vs. 0.62). AIMS65 was useless when predicting rebleeding compared to GBS or RS (0.56 vs. 0.70 vs. 0.71). GBS was better at predicting the need for transfusions. No patient with AIMS65 = 0, GBS ≤ 6, or RS ≤ 4 died. Considering the composite endpoint, an AIMS65 of 0 did not exclude high risk patients, but a GBS ≤ 1 or RS ≤ 2 did. The three scores were similar in predicting prolonged in-hospital stay. Delayed mortality was better predicted by AIMS65. CONCLUSION: AIMS65 is comparable to GBS and RS in essential endpoints such as inpatient mortality, the need for endoscopic intervention and length of stay. GBS is a better score predicting rebleeding and the need for transfusion, but AIMS65 shows a better performance predicting delayed mortality.

Evaluation of the efficacy and safety of Griflow(®) Dual, a pre-set two-flow infusion device for intravenous immunoglobulin (Flebogamma(®) 5%) administration.

PURPOSE: A single center, prospective study was performed to assess the efficacy and safety of Griflow(®) Dual a gravity-fed device for intravenous delivery of human immunoglobulin Flebogamma(®). METHODS: A total of 2 infusions in 2 visits per patient were assessed using G2 and G3 Griflow(®) Dual models that provide different flow rates adjusted to the patient's weight. To follow the most common method of intravenous immunoglobulin administration, infusion through the two-way device commenced with the low flow rate capillary (clamp closed) for 30 minutes and continued with the high flow rate, opening both ways. Reliability of flow delivery (average flow rates), adverse events, as well as functionality in daily practice (questionnaire to nurses) were assessed. RESULTS: twenty-five valid infusions were evaluated on 13 subjects. Except for the G2 model with closed clamp in which 14.5% deviation was observed, actual average flow rate values fell well into the maximum 10% deviation permitted with respect to expected charted values (G2: 55 and 142 mL/h; G3: 76 and 189 mL/h). Discrepancies could be explained by patient's arm movements or posture change during infusion. No adverse events related to the study device occurred. In the functionality questionnaires, nurse's comfort and safety of infusion with Griflow(®) Dual were rated higher than without Griflow(®) Dual but lower than with infusion pumps. CONCLUSIONS: Although it may not be as precise as an infusion pump, Griflow(®) Dual proved to be a reliable and suitable device to administer Flebogamma(®) 5%. Correct safety should be confirmed in a larger sample.

Prevalencia de caries y enfermedad periodontal en escolares del sector público de Cali, 2005 / Prevalence of cavities and periodontal diseases in school age children in Cali’s public school district in the year 2005

Introducción: La Secretaría de Salud Municipal de Cali, planteó la necesidad de conocer la situación bucodental en lapoblación escolar del sector público, mediante el perfil epidemiológico bucodental. Metodología: Estudio descriptivo. Se utilizó la metodología sugerida por la OMS para este tipo de trabajos con un muestreo en diversas etapas donde se combinan la técnica de conglomerados y el muestreo sistemático. Los autores examinaron 784 niños de 5 a 13 años de las 47 instituciones educativas seleccionadas. Se hizo una base de datos en el programa Access de donde se obtuvieron los porcentajes y los promedios. Resultados: La proporción de niños con historia de caries es 77%, la prevalencia de caries es 67.7%, el COE-d a los 5 años fue 0.64, el índice COP-D a los 12 años es 3.5. De los niños evaluados 46.1% se encuentran en condiciones de salud periodontal.En cuanto a la necesidad de asistencia y consulta, 91.5% requieren atención odontológica. Análisis: El porcentaje de niños sin historia de caries a los 12 años, para el presente perfil epidemiológico de la ciudad (2005),es superior al que suministran estudios similares en los niveles del departamento y de la nación (1998). A los 5 años de edad ocurre lo contrario, pues hay aumento en la incidencia de caries en los niños que ingresan a las instituciones educativas. El índice COP-D a los 12 años, clasifica al municipio en riesgo intermedio para caries. Conclusiones: Cali debe definir una estrategia en salud bucal donde se logre la concurrencia de los actores responsables a fin de alcanzar una acción efectiva que repercuta de modo positivo en la situación bucodental de la población. El índice COPD a los 12 años ubica al municipio en riesgo intermedio para caries, ello asociado con las condiciones periodontales identificadas, enfatizan que la salud bucal es una de las metas que se han de trabajar en los programas de salud pública del municipio.

