Investigation of Copy Number Variation in Children with Conotruncal Heart Defects

Background:Congenital heart defects (CHD) are the most prevalent group of structural abnormalities at birth and one of the main causes of infant morbidity and mortality. Studies have shown a contribution of the copy number variation in the genesis of cardiac malformations.Objectives:Investigate gene copy number variation (CNV) in children with conotruncal heart defect.Methods:Multiplex ligation-dependent probe amplification (MLPA) was performed in 39 patients with conotruncal heart defect. Clinical and laboratory assessments were conducted in all patients. The parents of the probands who presented abnormal findings were also investigated.Results:Gene copy number variation was detected in 7/39 patients: 22q11.2 deletion, 22q11.2 duplication, 15q11.2 duplication, 20p12.2 duplication, 19p deletion, 15q and 8p23.2 duplication with 10p12.31 duplication. The clinical characteristics were consistent with those reported in the literature associated with the encountered microdeletion/microduplication. None of these changes was inherited from the parents.Conclusions:Our results demonstrate that the technique of MLPA is useful in the investigation of microdeletions and microduplications in conotruncal congenital heart defects. Early diagnosis of the copy number variation in patients with congenital heart defect assists in the prevention of morbidity and decreased mortality in these patients.

The Benefits of Prone SPECT Myocardial Perfusion Imaging in Reducing Both Artifact Defects and Patient Radiation Exposure

AbstractBackground:Prone imaging has been demonstrated to minimize diaphragmatic and breast tissue attenuation.Objectives:To determine the role of prone imaging on the reduction of unnecessary rest perfusion studies and coronary angiographies performed, thus decreasing investigation time and radiation exposure.Methods:We examined 139 patients, 120 with an inferior wall and 19 with an anterior wall perfusion defect that might represented attenuation artifact. Post-stress images were acquired in both the supine and prone position. Coronary angiography was used as the “gold standard” for evaluating coronary artery patency. The study was terminated and rest imaging was obviated in the presence of complete improvement of the defect in the prone position. Quantitative interpretation was performed. Results were compared with clinical data and coronary angiographic findings.Results:Prone acquisition correctly revealed defect improvement in 89 patients (89/120) with inferior wall and 12 patients (12/19) with anterior wall attenuation artifact. Quantitative analysis demonstrated statistically significant difference in the mean summed stress scores (SSS) of supine and mean SSS of prone studies in patients with disappearing inferior wall defect in the prone position and patent right coronary artery (true negative results). The mean difference between SSS in supine and in prone position was higher with disappearing than with remaining defects.Conclusion:Technetium-99m (Tc-99m) tetrofosmin myocardial perfusion imaging with the patient in the prone position overcomes soft tissue attenuation; moreover it provides an inexpensive, accurate approach to limit the number of unnecessary rest perfusion studies and coronary angiographies performed.

Palliative Senning in the Treatment of Congenital Heart Disease with Severe Pulmonary Hypertension

Background:Transposition of the great arteries (TGA) is the most common cyanotic cardiopathy, with an incidence ranging between 0.2 and 0.4 per 1000 live births. Many patients not treated in the first few months of life may progress with severe pulmonary vascular disease. Treatment of these patients may include palliative surgery to redirect the flow at the atrial level.Objective:Report our institutional experience with the palliative Senning procedure in children diagnosed with TGA and double outlet right ventricle with severe pulmonary vascular disease, and to evaluate the early and late clinical progression of the palliative Senning procedure.Methods:Retrospective study based on the evaluation of medical records in the period of 1991 to 2014. Only patients without an indication for definitive surgical treatment of the cardiopathy due to elevated pulmonary pressure were included.Results:After one year of follow-up there was a mean increase in arterial oxygen saturation from 62.1% to 92.5% and a mean decrease in hematocrit from 49.4% to 36.3%. Lung histological analysis was feasible in 16 patients. In 8 patients, pulmonary biopsy grades 3 and 4 were evidenced.Conclusion:The palliative Senning procedure improved arterial oxygen saturation, reduced polycythemia, and provided a better quality of life for patients with TGA with ventricular septal defect, severe pulmonary hypertension, and poor prognosis.

