Miracle-dz/UniFAR-FTUs huggingface dataset:
Drosophila CG8422 encodes a functional diuretic hormone receptor

SUMMARY Diuretic hormone 44 (DH) is a bioactive neuropeptide that mediates osmotic balance in a wide variety of insects through increases in cAMP. It is structurally similar to mammalian corticotrophin releasing factor (CRF) peptides. In the moth Manduca and the cricket Acheta, functional studies have shown that its cognate receptor (DH-R) is related to the mammalian CRF receptor. The Drosophila genome contains two genes (CG8422 and CG12370) orthologous to Manduca and Acheta DH-Rs. Here, we present multiple lines of evidence to support the hypothesis that the orphan CG8422 G-protein-coupled receptor is a functional DH-R. When expressed in mammalian cells, CG8422 conferred selective sensitivity to DH, as indicated by translocation of aβ -arrestin-2-GFP reporter from the cytoplasm to the cell membrane. Consistent with its in vivo activities in other insects, DH activation of CG8422 elicited increases in a cAMP reporter system (CRE-luciferase), with an EC50 of 1.7 nmol l-1. CG8422 activation by DH also led to increases in intracellular calcium but at substantially higher doses (EC50 ∼300 nmol l-1). By microarray analysis, the CG8422 transcript was detectable in Drosophila head mRNA of different genotypes and under different environmental conditions. The identification of a Drosophila receptor for the DH neuropeptide provides a basis for genetic analysis of this critical factor's roles in maintaining physiological homeostasis.
Lentiviral vector-mediated transduction of neural progenitor cells before implantation into injured spinal cord and brain to detect their migration, deliver neurotrophic factors and repair tissue.

PURPOSE Stem cells represent an attractive source for cell replacement therapy in neurological disorders due to their self-renewal and multi-potency. Genetic manipulation of these cells may allow controlled release of therapeutic proteins, suppress immune rejection, or produce essential neurotransmitters. Furthermore, when the expression cassette is incorporated into the host genome ex vivo, this technique also may be used as a method to trace cells following implantation into tissues of interest. METHODS We explored the possibility of transducing pluripotent fetal rat cortical neural progenitor cells (NPCs) using lentiviral vectors encoding the green fluorescent protein (GFP) or neurotrophic factors (BDNF, CNTF, D15A, GDNF, MNT and NT-3) prior to implanting these cells into the contused spinal cord or injured brain. RESULTS In vitro staining of these cells for neural markers (such as nestin, GFAP, Tuj-1 and RIP) after transduction did not reveal any significant difference from non-transduced cells. When they were transduced with a vector encoding CNTF or MNT, however, cells started expressing GFAP in vitro. Following delayed (1 week) implantation into the lesion site of the moderately contused rat spinal cord or the injured brain, transduced cells survived up to 12 weeks post-implantation (the longest time point examined) and most of the NPCs turned into an astrocytic phenotype in the spinal cord, but not in the brain. Nestin and GFP positive cells were detected in the brain, but not in the spinal cord lesion. GFP positive cells in the spinal cord migrated rostrally and caudally from the lesion/implantation site towards uninjured tissue. CONCLUSIONS Novel findings in this study are the longterm expression of a foreign gene in NPCs using lentiviral vectors; this enabled tracking of the cells following implantation. This expression also allowed the observation that NPCs developed differently in the injured spinal cord and brain. Moreover, NPCs could be transduced to overexpress neurotrophic factors. In sum, NPC survival and the long-term transgene expression that allows easy tracking of migrating cells make NPCs promising candidates for implantation into the injured spinal cord or brain and a potentially powerful tool to enhance regeneration when transduced ex vivo to produce therapeutic molecules.
1999 ARRS Executive Council Award. Creation of saccular aneurysms in the rabbit: a model suitable for testing endovascular devices. American Roentgen Ray Society.

