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Biomedical subjects

J Ruberte

Publications and source records attributed to J Ruberte.

At least 19 recordsLinked to original sources

Mophometric study of the aortic arch and its major branches in rat fetuses on the 21st day of gestation.

The anatomy and embryology of the aortic arch and its branching tributaries (brachiocephalic trunk, left common carotid artery and left subclavian artery) in man and animals are well substantiated. However, the anatomical variations and morphometry of the aortic arch and its branching tributaries in rat fetus at the 21st gestation day have not been studied. Pregnant rats were hysterectomized and the arterial systems of 114 fetuses were injected with a polymerisable resin through the umbilical artery. After maceration, the vascular casts were dissected out and prepared for observations under a scanning electron microscope (SEM). The resulting SEM pictures were studied with a picture analyser and different vessel parameters (diameters, lengths and angles) were measured. The success rate of the microvascular cast injection was 46.5%. Out of the 53 observed aortic arch casts, 98.1% showed the classical branching pattern and one (1.9%) had no brachiocephalic trunk. Morphological analysis showed many differences, which were not linked to the litter. The statistical processing of the measurements enabled us to determine that the aorta diameter after the branching of the left subclavian artery was the most replicable parameter. Moreover, the results revealed some strong correlations between different parameters. There are probably no discrete categories among the various observed parameters as diameters and angles. Some parameters show very little variability and can thus be used as reference points for further studies such as the comparison of a control population with a population treated with a relevant xenobiotic.

Age Factors↗

Albendazole sulphoxide enantiomers in pregnant rats' embryo concentrations and developmental toxicity.

Three single oral doses (8.5, 10, and 14 mg/kg) of a racemic formulation of albendazole sulphoxide (ABZSO) were administered to pregnant rats on day 10 of gestation. Mother plasma and embryo concentrations of ABZSO enantiomers and albendazole sulphone (ABZSO(2)) were determined 9 h after administration. The (-)-ABZSO enantiomer showed higher peak concentrations in both maternal plasma and embryo than the (+) enantiomer. An increase in embryo concentrations of ABZSO enantiomers and ABZSO(2) was only observed when dose rose to 14 mg/kg. There was an increase in resorption when the dose increased, but significant differences were only found in the higher dose group when compared with the other groups. The incidence of external and skeletal malformations (mostly of the tail, vertebrae and ribs) rose significantly in the 10 mg/kg group, producing almost 20% and 90% of malformed fetuses, respectively, and gross external and skeletal abnormalities in the thoracic region and limbs were also found.

Abnormalities, Multiple↗

Vasculogenesis and angiogenesis in the subcardinal venous plexus of quail mesonephros: spatial and temporal morphological analysis.

Vasculogenesis and angiogenesis are involved in a coordinated program for the development of the mesonephric subcardinal venous plexus of quail embryo. Vasculogenesis occurs between days 3 and 4 of incubation, while angiogenesis takes place from day 5 to day 7. Examination of vascular corrosion casts and whole mounts, and tissue sections labelled with specific markers to hemangioblast lineage (QH1, LEP100 and AcPase activity), allowed us to distinguish six phases in the formation of subcardinal plexus. (1) Appearance of isolated angioblast-like cells where the subcardinal plexus will form. (2) Alignment of angioblast-like cells into cellular strands. (3) Formation of compact vascular cords by association of angioblast-like strands. (4) Polygonal interconnection of vascular cords to constitute the primary subcardinal plexus. In this stage, isolated angioblast-like cells were present inside inter-vascular spaces. (5) The splitting of primary inter-vascular spaces by angiogenic sprouts to form secondary subcardinal plexus (outward angiogenesis). Isolated angioblast-like cells were not present in this stage. (6) Expansion of the secondary subcardinal plexus by insertion of slender transcapillary tissue pillars (inward angiogenesis) and angiogenic sprouts. We also describe three morphogenetic gradients during the development of the subcardinal plexus: ventral-to-dorsal, cranial-to-caudal and lateral-to-medial.

Animals↗

Anthelmintic induced congenital malformations in sheep embryos using netobimin.

