The effect of thyroxine and triiodothyronine on fatigue, depression, and memory in a hypothyroid population

Background. This study was planned at a time when important questions were being raised about the adequacy of using one hormone to treat hypothyroidism instead of two. Specifically, this trial aimed to replicate prior findings which suggested that substituting 12.5 μg of liothyronine for 50 μg of levothyroxine might improve mood, cognition, and physical symptoms. Additionally, this trial aimed to extend findings to fatigue. ^ Methods. A randomized, double-blind, two-period, crossover design was used. Hypothyroid patients stabilized on levothyroxine were invited to participate. Thirty subjects were recruited and randomized. Sequence one received their standard levothyroxine dose in one capsule and placebo in another during the first six weeks. Sequence two received their usual levothyroxine dose minus 50 μg in one capsule and 10 μg of liothyronine in another. At the end of the first six week period, subjects were crossed over. T tests were used to assess carry-over and treatment effects. ^ Results. Twenty-seven subjects completed the trial. The majority of completers had an autoimmune etiology. Mean baseline levothyroxine dose was 121 μg/d (±26.0). Subjects reported small increases in fatigue as measured by the Piper Fatigue Scale (0.9, p = 0.09) and in symptoms of depression measured by the Beck Depression Inventory-II (2.3, p = 0.16) as well as the General Health Questionnaire-30 (4.7, p = 0.14) while treated with substitution treatment. However, none of these differences was statistically significant. Measures of working memory were essentially unchanged between treatments. Thyroid stimulating hormone was about twice as high during substitution treatment (p = 0.16). Free thyroxine index was reduced by 0.7 (p < 0.001), and total serum thyroxine was reduced by 3.0 (p < 0.001) while serum triiodothyronine was increased by 20.5 (p < 0.001) on substitution treatment. ^ Conclusions. Substituting an equivalent amount of liothyronine for a portion of levothyroxine in patients with hypothyroidism does not decrease fatigue, symptoms of depression, or improve working memory. However, due to changes in serum hormone levels and small increments in fatigue and depression symptoms on substitution treatment, a question was raised about the role of T3 in the serum. ^

Alpha C protein of group B Streptococcus as a potential vaccine candidate

Group B Streptococcus (GBS) is a leading cause of life-threatening infection in neonates and young infants, pregnant women, and non-pregnant adults with underlying medical conditions. Immunization has theoretical potential to prevent significant morbidity and mortality from GBS disease. Alpha C protein (α C), found in 70% of non-type III capsule polysaccharide group B Streptococcus, elicits antibodies protective against α C-expressing strains in experimental animals and is an appealing carrier for a GBS conjugate vaccine. We determined whether natural exposure to α C elicits antibodies in women and if high maternal α C-specific serum antibody at delivery is associated with protection against neonatal disease. An ELISA was designed to measure α C-specific IgM and IgG in human sera. A case-control design (1:3 ratio) was used to match α C-expressing GBS colonized and non-colonized women by age and compare quantified serum α C-specific IgM and IgG. Sera also were analyzed from bacteremic neonates and their mothers and from women with invasive GBS disease. Antibody concentrations were compared using t-tests on log-transformed data. Geometric mean concentrations of α C-specific IgM and IgG were similar in sera from 58 α C strain colonized and 174 age-matched non-colonized women (IgG 245 and 313 ng/ml; IgM 257 and 229 ng/ml, respectively). Delivery sera from mothers of 42 neonates with GBS α C sepsis had similar concentrations of α C-specific IgM (245 ng/ml) and IgG (371 ng/ml), but acute sera from 13 women with invasive α C-expressing GBS infection had significantly higher concentrations (IgM 383 and IgG 476 ng/ml [p=0.036 and 0.038, respectively]). Convalescent sera from 5 of these women 16-49 days later had high α C-specific IgM and IgG concentrations (1355 and 4173 ng/ml, respectively). In vitro killing of α C-expressing GBS correlated with total α C-specific antibody concentration. Invasive disease but not colonization elicits α C-specific IgM and IgG in adults. Whether α C-specific IgG induced by vaccine would protect against disease in neonates merits further investigation. ^

