Analysis of Stress Indicators During Cryopreservation of Seeds of Landrace Maize (Zea mays).

BACKGROUND: Maize breeding programs focus on the development of hybrid varieties and the cultivation of landrace materials is discouraged; however, they are a valuable source of genes and their conservation is advisable. OBJECTIVE: Analyzing some stress indicators during cryopreservation of maize landrace seeds. MATERIALS AND METHODS: Seeds of 35 accessions of landrace maize were collected in two regions of Costa Rica and cryopreserved by direct immersion in liquid nitrogen (LN). Membrane integrity, germination of seeds and DNA methylation in tissues were analyzed 5, 7 and 9 days after rewarming. RESULTS: Germination of landrace maize seeds was near 100 % for most accessions. No statistically significant differences in germination were observed between non-cryopreserved controls and seeds stored in LN for 1 h or 1 year. Membrane integrity, number of leaves and root and shoot length of plantlets were similar after cryostorage of seeds for 1 h and 1 year. A short delay in growth of cryostored seed compared to non-frozen controls was observed. Changes in the proportion of DNA methylation were noted from 0 to day 9 in the organs studied depending on the germination stage and cryopreservation treatment. CONCLUSION: It may be inferred that many of the methylated genes were related to growth and development. In addition, a cryobank of maize landraces from two regions of Costa Rica was established.

Transmission of lymphocystis disease virus to cultured gilthead seabream, Sparus aurata L., larvae.

The transmission of lymphocystis disease virus (LCDV) to gilthead seabream, Sparus aurata L., larvae was investigated using fertilized eggs from a farm with previous reports of lymphocystis disease. LCDV genome was detected by PCR-hybridization in blood samples from 17.5% of the asymptomatic gilthead seabream broodstock analysed. Using the same methodology, eggs spawned from these animals were LCDV positive, as well as larvae hatched from them. The presence of infective viral particles was confirmed by cytopathic effects development on SAF-1 cells. Whole-mount in situ hybridization (ISH) and immunohistochemistry (IHC) showed the presence of LCDV in the epidermis of larvae hatched from LCDV-positive eggs. When fertilized eggs were disinfected with iodine, no viral DNA was detected either in eggs (analysed by PCR-hybridization) or in larvae (PCR-hybridization and ISH). These results suggest the vertical transmission of LCDV, the virus being transmitted on the egg surface. Larvae hatched from disinfected eggs remain LCDV negative during the endotrophic phase, as showed by PCR-hybridization, ISH and IHC. After feeding on LCDV-positive rotifers, viral antigens were observed in the digestive tract, which suggests that viral entry could be achieved via the alimentary canal, and that rotifers can act as a vector in LCDV transmission to gilthead seabream larvae.

Ten years since the introduction of therapeutic hypothermia in neonates with perinatal hypoxic-ischaemic encephalopathy in Spain.

INTRODUCTION: More than a decade has passed since therapeutic hypothermia (TH) was introduced in Spain; this is the only neuroprotective intervention that has become standard practice in the treatment of perinatal hypoxic-ischaemic encephalopathy (HIE). This article aims to provide a current picture of the technique and to address the controversies surrounding its use. DEVELOPMENT: In the last 10 years, TH has been successfully implemented in the vast majority of tertiary hospitals in Spain, and more than 85% of newborns with moderate or severe HIE currently receive the treatment. The factors that can improve the efficacy of TH include early treatment onset (first 6 h of life) and the control of comorbid factors associated with perinatal asphyxia. In patients with moderate HIE, treatment onset after 6 h seems to have some neuroprotective efficacy. TH duration longer than 72 h or deeper hypothermia do not offer greater neuroprotective efficacy, but instead increase the risk of adverse effects. Controversy persists around the sedation of patients during TH, the application of the treatment in infants with mild HIE, and its application in other scenarios. Prognostic information and time frame are one of the most challenging aspects. CONCLUSIONS: TH is universal in countries with sufficient economic resources, although certain unresolved controversies remain. While the treatment is widespread in Spain, there is a need for devices for the transfer of these patients and their centralisation.