Benzimidazole compounds have teratogenic effects in domestic and experimental animals. In this study, 14 Manchega ewes were treated orally, under controlled conditions, with 20 mg netobimin (a prodrug of a benzimidazole compound) per/kg bodyweight on the 17th day of pregnancy. Congenital malformations and abortions affected 60 per cent of the lambs. The main malformations were skeletal and renal, but vascular malformations were observed for the first time. The abnormalities were investigated using radiological, dissection and vascular injection techniques, and associations among them were recorded. The anomalies are discussed in terms of embryological considerations.

Abnormalities, Drug-Induced↗

The lack of genital ridge vascularization in the early chick embryo: implications in the migration of the primordial germ cells.

It is known that chick primordial germ cells (PGCs) in early embryonic development migrate via the blood vascular system to colonize the gonadal anlagen. Classically, two factors have been involved in the extravasation of PGCs from the blood stream: chemotactic and mechanical factors. An accurate knowledge of the vascular system of the genital ridge is therefore necessary. However, development of gonadal vascularization in bird embryos has been scarcely studied. Our previous studies have shown that the gonadal arteries develop from the mesonephric arteries. The purpose of this work was to study the implications of the development of the vascular system of the chick genital ridge on PGCs colonization. We selected the Hamburger and Hamilton (H-H) stage 18, since the genital ridge is well developed and PGCs actively extravasate. Forty chick embryos of this stage were processed for scanning electron microscopy of vascular corrosion casts and of critical point-dried specimens as well as light microscopy. Our results are conclusive. We could not find any vessel or capillary network supplying the genital ridge; the dorsal aorta and the primordia of the mesonephric arteries were the closest vessels. However, numerous interendothelial spaces were found in the dorsal aorta at the level of the genital ridge. It is suggested that the interendothelial gaps may be very important in the exchange of substances between the avascular genital ridge and the aortic endothelium at this developmental stage. Two different routes are thought to be involved in PGC migration to the gonadal anlage at this stage: the aortic endothelium and the mesonephric arteries. Whereas mechanical factors may be important for extravasation of PGCs in the mesonephric arteries, no reasons have been found from the morphological point of view to support a slowness of the blood flow in the dorsal aorta at the level of the genital ridge facilitating the extravasation.

Animals↗

Injection technique and scanning electron microscopic study of the arterial pattern of the 20 gestation days (G20) rat fetus.

A technique to obtain microvascular corrosion casts of the G20 rat fetus and the normal pattern of the main arteries of the G20 rat fetus are described. The casts were studied by means of scanning electron microscopy (SEM). The arterial pattern is similar to that described in the adult; however, several variations have been found. It is concluded that the use of vascular corrosion casts studied by SEM may be particularly helpful to observe the extremely small arteries of rat fetuses. Moreover, we suggest that this technique may be useful in practical teratological studies.

Animals↗

Afferent portal venous system in the mesonephros and metanephros of chick embryos: development and degeneration.

BACKGROUND: In the chick embryo, both mesonephros and metanephros have a renal portal system. The classical literature gives uncertain answers about the development and degeneration of the meso- and metanephric portal venous system. Some mesonephric vessels present angiogenic processes to colonize the metanephros, while others show signs of degeneration and disappear together with the mesonephros. The adult avian kidney has a conspicuously placed valve, the renal portal valve. The development of this functionally important renal portal valve has not yet been studied in detail. METHODS: Scanning electron microscopy of vascular corrosion casts has been used in this study. Strong mesonephric degeneration as well as metanephric growth and maturation occur in the developmental stages selected for this investigation (7.5, 9, 11, 14, and 21 days of incubation). RESULTS: The mesonephric afferent venous system in the chick embryo is supplied by two vessels, the posterior and the anterior mesonephric portal veins. The posterior mesonephric portal veins show a similar pattern to the anuran (amphibian) kidney. The anterior mesonephric portal vein has not previously been described. Constrictions were found in this vessel, a probable sign of subsequent degeneration. The metanephric afferent venous system is also supplied by two vessels: the caudal and cranial metanephric portal veins. The caudal metanephric portal vein is derived from the postcardinal vein. The cranial metanephric portal vein grows independently throughout the development of the mesonephric vascular system. It is connected to the vertebral venous sinus already at the beginning of its development. The renal portal valve first appears as a capillary network that communicates with the developing afferent and efferent metanephric venous systems. This capillary network later develops to a venous valve. The metanephric afferent venous system shows typical angiogenic signs in corrosion cast, such as nodular protrusions, holes, and enlarged vessels. CONCLUSIONS: The postcardinal vein first supplies only the mesonephric tissue as a portal vessel. Then it becomes a common source for both kidney generations. Finally it supplies only the metanephric tissue with venous blood. However, two independent vessels were found to supply the cranial renal regions: the anterior mesonephric portal vein and the cranial metanephric portal vein.

