bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of novel Y chromosome encoded transcripts by testis transcriptome analysis of mice with deletions of the Y chromosome long arm
Authors: Touré Aminata , Clemente Emily J , Ellis Peter JI , Mahadevaiah Shantha K , Ojarikre Obah A , Ball Penny AF , Reynard Louise , Loveland Kate L , Burgoyne Paul S , Affara Nabeel A
Published: December 2005

Background

The male-specific region of the mouse Y chromosome long arm (MSYq) is comprised largely of repeated DNA, including multiple copies of the spermatid-expressed Ssty gene family. Large deletions of MSYq are associated with sperm head defects for which Ssty deficiency has been presumed to be responsible.

Results

In a search for further candidate genes associated with these defects we analyzed changes in the testis transcriptome resulting from MSYq deletions, using testis cDNA microarrays. This approach, aided by accumulating mouse MSYq sequence information, identified transcripts derived from two further spermatid-expressed multicopy MSYq gene families; like Ssty, each of these new MSYq gene families has multicopy relatives on the X chromosome. The Sly family encodes a protein with homology to the chromatin-associated proteins XLR and XMR that are encoded by the X chromosomal relatives. The second MSYq gene family was identified because the transcripts hybridized to a microarrayed X chromosome-encoded testis cDNA. The X loci ('Astx') encoding this cDNA had 92-94% sequence identity to over 100 putative Y loci ('Asty') across exons and introns; only low level Asty transcription was detected. More strongly transcribed recombinant loci were identified that included Asty exons 2-4 preceded by Ssty1 exons 1, 2 and part of exon 3. Transcription from the Ssty1 promotor generated spermatid-specific transcripts that, in addition to the variable inclusion of Ssty1 and Asty exons, included additional exons because of the serendipitous presence of splice sites further downstream.

Conclusion

We identified further MSYq-encoded transcripts expressed in spermatids and deriving from multicopy Y genes, deficiency of which may underlie the defects in sperm development associated with MSYq deletions.
Management of self-harm in adults: which way now?
Authors: Kapur Navneet
Published: November 2005

Self-harm remains an important public health problem and two sets of clinical guidelines have been published recently. While these include elements of accepted good practice they are not evidence-based. Further research might concentrate on either very large trials of low-intensity interventions or smaller trials of longer-term psychological treatments. The current management of self-harm may be improved by shifting professionals' views, involving users in staff training, and changing service provision – perhaps moving from risk assessment to needs assessment.
Cognitive heterogeneity in first-episode schizophrenia
Authors: Joyce Eileen M. , Hutton Sam B. , Mutsatsa Stanley H. , Barnes Thomas R. E.
Published: November 2005

Background

Studies of chronic schizophrenia suggest that there are subgroups with different profiles of cognitive impairment.

Aims

To determine whether such heterogeneity is present at illness onset and any relationship to clinical variables.

Method

Ninety-three community patients with first-episode schizophrenia and 50 healthy volunteers were assessed for premorbid (Revised National Adult Reading Test) and current IQ, memory and executive function.

Results

Half of those with schizophrenia had preserved IQ in the normal range but there was evidence of a specific impairment in spatial working memory even in those with high/average IQ; 37 out of 93 (40%) had generalised cognitive decline. Those with low premorbid IQ were significantly younger at illness onset. For the entire group, age at onset correlated positively with premorbid but not current IQ.

Conclusions

At illness onset, cognitive heterogeneity is present in people with schizophrenia, with a high proportion having undergone general cognitive decline. However, working memory impairment may be a common feature. Lower premorbid IQ is a risk factor for an earlier onset.
Alcohol consumption as a risk factor for anxiety and depression
Authors: Haynes Jonathan C. , Farrell Michael , Singleton Nicola , Meltzer Howard , Araya Ricardo , Lewis Glyn , Wiles Nicola J.
Published: November 2005

Background

Longitudinal studies have been inconclusive in identifying alcohol as a risk factor for anxiety and depression.

Aims

To examine whether excessive alcohol consumption is a risk factor for anxiety and depression in the general population, and whether anxiety and depression are risk factors for excessive alcohol consumption.

Method

Data were analysed from the 18-month follow-up of the Psychiatric Morbidity Among Adults Living in Private Households, 2000 survey.

