Chamber specific expression of Myosin heavy chain 7b in the heart of vertebrates

In extant vertebrates, myosin heavy chain (MyHC) 6 and 7 are the main isoforms of atrial and ventricular myocardium respectively, whereas MyHC7b has been proposed to be an ancient cardiac isoform only expressed during embryonic development in modern species. In preliminary immunohistochemical studies of the heart of the lesser spotted dogfish (Scyliorhinus canicula; Chondrichthyes), we have observed that while MF20 labels homogeneously all the myocardium, A4.1025 labels the inflow cardiac segments (sinus venosus and atrium) but not the outflow segments (ventricle and conus arteriosus). In order to interpret these results, we have performed western and slot blots from samples of dogfish and hamster (Mesocricetus auratus) hearts, HPLC-ESI-MS/MS from dogfish samples, and immunohistochemistry in hearts of representative species of vertebrates, namely elasmobranchs, polypteriforms, acipenseriforms, teleosts and mammals, using MF20 and A4.1025 antibodies. Western and slot blot results confirmed the specificity of MF20 and A4.1025 for MyHC in dogfish, as well as their differential reactivity against different myocardial segments. HPLC-ESI-MS/MS using protein databases from Callorhinchus milii (Chondrichthyes) and Chordata revealed the presence of MyHC6 and 7 in all the dogfish myocardial segments, and of MyHC7b only in the outflow segments. Immunohistochemistry showed that while MF20 signals were homogeneous in all the myocardial segments of all the species studied, A4.1025 signals were restricted to the inflow myocardial segments in elasmobranchs, homogeneous in teleosts and acipenseriforms, and low in the ventricle of polypteriforms. It can be inferred that the A4.1025 antibody, as opposed to MF20, has a low affinity for MyHC7b, at least in the dogfish. In addition, we show that MyHC distribution in the cardiac chambers has changed during the evolution of gnathostomes.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech.Study supported by grants CGL2017-85090-P and CGL2014-52356-P (Ministerio de Economía y Competitividad), FPU15/03209 (Ministerio de Educación, Cultura y Deporte), contract UMAJI75 (Junta de Andalucía, European Social Found), FEDER and Universidad de Málaga

Myocardial trabeculation in embryos of Scyliorhinus canicula (Elasmobranchii, Chondrichthyans)

Currently, three types of ventricular myoarchitecture are recognized in vertebrates, namely compact, spongy (trabeculated) and mixed myocardium. Mixed myocardium, which has been recently proposed as the primitive condition in gnathostomes, is composed of two myocardial layers: an inner trabeculated and an outer compact one. The trabeculation process has been studied in teleosts, showing exclusively spongy myocardium, and mammals and birds, characterized by a compact myocardial ventricular wall. In zebrafish, mouse and chicken embryos, the trabeculae develop as luminal myocardial ridges protruding into the lumen. In mammals and birds, further compactation of trabeculae leads to the formation of a compact layer. The potential mechanisms that may contribute to the formation of the ridges are under discussion and include myocardial proliferation, endocardial invagination, and bending of the entire myocardial layer. However, no description of the development of the mixed myocardium is available. To shed some light on this issue, we have studied the heart development of an elasmobranch species with mixed myocardium, the lesser spotted dogfish (Scyliorhinus canicula; Chondrichthyes), by means of histological and immunohistochemical techniques for light microscopy, semithin sections, scanning electron microscopy and transmission electron microscopy. Our results suggest that in the dogfish the intertrabecular spaces develop by connections between early intramyocardial spaces and the lumen of the ventricle through invaginations of the endocardial line. Chondrichthyans are the earliest diverged lineage of gnathostomes and, consequently, they have the most primitive cardiac design. Although chicken, mouse, and recently zebrafish have been considered powerful vertebrate models to study heart development, we propose that the trabeculation process in the dogfish is representative of the early steps of the ventricular morphogenesis in vertebrates.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech.Study supported by grant CGL2017-85090-P and CGL2014-52356-P (Ministerio de Economía y Competitividad), FPU15/03209 (Ministerio de Educación, Cultura y Deporte), FEDER, and Universidad de Málaga

