Exo-Ocean Exploration with Deep-Sea Sensor and Platform Technologies.

One of Saturn's largest moons, Enceladus, possesses a vast extraterrestrial ocean (i.e., exo-ocean) that is increasingly becoming the hotspot of future research initiatives dedicated to the exploration of putative life. Here, a new bio-exploration concept design for Enceladus' exo-ocean is proposed, focusing on the potential presence of organisms across a wide range of sizes (i.e., from uni- to multicellular and animal-like), according to state-of-the-art sensor and robotic platform technologies used in terrestrial deep-sea research. In particular, we focus on combined direct and indirect life-detection capabilities, based on optoacoustic imaging and passive acoustics, as well as molecular approaches. Such biologically oriented sampling can be accompanied by concomitant geochemical and oceanographic measurements to provide data relevant to exo-ocean exploration and understanding. Finally, we describe how this multidisciplinary monitoring approach is currently enabled in terrestrial oceans through cabled (fixed) observatories and their related mobile multiparametric platforms (i.e., Autonomous Underwater and Remotely Operated Vehicles, as well as crawlers, rovers, and biomimetic robots) and how their modified design can be used for exo-ocean exploration.

Hierarchical classification of G-protein-coupled receptors with data-driven selection of attributes and classifiers.

We address the important bioinformatics problem of predicting protein function from a protein's primary sequence. We consider the functional classification of G-Protein-Coupled Receptors (GPCRs), whose functions are specified in a class hierarchy. We tackle this task using a novel top-down hierarchical classification system where, for each node in the class hierarchy, the predictor attributes to be used in that node and the classifier to be applied to the selected attributes are chosen in a data-driven manner. Compared with a previous hierarchical classification system selecting classifiers only, our new system significantly reduced processing time without significantly sacrificing predictive accuracy.

Blood pressure estimation in the human fetal descending aorta.

OBJECTIVES: The objectives of this study were to estimate fetal blood pressure non-invasively from two-dimensional color Doppler-derived aortic blood flow and diameter waveforms, and to compare the results with invasively derived human fetal blood pressures available from the literature. METHODS: Aortic pressures were calculated from digitally recorded color Doppler cineloops of the fetal descending aorta by applying the Womersley model in combination with the two-element Windkessel model, assuming constant pulse wave velocity during the second half of pregnancy. The results were compared with invasively derived human fetal blood pressures obtained from the literature. RESULTS: In 21 normal pregnancies the estimated mean aortic pressure regression line increased linearly from 28 mmHg at 20 weeks of gestation to 45 mmHg at 40 weeks of gestation. The pulse pressure based on the regression line increased linearly from 21 mmHg at 20 weeks of gestation to 29 mmHg at 40 weeks of gestation. The aortic compliance exhibited a log linear relationship with the gestational age and a statistically significant eightfold increase was observed between 20 and 40 weeks. The aortic downstream peripheral resistance exhibited an exponentially decaying relationship across the same gestational age range. Non-invasively derived aortic systolic and diastolic aortic pressures were comparable with previously reported invasively derived systolic and diastolic umbilical arterial pressures; however, the mean pressures differed significantly from those reported in the umbilical artery in a separate study. The aortic systolic pressures calculated in this study were significantly higher than invasively derived left ventricular systolic pressures that have been previously reported in the literature. CONCLUSIONS: This study demonstrates the feasibility of estimating arterial blood pressure in the human fetus. The method described is of potential use in assessing fetal blood pressure non-invasively, particularly for studying relative changes with time.

Fetal heart rate and umbilical artery velocity variability in fetuses with congenital cardiac defects: a preliminary study.

OBJECTIVE: To examine whether variabilities in fetal heart rate and umbilical artery flow velocity are possible markers for hemodynamic dysfunction in fetuses with a congenital heart defect. METHODS: Doppler studies of the umbilical artery velocity waveform were performed at 20-35 weeks of gestation in 13 patients with a congenital heart defect. We determined absolute and variability values for heart rate and flow velocities from umbilical artery velocity waveforms of at least 18 s duration. We compared these findings with normal controls matched for gestational age. RESULTS: Fetuses with a congenital heart defect displayed decreased umbilical artery peak systolic and time-averaged velocities. However, variability in peak systolic and time-averaged velocities and fetal heart rate variability were increased compared with normal controls. Absolute fetal heart rates were similar between the two groups. CONCLUSIONS: Marked cardiovascular changes occur in the fetus with a congenital heart defect compared with the normal healthy fetus. We propose that variability in fetal heart rate and umbilical artery blood flow velocity could be additional markers for impaired homeostasis in the presence of fetal congenital heart disease.