Ten years since the introduction of therapeutic hypothermia in neonates with perinatal hypoxic-ischaemic encephalopathy in Spain. / Una década después de la implantación en España de la hipotermia terapéutica en el recién nacido con encefalopatía hipóxico-isquémica perinatal.

INTRODUCTION: More than a decade has passed since therapeutic hypothermia (TH) was introduced in Spain; this is the only neuroprotective intervention that has become standard practice in the treatment of perinatal hypoxic-ischaemic encephalopathy (HIE). This article aims to provide a current picture of the technique and to address the controversies surrounding its use. DEVELOPMENT: In the last 10 years, TH has been successfully implemented in the vast majority of tertiary hospitals in Spain, and more than 85% of newborns with moderate or severe HIE currently receive the treatment. The factors that can improve the efficacy of TH include early treatment onset (first 6hours of life) and the control of comorbid factors associated with perinatal asphyxia. In patients with moderate HIE, treatment onset after 6hours seems to have some neuroprotective efficacy. TH duration longer than 72hours or deeper hypothermia do not offer greater neuroprotective efficacy, but instead increase the risk of adverse effects. Unclarified aspects are the sedation of patients during TH, the application of the treatment in infants with mild HIE, and its application in other scenarios. Prognostic information and time frame are one of the most challenging aspects. CONCLUSIONS: TH is universal in countries with sufficient economic resources, although certain unresolved controversies remain. While the treatment is widespread in Spain, there is a need for cooling devices for the transfer of these patients and their centralisation.

Rapid TB diagnostic service and community action to FIND.TREAT.ALL#EndTB, Maputo, Mozambique, 2013-2018.

Finding and treating all tuberculosis (TB) patients is crucial for ending TB. We investigated whether rapid diagnostic turnaround time (TAT) and patient tracking could increase TB treatment initiation in Maputo, Mozambique. Among 3329 TB patients newly diagnosed by the University Eduardo Mondlane-Anti-Persoonsmijnen Ontmijnende Product Ontwikkeling/Anti-Personnel Landmines Detection Product Development (APOPO) Laboratory between 2013 and 2018, on average 61% were verifiably linked to care. This proportion increased from 54% (first half 2013) to 79% (second half 2018) after introducing a 24-hour TAT in 2015 and patient tracking conducted by a community-based partner, Associação Kenguelekezé, in 2017. Rapid, well-connected TB diagnostic services can reduce pre-treatment loss to follow-up and support the joint initiative of WHO, Stop TB and Global Fund to 'FIND.TREAT.ALL.#EndTB'.

Identifier et traiter tous les patients atteints de tuberculose (TB) est crucial pour mettre fin à la TB. Nous avons vérifié si un délai plus court de diagnostic (TAT) et un système de recherche des patients augmentait la mise en œuvre du traitement de la TB à Maputo, Mozambique. Parmi 3329 patients TB nouvellement diagnostiqués par le laboratoire UEM-APOPO (2013­2018), en moyenne 61% ont été réellement connectés à la prise en charge. Le pourcentage a augmenté de 54% (première moitié de 2013) à 79% (deuxième moitié de 2018) après l'introduction du TAT en 24h en 2015 et la surveillance des patients effectuée par notre partenaire l'association à base communautaire Kenguelekezé en 2017. Notre expérience suggère que des services de diagnostic de TB rapides, bien connectés peuvent réduire les abandons avant le traitement et soutenir les tentatives mondiales « FIND.TREAT.ALL.#EndTB ¼.