Introduction: The Health Secretary of Cali wanted to know the buccodental status among school children to establish if necessary public health measures. Objectives: To determine the epidemiological oral health profile on scholars at the public schools of Cali, 2005. Methodology: This descriptive study used who oral index systematic and conglomerate multiestrata were performed a total of 784 children were examined for determining their Oral Health Status. Range of age was 5-13 years old. Study children camefrom 47 public schools. Results: Caries history was detected in 77% of children and actual caries prevalence was 67.7% DMF-d index was 0.64 at five y/o and increase to 3.5 at 12 years/old children. 46.1 % of the evaluated children were periodontally healthy, 34.3% had gingivitis and bleed on probing and 19.6% had dental calculus. Periodontal disease prevalence has 53.9%. Analysis: Children without caries history were higher here when compared the results of National Oral Health morbility study, 1998. Cali level of caries risk were intermediary as compares to National standing DMF-d indicators. Conclusions: It is necessary that the city of Cali establish a strategy to improve the standards in Oral Health among their young populatio. DMF-d indicator at 12 years/old childrenset them at an intermediary caries level of risk. These might be priority of Public Health Policy.

Distribution of somatostatin-like immunoreactivity in the brain of the caecilian Dermophis mexicanus (Amphibia: Gymnophiona): comparative aspects in amphibians.

The organization of the somatostatin-like-immunoreactive (SOM-ir) structures in the brain of anuran and urodele amphibians has been well documented, and significant differences were noted between the two amphibian orders. However, comparable data are not available for the third order of amphibians, the gymnophionans (caecilians). In the present study, we analyzed the anatomical distribution of SOM-ir cells and fibers in the brain of the gymnophionan Dermophis mexicanus. In addition, because of its known relationship with catecholamines in other vertebrates, double immunostaining for SOM and tyrosine hydroxylase was used to investigate this situation in the gymnophionan. Abundant SOM-ir cell bodies and fibers were widely distributed throughout the brain. In the telencephalon, pallial and subpallial cells were labeled, being most numerous in the medial pallium and amygdaloid region. Most of the SOM-ir neurons were found in the preoptic area and hypothalamus and showed a clear projection to the median eminence. Less conspicuously, SOM-ir structures were found in the thalamus, tectum, tegmentum, and reticular formation. Both SOM-ir cells and fibers were demonstrated in the spinal cord. The double-immunohistofluorescence technique revealed that catecholaminergic neurons and SOM-ir cells are largely intermingled in many brain regions but form totally separated populations. Many differences were found between the distribution of SOM-ir structures in Dermophis and that in anurans or urodeles. Some features were shared only with anurans, such as the abundant pallial SOM-ir cells, whereas others were common only to urodeles, such as the organization of the hypothalamohypophysial SOM-ir system. In addition, some characteristics were found only in Dermophis, such as the localization of the SOM-ir spinal cells and the lack of colocalization of catecholamines and SOM throughout the brain. Therefore, any conclusions concerning the SOM system in amphibians are incomplete without considering evidence for gymnophionans.

Spatiotemporal sequence of appearance of NPFF-immunoreactive structures in the developing central nervous system of Xenopus laevis.

Neuropeptide FF-like immunoreactive (NPFFir) cells and fibers were analyzed through development of Xenopus laevis. The first NPFFir cells appeared in the embryonic hypothalamus, which projected to the intermediate lobe of the hypophysis, the brainstem and spinal cord. Slightly later, scattered NPFFir cells were present in the olfactory bulbs and ventral telencephalon. In the caudal medulla, NPFFir cells were observed in the nucleus of the solitary tract only at embryonic and early larval stages. Abundant NPFFir cells and fibers were demonstrated in the spinal cord. The sequence of appearance observed in Xenopus shares many developmental features with mammals although notable differences were observed in the telencephalon and hypothalamus. In general, NPFF immunoreactivity developed earlier in amphibians than in mammals.

Lateral and medial amygdala of anuran amphibians and their relation to olfactory and vomeronasal information.