Spectrum of cardiac lesions associated with Isolated Cleft Mitral Valve and their impact on therapeutic choices

Abstract Background: Isolated cleft mitral valve (ICMV) may occur alone or in association with other congenital heart lesions. The aim of this study was to describe the profile of cardiac lesions associated with ICMV and their potential impact on therapeutic management. Methods: We conducted a descriptive study with data retrieved from the Congenital Heart Disease (CHD) single-center registry of our institution, including patients with ICMV registered between December 2008 and November 2014. Results: Among 2177 patients retrieved from the CHD registry, 22 (1%) had ICMV. Median age at diagnosis was 5 years (6 days to 36 years). Nine patients (40.9%) had Down syndrome. Seventeen patients (77.3%) had associated lesions, including 11 (64.7%) with accessory chordae in the left ventricular outflow tract (LVOT) with no obstruction, 15 (88.2%) had ventricular septal defect (VSD), three had secundum atrial septal defect, and four had patent ductus arteriosus. Thirteen patients (59.1%) required surgical repair. The decision to proceed with surgery was mainly based on the severity of the associated lesion in eight patients (61.5%) and on the severity of the mitral regurgitation in four patients (30.8%). In one patient, surgery was decided based on the severity of both the associated lesion and mitral regurgitation. Conclusion: Our study shows that ICMV is rare and strongly associated with Down syndrome. The most common associated cardiac abnormalities were VSD and accessory chordae in the LVOT. We conclude that cardiac lesions associated with ICMV are of major interest, since in this study patients with cardiac lesions were diagnosed earlier. The decision to operate on these patients must take into account the severity of both mitral regurgitation and associated cardiac lesions.