The bulbus arteriosus of the holocephalan heart

El resumen aparece en el Program & Abstracts of the 10th International Congress of Vertebrate Morphology, Barcelona 2013.Anatomical Record, Volume 296, Special Feature — 1: P-074.Previous work has shown that the outflow tract of the elasmobranch heart, namely the cardiac portion intercalated between the ventricle and the ventral aorta, does not consist of a single component, the conus arteriosus, as has classically been assumed, but two, the myocardial conus arteriosus and the non-myocardial bulbus arteriosus. From the evolutionary perspective, knowledge of the anatomy of the cardiac outflow tract of the holocephali is important, as they are the sister group of elasmobranchs. Our aim is to describe the cardiac outflow tract of four holocephalan species, two of them, Chimaera monstrosa and Hydrolagus affinis of the family Chimaeridae, and the other two, Harriotta raleighana and Rhinochimaera atlantica, of the family Rhinochimaeridae. The cardiac outflow tract of the four species consisted of a myocardial conus arteriosus, furnished with valves, and a bulbus arteriosus devoid of cardiac muscle. Both the bulbus and conus are tubular in shape. The length of the bulbus relative to the total length of the outflow tract is somewhat smaller in the rhinochimaerids (15%-19%) than in the chimaerids (19%-23%). The bulbus is covered by epicardium and is crossed by the main coronary artery trunks. Histologically, the bulbus is mainly composed of elastin and collagen, and, to a lesser extent, by smooth muscle. This suggests that in holocephalans, the bulbus actively helps to protect the gill vasculature from exposure to high-pressure pulses of blood. Our results prove that the bulbus arteriosus is common to chondrichthyans. In addition, they support the hypothesis that the cardiac outflow tract consisted of a conus arteriosus and a bulbus arteriosus from the beginning of the jawed vertebrate radiation, contributing to our understanding of the morphological changes that have occurred at the arterial pole of the heart in both actinopterygians and sarcopterygians.Proyecto CGL2010-16417/BOS; Fondos FEDE

MYOCARDIAL STRUCTURE AND VASCULARIZATION OF THE HEART VENTRICLE IN HOLOCEPHALI: IMPLICATIONS FOR HEART EVOLUTION

El resumen aparece en el Program & Abstracts of the 10th International Congress of Vertebrate Morphology, Barcelona 2013. Anatomical Record, Volume 296, Special Feature — 1: P-075.It has been classically assumed that the ventricle of the primitive vertebrate heart is composed of spongy myocardium, supplied exclusively by oxygen-poor, luminal blood. This idea is on two facts: (1) extant agnathans have a spongy ventricular myocardium, and (2) in avian and mammalian embryos, the formation of trabeculated myocardium precedes the appearance of compact myocardium. Recently, it has been proposed that, like elasmobranchs, the early gnathostomes possess a fully vascularised ventricle composed of mixed myocardium. We tested this idea by studying the structure and vascularisation of the ventricular myocardium in four holocephalan species of the families Chimaeridae and Rhinochimaeridae. Chimaera monstrosa, Hidrolagus affinis and Harriotta raleighana have a spongy myocardium covered by a thin layer of cardiac muscle. In H. raleighana, the compacta is reduced to an extremely fine rim. In all three species there is a well-developed coronary artery system consisting of subepicardial vessels which give off branches that penetrate the myocardial trabeculae. Rhinochimaera atlantica has no compacta and its ventricular coronary artery system is reduced to subepicardial vessels that do not enter the spongy layer. This report is the first to show that in wild living vertebrates, a coronary artery system supplying the whole myocardium exists in the absence of a well-developed compacta, which supports experimental work that shows that myocardial cell proliferation and coronary vascular growth rely on genetically separated programs. We conclude that the mixed ventricular myocardium is primitive for chondrichthyans, and that the lack of compacta in some holocephalans is a derived character. Moreover our results support the hypotheses that the mixed myocardium is the primitive condition in gnathostomes, and that the absence of a compacta in different actinopterygian taxa is the result of its repeated loss during evolution.Proyecto CGL2010-16417/BOS; Fondos FEDE

The Myosin Heavy Chain specific A4.1025 antibody discriminates different cardiac segments in ancient groups of gnathostomes: Morphological and evolutionary implications

El resumen aparece en el Program & Abstracts of the 11th International Congress of Vertebrate Morphology, Washington DC 2016. Anatomical Record, Volume 299, Special Feature: 263.The pan-Myosin Heavy Chain (pan-MyHC) marker MF20 have been reported to show similar, homogeneous signal in the myocardial segments of the heart of teleosts and tetrapods. However, in an ongoing study of the myocardial structure of the dogfish (Scyliorhinus canicula; Chondrichthyes), we observed differential immunostaining of the cardiac segments using another pan-MyHC, the A4.1025 antibody. In order to investigate the relevance of this finding for better understanding of the morphology and evolution of the vertebrate heart, we performed immunohistochemistry, slot blot and western blot in several species of chondrichthyans, actinopterygians and mammals using the above mentioned antibodies. In the dogfish heart, A4.1025 and MF20 specifically recognized MyHC isoforms, although with different degree of affinity. MF20 reactivity was homogeneous and high in all the myocardial segments. However, A4.1025 reactivity was heterogeneous. It was high in the sinus venosus (external layer), atrium and atrioventricular region, low in the ventricle and conus arteriosus, and null in the internal layer of the sinus venosus. A heterogeneous pattern of A4.1025 immunoreactivity was also detected in two other elasmobranchs, a holocephalan, a polypteryform and an acipenseriform. In all of these species, MF20 immunoreactivity was homogeneous. In addition, both markers showed a homogeneous immunoreactivity pattern in teleosts and mammals. Our results indicate that in the hearts of ancient gnathostomes, in all of which a conspicuous conus arteriosus exists, one or more MyHC isoforms with low affinity for A4.1025 show segment-specific distributions. Thus, A4.1025 appears to be an appropriated marker to identify the cardiac segments and their boundaries. We propose that the segmentspecific distribution of MyHC isoforms may generate a particular type of myocardial contractility associated with the presence of a conus arteriosus.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. CGL2014-52356-P, CEIMAR, BIO 203, FEDE

