Stressful life events are associated with striatal dopamine receptor availability in alcohol dependence.

Stress plays a key role in modulating addictive behavior and can cause relapse following periods of abstinence. Common effects of stress and alcohol on the dopaminergic system have been suggested, although the precise mechanisms are unclear. Here, we investigated 20 detoxified alcohol-dependent patients and 19 matched healthy controls and assessed striatal D2/D3 availability using [18F]-fallypride positron emission tomography and stressful life events. We found a strong association between striatal D2/D3 availability and stress in patients, but not in healthy controls. Interestingly, we found increased D2/D3 receptor availability in patients with higher stress levels. This mirrors complex interactions between stress and alcohol intake in animal studies and emphasizes the importance to investigate stress exposure in neurobiological studies of addiction. CLINICAL TRIAL REGISTRATION: NCT01679145.

Activity studies of sesquiterpene oxides and sulfides from the plant Hyptis suaveolens (Lamiaceae) and its repellency on Ixodes ricinus (Acari: Ixodidae).

Hyptis suaveolens (Lamiaceae), a plant traditionally used as a mosquito repellent, has been investigated for repellent properties against nymphs of the tick Ixodes ricinus. Essential oils and volatile compounds of fresh and dried leaves, from plants originating from Laos and Guinea-Bissau, were identified by GC-MS and tested in a tick repellency bioassay. All the essential oils were strongly repellent against the ticks, even though the main volatile constituents differed in their proportions of potentially tick repellent chemicals. (+)/(-)-sabinene were present in high amounts in all preparations, and dominated the emission from dry and fresh leaves together with 1,8-cineol and α-phellandrene. 1,8-Cineol and sabinene were major compounds in the essential oils from H. suaveolens from Laos. Main compounds in H. suaveolens from Guinea-Bissau were (-)-sabinene, limonene and terpinolene. Among the sesquiterpene hydrocarbons identified, α-humulene exhibited strong tick repellency (96.8 %). Structure activity studies of oxidation or sulfidation products of germacrene D, α-humulene and ß-caryophyllene, showed increased tick repellent activity: of mint sulfide (59.4 %), humulene-6,7-oxide (94.5 %) and caryophyllene-6,7-oxide (96.9 %). The substitution of oxygen with sulfur slightly lowered the repellency. The effects of the constituents in the oils can then be regarded as a trade off between the subsequently lower volatility of the sesquiterpene derivatives compared to the monoterpenes and may thus increase their potential usefulness as tick repellents.

Rapid metabolism fosters microbial survival in the deep, hot subseafloor biosphere.

A fourth of the global seabed sediment volume is buried at depths where temperatures exceed 80 °C, a previously proposed thermal barrier for life in the subsurface. Here, we demonstrate, utilizing an extensive suite of radiotracer experiments, the prevalence of active methanogenic and sulfate-reducing populations in deeply buried marine sediment from the Nankai Trough subduction zone, heated to extreme temperature (up to ~120 °C). The small microbial community subsisted with high potential cell-specific rates of energy metabolism, which approach the rates of active surface sediments and laboratory cultures. Our discovery is in stark contrast to the extremely low metabolic rates otherwise observed in the deep subseafloor. As cells appear to invest most of their energy to repair thermal cell damage in the hot sediment, they are forced to balance delicately between subsistence near the upper temperature limit for life and a rich supply of substrates and energy from thermally driven reactions of the sedimentary organic matter.

Glutamate as a spectroscopic marker of hippocampal structural plasticity is elevated in long-term euthymic bipolar patients on chronic lithium therapy and correlates inversely with diurnal cortisol.

While an excess of glucocorticoids is associated with hippocampal pathology in mood disorders, lithium exerts robust neuroprotective and neurotrophic effects. Here, 21 stably remitted bipolar I patients who had been on chronic lithium maintenance therapy, on average, for more than a decade, and 19 carefully matched healthy controls were studied using 3 T (1)H-magnetic resonance spectroscopy of left and right hippocampus. Salivary cortisol samples were obtained to assess activity of the hypothalamus-pituitary-adrenal system. Absolute concentrations of N-acetylaspartate (NAA), choline-containing compounds and total creatine were similar in euthymic bipolar patients and healthy controls. Hippocampal glutamate concentrations were significantly increased as an effect of patient status (patients>controls) and laterality (left hippocampus>right hippocampus). Hippocampal glutamate content (Glu) was strongly correlated with NAA. Across groups and within the patient group, diurnal saliva cortisol levels showed a significant inverse relationship with both Glu and NAA. Taken together, these results add to the concept of bipolar disorder as an illness involving disturbed hippocampal structural plasticity under the opposing influences of lithium and glucocorticoids.

Scaling behavior of the reorientation kinetics of block copolymers exposed to electric fields.