Functional Characterization of AtxA, the Bacillus anthracis Virulence Regulator

Coordinated expression of virulence genes in Bacillus anthracis occurs via a multi-faceted signal transduction pathway that is dependent upon the AtxA protein. Intricate control of atxA gene transcription and AtxA protein function have become apparent from studies of AtxA-induced synthesis of the anthrax toxin proteins and the poly-D-glutamic acid capsule, two factors with important roles in B. anthracis pathogenesis. The amino-terminal region of the AtxA protein contains winged-helix (WH) and helix-turn-helix (HTH) motifs, structural features associated with DNA-binding. Using filter binding assays, I determined that AtxA interacted non-specifically at a low nanomolar affinity with a target promoter (Plef) and AtxA-independent promoters. AtxA also contains motifs associated with phosphoenolpyruvate: sugar phosphotransferase system (PTS) regulation. These PTS-regulated domains, PRD1 and PRD2, are within the central amino acid sequence. Specific histidines in the PRDs serve as sites of phosphorylation (H199 and H379). Phosphorylation of H199 increases AtxA activity; whereas, H379 phosphorylation decreases AtxA function. For my dissertation, I hypothesized that AtxA binds target promoters to activate transcription and that DNA-binding activity is regulated via structural changes within the PRDs and a carboxy-terminal EIIB-like motif that are induced by phosphorylation and ligand binding. I determined that AtxA has one large protease-inaccessible domain containing the PRDs and the carboxy-terminal end of the protein. These results suggest that AtxA has a domain that is distinct from the putative DNA-binding region of the protein. My data indicate that AtxA activity is associated with AtxA multimerization. Oligomeric AtxA was detected when co-affinity purification, non-denaturing gel electrophoresis, and bis(maleimido)hexane (BMH) cross-linking techniques were employed. I exploited the specificity of BMH for cysteine residues to show that AtxA was cross-linked at C402, implicating the carboxy-terminal EIIB-like region in protein-protein interactions. In addition, higher amounts of the cross-linked dimeric form of AtxA were observed when cells were cultured in conditions that promote toxin gene expression. Based on the results, I propose that AtxA multimerization requires the EIIB-like motif and multimerization of AtxA positively impacts function. I investigated the role of the PTS in the function of AtxA and the impact of phosphomimetic residues on AtxA multimerization. B. anthracis Enzyme I (EI) and HPr did not facilitate phosphorylation of AtxA in vitro. Moreover, markerless deletion of ptsHI in B. anthracis did not perturb AtxA function. Taken together, these results suggest that proteins other than the PTS phosphorylate AtxA. Point mutations mimicking phosphohistidine (H to D) and non-phosphorylated histidine (H to A) were tested for an impact on AtxA activity and multimerization. AtxA H199D, AtxA H199A, and AtxA H379A displayed multimerization phenotypes similar to that of the native protein, whereas AtxA H379D was not susceptible to BMH cross-linking or co-affinity purification with AtxA-His. These data suggest that phosphorylation of H379 may decrease AtxA activity by preventing AtxA multimerization. Overall, my data support the following model of AtxA function. AtxA binds to target gene promoters in an oligomeric state. AtxA activity is increased in response to the host-related signal bicarbonate/CO2 because this signal enhances AtxA multimerization. In contrast, AtxA activity is decreased by phosphorylation at H379 because multimerization is inhibited. Future studies will address the interplay between bicarbonate/CO2 signaling and phosphorylation on AtxA function.