La búsqueda y el tratamiento de todos los pacientes con tuberculosis (TB) son primordiales para poner fin a esta enfermedad. Se investigó si con un lapso corto de obtención del diagnóstico y el seguimiento de los pacientes aumentaría el inicio del tratamiento antituberculoso en Maputo, Mozambique. En promedio, en 61% de los 3329 casos nuevos de TB diagnosticados en el laboratorio UEM-APOPO (2013­2018) se confirmó la vinculación de los pacientes con los servicios de atención. El porcentaje aumentó de 54% (primer semestre del 2013) a 79% (segundo semestre del 2018), después de haber introducido un plazo de obtención del diagnóstico de 24 horas en el 2015 y la localización de los pacientes por parte de la asociación comunitaria Kenguelekezé en el 2017. Esta experiencia indica que los servicios diagnósticos de la TB que son rápidos y mantienen vínculos adecuados disminuyen la pérdida durante el seguimiento antes de comenzar el tratamiento y fortalecen los esfuerzos de la iniciativa 'FIND.TREAT.ALL.#EndTB'.

Extrinsic ileal occlusion secondary to appendiceal mucocele of unusual size.

Appendiceal mucocele due to papilar cystadenoma is a rare condition, characterized by a diffuse swelling, a distension of the lumen secondary to accumulation of mucoid substance and occasional mucinous intraperitoneal deposits. It is often asymptomatic (approximately 25%) and is found incidentally during abdominal imaging studies or surgical exploration. Pre-operative recognition is difficult but very important because of the possibility of rupture during surgery with the development of pseudomyxoma peritonei. A case of benign mucocele presenting acutely as an occlusive syndrome is reported. The surgical treatment consisted of an appendicectomy with partial caecectomy. The clinicopathological and imaging diagnostic features of this pathological entity, as well as the main therapeutic approaches suggested in the literature, are commented on.

Una década después de la implantación en España de la hipotermia terapéutica en el recién nacido con encefalopatía hipóxico-isquémica perinatal / Ten years since the introduction of therapeutic hypothermia in neonates with perinatal hypoxic-ischaemic encephalopathy in Spain

Introducción: Se cumple ahora más de una década del inicio de la hipotermia terapéutica (HT)en Espa ̃na, la única intervención neuroprotectora que ha venido a ser práctica estándar en eltratamiento de la encefalopatía hipóxico-isquémica perinatal (EHI). El objetivo de este artículoes ofrecer un panorama actual y presentar las controversias surgidas alrededor de la aplicaciónde esta terapia. Desarrollo: En esta década se ha implantado con éxito la HT en la gran mayoría de los hospitalesterciarios de Espa ̃na y más del 85% de los recién nacidos con EHI moderada-grave reciben estaterapia. Entre los aspectos que pueden mejorar la eficacia de la HT están su inicio precoz dentrode las primeras 6 h de vida y el control de factores comórbidos asociados a la asfixia perinatal. En los pacientes con EHI moderada el inicio después de las 6 h parece mantener cierta eficacianeuroprotectora. Una duración de la HT mayor de 72 horas o un enfriamiento más profundo noofrecen mayor eficacia neuroprotectora y aumentan el riesgo de efectos adversos. Aspectosno bien aclarados aún son la sedación durante la HT y la aplicación de esta intervención a losneonatos con EHI leve y en otros escenarios. La información pronóstica y su marco temporal esuno de los aspectos más desafiantes. Conclusiones: La HT es universal en países con recursos económicos, aunque existen puntos de controversia no resueltos. Si bien es un tratamiento generalizado en nuestro país, falta disponerde dispositivos para el traslado de estos pacientes y su centralización.(AU)

Introduction: More than a decade has passed since therapeutic hypothermia (TH) was introduced in Spain; this is the only neuroprotective intervention that has become standard practice inthe treatment of perinatal hypoxic-ischaemic encephalopathy (HIE). This article aims to providea current picture of the technique and to address the controversies surrounding its use. Development: In the last 10 years, TH has been successfully implemented in the vast majority of tertiary hospitals in Spain, and more than 85% of newborns with moderate or severeHIE currently receive the treatment. The factors that can improve the efficacy of TH includeearly treatment onset (first 6 hours of life) and the control of comorbid factors associated withperinatal asphyxia. In patients with moderate HIE, treatment onset after 6 hours seems to havesome neuroprotective efficacy. TH duration longer than 72 hours or deeper hypothermia do notoffer greater neuroprotective efficacy, but instead increase the risk of adverse effects. Unclarified aspects are the sedation of patients during TH, the application of the treatment in infantswith mild HIE, and its application in other scenarios. Prognostic information and time frame areone of the most challenging aspects. Conclusions: TH is universal in countries with sufficient economic resources, although certainunresolved controversies remain. While the treatment is widespread in Spain, there is a needfor cooling devices for the transfer of these patients and their centralisation.(AU)

Accuracy of giant African pouched rats for diagnosing tuberculosis: comparison with culture and Xpert® MTB/RIF.