Cardiac morphology and blood pressure in the adult zebrafish.

Zebrafish has become a popular model for the study of cardiovascular development. We performed morphologic analysis on 3 months postfertilization zebrafish hearts (n > or = 20) with scanning electron microscopy, hematoxylin and eosin staining and Masson's trichrome staining, and morphometric analysis on cell organelles with transmission electron photomicrographs. We measured atrial, ventricular, ventral, and dorsal aortic blood pressures (n > or = 5) with a servonull system. The atrioventricular orifice was positioned on the dorsomedial side of the anterior ventricle, surmounted by the single-chambered atrium. The atrioventricular valve was free of tension apparati but supported by papillary bands to prevent retrograde flow. The ventricle was spanned with fine trabeculae perpendicular to the compact layer and perforated with a subepicardial network of coronary arteries, which originated from the efferent branchial arteries by means of the main coronary vessel. Ventricular myocytes were larger than those in the atrium (P < 0.05) with abundant mitochondria close to the sarcolemmal. Sarcoplasmic reticulum was sparse in zebrafish ventricle. Bulbus arteriosus was located anterior to the ventricle, and functioned as an elastic reservoir to absorb the rapid rise of pressure during ventricular contraction. The dense matrix of collagen interspersed across the entire bulbus arteriosus exemplified the characteristics of vasculature smooth muscle. There were pressure gradients from atrium to ventricle, and from ventral to dorsal aorta, indicating that the valves and the branchial arteries, respectively, were points of resistance to blood flow. These data serve as a framework for structure-function investigations of the zebrafish cardiovascular system.

Power spectrum analysis of heart rate and blood flow velocity variability measured in the umbilical and uterine arteries in early pregnancy: a comparative study.

OBJECTIVE: To compare power spectral derived variability parameters from the fetal side of the placental circulation with those from the maternal side of the placental circulation, during early pregnancy. METHODS: Doppler velocity waveforms were obtained from both the umbilical and the uterine arteries in a study group of 40 pregnant women between 10 and 14 (n = 25) and 15 and 20 (n = 15) weeks of gestation. The coefficient of variation of both the beat-to-beat heart rate variability and the blood flow velocity variability was determined. The ratio of the integrated low-frequency components (< 0.2 Hz) and the integrated high-frequency components (> 0.2 Hz) from normalized power spectrum analysis (LH-ratio) was established, to reflect sympathovagal balance. RESULTS: The coefficient of variation and LH-ratio of fetal heart rate variability constitute only a fraction of the same maternal heart rate variability parameters. Nevertheless a highly significant increase (P < 0.001) in LH-ratio was demonstrated with advancing gestational age. The coefficient of variation and LH-ratio of blood flow velocity variability were significantly lower in the fetal umbilical artery only in the 10-14-weeks' gestation group. Due to a decrease of the maternal uterine blood flow velocity variability parameters with advancing gestational age, statistically equal fetal and maternal values for coefficient of variation and LH-ratio were found in the 15-20 weeks' gestation group. CONCLUSIONS: The increase in LH-ratio of fetal heart rate variability indicates functional development of the fetal autonomic nervous system at 15-20 weeks' gestation. The umbilical blood flow velocity variability may be secondary to maternal uterine arterial flow variability rather than due to primary changes in fetal cardiovascular function.

Structure and function of the developing zebrafish heart.

The combination of optical clarity and large scale of mutants makes the zebrafish vital for developmental biologists. However, there is no comprehensive reference of morphology and function for this animal. Since study of gene expression must be integrated with structure and function, we undertook a longitudinal study to define the cardiac morphology and physiology of the developing zebrafish. Our studies included 48-hr, 5-day, 2-week, 4-week, and 3-month post-fertilization zebrafish. We measured ventricular and body wet weights, and performed morphologic analysis on the heart with H&E and MF-20 antibody sections. Ventricular and dorsal aortic pressures were measured with a servonull system. Ventricular and body weight increased geometrically with development, but at different rates. Ventricle-to-body ratio decreased from 0.11 at 48-hr to 0.02 in adult. The heart is partitioned into sinus venosus, atrium, ventricle, and bulbus arteriosus as identified by the constriction between the segments at 48-hr. Valves were formed at 5-day post-fertilization. Until maturity, the atrium showed extensive pectinate muscles, and the atrial wall increased to two to three cell layers. The ventricular wall and the compact layer increased to three to four cell layers, while the extent and complexity in trabeculation continued. Further thickening of the heart wall was mainly by increase in cell size. The bulbus arteriosus had similar characteristics to the myocardium in early stages, but lost the MF-20 positive staining, and transitioned to smooth muscle layer. All pressures increased geometrically with development, and were linearly related to stage-specific values for body weight (P < 0.05). These data define the parameters of normal cardiac morphology and ventricular function in the developing zebrafish.