OBJECTIVE This study developed an animal model of intracranial aneurysms suitable for evaluating emerging endovascular devices for aneurysmal therapy. We characterized the short-, medium-, and long-term attributes of this endovascular technique for saccular aneurysmal creation in the rabbit. MATERIALS AND METHODS The right common carotid artery was surgically exposed in nine New Zealand white rabbits. Using endovascular techniques, we occluded the origin of the right common carotid artery with a pliable balloon. Elastase was incubated endoluminally in the proximal common carotid artery above the balloon. The common carotid artery was ligated distally. Animals were studied angiographically and sacrificed at 2 weeks (n = 3), 10 weeks (n = 3), and 24 weeks (n = 3) after aneurysm creation. Histology was obtained. RESULTS Saccular aneurysms formed in eight of the nine rabbits. The aneurysm projected from the apex of an approximately 90 degree curve of the parent vessel, the brachiocephalic artery. Mean aneurysm diameter was 4.5 mm (SD, 1.2 mm), and mean height was 7.5 mm (SD, 1.6 mm). All samples showed thinned elastic lamina and no evidence of inflammation. In four of eight aneurysms, unorganized thrombus was present in the dome of the aneurysm. CONCLUSION Arterial aneurysms with intact endothelium and deficient elastic lamina were reliably created in an area of high shear stress in New Zealand white rabbits. Three of these aneurysms remained patent for at least 6 months. We found a simple procedure that can be readily applied to the testing of new endovascular devices for a reliable creation of aneurysms in rabbits.
Back to work – evaluation of a multidisciplinary rehabilitation programme with emphasis on musculoskeletal disorders. A two-year follow-up

Purpose. The aim of this study was to evaluate the 2-year outcome of a multidisciplinary rehabilitation programme for working-age people, regarding sick leave and mental health. Method. The test persons consisted of 40 women and 20 men (mean age 46.8 ± SD 7.9) with musculoskeletal disorders, mainly neck and back pain. The rehabilitation programme was individually adapted and consisted of physical activity in several forms, relaxation, theoretical and practical education and individual guidance. Before, during and after the programme all participants were evaluated with the Global Self-Efficacy Index, Hospital Anxiety and Depression Scale, and stress test. Results. At the 2-year follow-up full-time sick leave had decreased by 37% (p < 0.0001) in the women, and by 25% (p < 0.05) in the men. Both women and men showed an increased quality of life (QoL) and decreased anxiety, depression and self-experienced stress at the 2-year follow-up compared with the start of the rehabilitation programme. Conclusions. The most important conclusion was that the effects of the rehabilitation programme persisted for up to 2 years. At 2 years the majority of the participants were still physically active, their QoL was increased, and most participants had returned to work.
Full‐matrix least‐squares refinement of lysozymes and analysis of anisotropic thermal motion

Crystal structures of turkey egg lysozyme (TEL) and human lysozyme (HL) were refined by full‐matrix least‐squares method using anisotropic temperature factors. The refinement converged at the conventional R‐values of 0.104 (TEL) and 0.115 (HL) for reflections with Fo > 0 to the resolution of 1.12 Å and 1.15 Å, respectively. The estimated r.m.s. coordinate errors for protein atoms were 0.031 Å (TEL) and 0.034 Å (HL). The introduction of anisotropic temperature factors markedly reduced the R‐value but did not significantly affect the main chain coordinates. The degree of anisotropy of atomic thermal motion has strong positive correlation with the square of distance from the molecular centroid. The ratio of the radial component of thermal ellipsoid to the r.m.s. magnitude of three principal components has negative correlation with the distance from the molecular centroid, suggesting the domination of libration rather than breathing motion. The TLS model was applied to elucidate the characteristics of the rigid‐body motion. The TLS tensors were determined by the least‐squares fit to observed temperature factors. The profile of the magnitude of reproduced temperature factors by the TLS method well fitted to that of observed Beqv. However, considerable disagreement was observed in the shape and orientation of thermal ellipsoid for atoms with large temperature factors, indicating the large contribution of local motion. The upper estimate of the external motion, 67% (TEL) and 61% (HL) of Beqv, was deduced from the plot of the magnitude of TLS tensors determined for main chain atoms which were grouped into shells according to the distance from the center of libration. In the external motion, the translational portion is predominant and the contribution of libration and screw motion is relatively small. The internal motion, estimated by subtracting the upper estimate of the external motion from the observed temperature factor, is very similar between TEL and HL in spite of the difference in 54 of 130 amino acid residues and in crystal packing, being suggested to reflect the intrinsic internal motion of chicken‐type lysozymes. Proteins 30:232–243, 1998. © 1998 Wiley‐Liss, Inc.
Biomechanics of unilateral compared with bilateral lumbar pedicle screw fixation for stabilization of unilateral vertebral disease.