SETTING: Enhanced tuberculosis (TB) case finding using detection rats in Tanzania. OBJECTIVES: To assess the diagnostic accuracy of detection rats compared with culture and Xpert® MTB/RIF, and to compare enhanced case-finding algorithms using rats in smear-negative presumptive TB patients. DESIGN: A fully paired diagnostic accuracy study in which sputum of new adult presumptive TB patients in Tanzania was tested using smear microscopy, 11 detection rats, culture and Xpert. RESULTS: Of 771 eligible participants, 345 (45%) were culture-positive for Mycobacterium tuberculosis, and 264 (34%) were human immunodeficiency virus (HIV) positive. The sensitivity of the detection rats was up to 75.1% (95%CI 70.1-79.5) when compared with culture, and up to 81.8% (95%CI 76.0-86.5) when compared with Xpert, which was statistically significantly higher than the sensitivity of smear microscopy. Corresponding specificity was 40.6% (95%CI 35.9-45.5) compared with culture. The accuracy of rat detection was independent of HIV status. Using rats for triage, followed by Xpert, would result in a statistically higher yield than rats followed by light-emitting diode fluorescence microscopy, whereas the number of false-positives would be significantly lower than when using Xpert alone. CONCLUSION: Although detection rats did not meet the accuracy criteria as standalone diagnostic or triage testing for presumptive TB, they have additive value as a triage test for enhanced case finding among smear-negative TB patients if more advanced diagnostics are not available.

Recursos y propuestas de mejora en el manejo de la Apnea Obstructiva del Sueño en Andalucía / Resources and proposals for improvement in the management of obstructive sleep apnea in Andalusia

Título abreviado: Se exponen los recursos asistenciales del Sistema Sanitario Público de Andalucía relacionados con el diagnóstico, tratamiento y seguimiento de los pacientes con apnea obstructiva del sueño. Igualmente se describen propuestas para una optimización en el manejo de esta entidad enfatizando la necesidad de recursos y colaboración entre diferentes niveles asistenciales. Objetivo: Determinar en el Sistema Sanitario Público de Andalucía (SSPA) los recursos actuales en la apnea obstructiva del sueño (AOS) e identificar propuestas de mejora. Método: Estudio transversal, basado en encuestas realizadas en 49 hospitales del SSPA. Se registraron variables relacionadas al diagnóstico, tratamiento y seguimiento de la AOS, distinguiendo entre centros que realizaban polisomnografía y los que no. Incluimos un apartado sobre propuestas de mejora. Resultados: Un 97% de los centros realizan poligrafía y un 29% polisomnografía, y sólo el 39,5% disponen de consulta específica. La demora diagnóstica para la poligrafía es de 169 ± 163,4 días y para la polisomnografía de 173 ± 152,5 días. Se realizan un total de 1.113 ± 1.004,6 pruebas por 100.000 habitantes y año, de ellas 235 ± 166,2 son poligrafías diagnósticas. La presión eficaz se titula en un 49% con auto-CPAP y el control terapéutico lo realiza frecuentemente la empresa suministradora (77%). Entre las propuestas de mejora, destacan la falta de un protocolo de derivación y coordinación entre diferentes niveles asistenciales (90% de los encuestados), y la demora diagnóstica, atribuible en un 63% de los casos al déficit de recursos físicos y de personal. Conclusión: El número de pruebas diagnósticas aún son insuficientes y la demora diagnóstica es excesiva. Sobre todo, en el control terapéutico la empresa suministradora tiene un papel relevante. Se propone la realización de protocolos entre diferentes unidades asistenciales y aumentar los recursos actuales. (AU)