Results

Hazardous and dependent drinking were not associated with onset of anxiety and depression at follow-up. Binge-drinking was non-significantly associated with incident anxiety and depression (adjusted OR= 1.36, 95% CI 0.74–2.50). Abstainers were less likely to have new-onset anxiety and depression at follow-up. Anxiety and depression or sub-threshold symptoms at baseline were not associated with incident hazardous or binge-drinking at follow-up, but there was weak evidence linking sub-threshold symptoms with onset of alcohol dependence (adjusted OR=2.04, 95% CI 0.84–4.97).

Conclusions

Excessive alcohol consumption was not associated with the onset of anxiety and depression but abstinence was associated with a lower risk. Sub-threshold symptoms were weakly associated with new-onset alcohol dependence.
Safety of women in mixed-sex and single-sex medium secure units: staff and patient perceptions
Authors: Mezey Gillian , Hassell Yonette , Bartlett Annie
Published: November 2005

Background

The development of single-sex medium secure units for women has been driven by concern about the vulnerability of women to sexual abuse and exploitation in mixed-sex secure settings. Less is known about how women patients and staff perceive gender segregation and their experiences in single-sex units.

Aims

To examine the impact of gender segregation on the safety of women patients detained in medium secure psychiatric facilities.

Method

A qualitative study was conducted involving individual interviews with 58 male and female staff and 31 women patients in single-sex and mixed-sex medium secure units throughout England and Wales.

Results

Women patients in both types of units reported high levels of actual and threatened physical and sexual violence. Women in single-sex units reported intimidation, threats and abuse by other women patients, although they were less vulnerable to sexual abuse and exploitation and serious physical assault.

Conclusions

Further development of single-sex secure units for women may not be justified on the grounds of safety issues alone. Risk assessment of forensic psychiatric patients must include a full assessment of their safety within the psychiatric setting.
The ground is in great shape, but can we field a kitted-out team? Commentary on … Mental health research system in England
Authors: Szmukler George
Published: November 2005

Chilvers & Clark make it clear that research and development (R&D) in the National Health Service (NHS) is now a managed process. There is a coherent strategy and much thought has been given to the use of resources. I welcome this development and the ‘Health Research System’ that forms its framework.
All-Trans Retinoic Acid Inhibits Vascular Endothelial Growth Factor Expression in a Cell Model of Neutrophil Activation
Authors: Tee Meng Kian , Vigne Jean-Louis , Taylor Robert N.
Published: December 2005

Infiltrating neutrophil granulocytes are a particularly rich source of vascular endothelial growth factor (VEGF) in the endometrium and may contribute to the angiogenesis of endometriosis lesions. The objective of this study is to evaluate the expression and regulation of VEGF in endometrial neutrophils and in a model of neutrophil differentiation relevant to endometriosis. Immunohistochemistry was performed on endometriosis patient biopsies and cultured neutrophil-like HL-60 cells were assessed. The study was set in a reproductive biology division within an academic medical center. Endometrial biopsies were performed on women with endometriosis and HL-60 cells were treated with all-trans retinoic acid (atRA) and dimethyl sulfoxide in vitro. Immunofluorescence histochemistry, VEGF mRNA and protein quantification, and transfection studies of VEGF gene promoter-luciferase constructs were all main outcome measures. Immunofluorescence studies verified the presence of neutrophils in eutopic endometrium from women with endometriosis. Examination of the regulation of VEGF using differentiated HL-60 cells as a model, revealed that atRA induced a dose- and time-dependent suppression of VEGF mRNA and protein. Transient transfection, truncation, EMSA, and site-directed mutagenesis of human VEGF promoter-luciferase constructs in HL-60 cells indicated that atRA repressed VEGF gene transcription via a direct repeat 1 element located between −443 and −431 bp relative to the transcription initiation site. Because retinoic acid is synthesized de novo in endometrial cells under the influence of progesterone, our findings suggest that the up-regulated VEGF and angiogenesis in tissue from women with endometriosis may reflect failure of neutrophil differentiation in these cases, and provide a rationale for retinoid therapy in this condition.
Adrenomedullin Regulates Cellular Glutathione Content via Modulation of γ-Glutamate-Cysteine Ligase Catalytic Subunit Expression
Authors: Kim Jee-Youn , Yim Ji-Hye , Cho Jin-Ho , Kim Jin-Hwan , Ko Jeong-Hun , Kim Su-Mi , Park Seungjoon , Park Jae-Hoon
Published: December 2005