We have followed the reorientation kinetics of various block copolymer solutions exposed to an external electric DC field. The characteristic time constants follow a power law indicating that the reorientation is driven by a decrease in electrostatic energy. Moreover, the observed exponent suggests an activated process in line with the expectations for a nucleation and growth process. When properly scaled, the data collapse onto a single master curve spanning several orders of magnitude both in reduced time and in reduced energy. The power law dependence of the rate of reorientation derived from computer simulations based on dynamic density functional theory agrees well with the experimental observations. First experiments in AC electric fields at sufficiently high frequencies confirm the notion that the reorientation process is dominated by differences in the dielectric constants rather than by mobile ions.

Hippocampal glutamate concentration predicts cerebral theta oscillations during cognitive processing.

RATIONALE: Brain waves reflect collective behavior of neurons and provide insight into distributed network processing. Frontal and hippocampal theta oscillations (4-7 Hz) were linked to cognitive tasks and animal studies have suggested an involvement of glutamatergic neurotransmission in integrative frontal-hippocampal processing. Human evidence for such relationships is lacking. METHODS: Here, we studied the associations between glutamate concentrations in the hippocampal region, measured by a 3-T proton magnetic resonance spectroscopy (1H-MRS), and EEG theta activity during an auditory target detection paradigm. RESULTS: A robust relationship between hippocampal glutamate and frontal theta activity during stimulus processing was found. Moreover, frontal theta oscillations were related to response speed. CONCLUSION: The results suggest a functional coupling between the frontal cortex and hippocampal region during stimulus processing and support the idea of the hippocampus as a neural rhythm generator driven by glutamatergic neurotransmission. These preliminary data show, for the first time, a relationship between in vivo measured glutamate and basic cerebral information processing in humans.

Three dimensional image-based simulation of ultrasonic wave propagation in polycrystalline metal using phase-field modeling.

When modeling ultrasonic wave propagation in metals, it is important to introduce mesoscopic crystalline structures because the anisotropy of the crystal structure and the heterogeneity of grains disturb ultrasonic waves. In this paper, a three-dimensional (3D) polycrystalline structure generated by multiphase-field modeling was introduced to ultrasonic simulation for nondestructive testing. 3D finite-element simulations of ultrasonic waves were validated and compared with visualization results obtained from laser Doppler vibrometer measurements. The simulation results and measurements showed good agreement with respect to the velocity and front shape of the pressure wave, as well as multiple scattering due to grains. This paper discussed the applicability of a transversely isotropic approach to ultrasonic wave propagation in a polycrystalline metal with columnar structures.

c-fos expression induces bone tumors in transgenic mice.

The proto-oncogene c-fos has been isolated as the cellular homolog of the v-fos gene found in the osteosarcoma inducing FBR- and FBJ-murine sarcoma viruses (MSV). Expression of the c-fos gene in transgenic mice leads to the development of bone lesions of which about half progress to bone tumors mainly chondrosarcomas. The tumors display a strong preference for males and have a latency with a mean of 9.5 months. However, also mice without visible lesions develop bone tumors with the same sex preference and latency. These consequences of c-fos expression are independent of the chosen promoter but dependent on a replacement of 3' noncoding sequences of c-fos by a long terminal repeat (LTR) of the FBJ-MSV virus.

Altered Pax gene expression in murine notochord mutants: the notochord is required to initiate and maintain ventral identity in the somite.

We have characterised the patterning capacity of the notochord on the somite using the murine Pax-1 gene as a ventral, and Pax-3 as a dorsal molecular marker. As model systems we chose the four mouse notochord mutants Brachyury curtailed (Tc), Danforth's short tail (Sd), Pintail (Pt) and truncate (tc). Their notochord either is initially absent or progressively degenerates. The use of these mutants enabled us to compare the effect of graded notochord deficiencies. All four mutants show premature termination of the vertebral column. This phenotype can be traced back to an impaired dorsoventral specification of the somites. In tc/tc and Tc/+ embryos the notochord in the affected regions is missing from the beginning. Consequently, Pax-1 is never activated, and Pax-3 remains to be expressed in the entire somite. In contrast, in Sd and Pt embryos the notochord secondarily degenerates. At the end of the prevertebral column Pax-1 expression is lost, while the Pax-3 signal occupies the former Pax-1 expressing zone. The altered pax gene expression in the notochord mutants suggests that the notochord is required for two processes in the dorsoventral patterning of the somite: first the induction of ventral structures, and second the maintenance of the ventral fate.

Expression of the novel murine homeobox gene Sax-1 in the developing nervous system.

