The brain-derived neurotrophic factor gene confers susceptibility to bipolar disorder: evidence from a family-based association study

Bipolar disorder (BP) is a severe psychiatric disease, with a strong genetic component, that affects 1% of the population worldwide and is characterized by recurrent episodes of mania and depression. Brain-derived neurotrophic factor (BDNF) has been implicated in the pathogenesis of mood disorders, and the aim of the present study was to test for the presence of linkage disequilibrium between two polymorphisms in the BDNF gene and BP in 283 nuclear families. Family-based association test (FBAT) results for the dinucleotide repeat (GT)(N) polymorphism at position -1040 bp showed that allele A3 was preferentially transmitted to the affected individuals (Z=2.035 and P=.042). FBAT results for the val66met SNP showed a significant association for allele G (Z=3.415 and P=.00064). Transmission/disequilibrium test (TDT) haplotype analysis showed a significant result for the 3-G allele combination (P=.000394), suggesting that a DNA variant in the vicinity of the BDNF locus confers susceptibility to BP. Given that there is no direct evidence that either of the polymorphisms we examined alters function, it is unlikely that the actual risk-conferring allele is from these two sites. Rather, the causative site is likely nearby and in linkage disequilibrium with the 3-G haplotype that we have identified.     

Divergent Effects of Short-Term,Very-Low-Calorie Diet on Insulin-Like Growth Factor-I and Insulin-Like Growth Factor Binding Protein-3 Serum Concentrations in Premenopausal Women with Obesity

DE PERGOLA, GIOVANNI, MAURO ZAMBONI, NICOLA PANNACCIULLI, EMANUELA TURCATO, FRANCESCO GIORGINO, FABIO ARMELLINI, FRANCESCO LOGOLUSO, MARCELLO SCIARAFFIA, OTTAVIO BOSELLO, RICCARDO GIORGINO. Divergent effects of short-term, very-low-calorie diet on insulinlike growth factor-I and insulin-like growth factor binding protein-3 serum concentrations in premenopausal women with obesity. Obes Res. 1998;6:408–415. Objective : Insulin-like growth factor-I (IGF-I) and insulinlike growth factor binding protein-3 (IGFBP-3) serum concentrations provide a good measure of the biological effects of growth, hormone. The aims of the present study were to: (1) investigate the associations of IGF-I and IGFBP-3 with body fat mass and distribution, and (2) evaluate the effects of 3 weeks of very-low-calorie diet (VLCD) (318 kcal/day, with 40 g protein, 35 g carbohydrate, and 2 g fat) on IGF-I and IGFBP-3 serum concentrations. Research Methods and Procedures : The study was performed in 21 nondiabetic premenopausal women with obesity (body mass index <27.0 kg/m²; age: ranging from 18 to 48 years). Body fat mass and distribution were measured by computed tomography. Results : Before dietary treatment, IGF-I and IGFBP-3 serum concentrations were inversely associated with visceral adipose tissue (VAT) area (p<0.005 and p<0.05, respectively), but not with either total body fat or subcutaneous adipose tissue area. VLCD produced a significant decrease of body mass index (p<0.001), total body fat (p<0.001), VAT (p<0.005), subcutaneous adipose tissue (p<0.001), IGF-I concentrations (p<0.05), and an increase of IGFBP-3 serum levels (p<0.001). The association of VAT with either IGF-I or IGFBP-3 serum concentrations was not maintained following VLCD. Discussion : Our study suggests that visceral adipose tissue, rather than adiposity per se, accounts for IGF-I and IGFBP-3 serum concentrations, and that rapid weight loss, possibly due to nutritional changes, results in lower IGF-I concentrations, higher IGFBP-3 concentrations, and abrogation of the inverse associations of VAT with IGF-I and IGFBP-3.     

Inactivation of the Regulatory Gene algU or gacA Can Affect the Ability of Biocontrol Pseudomonas fluorescens CHA0 To Persist as Culturable Cells in Nonsterile Soil

Rifampin-resistant Pseudomonas fluorescens CHA0-Rif and mutants in which the regulatory gene algU (encoding sigma factor σ^E) or gacA (encoding a global regulator of secondary metabolism) was inactivated were compared for persistence in three nonsterile soils. Functional algU and (particularly) gacA were needed for CHA0-Rif to maintain cell culturability in soil.     

Adeno-associated virus mediated gene delivery into coronary microvessels of chronically instrumented dogs.

The objective of this study was to assess the potential of adeno-associated virus (AAV)-mediated gene delivery into coronary microvessels in vivo in a large animal. Ten mongrel dogs were chronically instrumented and allowed to recover for 10 days. Dogs were reanesthetized, and the aorta was constricted by a hydraulic occluder, whereby left ventricular (LV) pressure increased by 30% and left circumflex coronary artery blood flow by 50%. Recombinant AAV (serotype 2, CMV enhancer/chicken beta-actin promoter) encoding for green fluorescent protein (GFP) was injected as a bolus into the left atrium during aortic constriction at total titers of 1010 or 1012 infectious units. Dogs were followed for 2 (n = 4)or4wk(n = 6). Hemodynamics or body weight did not change. In LV tissue slices, a fluorescein-labeled antibody to GFP stained endothelial and smooth muscle cells but was absent in myocytes. To quantify transduction, slices were then stained with antibodies against alpha-smooth muscle actin or von Willebrand factor. Approximately 4% of arterioles and 2% of microvessels stained positive for anti-GFP independent from viral titer or duration. By regression analyses, the percent of vessels transfected was proportional to the increase in LV systolic pressure during occlusion. AAV is a potential vector for gene transfer into the coronary microcirculation in large animals, including perhaps humans.     