Comportamento do Heterocromossômio em alguns Ortópteros do Brasil

In the present paper the behavior of the heterochromoso-mes in the course of the meiotic divisions of the spermatocytes in 15 species of Orthoptera belonging to 6 different families was studied. The species treated and their respective chromosome numbers were: Phaneropteridae: Anaulacomera sp. - 1 - 2n = 30 + X, n +15+ X and 15. Anaulacomera sp. - 2 - 2n - 30 + X, n = 15+ X and 15. Stilpnochlora marginella - 2n = 30 + X, n = 15= X and 15. Scudderia sp. - 2n = 30 + X, n = 15+ X and 15. Posldippus citrifolius - 2n = 24 + X, n = 12+X and 12. Acrididae: Osmilia violacea - 2n = 22+X, n = 11 + X and 11. Tropinotus discoideus - 2n = 22+ X, n = 11 + X and 11. Leptysma dorsalis - 2n = 22 + X, n = 11-J-X and 11. Orphulella punctata - 2n = 22-f X, n = 11 + X and 11. Conocephalidae: Conocephalus sp. - 2n = 32 + X, n = 16 + X and 16. Proscopiidae: Cephalocoema zilkari - 2n = 16 + X, n = 8+ X and 8. Tetanorhynchus mendesi - 2n = 16 + X, n = 8+X and 8. Gryliidae: Gryllus assimilis - 2n = 28 + X, n = 14+X and 14. Gryllodes sp. - 2n = 20 + X, n = 10- + and 10. Phalangopsitidae: Endecous cavernicola - 2n = 18 +X, n = 94-X and 9. It was pointed out by the present writer that in the Orthoptera similarly to what he observed in the Hemiptera the heterochromosome in the heterocinetic division shows in the same individual indifferently precession, synchronism or succession. This lack of specificity is therefore pointed here as constituting the rule and not the exception as formerly beleaved by the students of this problem, since it occurs in all the species referred to in the present paper and probably also m those hitherto investigated. The variability in the behavior of the heterochromosome which can have any position with regard to the autosomes even in the same follicle is attributed to the fact that being rather a stationary body it retains in anaphase the place it had in metaphase. When this place is in the equator of the cell the heterochromosome will be left behind as soon as anaphase begins (succession). When, on the contrary, laying out of this plane as generally happens (precession) it will sooner be reached (synchronism) or passed by the autosomes (succession). Due to the less kinetic activity of the heterochromosome it does not orient itself at metaphase remaining where it stands with the kinetochore looking indifferently to any direction. At the end of anaphase and sometimes earlier the heterochromosome begins to show mitotic activities revealed by the division of its body. Then, responding to the influence of the nearer pole it moves to it being enclosed with the autosomes in the nucleus formed there. The position of the heterochromosome in the cell is explained in the following manner: It is well known that the heterochromosome of the Orthoptera is always at the periphery of the nucleus, just beneath the nuclear membrane. This position may be any in regard of the axis of the dividing cell, so that if one of the poles of the spindle comes to coincide with it, the heterochromosome will appear at this pole in the metaphasic figures. If, on the other hand, the angle formed by the axis of the spindle with the ray reaching the heterochromosome increases the latter will appear in planes farther and farther apart from the nearer pole until it finishes by being in the equatorial plane. In this way it is not difficult to understand precession, synchronism or succession. In the species in which the heterochromosome is very large as it generally happens in the Phaneropteridae, the positions corresponding to precession are much more frequent. This is due to the fact that the probabilities for the heterochromosome taking an intermediary position between the equator and the poles at the time the spindle is set up are much greater than otherwise. Moreover, standing always outside the spindle area it searches for a place exactly where this area is larger, that is, in the vicinity of the poles. If it comes to enter the spindle area, what has very little probability, it would be, in virtue of its size, propelled toward the pole by the nearing anaphasic plate. The cases of succession are justly those in which the heterochromosome taking a position parallelly to the spindle axis it can adjust its large body also in the equator or in its proximity. In the species provided with small heterochromosome (Gryllidae, Conocephalidae, Acrididae) succession is found much more frequently because here as in the Hemiptera (PIZA 1945) the heterochromosome can equally take equatorial or subequatorial positions, and, furthermore, when in the spindle area it does offer no sereous obstacle to the passage of the autosomes. The position of the heterochromosome at the periphery of the nucleus at different stages may be as I suppose, at least in part a question of density. The less colourability and the surface irregularities characteristic of this element may well correspond to a less degree of condensation which may influence passive movements. In one of the species studied here (Anaulacomera sp.- 1) included in the Phaneropteridae it was observed that the plasmosome is left motionless in the spindle as the autosomes move toward the poles. It passes to one of the secondary spermatocytes being not included in its nucleus. In the second division it again passes to one of the cells being cast off when the spermatid is being transformed into spermatozoon. Thus it is regularly found among the tails of the spermatozoa in different stages of development. In the opinion of the present writer, at least in some cases, corpuscles described as Golgi body's remanents are nothing more than discarded plasmosomes.

Uma raça cromossômica natural de Tityus bahiensis (Scorpiones-Buthidae)