OBJECT An in vitro flexibility experiment was performed to compare the biomechanical stability of asymmetrical lumbar pedicle screw fixation (longer hardware attached ipsilaterally to a 1-sided lesion), short and long fixation, and fixation with and without interconnection to the involved vertebra. METHODS Seven human cadaveric specimens (T12-S1) were studied intact; after simulated unilateral lesions were created at L2-3 and L3-4, the segments were stabilized by 1) L2-4 unilateral fixation (L-3 excluded), 2) L2-4 bilateral fixation (L-3 included contralaterally), 3) L2-5 unilateral fixation (L-3 excluded), 4) L2-5 fixation ipsilateral (L-3 excluded) and L2-4 fixation contralateral (L-3 included), 5) L2-5 bilateral fixation (L-3 included contralaterally), and 6) L2-5 bilateral fixation (L-3 excluded). The testing order varied among specimens. Angular range of motion (ROM) and lax zone were recorded optically while loading to 6.0 Nm was created with nonconstraining pure moments. RESULTS Unilateral short fixation provided significantly worse stabilization than any other construct tested in all loading modes (p < 0.05, repeated-measures analysis of variance). There was a mean 56% reduction in ROM across the lesion after adding 1 additional level rostrally and caudally. Asymmetrical long/short stabilization provided similar stability to symmetrical long stabilization. Minimal additional stability was gained by including L-3 in the long bilateral fixation construct. CONCLUSIONS Unilateral fixation is inadequate for stabilizing a 2-level unilateral lesion. Bilateral fixation, whether symmetrical or asymmetrical, provides good stabilization for this injury. It is not important for stability to include the level of the lesion within the long construct contralaterally.
Role of nitric oxide in experimental obliterative bronchiolitis (chronic rejection) in the rat.

The role of nitric oxide in obliterative bronchiolitis development, i.e., chronic rejection, was investigated in the heterotopic rat tracheal allograft model. An increase in the intragraft inducible nitric oxide synthase (iNOS) mRNA and mononuclear inflammatory cell iNOS immunoreactivity was demonstrated during progressive loss of respiratory epithelium and airway occlusion in nontreated allografts compared to syngeneic grafts. In nontreated allografts, however, intragraft nitric oxide production was decreased, most likely because of loss of iNOS epithelial expression. Treatment with aminoguanidine, a preferential inhibitor of inducible nitric oxide synthase, was associated with enhanced proliferation of alpha-smooth muscle actin immunoreactive cells and the intensity of obliterative bronchiolitis early after transplantation. Aminoguanidine treatment did not affect iNOS mRNA synthesis or intragraft nitric oxide production, but decreased iNOS immunoreactivity in smooth muscle cells. Treatment with L-arginine, a precursor of nitric oxide, significantly reduced obliterative changes. L-arginine supplementation enhanced intragraft iNOS mRNA synthesis and iNOS immunoreactivity in capillary endothelial and smooth muscle cells as well as intragraft nitric oxide production. Immunohistochemical analysis of allografts showed that neither iNOS inhibition nor supplementation of the nitric oxide pathway affected the number of graft-infiltrating CD4+ and CD8+ T cells, ED1+ and ED3+ macrophages, immune activation with expression of IL-2R or MHC class II, or production of macrophage or Th1 cytokines. In contrast, L-arginine treatment was associated with increased staining for Th2 cytokines IL-4 and IL-10. In conclusion, this study demonstrates that nitric oxide has a protective role in obliterative bronchiolitis development in this model, and suggests that nitric oxide either directly or indirectly inhibits smooth muscle cell proliferation and modulates immune response towards Th2 cytokines.
Infarct size and nitric oxide synthase in murine myocardium.

