Collagen organization in canine myxomatous mitral valve disease: an x-ray diffraction study

Collagen fibrils, a major component of mitral valve leaflets, play an important role in defining shape and providing mechanical strength and flexibility. Histopathological studies show that collagen fibrils undergo dramatic changes in the course of myxomatous mitral valve disease in both dogs and humans. However, little is known about the detailed organization of collagen in this disease. This study was designed to analyze and compare collagen fibril organization in healthy and lesional areas of myxomatous mitral valves of dogs, using synchrotron small-angle x-ray diffraction. The orientation, density, and alignment of collagen fibrils were mapped across six different valves. The findings reveal a preferred collagen alignment in the main body of the leaflets between two commissures. Qualitative and quantitative analysis of the data showed significant differences between affected and lesion-free areas in terms of collagen content, fibril alignment, and total tissue volume. Regression analysis of the amount of collagen compared to the total tissue content at each point revealed a significant relationship between these two parameters in lesion-free but not in affected areas. This is the first time this technique has been used to map collagen fibrils in cardiac tissue; the findings have important applications to human cardiology.     

Osteopontin is associated with bioprosthetic heart valve calcification in humans

Calcification of non-osseous tissues such as heart valves or vessels is a major concern in clinical practice. The exact mechanism is still unknown. Numerous studies have shown that mineral deposits of crystalline hydroxyapatite within these tissues were associated with increased non-collagenous protein content. More recently osteopontin was found to be associated with calcification in living tissues such as vessels and native human aortic valves. The aim of this study was to determine whether or not non-collagenous proteins can also be found in non-living tissues such as glutaraldehyde-pretreatedporcine valves after implantation in humans. Thirty-eight glutaraldéhyde pretreated porcine bioprostheses were studied: 16 not implanted and 22 after 11 years of implantation in the aortic and mitral valve position in humans. In areas of calcification vizualized by Von Kossa staining and microradiography, immunostaining using polyclonal antibodies against calcium-bindingproteins showed osteopontin positive staining and no staining for osteocalcin, bone sialoprotein or osteonectin. In uncalcified areas and in non-implanted valves, staining for osteopontin or other calcium-binding proteins was negative. Western blot analysis of macroscopically calcified and uncalcified areas showed that several proteins were adsorbed in implanted valves and confirmed the presence of osteopontin in the calcified areas, while no immunolabelling was found in non-calcified areas, in uncalcified valves and in non-implanted valves. Thus the presence of osteopontin in the calcified areas of bioprosthetic heart valves implanted in human indicates that this protein is associated with bioprosthetic valvular calcification. Since these valves are made of non-living connective tissue, and no cell immunostained for osteopontin was found around the calcified area, this study suggests that a non-cellular mediated mechanism involving protein adsorption may play a role in bioprosthetic valvular calcification.     

Age-Dependent Changes in Geometry,Tissue Composition and Mechanical Properties of Fetal to Adult Cryopreserved Human Heart Valves

There is limited information about age-specific structural and functional properties of human heart valves, while this information is key to the development and evaluation of living valve replacements for pediatric and adolescent patients. Here, we present an extended data set of structure-function properties of cryopreserved human pulmonary and aortic heart valves, providing age-specific information for living valve replacements. Tissue composition, morphology, mechanical properties, and maturation of leaflets from 16 pairs of structurally unaffected aortic and pulmonary valves of human donors (fetal-53 years) were analyzed. Interestingly, no major differences were observed between the aortic and pulmonary valves. Valve annulus and leaflet dimensions increase throughout life. The typical three-layered leaflet structure is present before birth, but becomes more distinct with age. After birth, cell numbers decrease rapidly, while remaining cells obtain a quiescent phenotype and reside in the ventricularis and spongiosa. With age and maturation–but more pronounced in aortic valves–the matrix shows an increasing amount of collagen and collagen cross-links and a reduction in glycosaminoglycans. These matrix changes correlate with increasing leaflet stiffness with age. Our data provide a new and comprehensive overview of the changes of structure-function properties of fetal to adult human semilunar heart valves that can be used to evaluate and optimize future therapies, such as tissue engineering of heart valves. Changing hemodynamic conditions with age can explain initial changes in matrix composition and consequent mechanical properties, but cannot explain the ongoing changes in valve dimensions and matrix composition at older age.     