A natural chromosomal race of Tityus babiensis (Scorpiones Buthidae) is described in the present paper. Five males and seven females received from St. Joaquim, State of S. Paulo, gave the following interesting results: All the spermatogonia of the five males were provided with 9 chromosomes of different sizes. All primary spermatocytes showed at metaphase one independent bivalent of normal shape and a complex group formed by 7 chromosomes which have exchanged parts. Some of the chromosomes associated in the complex group, to Judge by their behavior, were composed of fragments of three different chromosomes, being thus paired with three other members of the compound group. The manner in which all the 7 components of the group have paired with each other showed to be very constant. They gave always origin to a double-cross configuration, the longst branch of which being formed by a long chromosome paired with two components of the group and with a third chromosome that did not belong to the group. The chromosomes of the independent bivalent separate regularly, going to different poles. From the 7 elements of the compound group, 4 go to one pole and 3 to the opposite one. Consequently, secondary spermatocytes with 4 and 5 chromosomes are produced. The females, so far as it can be inferred from the study of the follicular cells of the ovariuterus, have 10 chromosomes. These females are, therefore, considered as being monogametic, that is, as producing eggs with 5 chromosomes. A sex-determining mechanism arose in this manner, the spermatozoa with 5 chromosomes giving origin to females and those with 4 to males. The fact that the sex chromosome is one of the elements taking part in the formation of the group, seems highly interesting to the author. Tetraploid cysts have been occasionally found in the testis. In one individual the chromosomes of the tetraploid primary spermatocytes behaved as expected, forming a group of 14 elements, and two independent pairs or a tetravalent group In another individual, the chromosomes of the tetraploid cells have formed two independent groups of 7, and two independent pairs, as if both chromosomal sets were by their turn entirely independent frcm one another. This fact is certainly not devoid of special interest. The males as well as the females studied in this paper differed in nothing from the typical members of the species. The unique differential character of the new race is found in the umber and behavior of its chromosomes. It is highly remarkable that the occurrences which have transformed the 6 chromosomes normally present in the species into a new set of 9 elements, 7 of which have been profoun- dly altered in their structure, do not show any influence on the morphology of the organism. This fact, together with those found in the salivary-chromosomes races of Drosophila and Sciara. compromises strongly the genetical concept of position effects.

Breve notícia sôbre a espermatogênese de Lutosa brasiliensis Brunner (Tettigoniodea-Stenopelmatidae)

Lutosa brasiliensis, an Orthopteran Tettigonioidean belonging to the family Stenopelmatidae is referred to in this paper The spermatogonia are provided with 15 chromosomes, that is, 7 pairs of autosomes and a single sex chromosome. One pair of autosomes is much larger than the rest, two pairs are of median sized elements, and four pairs are of small ones. The daughter sex chromosomes show at anaphase great difficulty in reaching the poles, being left for a long while in the region of the equator where they are seen stretched one after the other on the same line or lying side by side in different positions. When the spermatogonium divides each daughter cell gets passively its sex chromosome. Though slowly, the sex chromosome finishes by beins enclosed in the nucleus. Its behavior may be attributed to a very weak kinetic activity of the centromere. In view of se pronouced an inertness of the sex chromosomes, two things may be expected : primary spermatocyte nuclei with two sex chromosomes, and primary spermatocytes with the sex chromosome lying outside the nucleus. Both situations have been discovered. The latter, together with the delay of the spermatogonial sex chromosome in reaching the poles suggested to the anther the mechanism which might have given origin to the cases in which the sex chromosome normally does not enter the nucleus to rejoin the autosomes, remaning outside in its own nucleus. It may well be supposed that accidents like that found in the present individual have turned to be a normal event in the course of the evolution of some species. Trie primary spermatocytes are provided with chromatoid bodies which remain visible all over the whole history of the cells and pass to one of the resulting secondary spermatocytes, the larger of them being found later in the area occupied by the tails of the spermatozoa. No relation of these bodies to nucleoli con?d be established. Pachytene and diplotene nuclei are normal Metaphase nuclei show 7 autosomal tetrads, one of which being much larger than the rest. At this stage the chromosomes have a pronounced tendency to form clumps. Even when they are separated from each other they generally appear competed by chromosomal substance. The sex chromosome Hes always in one of the poles, being enclosed in the nucleus formed there. The stickness of the chromosomes can also be noted at anaphase. Telophase chromosomes distend them- selves for giving origin to secondary spermatocyte nuclei in a state comparable to a beginning prophase. As the secondary spermatocytes approach metaphase the autosomes appear entirely divided except at the kinetochore where the chromatids remain united. In the division of the secondary spermatocytes nothing else merits special reference.

Mapping of the healthy immunoglobulin repertoire over the course of B cell maturation by deep pyrosequencing

Healthy immunoglobulin repertoire has not been extensively evaluated reflecting in part the challenge of generating sufficiently robust data sets by conventional clonal sequencing. Deep sequencing has revolutionized the capacity to evaluate the depth and breadth of the Ig repertoire along the B cell developmental pathway, and can be used to pin point defect(s) of primary or acquired B-cell associated diseases. In this study healthy IgM and IgG repertoires were studied by 454-pyrosequencing to establish the healthy controls for diseased repertoires. (...)