Control of the master virulence regulatory gene atxA in Bacillus anthracis

Transcription of the Bacillus anthracis structural genes for the anthrax toxin proteins and biosynthetic operon for capsule are positively regulated by AtxA, a transcription regulator with unique properties. Consistent with the role of atxA in virulence factor expression, a B. anthracis atxA-null mutant is avirulent in a murine model for anthrax. In batch culture, multiple signals impact atxA transcript levels, and the timing and steady state level of atxA expression is critical for optimal toxin and capsule synthesis. Despite the apparent complex control of atxA transcription, only one trans-acting protein, the transition state regulator AbrB, has been demonstrated to directly interact with the atxA promoter. The AbrB-binding site has been described, but additional cis-acting control sequences have not been defined. Using transcriptional lacZ fusions, electrophoretic mobility shift assays, and Western blot analysis, the cis-acting elements and trans-acting factors involved in regulation of atxA in B. anthracis strains containing either both virulence plasmids, pXO1 and pXO2, or only one plasmid, pXO1, were studied. This work demonstrates that atxA transcription from the major start site P1 is dependent upon a consensus sequence for the housekeeping sigma factor SigA, and an A+T-rich upstream element (UP-element) for RNA polymerase (RNAP). In addition, the data show that a trans-acting protein(s) other than AbrB negatively impacts atxA transcription when it binds specifically to a 9-bp palindrome within atxA promoter sequences located downstream of P1. Mutation of the palindrome prevents binding of the trans-acting protein(s) and results in a corresponding increase in AtxA and anthrax toxin production in a strain- and culture-dependent manner. The identity of the trans-acting repressor protein(s) remains elusive; however, phenotypes associated with mutation of the repressor binding site have revealed that the trans-acting repressor protein(s) indirectly controls B. anthracis development. Mutation of the repressor binding site results in misregulation and overexpression of AtxA in conditions conducive for development, leading to a marked sporulation defect that is both atxA- and pXO2-61-dependent. pXO2-61 is homologous to the sensor domain of sporulation sensor histidine kinases and is proposed to titrate an activating signal away from the sporulation phosphorelay when overexpressed by AtxA. These results indicate that AtxA is not only a master virulence regulator, but also a modulator of proper B. anthracis development. Also demonstrated in this work is the impact of the developmental regulators AbrB, Spo0A, and SigH on atxA expression and anthrax toxin production in a genetically incomplete (pXO1+, pXO2-) and genetically complete (pXO1+, pXO2+) strain background. AtxA and anthrax toxin production resulting from deletion of the developmental regulators are strain-dependent suggesting that factors on pXO2 are involved in control of atxA. The only developmental deletion mutant that resulted in a prominent and consistent strain-independent increase in AtxA protein levels was an abrB-null mutant. As a result of increased AtxA levels, there is early and increased production of anthrax toxins in an abrB-null mutant. In addition, the abrB-null mutant exhibited an increase in virulence in a murine model for anthrax. In contrast, virulence of the atxA promoter mutant was unaffected in a murine model for anthrax despite the production of 5-fold more AtxA than the abrB-null mutant. These results imply that AtxA is not the only factor impacting pathogenesis in an abrB-null mutant. Overall, this work highlights the complex regulatory network that governs expression of atxA and provides an additional role for AtxA in B. anthracis development.

Are anatomic brain locations important in determining the clinical outcomes in acute ischemic stroke? --- An exploratory analysis of the NINDS Stroke Trial data