Adrenomedullin (AM) participates in a wide range of physiological and pathological processes including vasorelaxation, angiogenesis, cancer promotion, and apoptosis. Recently, it has been reported that AM protects a variety of cells against oxidative stress induced by stressors such as hypoxia, ischemia/reperfusion, and hydrogen peroxide through the phosphatidylinositol 3-kinase (PI3K)-dependent pathway. However, the molecular mechanisms underlying the pathway of cell survival against hypoxic injury are largely unknown. In an effort to investigate the survival mechanism against hypoxic injury, we studied the effects of AM on cellular levels of reactive oxygen species, well-known mediators of cell death after oxidative stress, and the mechanism involved in the regulation of reactive oxygen species levels. Here, we show that AM increases γ-glutamate-cysteine ligase (γ-GCL) activity under both hypoxic and normoxic conditions, resulting in an up-regulation of cellular glutathione levels to more than 2-fold higher than basal expression. In addition, we demonstrate that AM induces concentration-dependent expression of the catalytic subunit of γ-GCL (γ-GCLC) at the mRNA and protein levels through the activation of the γ-GCLC promoter fragment sequence from −597 to −320. However, when treated with the PI3K inhibitors, the effects of AM on γ-GCLC expression were completely abrogated, suggesting that a PI3K pathway linked AM with the transcriptional activation of the γ-GCLC promoter. Taken together, our data suggests that AM participates in the regulation of cellular redox status via glutathione synthesis. These results may explain, in part, the mechanism by which AM protects cells against oxidative stress.
Estradiol Modulation of Kainic Acid-Induced Calcium Elevation in Neonatal Hippocampal Neurons
Authors: Hilton Genell D. , Bambrick Linda L. , Thompson Scott M. , McCarthy Margaret M.
Published: December 2005

The developing hippocampus of both males and females is exposed to high levels of the gonadal steroid estradiol. The impact of this estradiol exposure on developing hippocampal neurons is essentially unknown. In the rat, the newborn hippocampus is relatively insensitive to excitotoxic brain injury, which in adults is associated with the release of amino acids, in particular glutamate, resulting in a significant increase in intracellular calcium and eventual cell death. We have shown previously in the rat that administration of the glutamate agonist, kainic acid (KA), on the day of birth results in limited hippocampal damage, which is ameliorated by treatment with the gonadal steroid, estradiol. We now show that KA induces an increase in intracellular calcium through L-type voltage-sensitive calcium channels early in development and, later in development, through polyamine-sensitive α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors with a modest increase through N-methyl-d-aspartate receptors. Pretreatment with the gonadal steroid, estradiol, decreases the percentage of neurons responding to KA and decreases the peak amplitude of the calcium transient early in development but has no effect later in development. Taken together, these data suggest that there is a developmental shift in the route of KA-induced intracellular calcium and estradiol modulates KA-induced intracellular calcium to a time restricted to early development, but whether this is the basis of the neuroprotective effect of estradiol remains to be determined.
Corticotropin-Releasing Factor Is Cytoprotective in Xenopus Tadpole Tail: Coordination of Ligand, Receptor, and Binding Protein in Tail Muscle Cell Survival
Authors: Boorse Graham C. , Kholdani Cyrus A. , Seasholtz Audrey F. , Denver Robert J.
Published: December 2005

Upon metamorphosis, amphibian tadpoles lose their tails through programmed cell death induced by thyroid hormone (T3). Before transformation, the tail functions as an essential locomotory organ. The binding protein for the stress neuropeptide corticotropin-releasing factor (CRF; CRF-BP) is strongly up-regulated in the tail of Xenopus tadpoles during spontaneous or T3-induced metamorphosis. This finding led us to investigate physiological roles for CRF and CRF-BP in tadpole tail. We found CRF, CRF-BP, and functional CRF1 receptor in tail and CRF and functional CRF1 receptors, but not CRF-BP, in the tail muscle-derived cell line XLT-15. CRF, acting via the CRF1 receptor, slowed spontaneous tail regression in explant culture and caused a reduction in caspase 3/7 activity. CRF increased, but stable CRF-BP overexpression decreased, [3H]thymidine incorporation in XLT-15 cells. Overexpression of CRF-BP in vivo accelerated the loss of tail muscle cells during spontaneous metamorphosis. Lastly, exposure of tail explants to hypoxia increased CRF and urocortin 1 but strongly decreased CRF-BP mRNA expression. We show that CRF is expressed in tadpole tail, is up-regulated by environmental stressors, and is cytoprotective. The inhibitory binding protein for CRF is regulated by hormones or by environmental stressors and can modulate CRF bioactivity.
The Vitamin D Receptor Represses Transcription of the Pituitary Transcription Factor Pit-1 Gene without Involvement of the Retinoid X Receptor
Authors: Seoane Samuel , Perez-Fernandez Roman
Published: December 2005