Histologia del sistema reproductor masculino de Helobdella hyalina (Hirudinea, Glossiphoniidae) en Argentina

A detailed preliminary histological analysis of Helobdella hyalina Ringuelet, 1942 male system from Los Talas, Buenos Aires, Argentina is described. Six pairs of testisacs, located between the crop caeca, form the male reproductive system. Each testisac is clothed by the mesotelium. Inside it, the germinal cells are connected to the citophore and develop functional unit called poliplast. The spermatozoa are released into testisacs after the reabsortion of the citophore. Five stages of spermatogenesis are described taking into account the successive maturation stages of germinal cells and the changes in the citophore size. Lining cells and gland cells were found in the seminal vesicle. Five different types of gland cells are placed inside the ejaculatory ducts, as well as two kinds of cells are found in its distal portion: type 1, which produces eosinophilic granular secretion, type 2, with amorphous and slightly eosinophilic. Three distinct gland cells are located in the proximal portion of the duct: type 3, which produces a strongly eosinophilic granular secretion; type 4, with a negative eosinophilic amorphous secretion and type 5, with a basophilic granular secretion. Type 5 gland cells are described for the ducts of this species only.

Biologia do saguirú (Characidae, Curimatinae)

In Brazil all the fishes belonging to the sub-family Curimatinae are called « saguirú ». The present work gives a biological study of the Curimatus elegans Steind., a small fish without any economical importance, which is to be found along the whole brazilian coast, down till Paraguay. The specimens utilized for the present study come from Fortaleza (Ceará, north-eastern Brazil). The C. elegans is « ilyophagus », that means, it feeds itself exclusively with those organic materials to be found in mud, specially with microscopical algae. The intestines are very extent, some of them measuring about 9 to 11 times body's length. Studies have been made about growth and age of the C. elegans; the biggest sizes found were of 153 mm. for females and 88 mm. for males. The C. elegans shows developed sexual glands during a long period (April to September). The movements of the spermatozoa, in contact with water is of 40 to 50 seconds of intense movements, ceasing after 70 to 100 seconds. In contact with 0.5% NaCl-solution spermatozoa show a big increase in movements-time, that can last till about 25 minutes. The eggs' diameter measures 0.70 to 0.73 mm., mature and hydrated it attains 0.93 to 1,00 mm. There is a certain correlation between the size of the body and the quantity of eggs. Big specimens can produce a total of 200.000 eggs. The average quantity contained in 1 gr. and 1 cc. is 6018 and 6229 eggs, respectively. Maturity and spawning in laboratory has been obtained due to injections of suspension of fish-hypophysis. Three or four hours after the injection, fishes show more movement and evident signs of excitation, proceeding spawning after 5 to 6 hours. Males, persecuting females, describe successive circles (merry-go-round) - carroussel), swimming side by side with females up to water's surface, where sexual products are start beating dry, for there is no blood yet. Circulation-scheme is to be found on fig. 4 and 5. The swim-bladder and the stomach are but delineated; the intestine is formed by a cylindric tube, all closed. At the place, where later on there will open the mouth, we find a group of ciliary hairs that produce a liquid current, very evident by the semi-circle formed by attached solid particles. After 36 hours, opening of the mouth and formation of the gill slits begin. At the age of 90 hours (4 mm.) the larvas swim well and start to feed themselves; the digestive tube is now all open and the swimbladder works already. During the first days of life, larvas have an adhesive organ situated at their frontal region (fig. 7) in form of a crescent, by means of which they hang to surrounding vegetation (fig. 6). When the larva begins to swim and to feed itself and its yolk are having been absorbed. the adhesive organ retracts and disappears. While larvas and alevins feed themselves with plancton, they have small eye-teeth, which disappear,. when fishes become « ilyophagus ». There exist too, during their life as larvas, pharyngeal-teeth. The lateral line appears in the larva after 16 to 18 days; more or less at the same time all fins are completely developed. Shortly after, first scales appear (20 to 23 days). Evolution of intestines twisting followed (fig. 9). Larvas show at different parts of their bodies small of organs excretory functions, that are constituted by bottons in serial disposition, every one with an excretory canal that opens towards the outside. These formations disappear suddenly when larvas attain their phase of alevin. The existence of a great number of said formations at the caudal fin (fig. 12) is of great interest. In our experiences of breeding we have employed several thousands of C. elegans larvas in different environs and we made conditions of surrounding change (illumination), depth of water, temperature, presence of sand at bottom of aquariums and without sand, food). In this way we could compare the results obtained, estimate the action of each factor for the realisation of a good bring-up of larvas.