Background and purpose. Brain lesions in acute ischemic stroke measured by imaging tools provide important clinical information for diagnosis and final infarct volume has been considered as a potential surrogate marker for clinical outcomes. Strong correlations have been found between lesion volume and clinical outcomes in the NINDS t-PA Stroke Trial but little has been published about lesion location and clinical outcomes. Studies of the National Institute of Neurological Disorders and Stroke (NINDS) t-PA Stroke Trial data found the direction of the t-PA treatment effect on a decrease in CT lesion volume was consistent with the observed clinical effects at 3 months, but measure of t-PA treatment benefits using CT lesion volumes showed a diminished statistical significance, as compared to using clinical scales. ^ Methods. We used the global test to evaluate the hypothesis that lesion locations were strongly associated with clinical outcomes within each treatment group at 3 months after stroke. The anatomic locations of CT scans were used for analysis. We also assessed the effect of t-PA on lesion location using a global statistical test. ^ Results. In the t-PA group, patients with frontal lesions had larger infarct volumes and worse NIHSS score at 3 months after stroke. The clinical status of patients with frontal lesions in t-PA group was less likely to be affected by lesion volume, as compared to those who had no frontal lesions in at 3 months. For patients within the placebo group, both brain stem and internal capsule locations were significantly associated with a lower odd of having favorable outcomes at 3 months. Using a global test we could not detect a significant effect of t-PA treatment on lesion location although differences between two treatment groups in the proportion of lesion findings in each location were found. ^ Conclusions. Frontal, brain stem, and internal capsule locations were significantly related to clinical status at 3 months after stroke onset. We detect no significant t-PA effect on all 9 locations although proportion of lesion findings in differed among locations between the two treatment groups.^

Contents of Ag, Bi, In, and Tl in suspended and aerosol matter at Station DM8-579 in the Peru Basin, Southeast Pacific

A new electrothermal atomizer for use in direct determination of Ag, Bi, In, and Tl in marine, riverine, and aeolian particulate matter on membrane filters is described. A sample capsule and atomization cell are heated separately. That is why it is possible to separate and optimize decomposition of a sample, vaporization of elements and atomization of their vapors. Noise reduction and design, which localizes the vapors in a light absorption zone, decrease detection limits of these four elements by factor of at least 3 to 10. Some analytical results are given.

Pollen profiles from three former lakes in the Lake Garda area

1. Late glacial and postglacial sediments from three former lakes in the Lake Garda area (Southern Alps) were investigated. 2. The pollen diagram from Bondone (1550 m) shows an older phase rich in NAP. A younger one corresponds with the Younger Dryas time according to two radiocarbon determinations. In the Preboreal no climatic deterioration could be found. 3. At first plants, which are nowadays typical for snow-ground, pioneer and dwarf shrub associations, immigrated into the surroundings of Bondone. In Alleröd times larch and pine appeared as the first trees. At the beginning of the Preboreal dense forest existed in that region. During the Alleröd timber line was at about 1500 m. 4. In the pollen diagrams from Saltarino (194 m) and Fiavè (654 m) an oldest period rich in NAP is followed by two stadial and two interstadial phases. Tree birches and larches immigrated during the oldest interstadial phase. 5. In the case of Saltarino and Fiavè only a preliminary dating could be made. A correlation seems to be possible with diagrams published by Zoller as well as with the diagram of Bondone. Discrepances in dating, which arise then, are discussed. According to the two possibilities of dating the youngest stadial is synchronous either with the so-called Piottino stadial or the Younger Dryas time. Consequently the oldest interstadial phase of Saltarino corresponds either with the Bölling or with a pre-Bölling interstadial. The last possibility seems to be more probable. 6. In the southern part of the Lake Garda area reforestation was preceded by a long shrub phase mainly with Juniperus. At about 650 m there was a period with Pinus mugo and only with a small amount of Juniperus before reforestation. A phase with Betula nana well known from areas north of the Alps could nowhere be found. 7. In the area under study larch appeared as the first tree. Lateron it has been the most important constituent of the forests near timber line. Birch, which plays an important role as a pioneer tree in Denmark - for instance at the transition of the pollen zones III/IV - as well as in Southern Germany during Bölling time, was of less importance at the southern border of the Alps. In that area the spreading of Pinus occurred very early causing dense forests. 8. During the last stadial phase (probably Younger Dryas time) dense forests with Pinus and Larix existed at 650 m. In the lower part of the Lake Garda area, however, both thermophilous trees as Quercus and herbs frequently occurred. This leads to the conclusion that during this time tree growth was limited by dryness in lower altitudes of the border of the Southern Alps. Pinus and Juniperus, however, do not show higher values in this period, a fact which cannot yet be explained. 9. A list of plants, which were found in the sediments, is compiled. Helodium lanatum, Dictamnus albus, Mercurialis cf. ovata, Buxus, Cerinthe cf. minor, Onosma, Anthericum and Asphodelus albus are findings, which are of special interest for the history of the flora of that region.