Short title: The healthcare resources of the Andalusian Public Health System related to the diagnosis, treatment and follow-up of patients with obstructive sleep apnea are presented. Proposals for optimizing the management of this entity are also described, emphasizing the need for resources and collaboration between different levels of care. Objective: Determine the current resources in the Public Health System of Andalusia (SSPA) in obstructive sleep apnea (OSA) and identify proposals for improvement. Method: Cross-sectional study, based on surveys conducted in 49 SSPA hospitals. Variables related to the diagnosis, treatment, and follow-up of OSA were recorded, distinguishing between centers that performed polysomnography and those that did not. We include a section on proposals for improvement. Results: 97% of the centers perform polygraphy and 29% polysomnography, and only 39.5% have a specific consultation. The diagnostic delay for polygraphy is 169 ± 163.4 days and for polysomnography 173 ± 152.5 days. A total of 1,113 ± 1,004.6 tests are performed per 100,000 inhabitants per year, of which 235 ± 166.2 are diagnostic polygraphs. The effective pressure is titrated in 49% with auto-CPAP and therapeutic control is frequently carried out by the supplying company (77%). Among the proposals for improvement, the lack of a protocol for referral and coordination between different levels of care (90% of those surveyed), and the diagnostic delay, attributable in 63% of cases to the lack of physical and personnel resources, stand out. Conclusion: The number of diagnostic tests are still insufficient and the diagnostic delay is excessive. Above all, in therapeutic control, the supplier company has a relevant role. It is proposed to carry out protocols between different care units and increase current resources

[Medication reconciliation: From admission to primary care]. / Conciliación de la medicación: desde el ingreso hasta la atención primaria.

OBJECTIVES: To implement a medication reconciliation circuit of inter-level, comprehensive and multidisciplinary approach in an integrated health organization. To measure the discrepancies detected in each of the steps studied. METHOD: A prospective intervention study of one-year duration. The medication is reconciled at admission to the hospital, at discharge and when the patient goes to his Primary Care physician. The number and type of discrepancies detected each time the medication is reconciled are collected and resolved, as well as the total number of drugs before and after each reconciliation process quantified. RESULTS: Between November 1, 2013 and October 31, 2014 the medication had been reconciled to 77 patients, 63% male, mean age 69,5 years. Mean admission discrepancy per patient was 7,85, 3,67 at discharge and 2,19 at Primary Care. CONCLUSIONS: This program of medication reconciliation, in addition to detect and resolve discrepancies, has been a starting point for establishing new channels of communication between the different health professionals who have participated in the program and disseminate the safety culture within the organization.

Obstetric antiphospholipid syndrome.

Obstetric antiphospholipid syndrome is an acquired autoimmune disorder that is associated with various obstetric complications and, in the absence of prior history of thrombosis, with the presence of antiphospholipid antibodies directed against other phospholipids, proteins called cofactors or PL-cofactor complexes. Although the obstetric complications have been related to the procoagulant properties of antiphospholipid antibodies, pathological studies of human placenta have shown the proinflammatory capacity of antiphospholipid antibodies via the complement system and proinflammatory cytokines. There is no general agreement on which antiphospholipid antibodies profile (laboratory) confers the greatest obstetric risk, but the best candidates are categories I and IIa. Combined treatment with low doses of aspirin and heparin achieves good obstetric and maternal outcomes. In this study, we also review the therapeutic possibilities in refractory cases, although the likelihood of progressing to other autoimmune diseases is low. We briefly comment on incomplete obstetric antiphospholipid syndrome, also known as antiphospholipid antibody-mediated pregnancy morbidity syndrome.

Lumbar ontogenetic allometry and dimorphism in humans. A case for comparison between interspecific and intraspecific scaling.