Análise citológica e cariométrica da ação da colchina sôbre a espermatogênese dos hemípteros

The action of colchicine upon the spermatogenesis of Triatoma infestans, (Hemipt. Heteroptera), has been studied and the different categories of giant spermatids that appear during the treatment have been compared with the nuclear volumes of the whole series of normal spermatogenetic stages. The following facts have been ascertained: 1) 4 hours after the treatment the gonial mitotic metaphases, and the 1st. and 2nd. metaphases of meiosis are stopped. The prophasic stages of meiosis and diakynesis appear to be normal. After 9 days of treatment, all the tetrads are broken in the meiotic metaphases and the cells appear with 44 and 22 chromosomes respectively, scattered in the cytoplasm. 2) At 9 days, practically all spermatogenetic stages have disappeared except for a few cysts of spermatogonia, and practically the whole testicle is full of cysts of spermatozoa and spermatid, with some large zones of necrosis with pycnotic nuclei. The spermatids appear to be of different sizes and the statistical analysis of the nuclear volumes gives a polymodal hystogram with 4 modes, whose volumes are in the ratio of 1:2:4:8. Ripe spermatozoa seem to have a certain volume variability, that has not been possible to analyse quantitatively. All these facts confirm what DOOLEY found in the colchicinized Orthoptera testicle. 3) The caryometric analysis conducted statistically on the normal stages of the spermatogenesis (resting spermatogonia, gonial prophases, leptotene, "confused stage", diakynesis, and spermatid) revealed the following facts: a) Considering the volume of the resting, spermatogonia as 1, their mitotic prophases have a volume of 2. Some rare prophases appear to have a volume of 4 and probably belong to tetraployd spermatogonia normally present in the testicle of Hemiptera. b) The first spermatocyte at the beginning of the auxocitary growth (leptotene) has a volume of 2, which is equal to that of them gonial prophase. It grows further during the "confused stage" and reduplicates, reaching thus the volume of 4. Diakynesis has a rather variable nuclear volume and it is higher than volume 4. This is probably of physico-chemical nature and not a growth increase. c) The spermatid at the beginning of the spermiogenetic process has a volume of 1 which is very constant and homogeneous. 4) These results can be summarized concluding that the meiotic process begins from a spermatogonium at the end of his mitotic interphasic growth (vol. 2) and instead of entering into the mitotic prophase transforms itself into the leptotene spermatocyte. During the diplotene ("confused stage") the volume of the nucleus doubles once more and reaches volume 4. In consequence of the two successive meiotic divisions the spermatid, although having an haploid number of chromosomes, has a nuclear volume of 1, just like the diploid spermatogonium. The interpretation of this strange result probably comes from the existence of the "tertiary split" in the chromosomes of the haploid set, that has been illustrated in the Hemiptera by HUGUES SCHRADER and in Orthoptera by MICKEY and co-workers. The tertiary split indicates that the chromosomes of the haploid set are constituted from almost two chromonemata, and this double constitution corresponds to the double cycle of reduplication that takes place during the spermatogenesis starting from the resting gonia. In Triatoma infestans the tertiary split appears in the chromosomes in the 1st. and 2nd. metaphases and in the diakynesis. In the blocked metaphases at the 9th. day of colchicinization some of the 44 elements scattered in the cytoplasm, show, when properly oriented, the split very clearly. Some new and strange facts revealed by SCHRADER and LEUCHTEMBERGER in Arvelius suggest the possibility of other interpretations of the rhythmic growth in special cases. There appears the necessity of more knowledge about the multiple or simple constitution of the chromosomes in somatic and spermatogonial mitosis.