Celiotomía en una tortuga mora

En el presente trabajo se describe la técnica quirúrgica empleada para resolver una obstrucción de oviducto en una tortuga mora debido a una alteración ovulatoria.In this work it is described the surgical technique used to resolve an oviduct obstruction in a Greek tortoise (Testudo graeca) caused by the retention of eggs

Anatomical, histochemical and immunohistochemical characterization of the outflow tract of ray hearts (Rajiformes; Chondrichthyes)

El resumen aparece en el Program & Abstracts of the 11th International Congress of Vertebrate Morphology, Washington DC 2016. Anatomical Record, Volume 299, Special Feature: 264.Recent work has shown that the cardiac outflow tract of sharks and chimaeras does not consist of a single myocardial component, the conus arteriosus, as classically accepted, but two, namely, the myocardial conus arteriosus and the non-myocardial bulbus arteriosus. However, the anatomical composition of the outflow tract of the batoid hearts remains unknown. The present study was designed to fill this gap. The material examined consisted of hearts of two species of rays, namely, the Mediterranean starry ray (Raja asterias) and sandy ray (Leucoraja circularis). They were studied using scanning electron microscopy, and histochemical and inmunohistochemical techniques. In both species, the outflow tract consists of two components, proximal and distal with regard to the ventricle. The proximal component is the conus arteriosus; it is characterized by the presence of compact myocardium in its wall and several transverse rows of pocket-shaped valves at its luminal side. Each valve consists of a leaflet and its supporting sinus. Histologically, the leaflet has two fibrosas, inner and outer, and a middle coat, the spongiosa. The distal component lacks myocardium. Its wall consists of smooth muscle cells, elastic fibers and collagen. Thus, it shows an arterial-like structure. However, it differs from the aorta because it is covered by the epicardium and crossed by coronary arteries. These findings indicate that the distal component is morphologically equivalent to the bulbus arteriosus of sharks and chimaeras. In contrast to foregoing descriptions, the valves of the first transverse row are distally anchored to the bulbus arteriosus and not to the ventral aorta. Our findings give added support to the notion that presence of a bulbus arteriosus at the arterial pole of the heart is common to all chondrichtyans, and not an apomorphy of actinopterygians as classically thought.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. CGL2014-52356-P, CEIMAR, BIO 203, FEDE

Fractura de quinta vértebra lumbar : nueva técnica de reducción

En este trabajo pretendemos describir una técnica de fijación externa, con la que se reduce e inmoviliza una fractura en la quinta vertebral lumbar. Usamos un fijador externo tipo II doble de Meynar.In this essay we want to describe a method of external fixation by which you can fixate and inmobilize afracture in the 5th lumbar vertebrae. We use the double external fixators type II of Meynar

Aportes al diagnóstico ecográfico : 17 casos

En este trabajo se relacionan diecisiete casos, con los cuales queremos mostrar las grandes posibilidades de la ecografía como medio de diagnóstico, en las distintas patologías que en él se citan.In this work we find seventeen cases correlated, with which we want to show the great diagnostic possibilities of the echography as well as the source of information it proves to be for the different pathologies mentioned

The utility of the MMPI-2-RF to predict the outcome of a smoking-cessation treatment

People with psychopathology experience high rates of smoking and have more trouble quitting. The Minnesota Multiphasic Personality Inventory-2 Restructured Form (MMPI-2-RF; Tellegen & Ben-Porath, 2008, 2009) is a valid and reliable instrument for the assessment of psychopathology. In this study, we examined the ability of the MMPI-2-RF to assess psychopathology and to predict smoking cessation outcomes in a sample of 281 smokers seeking psychological treatment to stop smoking at the end of treatment and at 6- and 12-month follow-ups. Results showed that T-scores < 65 on Disaffiliativeness (DSF) scale were associated with a higher likelihood of smoking at the end of treatment, and T-scores = 65 on Neurological Complaints (NUC) scale were associated with a higher likelihood of smoking at the 12-month follow-up, after controlling for the effect of age and initial levels of nicotine dependence. The results highlight the usefulness of the MMPI-2-RF in the field of smoking cessation treatment