The ontogenetic allometry of the lumbar region of 1913 humans (1228 females and 685 males), ranging from newborn to 21-year-old individuals, was studied by means of length, width, projected surface area and bone mineral density of the segment L2 - L4, obtained by dual X-ray absorptiometry (DXA). All these parameters were regressed to body mass and height of the individuals, considered alternatively as the independent variable. Firstly, we addressed the comparison between the results obtained on both sexes in order to elucidate whether ontogenetic differences existed. Length of the segments increased significantly faster in females than in males, independently whether the regression was made against body mass or height, while in both types of regression width scaled in males faster than in females. Regarding bone mineral density, although males increased bone mineral density faster than females, slope differences were not significant. However, y-interception was significantly higher in females than in males when bone mineral density was regressed to body mass. Results on length and width are compared with others from previous research on allometry. Finally, global results are discussed as regards the slope predictions for interspecific scaling.

Recomendaciones para la normalización gradual en la enfermedad COVID-19 en las unidades de trastornos respiratorios del sueño de Andalucía / No disponible

No disponible

Sex differences on the electrocardiographic pattern of cardiac repolarization: possible role of testosterone.

BACKGROUND: Cardiac repolarization has been shown to be shorter and faster in men compared with women. In this study, we examined the electrocardiographic pattern of repolarization in patients with abnormal plasma levels of testosterone to gain insight into the role that this hormone plays in modulating repolarization. METHODS AND RESULTS: Repolarization was measured in 27 castrated men, 26 women with virilization, and 53 control subjects pair-matched for age and sex. Repolarization in castrated men was slower and longer than that of normal men. Women with virilization exhibited a shorter and faster repolarization than normal women and castrated men. These differences are the opposite of those found in the normal population. Finally, the changes observed in castrated men may be reverted by testosterone. CONCLUSIONS: We conclude that testosterone plays an important role in modulating cardiac repolarization.

Informativeness of polymorphic markers for prenatal diagnosis of recessive dystrophic epidermolysis bullosa in Spanish families at risk.

DNA polymorphic markers are useful for the prenatal diagnosis of generalized recessive dystrophic epidermolysis bullosa (GRDEB) in families with at least one child affected with the disease. It is our policy to investigate families at risk using DNA polymorphic markers before a new pregnancy is intended, to inform about the real chances of offering an accurate prenatal diagnosis based on such DNA markers. We have analysed 10 Spanish families with at least one child affected with GRDEB with three different markers linked to the type VII collagen gene: the intragenic PvuII RFLP, and two markers mapped close to the COL7A1 gene, an MspI polymorphism belonging to the anonymous marker D3S2, and the microsatellite D3S1100. The PvuII marker was partially informative in two of 10 families, and was fully informative in three additional families; The MspI marker was partially informative in two of 10 families, and was fully informative in three additional families; it was not informative in five families. The D3S1100 microsatellite was partially informative in two out of 10 families, and fully informative in the other eight families. Combination of all three markers was shown to be partially informative in one family and fully informative in the remaining nine families. Using this combination of markers, we have inferred an accurate linkage-based prenatal diagnosis of GRDEB in four pregnancies. Surprisingly, the intragenic marker PvuII had a very limited usefulness, despite the results of previous studies; the microsatellite D3S1100 offered the best results for an eventual prenatal diagnosis of GRDEB. However, families at risk should be informed that extragenic markers may fail due to the possibility of recombination, estimated as 5% for D3S1100.

Central nervous system vasculopathy in neonatal lupus erythematosus.

Central nervous system involvement in neonatal lupus erythematosus (NLE) has not been previously reported. We report four patients with NLE, all with complete congenital heart block and three with cerebral ultrasound and color Doppler flow imaging (CDFI) studies demonstrating evidence of associated vasculopathy in the gangliothalamic vasculature. CDFI confirmed blood flow through the affected vessels, indicating that blood flow was not compromised at this early stage. Short-term follow-up revealed no signs of progression of the vasculopathy, focal ischemia, gangliothalamic atrophy, or neurological impairment. Nevertheless, the implications of this finding with respect to the natural history of NLE remain to be defined, particularly in cases in which the disease develops into systemic lupus erythematosus later in life. Besides specific diagnostic studies for NLE, cerebral ultrasound, and CDFI studies are mandatory in all cases of complete congenital heart block, regardless of whether mothers are diagnosed as having connective-tissue disease or not. Neonates with signs of vasculopathy in the gangliothalamic region should be examined for NLE.