Pollen and macroremains from four profiles from site Samerberg 1

Pollen and macrofossil analysis of lake sediments revealed the complete development of vegetation from Riss late-glacial to early Würm glacial times at Samerberg (12°12' E, 47°45' N, 600 m a.s.l) on the northern border of the Alps. The pollen bearing sediments overlie three stratigraphic units, at the base a ground-moraine, then a 13 m thick layer of pollen free silt and clay, and then a younger moraine; all the sediments including the pollen bearing sediments, lie below the Würm moraine. The lake, which had developed in an older glacial basin, became extinct, when the ice of the river Inn glacier filled its basin during Würm full-glacial time at the latest. One interglacial, three interstadials, and the interdigitating treeless periods were identified at Samerberg. Whereas the cold periods cannot be distinguished from one another pollenanalytically, the interglacial and the two older interstadials have distinctive characteristics. A shrub phase with Juniperus initiated reforestation and was followed by a pine phase during the interglacial and each of the three interstadials. The further development of the interglacial vegetation proceeded with a phase when deciduous trees (mainly Quercus, oak) and hazel (Corylus) dominated, though spruce (Picea) was present at the same time in the area. A phase with abundant yew (Taxus) led to an apparently long lasting period with dominant spruce and fir (Abies) accompanied by some hornbeam (Carpinus). The vegetational development shows the main characteristics of the Riss/Würm interglacial, though certain differences in the vegetational development in the northern alpine foreland are obvious. These differences may result from the existence of an altitudinal zonation of the vegetation in the vicinity of the site and are the expression of its position at the border of the Alps. A greater age (e.g. the Holsteinian) can be excluded by reason of the vegetational development, and is also not indicated at first sight from the geological and stratigraphical data of the site. Characteristic of the Riss/Würm vegetational development in southern Germany - at least in the region between Lake Starnberg/Samerberg/Salzach - is the conspicuous yew phase. According to absolute pollen counts, yew not only displaced the deciduous species, but also displaced spruce preferentially, thus indicating climatic conditions less favourable for spruce, caused by mild winters (Ilex spreading!) and by short-term low precipitation, indicated by the reduced sedimentation rate. The oldest interstadials is bipartite, as due to the climatic deterioration the early vegetational development, culminating in a spruce phase, had been interrupted by another expansion of pine. A younger spruce-dominated period with fir and perhaps also with hornbeam and beech (Fagus) followed. An identical climatic development has been reported from other European sites with long pollen sequences (see chapter 6.7). However, different tree species are found in the same time intervals in Middle Europe during Early Würm times. Sediments of the last interglacial (Eem or Riss/Würm) have been found in all cases below the sediments of the bipartite interstadial, and in addition one more interstadial occurs in the overlying sediments. This proves that Eem and Riss/Würm of the north-european plain resp. of the alpine foreland are contemporaneous interglacials although this has been questioned by some authors. The climax vegetation of the second interstadial was a spruce forest without fir and without more demanding deciduous tree species. The vegetational development of the third interstadial is recorded fragmentary only. But it has been established that a spruce forest was present. The oldest interstadial must correspond to the danish Brørup interstadial as it is expressed in northern Germany, the second one to the Odderade interstadial. A third Early Würm interstadial, preserved fragmentarily at Samerberg, is known from other sites. The dutch Amersfoort interstadial most likely is the equivalent to the older part of the bipartite danish Brørup interstadial.