Animals↗

Disposition of netobimin, albendazole, and its metabolites in the pregnant rat: developmental toxicity.

Netobimin (NTB), a benzimidazole prodrug with a good anthelmintic spectrum, was administered orally to female rats at a dose of 59.5 mg NTB/kg, to study its pharmacokinetic behavior and the disposition of its most important metabolites, albendazole (ABZ), albendazole sulfoxide (ABZSO), and albendazole sulfone (ABZSO2). ABZ was found in plasma after 6 hr. Peak plasma concentrations (Cmax) and areas under curves (AUC) of ABZSO were eight- and fourfold higher, respectively, than those of ABZSO2. To study NTB disposition in pregnant rats, three different drug doses (50, 59.5, and 70.7 mg/kg) were given. No significant differences were found between plasma concentrations for each metabolite at the three doses studied. Only ABZ concentrations rose slightly as dose increased. ABZ, ABZSO, and ABZSO2 were found in amniotic sacs and embryos at concentrations that were higher than plasma at the same times. The fetuses obtained after administration of each of the doses of NTB were studied to detect developmental toxicity. A significant correlation was found between rate of developmental toxicity and metabolite concentration. ABZSO embryo concentrations could be the main factor accounting for toxicity.

Albendazole↗

Study of the distribution of albendazole-sulphoxide (ABZ-SO) in fertilized egg compartments.

In order to use the chicken embryo in teratogenic studies, it is necessary to know the internal volume in which a xenobiotic distributes. The inoculation of a xenobiotic in one of the compartments of the fertilized egg is the usual technique used in these studies. Neither the concentration nor the moment in which the xenobiotic comes into contact with the chicken embryo have been considered. Predicting the internal volume of distribution in the egg from some of the external parameters that do not interfere with the normal development is necessary. A simple method to calibrate these external parameters and their correlation with the different compartments of the fertilized eggs as well as the different distribution of the xenobiotic in these compartments has been successfully demonstrated. After injection of ABZ-SO, the maximum concentration in the embryo is reached by 36 h. The mean AUC for the albumen (sharp and obtuse end), yolk, and embryo were 78.4, 40.7, 79.2, and 10.8 micrograms.h/ml respectively. The results obtained about the kinetics of the diffusion of ABZ-SO indicate that this compound does not have a homogeneous distribution in all the compartments of the fertilized egg. These results highlight that whenever fertilized eggs are used as a screening for the possible toxicity of a drug or other substances, the dose of the xenobiotic to be injected has to be precisely determined in accordance with the total volume and the stage of embryonic development selected to be affected, starting from the previous knowledge of when and how much substance accedes to the embryo.

Albendazole↗

The morphometric study of the 19-day chicken embryo skeleton (gallus domesticus).

A series of 40 chicken embryos were processed after 19 d of incubation by dissection, diaphanisation, skeleton stained with alizarin red and examined by image analysis. Six bones (femur, tibiotarsus, tarsometatarsus, humerus, ulna and carpometacarpus) and 4 parameters (perimeter, area, the largest and the smallest Feret diameter) were evaluated. The smallest Feret diameter proved unusable. The coefficients of variation for the other 3 parameters were comparable for the 6 bones. All the bones were valuable for comparing morphometric values between a control population and a population treated with a xenobiotic in order to quantify the chemical's influence on ossification.