Complete genome sequence of Neisseria meningitidis serogroup B strain MC58.

The 2,272,351-base pair genome of Neisseria meningitidis strain MC58 (serogroup B), a causative agent of meningitis and septicemia, contains 2158 predicted coding regions, 1158 (53.7%) of which were assigned a biological role. Three major islands of horizontal DNA transfer were identified; two of these contain genes encoding proteins involved in pathogenicity, and the third island contains coding sequences only for hypothetical proteins. Insights into the commensal and virulence behavior of N. meningitidis can be gleaned from the genome, in which sequences for structural proteins of the pilus are clustered and several coding regions unique to serogroup B capsular polysaccharide synthesis can be identified. Finally, N. meningitidis contains more genes that undergo phase variation than any pathogen studied to date, a mechanism that controls their expression and contributes to the evasion of the host immune system.

Do heart rate and velocity variability derived from umbilical artery velocity waveforms change prior to clinical pregnancy-induced hypertension?

OBJECTIVE: To investigate the hypothesis that alterations in heart rate variability, peak systolic velocity variability and time-averaged velocity variability in the human umbilical artery may predict early signs of dysfunctional fetal-placental coupling in pregnancies that later develop pregnancy-induced hypertension. METHODS: Doppler flow velocity recordings from the umbilical artery were performed at 10-20 weeks of gestation in 12 nulliparous women who subsequently developed pregnancy-induced hypertension. From umbilical artery velocity waveforms of at least 12 s in duration we determined absolute values and beat-to-beat variability in fetal heart rate, peak systolic and time-averaged velocity and compared these findings with those in normal nulliparous pregnant women matched for gestational age. RESULTS: Absolute values for fetal heart rate, peak systolic and time-averaged velocity as well as beat-to-beat variability in fetal heart rate did not differ significantly between women later developing pregnancy-induced hypertension and normal controls. However, variability in peak systolic velocity and time-averaged velocity were decreased in women who subsequently developed pregnancy-induced hypertension. CONCLUSIONS: Whereas fetal heart rate variability was similar, umbilical artery flow velocity variability was reduced in women developing pregnancy-induced hypertension compared with controls. It is proposed from this study that variability of the umbilical artery flow velocity is associated with mechanical changes in the vascular bed of women who later develop pregnancy-induced hypertension.

Rate of coronary vascularization during embryonic chicken development is influenced by the rate of myocardial growth.

OBJECTIVE: We tested the hypothesis that the degree of coronary microvessel formation in the embryonic heart is regulated by the magnitude of myocardial growth. METHODS: The outflow tract of Hamburger-Hamilton stage 21 chicken hearts (prior to the onset of coronary vasculogenesis) was constricted in ovo with a loop of 10-0-nylon suture, and the hearts were studied at stages 29 and 36. RESULTS: At stage 29 ventricular mass was 64% greater in the pressure-overloaded than in the hearts of sham-operated controls, but vascular volume density and numerical density, determined by electron microscopic morphometry, were identical. As demonstrated by histological morphometric evaluation, the compact region of the left ventricle at stage 29 was 43% thicker than the shams. However, by stage 36 heart mass, thickness of the compact region, and overall wall thickness (demonstrated by scanning electron microscopy) were significantly less than in the sham group of this stage, but vascular volume density was virtually identical in the two groups. Formation of the two main coronary arteries was clearly impeded in the banded hearts, i.e., the coronaries were stunted in their development or failed to completely form coronary ostia. CONCLUSIONS: Vascular growth is proportional to myocardial growth in the embryonic, overloaded heart, but the persistence of the pressure overload results in a failure of or severe limitations in coronary artery development. These data support the hypothesis that vascular growth during this period of development is regulated, at least in part, by the rate and magnitude of myocardial growth.

Fetal heart rate and umbilical artery velocity variability in pregnancies complicated by insulin-dependent diabetes mellitus.