Proteomic and Metabolomic Analyses of Mitochondrial Complex I-deficient Mouse Model Generated by Spontaneous B2 Short Interspersed Nuclear Element (SINE) Insertion into NADH Dehydrogenase (Ubiquinone) Fe-S Protein 4 (Ndufs4) Gene.

Eukaryotic cells generate energy in the form of ATP, through a network of mitochondrial complexes and electron carriers known as the oxidative phosphorylation system. In mammals, mitochondrial complex I (CI) is the largest component of this system, comprising 45 different subunits encoded by mitochondrial and nuclear DNA. Humans diagnosed with mutations in the gene NDUFS4, encoding a nuclear DNA-encoded subunit of CI (NADH dehydrogenase ubiquinone Fe-S protein 4), typically suffer from Leigh syndrome, a neurodegenerative disease with onset in infancy or early childhood. Mitochondria from NDUFS4 patients usually lack detectable NDUFS4 protein and show a CI stability/assembly defect. Here, we describe a recessive mouse phenotype caused by the insertion of a transposable element into Ndufs4, identified by a novel combined linkage and expression analysis. Designated Ndufs4(fky), the mutation leads to aberrant transcript splicing and absence of NDUFS4 protein in all tissues tested of homozygous mice. Physical and behavioral symptoms displayed by Ndufs4(fky/fky) mice include temporary fur loss, growth retardation, unsteady gait, and abnormal body posture when suspended by the tail. Analysis of CI in Ndufs4(fky/fky) mice using blue native PAGE revealed the presence of a faster migrating crippled complex. This crippled CI was shown to lack subunits of the "N assembly module", which contains the NADH binding site, but contained two assembly factors not present in intact CI. Metabolomic analysis of the blood by tandem mass spectrometry showed increased hydroxyacylcarnitine species, implying that the CI defect leads to an imbalanced NADH/NAD(+) ratio that inhibits mitochondrial fatty acid β-oxidation.

Extracorporeal membrane oxygenation support after heart explantation: a proposal on how to deal with hyperacute rejection of heart transplant

Purpose: In extreme situations, such as hyperacute rejection of heart transplant or major bleeding per-operating complications, an urgent heart explantation might be the only means of survival. The aim of this experimental study was to improve the surgical technique and the hemodynamics of an Extracorporeal Membrane Oxygenation (ECMO) support through a peripheral vascular access in an acardia model. Methods: An ECMO support was established in 7 bovine experiments (59±6.1 kg) by the transjugular insertion to the caval axis of a self-expanded cannula, with return through a carotid artery. After baseline measurements of pump flow and arterial and central venous pressure, ventricular fibrillation was induced (B), the great arteries were clamped, the heart was excised and right and left atria remnants, containing the pulmonary veins, were sutured together leaving an atrial septal defect (ASD) over the cannula in the caval axis. Measurements were taken with the pulmonary artery (PA) clamped (C) and anastomosed with the caval axis (D). Regular arterial and central venous blood gases tests were performed. The ANOVA test for repeated measures was used to test the null hypothesis and a Bonferroni t method for assessing the significance in the between groups pairwise comparison of mean pump flow. Results: Initial pump flow (A) was 4.3±0.6 L/min dropping to 2.8±0.7 L/min (P B-A= 0.003) 10 minutes after induction of ventricular fibrillation (B). After cardiectomy, with the pulmonary artery clamped (C) it augmented not significantly to 3.5±0.8 L/min (P C-B= 0.33, P C-A= 0.029). Finally, PA anastomosis to the caval axis was followed by an almost to baseline pump flow augmentation (4.1±0.7 L/min, P D-B= 0.009, P D-C= 0.006, P D-A= 0.597), permitting a full ECMO support in acardia by a peripheral vascular access. Conclusions: ECMO support in acardia is feasible, providing new opportunities in situations where heart must urgently be explanted, as in hyperacute rejection of heart transplant. Adequate drainage of pulmonary circulation is pivotal in order to avoid pulmonary congestion and loss of volume from the normal right to left shunt of bronchial vessels. Furthermore, the PA anastomosis to the caval axis not only improves pump flow but it also permits an ECMO support by a peripheral vascular access and the closure of the chest.