Animals↗

Angiogenesis in the gonadal capillary network of the chick embryo.

Seventy-one chick embryos of both sexes at the 35 Hamburger and Hamilton (H-H) developmental stage were processed for scanning electron microscopy of vascular corrosion casts and of critical point dried specimens, as well as transmission electron- and light microscopy, in order to study the angiogenic structures. The gonadal subepithelial capillary network was located at the level of the tunica albuginea under the covering epithelium. The casts showed a densely-meshed capillary network and numerous sprouting (nodular protrusions or capillary sprouts) and non-sprouting (enlarged vessels and angiogenic holes) angiogenic structures that were randomly distributed and mixed. Four types of angiogenic holes were encountered in the casts: primary (diameter < 2.5 microns), secondary (diameter > 2.5 microns), tertiary (variable diameter and circular narrowings on one side), and open angiogenic holes. We suggest that the different morphologies reflect evolution of these holes. Furthermore, the open angiogenic hole would probably either form nodular protrusions at its open ends, which tend to join with other nodular protrusions and neighboring capillaries and form new vessels; or there would be fusion with two or more neighboring open holes. Correlative critical point dried sections showed fenestrations in the capillary walls and transcapillary pillars that corresponded to the angiogenic holes found in the casts. Ultrathin sections of the vessels presented typical characteristics of growing endothelium: large nuclei with loosely textured chromatin, abundant cytoplasm rich in cell organelles and intraluminal endothelial processes.

Animals↗

Development and degeneration of the arterial system in the mesonephros and metanephros of chicken embryos.

BACKGROUND: The general morphology of the mesonephric and metanephric arteries in chicken embryos has already been described previously. Moreover, the general basis of glomerulogenesis has also been established. However, the degeneration of the mesonephric vascular system, and especially glomerular degeneration, have not been well established yet. Also the morphology of the metanephric angiogenic buds has not been studied yet. METHODS: Scanning electron microscopy of vascular corrosion casts and critical point dried specimens as well as light microscopy of serially sectioned material has been used in this study. Mesonephric degeneration coincides in time with metanephros growth and maturation in the developmental stages of chicken embryos chosen for this investigation (7.5, 9, 11, and 14 days of incubation). RESULTS: The arterial system of the mesonephros in embryonic chicken is similar to that of the anuran kidney, as described in the literature. The morphology of the degenerating mesonephric glomeruli shows that the glomerular capillaries are more thick, tortuous, and numerous than those in normal glomeruli. The podocytes also show degeneration. The arterial system of the metanephros grows directly from the aorta and from the mesonephric arterial system. During these stages of rapid growth, the metanephros shows angiogenic buds. These angiogenic buds can be either pointed or round blind endings. CONCLUSIONS: The distribution and topography of the mesonephric and metanephric arteries is in general accordance with the literature. The process of glomerular degeneration in the mesonephros seems similar to glomerular senescence in man but is different from that of the aged rat glomeruli. The round angiogenic buds observed in the metanephros resemble tumoral angiogenic buds in some aspects. However, both angiogenesis and the degenerative phenomena are part of the normal developmental process. Consequently, the involved mechanisms are probably under sole genetic control. The system studied here offers therefore the possibility to study vascular growth and degeneration on the same model in physiological conditions without application of vasoactive or pathological agents.

Animals↗

Malformations of the vertebral bodies and the ribs associated to spinal dysraphism without spina bifida in a Pekingese dog.

A Pekingese dog with hemivertebrae, rib malformations and spinal cord dysraphism without spina bifida was presented. Two types of hemivertebrae were seen: bilateral and unilateral. Thoracic hemivertebrae were associated with fused ribs and rudimentary ribs. Spinal dysraphism consisted of polycavitary syringomyelia in the dorsal and lateral funiculi, hydromyelia and anomalies of the dorsal median septum and median fissure, associated to lumbar and sacral hemivertebrae. Cauda equina agenesia was also present. To conclude, the dog showed two malformations from different embryonic origins. Vertebral and rib malformations are of mesodermic origin and spinal dysraphism is of ectodermic origin. A possible common mechanism responsible for both anomalies is discussed.