The structure of the intrapigmented aesthetes and the properiostracum layer in Callochiton achatinus (Mollusca: Polyplacophora)

The ultrastructure of the intrapigmented aesthetes which occur in specific areas of the different valves of Callochiton achatinus are described. The possible functions of the aesthete and the pigmented body are discussed in the light of their ultrastructure and the ecological requirements of the animal. The properiostracum in C. achatinus is unusually thick and has a structure very similar in appearance to that of collagen.     

Evaluation of "unique" aspects of human vertebral bodies and pedicles with a consideration of Australopithecus africanus

Recent functional studies of human vertebrae have revealed that loads borne by the axial skeleton during bipedal postures and locomotion pass through the pedicles and posterior elements as well as through the bodies and discs. Accordingly, particular morphological attributes of these vertebral elements have been linked exclusively with bipedalism. In order to test the validity of current form-function associations in human vertebral anatomy, this study considers the morphology of human thoracolumbar vertebral bodies and pedicles in the context of a wide comparative primate sample. The last lumbar vertebra of STS 14 (Australopithecus africanus) is also included in the analysis. Results indicate that certain features of human vertebrae previously thought to reflect bipedalism are characteristic of several nonhuman primates, including those whose posture is habitually pronograde. These features include the decrease in vertebral body surface area and the increase in cross-sectional area of the pedicle between the penultimate and last lumbar vertebra. In addition, although humans have relatively large and wide last lumbar pedicles, the enlargement and widening of the pedicle between the penultimate and last lumbar vertebra is not unique to humans. On the other hand, human vertebrae do exhibit several unique adaptations to bipedal posture and locomotion: (1) the vertebral body surface areas of the lower lumbar vertebrae and the cross-sectional areas of the last lumbar pedicles are large relative to body size, and (2) the last lumbar pedicles are wider relative to length and to body size than are those of nonhuman primates. The last lumbar vertebra of STS 14 does not exhibit any of these human-like vertebral features—its pedicles and body surface areas are relatively small, and its pedicles are not relatively wide, but relatively short.     

Venous valves in the large branches of superior vena cava

In 97 human cadavers the valves of the following blood vessels were investigated with regard to their cusps and their sizes and positions: the internal jugular veins, the subclavian veins, the brachiocephalic veins, and the superior vena cava. The cusps of each of the valves, which consist of two or three parts, are neither always of equal size nor obligatorily sufficient. Unipartite valves may be sufficient as well as insufficient. Internal jugular veins: The inferior bulb of the internal jugular vein is provided with valves which in 6% of the cases consist of three parts, in 66% of two parts, and in 15% of only one cusp. The concave margins of most of them go down as far as the venous angle. The convex edges attached to the wall of the vein extend to a higher level on the right side than on the left. In 13% there do not exist any valves. Varieties are described separately in this paper. Subclavian veins: Valves are found along the length of the vessel. Only few of them reach the venous angle. In rare cases there exist two valves: one at the beginning, the other at the end of the subclavian vein. In 4% of the cases the valves consist of three, in 75% of two cusps. In 12% they are unipartite. In 9% there are no valves to be found. The right side is more often without valves than the left. Brachiocephalic veins: Only a minority of these vessels is provided with valves. Most of these consist of one cusp, are insufficient, and are situated in the left innominate vein.(ABSTRACT TRUNCATED AT 250 WORDS)     