Stochastic homogenization of the laser intensity to improve the irradiation uniformity of capsules directly driven by thousands laser beams

Illumination uniformity of a spherical capsule directly driven by laser beams has been assessed numerically. Laser facilities characterized by ND = 12, 20, 24, 32, 48 and 60 directions of irradiation with associated a single laser beam or a bundle of NB laser beams have been considered. The laser beam intensity profile is assumed super-Gaussian and the calculations take into account beam imperfections as power imbalance and pointing errors. The optimum laser intensity profile, which minimizes the root-mean-square deviation of the capsule illumination, depends on the values of the beam imperfections. Assuming that the NB beams are statistically independents is found that they provide a stochastic homogenization of the laser intensity associated to the whole bundle, reducing the errors associated to the whole bundle by the factor  , which in turn improves the illumination uniformity of the capsule. Moreover, it is found that the uniformity of the irradiation is almost the same for all facilities and only depends on the total number of laser beams Ntot = ND × NB.

Systematic Analysis of Direct-Drive Baseline Designs for Shock-Ignition with the Laser Megajoule

Direct-drive inertial confinement thermonuclear fusion consists in illuminating a shell of cryogenic Deuterium and Tritium (DT) mixture with many intense beams of laser light. Capsule is composed of DT gassurrounded by cryogenic DT as combustible fuel. Basic rules are used to define shell geometry from aspect ratio, fuel mass and layers densities. We define baseline designs using two aspect ratio (A=3 and A=5) who complete HiPER baseline design (A=7.7). Aspect ratio is defined as the ratio of ice DT shell inner radius over DT shell thickness. Low aspect ratio improves hydrodynamics stabilities of imploding shell. Laser impulsion shape and ablator thickness are initially defined by using Lindl (1995) pressure ablation and mass ablation formulae for direct-drive using CH layer as ablator. In flight adiabat parameter is close to one during implosion. Velocitie simplosions chosen are between 260 km/s and 365 km/s. More than thousand calculations are realized for each aspect ratio in order to optimize the laser pulse shape. Calculations are performed using the one-dimensional version of the Lagrangian radiation hydrodynamics FCI2. We choose implosion velocities for each initial aspect ratio, and we compute scaled-target family curves for each one to find self-ignition threshold. Then, we pick points on each curves that potentially product high thermonuclear gain and compute shock ignition in the context of Laser MegaJoule. This systematic analyze reveals many working points which complete previous studies ´allowing to highlight baseline designs, according to laser intensity and energy, combustible mass and initial aspect ratio to be relevant for Laser MegaJoule.

Symmetry Issues in Directly Irradiated Targets

In direct drive Inertial Confinement Fusion (ICF), the typical laser beam to laser beam angle is around 30o. This fact makes the study of the irradiation symmetry agenuine 3D problem. In this paper we use the three dimensional version of the MULTI hydrocode to assess the symmetry of such ICF implosions. More specifically, we study a shock-ignition proposal for the Laser-M´egajoule facility (LMJ) in which two of the equatorial beam cones are used to implode and pre compress a spherical capsule (the “reference” capsule of HiPER project) made of 0.59 mg of pure Deuterium-Tritium mixture. The symmetry of this scheme is analysed and optimized to get a design inside the operating limits of LMJ. The studied configuration has been found essentially axial-symmetric, so that the use of 2D hydrocodes would be appropriate for this specific situation.

Ignition conditions for inertial confinement fusion targets with a nuclear spin-polarized DT fuel

The nuclear fusion cross-section is modified when the spins of the interacting nuclei are polarized. In the case of deuterium?tritium it has been theoretically predicted that the nuclear fusion cross-section could be increased by a factor d = 1.5 if all the nuclei were polarized. In inertial confinement fusion this would result in a modification of the required ignition conditions. Using numerical simulations it is found that the required hot-spot temperature and areal density can both be reduced by about 15% for a fully polarized nuclear fuel. Moreover, numerical simulations of a directly driven capsule show that the required laser power and energy to achieve a high gain scale as d-0.6 and d-0.4 respectively, while the maximum achievable energy gain scales as d0.9.