Animals↗

Vascular architecture of the lactating and non-lactating teat of the bitch: a scanning electron- and light microscope study.

Tissues from fourteen mammary glands of eight bitches were prepared for scanning electron microscopy of vascular corrosion casts and for histology to study the vasculature of the lactating and non-lactating teats. The densely-meshed mammary dermal capillary network formed ridges and troughs. The teat ducts were vascularized by a relatively densely-meshed capillary network which drained into veins longitudinally oriented to the ducts. Between eight and fifteen teat duct openings were seen on the tip of the teat, that were sometimes divided by a septum. The inner vascularization of the teat showed that the main papillary arteries divided into undulating secondary papillary arteries which presented numerous semi-constrictions and loops. Their structure may help during erection of the teat. Arteriovenous anastomoses found at different points may participate in blood flow maintenance during suckling, heat regulation and teat erection. Veins freely anastomosed and ran longitudinally to the axis of the teat. They exhibited numerous bicuspid valves. In non-lactating teats, vessels showed the same main architecture and characteristics mentioned above, although these were considerably less marked. The structure of the vascular elements in the teat of the bitch could favor blood flow during suckling and suggest that vessels adapt to the physiological situation.

Animals↗

Histochemical and morphometric study of the cricoarytenoideus lateralis muscle in the horse.

Histochemical and morphometric parameters of the cricoarytenoideus lateralis muscle of the horse are presented. Using myosin ATPase staining after acid preincubation, 3 fibre types (I, IIA and IIC) were identified. Using NADH-TR staining, type I fibres showed high oxidative capacity, whereas type II fibres had high or low oxidative capacity. The type I to type II ratio was of 35:65. This ratio remained constant in the age range examined. Statistically significant (p < 0.01) differences were found in values for fibre size between groups of horses weighing more than 500 kg and less than 400 kg. Mean area of type II fibres was greater (p < 0.001) than that of type I fibres. There were no significant differences in mean area between left and right muscles in the group of animals with less weight. In contrast, significant differences (p < 0.05) in mean area between left and right muscles were found for type I fibres in the group of animals exhibiting a higher weight. The histographical distribution of fibre type areas was unimodal. Most adult horses showed muscle fibre type grouping in the left muscle.

Age Factors↗

Muscular pathology in equine laryngeal neuropathy.

Three adductor muscles of the larynx, the cricoarytenoideus lateralis (CAL), the arytenoideus transversus (AT) and the ventricularis (Ve), from 36 horses were examined histologically. The neurogenic changes seen in each muscle were evaluated qualitatively. In addition, in 6 horses with clinical and subclinical signs of neurogenic atrophy, measurements of muscle fibre area were performed. Neurogenic changes observed in the Ve were less than in CAL and AT. Measurements of muscle fibre area also demonstrated that CAL and AT showed a wider range of pathological changes than did Ve. The results show that denervation does not uniformly affect all adductor muscles of the larynx. On the other hand, the appearance in some animals of fibre type grouping in the right AT to the same or to a greater extent than in the left AT supports the classification of equine laryngeal neuropathy as a distal axonopathy.

Animals↗

Technical improvements in corrosion casting of small specimens: a study on mesonephric tubules and vessels of chicken embryos.

The injection technique for corrosion casting of small, embryonic material can be improved by using a "chemical ligature" (cyanoacrylate). With this simple method, leakage of the resin at the injection site is prevented and the mechanical stability of the cannula-vessel coupling is improved. The blood vascular system of chicken embryos as small as Hamburger-Hamilton stage 24 (approximately 4.5 days of incubation) has successfully been injected using this procedure. Corrosion casts of the mesonephric tubular system have been made in a similar manner. Additionally, a simple way for secure transport of the fragile casts by immersion in 10% gelatin is suggested.

Animals↗