We have isolated the novel murine Sax-1 gene, a member of the NK-1 class of homeobox genes, and report its expression pattern in the developing central nervous system (CNS) in comparison to two other homeobox genes, Evx-1 and Pax-6. Sax-1 was found to be transiently expressed in the developing posterior CNS. First seen in the ectoderm lateral to the primitive streak, the signal later encompassed the neural plate. Posteriorly, the expression domain overlapped with the Evx-1 expression in the streak, while anteriorly it was delimited by the Pax-6 signal in the neural tube. This early phase starting at day 9.5 pc, Sax-1 was expressed in distinct areas of spinal cord, hindbrain and forebrain. Particularly strong signals were detected in rhombomere 1 and in the pretectum. In these areas, subsets of neurons may be marked and specified. In addition to the normal pattern of Sax-1 during development, the expression in different mouse mutants was analysed. In Brachyury curtailed homozygotes, the expression of Sax-1 was found to be reduced during neurulation and even lost at day 9.0 pc. Ventral shift and finally loss of the signal in the ventral spinal cord was observed in Danforth's short tail homozygotes.

Lbx1 marks a subset of interneurons in chick hindbrain and spinal cord.

The putative transcription factor Lbx1 is expressed in the mantle zone of the hindbrain and spinal cord caudal to rhombomere 1, in a specific domain of the alar plate. The Lbx1 domain overlaps with the expression domains for Tlx3 and partially with the domains for Pax2/Lim1. The ventral border of the Lbx1 domain coincides with the ventral border of the dorsalmost Serrate1 stripe in the ventricular zone. The latter borders the intermediate stripe of both Delta and Lunatic fringe expression. The Lbx1 domain contains differentiated interneurons that project into the lateral longitudinal fasciculus.

Laparoscopically assisted peritoneal shunt insertion in hydrocephalus: a prospective controlled study.

BACKGROUND: Shunting of cerebrospinal fluid to the peritoneal cavity is standard therapy for the management of hydrocephalus. Common problems, however, are infection and shunt malfunction, which frequently is related to the peritoneal end of the catheter. Laparoscopic revision of distal shunt malfunction has become popular, but endoscopic techniques for primary placement of the peritoneal catheter are not performed often. This study aimed to compare laparoscopically assisted peritoneal catheter placement with the conventional minilaparotomy technique. METHODS: In the prospective arm of the study, 50 patients underwent laparoscopic distal shunt placement. The findings were compared with those for another group of 50 patients who underwent surgery by the standard transrectal or pararectal approach. Both groups were similar with regard to age, gender, American Society of Anesthesiologists (ASA) scores, indications for surgery, and frequency of previous abdominal operations. RESULTS: No intraoperative complications occurred. The mean time for surgery was 59 min in the laparoscopically assisted treatment group and 49 min in the standard group. During follow-up assessment, 3 instances of distal catheter malfunction or infection (2 malfunctions and 1 infection) occurred in the endoscopic group, and 12 instances (6 malfunctions and 6 infections) occurred in the control group. This difference was statistically significant. CONCLUSIONS: The findings from this prospective controlled study indicate that the risk for long-term complications attributable to distal shunt malfunction is reduced when laparoscopic techniques are used to place the peritoneal end of the shunt catheter.

Enhancing biosensor performance using multienzyme systems.

Enhancing the performance of biosensors, in terms of increasing the range of analytes that may be detected, and the sensitivity and specificity of the detection event, would improve the prospects for commercializing this technology. Coupling the catalytic activities of several enzymes is one approach being used to address these issues. Sequences of enzymes, where ligand binding triggers the activation of enzymes, or where biocatalytic pre-concentration of intermediates permits augmentation of the signal, may be used. In addition, enzymatic recycling of the analyte can be used to increase the sensitivity by several orders of magnitude.

The UVA light used during the fluorescence microscopy assay affects the level of intracellular calcium being measured in experiments with electric-field exposure.

In the present paper, the induction of calcium signals in neuroblastoma cells, cells of T-cell leukemia, and osteogenic sarcoma cells were investigated in relation to the UVA irradiation used in fluorescence microscopy. Methods were developed to measure both the mean UVA irradiance and the intensity profile in the UVA-illuminated area of the microscope. This allowed us to calculate the applied UVA radiant exposure of the cells during each experiment. This investigation was undertaken because of the conflicting results in the literature on the effects of electromagnetic fields on the signals of the calcium-sensitive fluorescence probe FURA-2 in lymphocytes. Taking into account that each group used a different system with different optics and lamps, these conflicting results are now at least partially understandable. Our measurements indicate that in a typical experiment with FURA-2 the cells were irradiated with up to 776 kJ m(-2) during 25 min of exposure to UVA light. This causes changes in intracellular free Ca(2+) concentrations ([Ca(2+)](i)). Designating cells in which the [Ca(2+)](i) was distinctly increased during the experiment as "responding", we found Hill-type dependences on the irradiance. Jurkat cells showed a 50% response even at 10 kJ m(-2) and osteosarcoma cells at about 60 kJ m(-2), whereas neuroblastoma cells even at the maximum possible dose responded only minimally. In the case of neuroblastoma cells, we found a dependence of this effect on the CO(2) partial pressure during the preincubation. An electrical treatment with an a.c. field (5 kHz sinusoidal, amplitude modulation 16 Hz 100%, 800 V m(-1), 5 min) had a significant effect on intracellular calcium in neuroblastoma cells only in the case of cells that were not pretreated with CO(2) with high fluences of UVA irradiation. In conclusion, these results indicate that the possibility of UVA artifacts must be considered in all experiments using fluorescence microscopy. Furthermore, our results lead to the hypothesis that oxidative stress could be the link between UVA and electric-field effects.