Postnatal adaptation of brain circulation in preterm infants.

Global and regional postnatal cerebral circulatory changes in stable preterm infants were studied, and their relation to brain injury was assessed. Thirty-five preterm infants were studied on the first and second days of age. Cerebral blood flow (CBF) (mL/hg per min) and cerebral blood volume (CBV) (mL/hg) were measured using near-infrared spectroscopy. The cerebral blood flow velocity (cm/second) (peak systolic, diastolic flow, mean flow) and resistance index (RI) were determined in the internal carotid, anterior cerebral, and striate arteries by color Doppler flow imaging. Serial cerebral ultrasound studies were performed to detect changes in brain parenchymal echogenicity or intraventricular hemorrhage (IVH); the maximum severity of these findings was considered. CBF and cerebral blood flow velocity increased significantly with time, and such changes were independent of mean blood pressure, PO(2), PCO(2), hematocrit, or glycemia. In contrast, CBV and RI remained unchanged. According to the results of sonograms, no differences were found in postnatal CBF and cerebral blood flow velocity changes, regardless of whether patients had or did not have parenchymal lesions or IVH. However, higher CBV values were found on the second day in infants with IVH compared with infants without IVH. Early coupling of CBF and metabolic demands is independent of blood pressure. Improved venous return, instead of vasodilation, could be important in this adaptation.

Immunocytochemical and ultrastructural characterization of endocrine cells and nerves in the intestine of Rana temporaria.

Endocrine cells have been identified in the intestine of the frog Rana temporaria after application of the Grimelius and Masson-Fontana techniques. These endocrine cells were examined using immunocytochemical techniques on paraffin and semithin sections for light microscopy. After testing 19 antisera, 12 immunoreactivities were identified. Numerous serotonin-, somatostatin- and GLP-1-immunoreactive cells; a moderate number of PYY-, glucagon-, VIP-, gastrin/CCK-immunoreactive cells and few human PP-, bombesin-, substance P- and neurotensin-immunoreactive cells were found. VIP- and met-enkephalin were identified in nerve fibers of the muscular layer. Using semithin-thin sections five types of endocrine cells (serotonin-, somatostatin-, gastrin/CCK-, glucagon- and bombesin-immunoreactive cells) have been characterized according to their immunocytochemical reaction and the ultrastructure of the secretory granules.

Structural study of the frog Rana temporaria larval stomach.

The gastric wall of Rana temporaria tadpoles consists of a well-developed mucosa and thin muscular and serosa layers. Three cellular types--mucous, ciliated and endocrine cells--make up the lining epithelium. Different types of endocrine cells exist. Argyrophylic endocrine cells can be recognized in semithin sections of plastic-embedded material while non-argyrophylic endocrine cells can only be identified under the electron microscope. Glands are composed mainly of well-differentiated oxyntic cells and, occasionally, scarce endocrine cells. Oxyntic cells show abundant mitochondria and smooth endoplasmic reticulum, but do not contain zymogen granules as do those present in adults. Secretory canaliculi with microvilli are also well-developed. The lamina propria contains numerous vascular sinuses and nerve bundles which innervate the endothelium and some endocrine cells. The neuroendocrine regulation of frog gastric functions seems therefore to have developed in young tadpoles. Nerve fibers also innervate the muscular propria, which is composed of a single layer of smooth muscle cells. Underlying the muscle, connective fibers and a flattened layer of mesothelial cells make up the serosa. In summary, the structure of the frog larval stomach shows a well-differentiated histological pattern, especially referring to surface epithelium and glands. Some of the histological traits will also be present in adult frogs while others are characteristic of the tadpole's stage.