Unexpected differences between D- and L- tyrosine lead to chiral enhancement in racemic mixtures.

We report here an unexpected difference in the solubilities of D- and L-tyrosine in water, which could be discerned by their rate of crystallization and the resulting concentrations of their saturated solutions. A supersaturated solution of 10 mM L-tyrosine at 20 degrees C crystallized much more slowly than that of D-tyrosine under the same conditions, and the saturated solution of L-tyrosine was more concentrated than that of D-tyrosine. Supersaturated solutions of 10 mM DL-tyrosine in water formed precipitates of predominantly D-tyrosine and DL-tyrosine, resulting in an excess of L-tyrosine in the saturated solution. The experimental setups were monitored independently by UV-absorption, radioactivity tracing, optical rotation and X-ray diffraction. The process of nucleation and crystallization of D- and L-tyrosine is characterized by an exceptionally high cooperativity. It is possible that minute energy differences between D- and L-tyrosine, originating from parity violation or other non-conservative chiral discriminatory rules, could account for the observations. The physical process that initiated chiral selection in biological systems remains a challenging problem in understanding the origin of life, and it is possible that chiral compounds were concentrated from supersaturated racemic mixtures by preferential crystallization.     

Anti-CEA monoclonal antibody: technetium-99m labeling and the validation process of a scintigraphic animal model with a non-cellular antigenic implant.

Animal models are currently used to verify the biodistribution of different radiopharmaceuticals before its clinical application in Nuclear Medicine; however, there may be some limitations. The utilization of labelled anti-tumor monoclonal antibodies (MoAb) in experimental models often requires implant of human antigens (usually a cellular implant), which cannot be achieved in immunocompetent animals. Our purpose was to label an anti-CEA MoAb with technetium-99m (99Tc) and to validate a simplified animal model using a noncellular antigenic implant. MoAb was directly labelled with 99mTc, after reduction with 2-mercaptoethanol. Labeling efficiency was checked by ascending chromatography and immunoreactive fraction was measured in plastic wells sensitized with the antigen. Radiopharmaceutical biodistribution was evaluated by dissection and scintigraphy in 5 mice groups; following the subcutaneous administration of Al(OH)3, CEA adsorbed Al(OH)2 and a control group evaluation. Labeling efficiency was 94+/-3%, which showed to be stable for 24 hr, with immunoreactive fraction above 50%. Invasive biodistribution evaluation showed prolonged blood retention, hepatic and renal uptake. A significant increase in uptake was observed in scintigraphic studies of animals with CEA-adsorbed Al(OH)3 implants compared with the other groups (p<0.05). The non-cellular antigenic implant model simplifies the pre-clinical evaluation of labelled MoAb.     

DIVERSITY,LIFE HISTORY,AND ECOLOGY OF TRENTEPOHLIA AND PRINTZINA (TRENTEPOHLIALES,CHLOROPHYTA) IN URBAN HABITATS IN WESTERN IRELAND 1

On the basis of field and culture investigations, five species of the genera Trentepohlia and Printzina were found to occur in urban habitats in western Ireland: Trentepohlia abietina (Flotow) Hansgirg, T. aurea (Linnaeus) Martius, T. iolithus (Linnaeus) Wallroth, T. cf. umbrina (Kützing) Bornet, and Printzina lagenifera (Hildebrandt) Thompson et Wujek. These species formed perennial populations on a variety of substrata. T. abietina occurred on bark of trees; T. cf. umbrina occurred on stone walls; and P. lagenifera grew on several substrata, mainly cement and asbestos sheeting. T. aurea and T. iolithus were found on old concrete and cement walls; in particular, the latter species formed characteristic, extensive, deep‐red patches on many buildings. In culture, best growth and reproduction of these species were observed at 10 and 15° C, 16:8 h light:dark. Both in culture and in the field, reproduction took place by release of biflagellate swarmers behaving as asexual spores, germinating to produce new plants without any evidence of sexual fusion; release of biflagellate swarmers in the field was generally observed in all seasons throughout a whole annual cycle. Confirmation of the occurrence of sexual reproduction in Trentepohlia was not obtained.     

5-enol-Pyruvyl-Shikimate-3-Phosphate Synthase from Zea mays Cultured Cells (Purification and Properties)

The shikimate pathway enzyme 5-enol-pyruvyl-shikimate-3-phosphate (EPSP) synthase (3-phosphoshikimate-1-carboxyvinyl transferase, EC 2.5.1.19) was purified from cultured maize (Zea mays L. var Black Mexican Sweet) cells. Homogeneous enzyme preparations were obtained by a four-step procedure using ammonium sulfate fractionation, anion- and cation-exchange chromatography, and substrate elution from a cellulose phosphate column. The last step resulted in two well-separated activities of about the same molecular weight. A 2000- to 3000-fold purification, with an overall recovery of one-fourth of the initial activity, was achieved. Both EPSP synthase isoforms were characterized with respect to structural, kinetic, and biochemical properties. Only slight differences are seen in molecular mass, activation energy, and apparent affinities for the two substrates. A more pronounced difference was found between their thermal inactivation rates. Two EPSP synthase isoforms were also elucidated in crude homogenates by anion-exchange fast protein liquid chromatography. This allowed us to follow their expression during a culture growth cycle. One form was found at substantial levels throughout, whereas the other increased in exponentially growing cells and declined in late-logarithmic phase. The analysis of highly purified plastid preparations demonstrated a plastidial localization of both proteins. Possible functional roles for maize EPSP synthase isozymes, with regard to the dual-pathway hypothesis and to the recent findings on defense-related aromatic biosynthesis in higher plants, are discussed.