OBJECTIVES: To examine the variability in fetal heart rate and absolute flow velocity, which are possible hemodynamic markers of cardiovascular homeostasis in pregnancies complicated by diabetes mellitus. METHODS: Doppler studies of umbilical artery velocity waveforms were performed at 12-21 weeks of gestation in 16 women with well-controlled type I (insulin-dependent) diabetes mellitus. From umbilical artery velocity waveforms of at least 13 s in duration, we determined absolute values and beat-to-beat variability for fetal heart rate and umbilical artery flow velocities and compared these findings with normal controls matched for gestational age. RESULTS: Fetuses of diabetic women displayed increased fetal heart rate variability and umbilical artery peak systolic velocity. Fetal heart rate, umbilical artery time-averaged velocity and variability in umbilical artery flow velocity were not essentially different between the two groups. CONCLUSION: Fetal heart rate variability and umbilical artery peak systolic velocity may be markers for fetal cardiovascular homeostasis in pregnancies complicated by insulin-dependent diabetes mellitus.

The Paediatric Cardiology Hall of Fame: Helen Brooke Taussig MD. May 24, 1898 to May 21, 1986.

The life and accomplishments of Helen Taussig are reviewed at the centennial of her birth in 1898. Now, a little over 50 years since the first Blalock Taussig shunt in 1944, her legacy remains a model of brilliant scientific innovation and loving patient care.

Remodeling of chick embryonic ventricular myoarchitecture under experimentally changed loading conditions.

Adult myocardium adapts to changing functional demands by hyper- or hypotrophy while the developing heart reacts by hyper- or hypoplasia. How embryonic myocardial architecture adjusts to experimentally altered loading is not known. We subjected the chick embryonic hearts to mechanically altered loading to study its influence upon ventricular myoarchitecture. Chick embryonic hearts were subjected to conotruncal banding (increased afterload model), or left atrial ligation or clipping, creating a combined model of increased preload in right ventricle and decreased preload in left ventricle. Modifications of myocardial architecture were studied by scanning electron microscopy and histology with morphometry. In the conotruncal banded group, there was a mild to moderate ventricular dilatation, thickening of the compact myocardium and trabeculae, and spiraling of trabecular course in the left ventricle. Right atrioventricular valve morphology was altered from normal muscular flap towards a bicuspid structure. Left atrial ligation or clipping resulted in hypoplasia of the left heart structures with compensatory overdevelopment on the right side. Hypoplastic left ventricle had decreased myocardial volume and showed accelerated trabecular compaction. Increased volume load in the right ventricle was compensated primarily by chamber dilatation with altered trabecular pattern, and by trabecular proliferation and thickening of the compact myocardium at the later stages. A ventricular septal defect was noted in all conotruncal banded, and 25% of left atrial ligated hearts. Increasing pressure load is a main stimulus for embryonic myocardial growth, while increased volume load is compensated primarily by dilatation. Adequate loading is important for normal cardiac morphogenesis and the development of typical myocardial patterns.

Assessment of fetal heart rate variability and velocity variability by Doppler velocimetry of the descending aorta at 10-20 weeks of gestation.

OBJECTIVES: Determination of gestational age-related modulations in fetal heart rate and descending aorta blood flow velocity in the early human fetus and comparison of aortic variability data with data obtained from the umbilical artery. It is hypothesized that these modulations present in the umbilical artery also occur in the descending aorta. METHODS: Doppler studies of descending aorta velocity waveforms were performed at 10-20 weeks in 55 normal pregnant women. In 24 of the 55 women, Doppler recordings from both the descending aorta and the umbilical artery were collected. Absolute values and variability of fetal heart rate, peak systolic and time-averaged velocities were determined from flow velocity waveforms of at least 18 s in duration. RESULTS: From 10 to 20 weeks of gestation, the descending aorta peak systolic and time-averaged velocities increased, whereas the fetal heart rate decreased. The descending aorta peak systolic variability also increased. However, the time-averaged velocity variability and fetal heart rate variability remained constant during the study period. In the subset of 24 women, the fetal heart rate variability and velocity variability data from the descending aorta and umbilical artery were not significantly different. CONCLUSIONS: Reproducible fetal heart rate and velocity variability data can be derived from the descending aorta and umbilical artery. The increase in heart rate variability observed in the umbilical artery was not seen in recordings obtained from the descending aorta. Different fetal activity states may be the underlying mechanism for these heart rate variability discrepancies.

Heart rate and flow velocity variability as determined from umbilical Doppler velocimetry at 10-20 weeks of gestation.