Controversy surrounds the involvement of nitric oxide (NO) in myocardial ischaemia-reperfusion injury and the balance between deleterious and beneficial effects. NO synthase (NOS) is expressed constitutively as two isoforms: endothelial (eNOS) and neuronal (nNOS). Knockout mice lacking the gene for either eNOS (eNOS KO) or nNOS (nNOS KO), were compared with wild-types (WT) during a protocol of global ischaemia-reperfusion injury. Thirty-six mouse hearts (12 from each group) were isolated and the aorta cannulated for Langendorff perfusion with modified Krebs solution at constant pressure. An apical suture connected the left ventricle to a force transducer via a light weight coupling rod. Following stabilization hearts were subjected to 30 min of global ischaemia at 37 degrees C. During 30 min reperfusion, the recovery of baseline force-rate product (F%) was recorded. Hearts were then stained with tetrazolium, frozen, sliced, and fixed with formalin. Slices were compressed between plexiglas plates, and a magnified video image digitized to allow planimetry for infarct size (as percentage of ventricular volume I/R). Although recovery of contractile function did not differ between groups, eNOS KOs suffered significantly larger infarcts than WT or nNOS KOs (41 v 33 and 30% respectively, P<0.05 for both comparisons). A protective role for eNOS against global ischaemia-reperfusion injury has been demonstrated for the first time in murine myocardium. This may have important clinical implications for future pharmacotherapy to enhance myocardial protection.
X chromosome inactivation in women with alcoholism.

BACKGROUND All female mammals with 2 X chromosomes balance gene expression with males having only 1 X by inactivating one of their X chromosomes (X chromosome inactivation [XCI]). Analysis of XCI in females offers the opportunity to investigate both X-linked genetic factors and early embryonic development that may contribute to alcoholism. Increases in the prevalence of skewing of XCI in women with alcoholism could implicate biological risk factors. METHODS The pattern of XCI was examined in DNA isolated in blood from 44 adult women meeting DSM-IV criteria for an alcohol use disorder and 45 control women with no known history of alcohol abuse or dependence. XCI status was determined by analyzing digested and undigested polymerase chain reaction (PCR) products of the polymorphic androgen receptor (AR) gene located on the X chromosome. Subjects were categorized into 3 groups based upon the degree of XCI skewness: random (50:50 to 64:36%), moderately skewed (65:35 to 80:20%), and highly skewed (>80:20%). RESULTS XCI status from informative women with alcoholism was found to be random in 59% (n = 26), moderately skewed in 27% (n = 12), or highly skewed in 14% (n = 6). Control subjects showed 60, 29, and 11%, respectively. The distribution of skewed XCI observed among women with alcoholism did not differ statistically from that of control subjects (χ(2) test = 0.14, 2 df, p = 0.93). CONCLUSIONS Our data did not support an increase in XCI skewness among women with alcoholism or implicate early developmental events associated with embryonic cell loss or unequal (nonrandom) expression of X-linked gene(s) or defects in alcoholism among women.
The development of emotion perception in face and voice during infancy.

PURPOSE Interacting with others by reading their emotional expressions is an essential social skill in humans. How this ability develops during infancy and what brain processes underpin infants' perception of emotion in different modalities are the questions dealt with in this paper. METHODS Literature review. RESULTS The first part provides a systematic review of behavioral findings on infants' developing emotion-reading abilities. The second part presents a set of new electrophysiological studies that provide insights into the brain processes underlying infants' developing abilities. Throughout, evidence from unimodal (face or voice) and multimodal (face and voice) processing of emotion is considered. The implications of the reviewed findings for our understanding of developmental models of emotion processing are discussed. CONCLUSIONS The reviewed infant data suggest that (a) early in development, emotion enhances the sensory processing of faces and voices, (b) infants' ability to allocate increased attentional resources to negative emotional information develops earlier in the vocal domain than in the facial domain, and (c) at least by the age of 7 months, infants reliably match and recognize emotional information across face and voice.
Distinct temporal filters in mitral cells and external tufted cells of the olfactory bulb