A Starling resistor regulates cerebral venous outflow in dogs

This investigation was undertaken to determine whether a Starling resistor or venous waterfall effect exists between the sagittal sinus and the cerebral veins such that increases in sagittal sinus pressure (Pss) do not abolish cerebral venous outflow and to examine two possible contributions of extracranial venous valves in regulating outflow. Anesthetized dogs were subjected to positive end-expiratory pressure (PEEP) before and after intracranial pressure (Pic) was elevated by inflation of an epidural balloon. PEEP raised Pss equally in all animals, but Pic and cerebral venous pressure (Pcv) increased less in the presence of intracranial hypertension. When Pss was low, passage of a catheter in the cerebral vein in and out of the sagittal sinus demonstrated an abrupt drop in pressure as the sinus was entered. When Pss was raised and lowered independently of superior vena caval pressure (Psvc) the changes in Pic and Pcv were less when Pss was decreased than when it was increased. Sustained increases and decreases in Psvc caused increases and decreases in Pcv, Pic, Pss, and external jugular venous pressure (Pejv) regardless of whether external jugular venous valves were present or absent. We conclude that a Starling resistor between the sagittal sinus and the cerebral veins regulates cerebral venous outflow when Pss is increased by PEEP and other maneuvers that raise Psvc. The waterfall maintains Pcv and Pic at normal levels when Psvc and Pss are reduced. Extracranial venous valves are not essential to this mechanism.     

Growth and morphological changes in the stomach of newborn pigs during the first three days after birth.

Growth and morphological changes in the stomach of newborn pigs were examined during the first 3 days after birth. The stomach grew disproportionately faster than the body as a whole during this period. The growth was due to hyperplasia and hypertrophy during the first day and mainly to hyperplasia thereafter as gastric DNA content increased progressively after birth, and the protein:DNA and RNA:DNA ratios increased only on the first day. Histological and morphometric analyses revealed that the growth was more pronounced in the gastric body region than in the cardiac and pyloric regions, and more pronounced in the mucosal layer than in other layers. The percentage of mucosal volume occupied by parietal cells (volume density) and the number of parietal cells per unit volume of gastric mucosa (numerical density) increased significantly 3 days after birth in the cardiac and body regions, but not in the pyloric region, of the stomach. The observed morphological changes coincide with the known pattern of functional maturation during the immediate postnatal period. It is suggested that a high level of circulating gastrin and oral ingestion of milk-derived growth factors in the newborn pig contribute to these changes.     

The venous territories (venosomes) of the human body: experimental study and clinical implications

The venous architecture of the integument and the underlying deep tissues was studied in six total-body human fresh cadavers and a series of isolated regional studies of the limbs and torso. A radiopaque lead oxide mixture was injected, and the integument and deep tissues were dissected and radiographed. The sites of the venous perforators were plotted and traced to their underlying parent veins that accompany the source (segmental) arteries. A series of cross-sectional studies were made in one subject to illustrate the course of the perforators between the integument and the deep tissues. The veins were dissected under magnification to identify the site and orientation of the valves. Results revealed a large number of valveless (oscillating) veins within the integument and deep tissues that link adjacent valved venous territories and allow equilibration of flow and pressure throughout the tissue. Where choke arteries define the arterial territories, they are matched by boundaries of oscillating veins in the venous studies. The venous architecture is a continuous network of arcades that follow the connective-tissue framework of the body. The veins converge from mobile to fixed areas, and they "hitchhike" with nerves. The venous drainage mirrors the arterial supply in the deep tissues and in most areas of the integument in the head, neck, and torso. In the limbs, the stellate pattern of the venous perforators is modified by longitudinal channels in the subdermal network. However, when an island flap is raised, these longitudinal channels are disconnected, and once again the arterial and venous patterns match. Our venous studies add strength to the angiosome concept. Where source arteries supply a composite block of tissue, we have demonstrated radiologically and by microdissection that the branches of these arteries are accompanied by veins that drain in the opposite direction and return to the same locus. Hence each angiosome consists of matching arteriosomes and venosomes. The clinical implications of these results are discussed with particular reference to the design of flaps, the delay phenomenon, venous free flaps, the pathogenesis of flap necrosis, the "muscle pump," varicose veins, and venous ulceration.     

Development of a middle cerebral artery occlusion model in the nonhuman primate and a safety study of i.v. infusion of human mesenchymal stem cells

Background

Most experimental stroke research is carried out in rodents, but given differences between rodents and human, nonhuman primate (NHP) models may provide a valuable tool to study therapeutic interventions. The authors developed a surgical method for transient occlusion of the M1 branch of middle cerebral artery (MCA) in the African green monkey to evaluate safety aspects of intravenous infusion of mesenchymal stem cells (hMSCs) derived from human bone marrow.