The amygdala of anurans is currently considered as a complex of nuclei that share many features with their counterparts in amniotes. In the present study, the subdivisions of the amygdala that are directly related to olfactory and vomeronasal information, were investigated in the anurans Rana perezi and Xenopus laevis. In particular, the connectivity of the main and accessory olfactory bulbs and their related amygdaloid nuclei was studied by means of in vivo and in vitro tract-tracing with dextran amines. The projections observed from the main olfactory bulb clearly innervate the newly redefined lateral amygdala within the ventral pallium and, to a lesser extent, the rostral portion of the medial amygdala. Injections into the accessory olfactory bulb exclusively revealed projections to the medial amygdala. Tracer applications into the lateral and medial nuclei revealed abundant intra-amygdaloid connections. The dual flow of olfactory and vomeronasal projections throughout the telencephalon was not strictly segregated since the lateral pallium and the lateral amygdala, both receiving olfactory information, were found to project to the medial amygdala (the only target of vomeronasal information), which in turn projects to the lateral amygdala. Additionally, both the lateral and the medial amygdala strongly project to the hypothalamus through the anuran equivalent of the stria terminalis. The main hodological features found in the present study suggest that forerunners of the olfactory and vomeronasal amygdaloid nuclei can be distinguished in anurans. This supports the notion that all tetrapods share a common pattern of organization of the amygdaloid complex, which links environmental (olfactory/vomeronasal) information and the behavioural response of the animal.

Calbindin-D28k immunoreactivity in the spinal cord of Xenopus laevis and its participation in ascending and descending projections.

Immunohistochemistry for calbindin-D28k (CB) revealed that the spinal cord of Xenopus laevis possess a large number of CB-containing neurons widely distributed in both the dorsal and ventral horns, including areas which possess long ascending projections to supraspinal structures. In addition, the presence of CB-immunoreactive axons in the spinal funiculi suggested that descending projections containing this calcium binding protein may originate in different brainstem nuclei. Apart from mapping CB-containing elements in the spinal cord, a double labeling approach was used that combined the retrograde transport of dextran amines with CB immunohistochemistry. Thus, dextran amine injections into the lateral reticular region of the rhombencephalon, the parabrachial region, the mesencephalon and the dorsal thalamus revealed many retrogradely labeled cells in the spinal cord, a few number of which were double labeled for CB and found in the superficial dorsal horn and in the ventral medial region of the ventral horn. Their axons passed mainly via the lateral funiculus. Tracer application into the cervical spinal cord, combined with CB immunohistochemistry, resulted in retrogradely labeled cells throughout the brain, five groups of which showed CB immunoreactivity: (1) the mesencephalic trigeminal nucleus, (2) the laterodorsal tegmental nucleus, (3) the raphe nucleus, (4) the middle reticular nucleus and (5) the inferior reticular nucleus. The presence of CB in spinal pathways suggests that CB may play a role in controlling spinal cells, mainly subserving visceroceptive and nociceptive information to supraspinal levels, and might also modulate reticulospinal pathways.

Colocalization of nitric oxide synthase and monoamines in neurons of the amphibian brain.

By means of double immunohistofluorescence techniques, we have investigated the colocalization of nitric oxide synthase and tyrosine hydroxylase (TH) or serotonin (5-HT) in the central nervous system of the anurans Rana perezi and Xenopus laevis and the urodele Pleurodeles waltl. A wide codistribution of neuronal populations, expressing these markers, was found throughout the brain and spinal cord. In contrast, colocalization of these markers was rather restricted. Only in the caudal portion of the brainstem raphe column in anurans, approximately 80% of the 5-HT-positive cells were also NOS-immunoreactive, whereas in the urodele brain, about 40% of the serotonergic cells at the level of the glossopharyngeal motor nucleus were simultaneously NOS-positive. In various brain regions, a wide codistribution of NOS- and TH-containing neurons was observed, but real colocalization of nitrergic and catecholaminergic cells was only found in a small neuron population in the posterior tubercle of anuran amphibians. Therefore, in amphibians, only a distinct and small cell population within the serotonergic raphe column (anurans and urodele) and in the catecholaminergic posterior tubercle (anurans) seem to produce simultaneously nitric oxide.