The protein phosphatase 7 regulates phytochrome signaling in Arabidopsis.

The psi2 mutant of Arabidopsis displays amplification of the responses controlled by the red/far red light photoreceptors phytochrome A (phyA) and phytochrome B (phyB) but no apparent defect in blue light perception. We found that loss-of-function alleles of the protein phosphatase 7 (AtPP7) are responsible for the light hypersensitivity in psi2 demonstrating that AtPP7 controls the levels of phytochrome signaling. Plants expressing reduced levels of AtPP7 mRNA display reduced blue-light induced cryptochrome signaling but no noticeable deficiency in phytochrome signaling. Our genetic analysis suggests that phytochrome signaling is enhanced in the AtPP7 loss of function alleles, including in blue light, which masks the reduced cryptochrome signaling. AtPP7 has been found to interact both in yeast and in planta assays with nucleotide-diphosphate kinase 2 (NDPK2), a positive regulator of phytochrome signals. Analysis of ndpk2-psi2 double mutants suggests that NDPK2 plays a critical role in the AtPP7 regulation of the phytochrome pathway and identifies NDPK2 as an upstream element involved in the modulation of the salicylic acid (SA)-dependent defense pathway by light. Thus, cryptochrome- and phytochrome-specific light signals synchronously control their relative contribution to the regulation of plant development. Interestingly, PP7 and NDPK are also components of animal light signaling systems.

CD14 expression on monocytes and TNF alpha production in patients with septic shock, cardiogenic shock or bacterial pneumonia.

OBJECTIVES: In patients with septic shock, circulating monocytes become refractory to stimulation with microbial products. Whether this hyporesponsive state is induced by infection or is related to shock is unknown. To address this question, we measured TNF alpha production by monocytes or by whole blood obtained from healthy volunteers (controls), from patients with septic shock, from patients with severe infection (bacterial pneumonia) without shock, and from patients with cardiogenic shock without infection. MEASUREMENTS: The numbers of circulating monocytes, of CD14+ monocytes, and the expression of monocyte CD14 and the LPS receptor, were assessed by flow cytometry. Monocytes or whole blood were stimulated with lipopolysaccharide endotoxin (LPS), heat-killed Escherichia coli or Staphylococcus aureus, and TNF alpha production was measured by bioassay. RESULTS: The number of circulating monocytes, of CD14+ monocytes, and the monocyte CD14 expression were significantly lower in patients with septic shock than in controls, in patients with bacterial pneumonia or in those with cardiogenic shock (p < 0.001). Monocytes or whole blood of patients with septic shock exhibited a profound deficiency of TNF alpha production in response to all stimuli (p < 0.05 compared to controls). Whole blood of patients with cardiogenic shock also exhibited this defect (p < 0.05 compared to controls), although to a lesser extent, despite normal monocyte counts and normal CD14 expression. CONCLUSIONS: Unlike patients with bacterial pneumonia, patients with septic or cardiogenic shock display profoundly defective TNF alpha production in response to a broad range of infectious stimuli. Thus, down-regulation of cytokine production appears to occur in patients with systemic, but not localised, albeit severe, infections and also in patients with non-infectious circulatory failure. Whilst depletion of monocytes and reduced monocyte CD14 expression are likely to be critical components of the hyporesponsiveness observed in patients with septic shock, other as yet unidentified factors are at work in this group and in patients with cardiogenic shock.