Methods

The left Sylvian fissure was exposed by a small fronto-temporal craniotomy. The M1 branch of the MCA was exposed by microsurgical dissection and clipped for 2 to 4 hours. Neurological examinations and magnetic resonance imaging (MRI) were carried out at regular post-operative course. hMSCs were infused 1 hour after reperfusion (clip release) in the 3-hour occlusion model.

Results

During M1 occlusion, two patterns of changes were observed in the lateral hemisphere surface. One pattern (Pattern 1) was darkening of venous blood, small vessel collapse, and blood pooling with no venous return in cortical veins. Animals with these three features had severe and lasting hemiplegia and MRI demonstrated extensive MCA territory infarction. Animals in the second pattern (Pattern 2) displayed darkening of venous blood, small vessel collapse, and reduced but incompletely occluded venous flow and the functional deficit was much less severe and MRI indicated smaller infarction areas in brain. The severe group (Pattern 1) likely had less extensive collateral circulation than the less severe group (Pattern 2) where venous pooling of blood was not observed. The hMSC infused animals showed a trend for greater functional improvement that was not statistically significant in the acute phase and no additive negative effects.

Conclusions

These results indicate inter-animal variability of collateral circulation after complete M1 occlusion and that hMSC infusion is safe in the developed NHP stroke model.     

Age-related hematological changes in normal F344 rats: during the neonatal period.

In order to clarify age-related changes in hematological values of normal rats after birth, blood samples from neonatal F344 rats of both sexes were examined periodically during the period from 0 to 40 days postpartum. The erythrocyte count (RBC) increased with time after birth as a function of age. In contrast, the reticulocyte count (Retics) continuously decreased with time after birth. Hemoglobin (Hb), hematocrit (Ht), and the mean corpuscular hemoglobin concentration (MCHC) tended to decrease after birth until weaning (about 21 days postpartum), but they began to increase after weaning. Mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) also gradually decreased after birth until weaning, but they were unchanged thereafter. The platelet count (PLT) gradually increased after birth and reached a plateau at weaning. Microscopic examination of blood smears revealed that erythrocytes at birth had characteristic morphological features such as anisocytosis, polychromasia, basophilic stippling, Howell-Jolly body, and erythroblastosis. These characteristic features, however, disappeared by 30 days after birth. The total leukocyte count (WBC) gradually increased with time after birth, due to an increase in the number of lymphocytes. The lymphocyte count started to rapidly increase within several days after birth and the increase continued thereafter. Other differential leukocyte counts also showed various characteristic patterns of changes during the neonatal period. There were no apparent differences between males and females in these changes in hematological values.     

Reverse-flow island flap: clinical report and venous drainage

Twenty-two reverse-flow island flaps were transferred. These included peroneal, forearm, anterior tibial, and temporal flaps. Sixteen of 22 flaps survived completely. We encountered partial necrosis in 4 flaps and total necrosis in only 2 flaps. We credit this success to the reliability and availability of the peroneal, forearm, and temporal flaps; but we do not encourage use of the anterior tibial flap. The flaps that survived well did not show any signs of venous congestion. The advantage of the reverse-flow island flap is that it can be transferred from a proximal to a distal location. Using cadavers and fresh amputated limbs, studies on venous drainage of the reverse-flow island flap were performed. The venae comitantes had numerous venous valves and communicating branches, but more than sufficient reflux of the venous blood occurred through the valves at pressures of 90 to 105 cmH2O. We believe that the venous drainage of the reverse-flow island flap occurs as a result of reflux actions at the valve, communicating branches between the venae comitantes, and bypass vessels around the valves.     