Second generation biosensors.

Enzyme-membrane electrodes using glucose oxidase in combination with peroxide detection dominate in the field of laboratory analyzers for diluted samples. Using the same indication principle, extremely fast responding glucose sensors have been fabricated by covering thin metal electrodes with a porous enzyme layer. In the second generation auxiliary enzymes and/or co-reactants are coimmobilized with the analyte converting enzyme in order to improve the analytical quality and to simplify the performance. Following this line oxidizable interferences are suppressed by using a glucose oxidase/peroxidase complex which communicates with the electrode at a low working potential. Furthermore, fluctuations of pH or buffer capacity are ineffective when using a glucose oxidase/peroxidase layer covered fluoride FET in the potentiometric glucose determination. Enzymatic recycling of the analyte and/or accumulation of intermediates increase the sensitivity by several orders of magnitude. Inclusion of NAD bound to PEG in the glucose dehydrogenase layer allows a reagentless glucose measurement.

Vasopressin and oxytocin content of human fetal brain at different stages of gestation.

Pituitaries from 38 human fetuses and 5 microdissected hypothalamic areas from each of 14 fetuses were assayed for vasopressin and oxytocin. The fetuses were between 15 and 34 weeks of gestation. Vasopressin and oxytocin were found in all the pituitary glands and all the hypothalamic areas studied. A trend of increasing content of both hormones was found in the pituitary gland towards the end of gestation with a marked increase in vasopressin/oxytocin ratio. The human fetus synthesizes oxytocin and vasopressin at least from 15 weeks of gestation and the presence of both hormones in the hypothalamus is not limited to the supraoptic and paraventricular nuclei.

Enzyme sensor-FIA-system for on-line monitoring of glucose, lactate and glutamine in animal cell cultures.

Enzyme sensors for glucose, lactate and glutamine were connected via flow-injection analysis (FIA) devices to two different bioprocesses. They were used for on-line process control of perfused bioreactor systems containing mammalian cell lines producing a monoclonal antibody and recombinant interleukin-2. The biosensor system gives direct access to important process data which can be used as control parameters for long term cell cultivation systems.

Canadian guidelines for economic evaluation of pharmaceuticals. Canadian Collaborative Workshop for Pharmacoeconomics.

In 1994, Canada became the second country to release national guidelines for the economic evaluation of pharmaceuticals. The guidelines were developed over a period of 18 months through an elaborate process of broad consultation with a wide variety of relevant stakeholders. The intent of the guidelines is to provide guidance to doers and users of studies, by laying out the general 'state of the art' regarding methods, and by providing specific methodological advice on many matters. The aim is to improve the scientific quality and integrity of studies, and to enhance consistency and comparability across studies. This article presents the Canadian guidelines, both in summary and in detail. Because the techniques of economic evaluation are widely applicable beyond pharmaceuticals, the guidelines will be of interest to researchers and decision makers in all fields of healthcare. Because the methods are not country specific, the guidelines will be of interest to those in other countries as well as in Canada.

Preparation of solid phantoms with defined scattering and absorption properties for optical tomography.

We have developed diffusely scattering solid phantoms with optical (scattering) properties amenable to theoretical calculations. Monodisperse quartz glass spheres were used as scatterers embedded in polyester resin. An infrared dye was added to simulate absorption by biological tissue. Solid phantoms were tested for their macroscopic homogeneity. Several phantoms were built with well-defined spatial variations in their transport scattering and absorption coefficients to be used for optical tomography. Scattering, transport scattering, and absorption coefficients of solid, homogeneous phantoms and of aqueous suspensions of monodisperse quartz glass spheres were derived from measurements of time-integrated collimated transmittance and time-resolved diffuse transmittance. For aqueous suspensions of monodisperse quartz glass spheres at known number density scattering and transport scattering coefficients calculated by Mie theory are in quantitative agreement with experimentally derived values. In addition, diffuse reflectance and diffuse transmittance of aqueous suspensions at various number densities were measured and found to be in excellent agreement with results of Monte Carlo calculations using theoretical values for the scattering coefficients and anisotropy parameters.