1. The aim of this study was to define from umbilical artery flow velocity waveforms absolute peak systolic and time-averaged velocity, fetal heart rate, fetal heart rate variability and flow velocity variability, and the relation between fetal heart rate and velocity variables in early pregnancy.2.A total of 108 women presenting with a normal pregnancy from 10 to 20 weeks of gestation consented to participate in a cross-sectional study design. Doppler ultrasound recordings were made from the free-floating loop of the umbilical cord.3. Umbilical artery peak systolic and time-averaged velocity increased at 10-20 weeks, whereas fetal heart rate decreased at 10-15 weeks of gestation and plateaued thereafter. Umbilical artery peak systolic velocity variability and fetal heart rate variability increased at 10-20 and 15-20 weeks respectively.4. The inverse relationship between umbilical artery flow velocity and fetal heart rate at 10-15 weeks of gestation suggests that the Frank-Starling mechanism regulates cardiovascular control as early as the late first and early second trimesters of pregnancy. A different underlying mechanism is suggested for the observed variability profiles in heart rate and umbilical artery peak systolic velocity. It is speculated that heart rate variability is mediated by maturation of the parasympathetic nervous system, whereas peak systolic velocity variability reflects the activation of a haemodynamic feedback mechanism.

An estimate of fetal autonomic state by spectral analysis of human umbilical artery flow velocity waveforms.

OBJECTIVE: Determination of gestational age-related fluctuations in heart rate in the umbilical artery of the early human fetus. METHODS: Doppler velocity recordings from human umbilical artery were obtained, in a cross-sectional study design in 137 singleton pregnancies at 10-20 weeks of gestation. After exclusion criteria were applied, data on 117 normal pregnancies were available and subdivided into group I: 10-12 weeks (n = 49); group II: 13-16 weeks (n = 43); and group III: 17-20 weeks (n = 25). Blood flow velocity waveforms were reconstructed from Doppler audio signals. Variability in heart rate was calculated using Fast Fourier Transforms (FFT). Individual heart rate variability power spectra were subdivided into frequency bands. RESULTS: Fetal heart rate variability decreases at 10-20 weeks and demonstrates a shift to lower frequencies at 17-20 weeks. CONCLUSIONS: Fetal heart rate variability is related to gestational age and shows a shift to lower frequencies which may reflect autonomic functional development.

A quantitative study of the ventricular myoarchitecture in the stage 21-29 chick embryo following decreased loading.

During the early developmental period, ventricular myoarchitecture undergoes a transition from a smooth-walled cardiac tube, to left and right ventricular chambers filled with a sponge-like network of trabecular struts. We measured the quantitative changes of ventricular myocardium properties in normal stage 21-29 chick embryos and after chronic verapamil suffusion, which is known to decrease work load and decelerate ventricular growth. The morphologic parameters (compact layer thickness, ventricular wall composition, porosity of different layers and trabecular orientation) were determined from scanning electron micrographs of transversely dissected perfusion-fixed hearts. A vascular bed of stage 21 chick embryos was suffused with 1 ng of verapamil at 1 microliter per hour up to stages 24, 27 and 29 via a miniosmotic pump. From stage 24, the thickness of the compact myocardium in the left ventricle was greater than that of the right. The increase in thickness was minimal between stages 24 and 27, while the predominantly radially arranged trabeculae comprised up to 75% to total myocardial mass. The ratio of intertrabecular spaces to trabeculae (local porosity) decreased form the ventricular center (70%) towards the compact myocardium (0%). In verapamil-treated embryos, the hearts were smaller and showed delayed development. The compact myocardium was thinner than normal, and the proportion of trabeculae was higher than in controls. The local porosity values were similar in control and experimental groups. Decreased load resulted in delayed growth and morphogenesis, expressed as persistence of trabeculae and a thinner compact myocardium. Embryonic heart pumping function is largely based on extensively developed trabeculation with regionally different properties.

VEGF and bFGF stimulate myocardial vascularization in embryonic chick.

We tested the hypothesis that early vascularization of the embryonic heart is enhanced after bolus injections of vascular, endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) into the vitelline vein before the onset of myocardial vasculogenesis (3.5 days, stage 21). Electron and light microscopy were utilized to obtain morphometric data. At stages 29 and 31, myocardial vessel volume or numerical density were higher in embryos injected with 50 ng bFGF than in the saline-injected controls. A VEGF injection increased vascular volume density at stage 29 and both volume and numerical, density at stage 31, bFGF, but not VEGF, was associated with an enhancement of the sinusoidal system (spongy layer of the ventricle) at stage 29. This effect disappeared by stage 31. In conclusion, 1) enhancement of bFGF or VEGF before myocardial vascularization increases vascular growth, but the initial effect of bFGF is greater; 2) the effects of these growth factors on vascular volume and numerical density are temporally dependent; and 3) bFGF, in addition to its effects on the coronary vasculature, influences ventricular modeling by apparently acting on myocytes as well as endothelial cells.