Ultrastructural analysis of the mouse nasal septum. Respiratory cilia prior to an after birth. A quantitative freeze-fracture study

Respiratory cilia of mice from 3 days prior to birth till 21 days after birth were examined by freeze-fracture preparations for the determination of the frequency and size of the intramembranous particles (IMPs). Dependent on the age of the animals different numbers and sizes of IMPs were found. A significant drop in total number of IMPs in all diameter classes was most evident from the 19th day of gestation to the 20th day (1 day prior to birth), followed by a marked rise between embryonal day 20 and the 1st postnatal day. The results were discussed with regard to morphological and functional changes in the respiratory system prior to and after birth.     

Age and individual variability of aortic valve structure in man

In persons of both sex at the age beginning from birth up to 90 years. 275 aortal valves have been investigated. A morphological classification of the valves has been suggested depending on peculiarities of their structure and main dimentions: valves with the valvula surfaces looking as a part of a spheroid, ellipsoid, or having a stepwise, cochliowise form; according to the mode of the valvula closure when the valve is closed: with straight, arched and wavy lines of closure; according to the size: valves with predominant dimentions of one valvula when two others are equal, valves with two equal valvulae and both are larger than the third one, valves with equal valvulae, valves with three different valvulae. Distribution of various types of the valves in accordance with the given classification is determined, the most frequently occurring forms are demonstrated. The valvulae grow in different directions unevenly and asynchronously with the aortal ostium growth, therefore during certain age periods a probability on a nonhermetic valve increases, especially from 1 up to 3 and from 56 up to 70 years of age.     

Evolution of insect mushroom bodies: old clues,new insights

The mushroom bodies are a morphologically diverse sensory integration and learning and memory center in the brains of various invertebrate species, of which those of insects are the best described. Insect mushroom bodies are composed of numerous tiny intrinsic neurons (Kenyon cells) that form calyces with their dendrites and a pedunculus and lobes with their axons. The identities of conserved Kenyon cell subpopulations and the correlations between morphological and functional specializations of the mushroom bodies are just beginning to be elucidated, providing insight into mechanisms of mushroom body evolution. Comparisons of mushroom body organization in different insect lineages reveal trends in the evolution of subcompartments correlated with the elaboration, reduction, acquisition or loss of Kenyon cell subpopulations. Furthermore, these changes often appear correlated with variation in type and strength of afferent input and in behavioral ecology. These and other features of mushroom body organization suggest a striking convergence with mammalian cortex, with Kenyon cell subpopulations displaying evolutionary modularity in a manner reminiscent of cortical areas.     

Effects of captopril on the development of rat doxorubicin nephropathy.

The effects of a daily administration of an anti-converting enzyme inhibitor. Captopril (CPT) (100 mg/kg/orally), on the development of functional and morphological alterations induced in rats by a single injection (7.5 mg/kg/iv) of Doxorubicin (DXR) (Adriamycin*), were investigated. Twenty-four-hour protein excretion, urine output, food intake, water intake, and body weight gain were measured weekly for 30 days. Transmission and scanning electron microscopy observations were performed on kidney samples after 30 days. Four groups were studied. Group 1 were control rats. Group 2 were rats injected with DXR. Group 3 were rats injected with DXR and treated with CPT for 30 days. Group 4 were rats injected with DXR and treated with CPT for 15 days (CPT treatment started 15 days after DXR injection). Group 1 did not show significant functional or morphological changes. Group 2 showed severe proteinuria, significant increase in urinary volume within 2 weeks, significant body weight reduction and diffuse morphological changes. These changes mainly consisted of podocyte swelling, severe foot process fusion, and presence of casts within tubular lumen. Group 3, with respect to group 2, showed a significant reduction of the 24 h protein excretion and urine output. This group displayed morphological changes similar to those observed in group 2, but with a focal distribution. Group 4 showed functional and morphological changes comparable with those of group 2. It is concluded that CPT partially inhibits the development of the functional and morphological damage induced by DXR in the rat kidney. However, CPT did not influence the natural development of nephropathy when treatment started 15 days after DXR injection.     