Short-term synaptic plasticity is a critical regulator of neural circuits, and largely determines how information is temporally processed. In the olfactory bulb, afferent olfactory receptor neurons respond to increasing concentrations of odorants with barrages of action potentials, and their terminals have an extraordinarily high release probability (Sicard, 1986; Murphy et al., 2004). These features suggest that during naturalistic stimuli, afferent input to the olfactory bulb is subject to strong synaptic depression, presumably truncating the postsynaptic response to afferent stimuli. To examine this issue, we used single glomerular stimulation in mouse olfactory bulb slices to measure the synaptic dynamics of afferent-evoked input at physiological stimulus frequencies. In cell-attached recordings, mitral cells responded to high frequency stimulation with sustained responses, whereas external tufted cells responded transiently. Consistent with previous reports (Murphy et al., 2004), olfactory nerve terminals onto both cell types had a high release probability (0.7), from a single pool of slowly recycling vesicles, indicating that the distinct responses of mitral and external tufted cells to high frequency stimulation did not originate presyaptically. Rather, distinct temporal response profiles in mitral cells and external tufted cells could be attributed to slow dendrodendritic responses in mitral cells, as blocking this slow current in mitral cells converted mitral cell responses to a transient response profile, typical of external tufted cells. Our results suggest that despite strong axodendritic synaptic depression, the balance of axodendritic and dendrodendritic circuitry in external tufted cells and mitral cells, respectively, tunes the postsynaptic responses to high frequency, naturalistic stimulation. Key Points The release probability of the ORN is reportedly one of the highest in the brain (Murphy et al., 2004), which is predicted to impose a transient temporal filter on postsynaptic cells. Mitral cells responded to high frequency ORN stimulation with sustained transmission, whereas external tufted cells responded transiently. The release probability of ORNs (0.7) was equivalent across mitral and external tufted cells and could be explained by a single pool of slowly recycling vesicles. The sustained response in mitral cells resulted from dendrodendritic amplification in mitral cells, which was blocked by NMDA and mGluR1 receptor antagonists, converting mitral cell responses to transient response profiles. Our results suggest that although the afferent ORN synapse shows strong synaptic depression, dendrodendritic circuitry in mitral cells produces robust amplification of brief afferent input, thus the relative strength of axodendritic and dendrodendritic input determines the postsynaptic response profile.
Designer deletion strains derived from Saccharomyces cerevisiae S288C: A useful set of strains and plasmids for PCR‐mediated gene disruption and other applications

A set of yeast strains based on Saccharomyces cerevisiae S288C in which commonly used selectable marker genes are deleted by design based on the yeast genome sequence has been constructed and analysed. These strains minimize or eliminate the homology to the corresponding marker genes in commonly used vectors without significantly affecting adjacent gene expression. Because the homology between commonly used auxotrophic marker gene segments and genomic sequences has been largely or completely abolished, these strains will also reduce plasmid integration events which can interfere with a wide variety of molecular genetic applications. We also report the construction of new members of the pRS400 series of vectors, containing the kanMX, ADE2 and MET15 genes. © 1998 John Wiley & Sons, Ltd.
Physiological and psychometric variables for monitoring recovery during tapering for major competition.

PURPOSE This study attempted to identify variables that are useful in monitoring recovery during tapering. METHODS Changes in physiological variables, tethered swimming force, mood states, and self-ratings of well-being were measured in 10 elite swimmers from before to after 2 wk of tapering for national championships. Physiological measures included resting heart rate (HR); blood pressure (BP); blood lactate concentration; red blood cell, white blood cell, and differential counts; and plasma cortisol, free testosterone, and catecholamine concentrations. Measures taken after 100-m maximal and 200-m standardized submaximal swims included HR, BP, and blood lactate concentration. RESULTS Step-down regression analysis showed that changes in plasma norepinephrine concentration, heart rate after maximal effort swimming and confusion as measured by the Profile of Mood States (POMS) predicted the change in swimming time with tapering (r2 = 0.98); the change in plasma norepinephrine concentration predicted the change in swim time with tapering (r2 = 0.82) by itself. CONCLUSION These data suggest that recovery after intense training can be monitored during tapering and that an accurate prediction of performance changes may be possible if the changes in a range of physiological and psychological variables are measured.