Scaling of ctenes and consequences for swimming performance in the ctenophore Pleurobrachia bachei

Ctenophores coordinate large macrociliary structures called ctenes to propel themselves through the water. The morphology and kinematics of the ctenes mediate swimming performance. We investigated morphological and kinematic factors affecting swimming performance in free‐swimming ctenophores (Pleurobrachia bachei) using high speed videography. Our morphological results showed that the relationship between body size and ctene morphology and arrangement in P. bachei were well described using linear (i.e., isometric) relationships, which suggests functional limitations of ctenes that vary among individuals of different sizes. Our kinematic results showed that isometric constraints on swimming performance can potentially be overcome by alterations in kinematics: (a) swimming speed in P. bachei increased with ctene beat frequency over a range of body lengths, and (b) the separation of ctenes into clumps of cilia allowed the ctene to increase in width during the effective stroke and decrease in width during recovery. Separation increases the surface area of the ctene during the effective stroke, likely increasing the thrust produced. The finding that ctenes are not monoliths and instead are separated into clumps of cilia has not been previously described, and we subsequently observed this trait in three other ctenophore species: Euplokamis dunlapae, Bolinopsis infundibulum, and Beroe mitrata. Flexibility in function may be a necessary corollary to isometric development of the ctenes as propulsive structures.     

Mosaic Evolution of the Basicranium in <Emphasis Type="Italic">Homo</Emphasis> and its Relation to Modular Development

Mosaic evolution describes different rates of evolutionary change in different body units. Morphologically these units are described by more relationships within a unit than between different units which relates mosaic evolution with morphological integration and modularity. Recent evidence suggests mosaic evolution at the human basicranium due to different evolutionary rates of midline and lateral cranial base morphology but this hypothesis has not yet been addressed explicitly. We this hypothesis and explore how mosaic evolution relates to modular development. Evolutionary data sets on midline (N = 186) and lateral (N = 86) basicranial morphology are compared with 3D data on pre- and postnatal basicranial ontogeny (N = 71). Our results support the hypothesis of mosaic evolution and suggest a modular nature of basicranial development. Different embryological basicranial units likely became differently modified during evolution, with relatively stable midline elements and more variable lateral elements. In addition, developmental data suggests that modularity patterns change throughout ontogeny. During prenatal ontogeny lateral basicranial elements (greater sphenoid wings and petrosal pyramids) change together compared with the midline base. Close to birth the greater sphenoid wings keep a spatially stable position, while the petrosal pyramids become dissociated and shifted posteriorly. After birth the greater sphenoid wings and petrosal pyramids change again jointly and with respect to midline cranial base elements. This sequential pattern of integration and modularization and re-integration describes human basicranial ontogeny in a way that is potentially important for the understanding of evolutionary change. Phylogenetic modifications of this pattern during morphogenesis, growth, and development may underlie the mosaic evolution of the hominin basicranium.     

Absence of venous valves in mice lacking Connexin37

Venous valves play a crucial role in blood circulation, promoting the one-way movement of blood from superficial and deep veins towards the heart. By preventing retrograde flow, venous valves spare capillaries and venules from being subjected to damaging elevations in pressure, especially during skeletal muscle contraction. Pathologically, valvular incompetence or absence of valves are common features of venous disorders such as chronic venous insufficiency and varicose veins. The underlying causes of these conditions are not well understood, but congenital venous valve aplasia or agenesis may play a role in some cases. Despite progress in the study of cardiac and lymphatic valve morphogenesis, the molecular mechanisms controlling the development and maintenance of venous valves remain poorly understood. Here, we show that in valved veins of the mouse, three gap junction proteins (Connexins, Cxs), Cx37, Cx43, and Cx47, are expressed exclusively in the valves in a highly polarized fashion, with Cx43 on the upstream side of the valve leaflet and Cx37 on the downstream side. Surprisingly, Cx43 expression is strongly induced in the non-valve venous endothelium in superficial veins following wounding of the overlying skin. Moreover, we show that in Cx37-deficient mice, venous valves are entirely absent. Thus, Cx37, a protein involved in cell–cell communication, is one of only a few proteins identified so far as critical for the development or maintenance of venous valves. Because Cxs are necessary for the development of valves in lymphatic vessels as well, our results support the notion of common molecular pathways controlling valve development in veins and lymphatic vessels.