Pituitary transcription factor-1 (Pit-1) plays a key role in cell differentiation during organogenesis of the anterior pituitary, and as a transcriptional activator for the pituitary GH and prolactin genes. However, Pit-1 is also expressed in nonpituitary cell types and tissues. In breast tumors, Pit-1 mRNA and protein levels are increased with respect to normal breast, and in MCF-7 human breast adenocarcinoma cells, Pit-1 increases GH secretion and cell proliferation. We report here that 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] administration to MCF-7 cells induces a significant decrease in Pit-1 mRNA and protein levels. By deletion analyses, we mapped a region (located between −147 and −171 bp from the transcription start site of the Pit-1 gene) that is sufficient for the repressive response to 1,25-(OH)2D3. Gel mobility shift and chromatin immunoprecipitation assays confirmed the direct interaction between the vitamin D receptor (VDR) as homodimer (without the retinoid X receptor), and the Pit-1 promoter, supporting the view that Pit-1 is a direct transcriptional target of VDR. Our data also indicate that recruitment of histone deacetylase 1 is involved in this repressive effect. This ligand-dependent Pit-1 gene inhibition by VDR in the absence of the retinoid X receptor seems to indicate a new mechanism of transcriptional repression by 1,25-(OH)2D3.
Transcriptional Intermediary Factor 1α Mediates Physical Interaction and Functional Synergy between the Coactivator-Associated Arginine Methyltransferase 1 and Glucocorticoid Receptor-Interacting Protein 1 Nuclear Receptor Coactivators
Authors: Teyssier Catherine , Ou Chen-Yin , Khetchoumian Konstantin , Losson Régine , Stallcup Michael R.
Published: December 2005

In previous studies transcriptional intermediary factor 1α (TIF1α) was identified as a direct binding partner and potential transcriptional coactivator for nuclear receptors (NRs) but its overexpression inhibited, rather than enhanced, transcriptional activation by NRs. Here we show that TIF1α bound to and enhanced the function of the C-terminal activation domain (AD) of coactivator associated arginine methyltransferase 1 (CARM1) and the N-terminal AD of glucocorticoid receptor-interacting protein 1 (GRIP1). Furthermore, although TIF1α had little or no NR coactivator activity by itself, it cooperated synergistically with GRIP1 and CARM1 to enhance NR-mediated transcription. Inhibition of endogenous TIF1α expression reduced transcriptional activation by the GRIP1 N-terminal domain but not by the CARM1 C-terminal domain, suggesting that TIF1α may be more important for mediating the activity of the former than the latter. Reduction of endogenous TIF1α levels also compromised the androgen-dependent induction of an endogenous target gene of the androgen receptor. Finally, TIF1α formed a ternary complex with the GRIP1 N-terminal and CARM1 C-terminal domains. Thus, we conclude that TIF1α cooperates with NR coactivators GRIP1 and CARM1 by forming a stable ternary complex with them and enhancing the AD function of one or both of them.
Ace2p contributes to fission yeast septin ring assembly by regulating mid2+ expression
Authors: Petit Claudia S. , Mehta Sapna , Roberts Rachel H. , Gould Kathleen L.
Published: November 2005

The fission yeast Schizosaccharomyces pombe divides through constriction of an actomyosin-based contractile ring followed by formation and degradation of a medial septum. Formation of an organized septin ring is also important for the completion of S. pombe cell division and this event relies on the production of Mid2p. mid2+ mRNA and protein accumulate in mitosis. Recent microarray analyses identified mid2+ as a target of the Ace2p transcription factor, and ace2+ as a target of the Sep1p transcription factor. In this study, we find that Mid2p production is controlled by Ace2p functioning downstream of Sep1p. Consequently, both Sep1p and Ace2p are required for septin ring assembly and genetic analyses indicate that septin rings function in parallel with other Ace2p targets to achieve efficient cell division. Conversely, forced overproduction of Sep1p or Ace2p prevents septin ring disassembly. We find that Ace2p levels peak during anaphase and Ace2p is post-translationally modified by phosphorylation and ubiquitylation. Ace2p localizes symmetrically to dividing nuclei and functions independently of the septation initiation network.
Male song quality affects circulating but not yolk steroid concentrations in female canaries (Serinus canaria)
Authors: Marshall Rupert C. , Leisler Bernd , Catchpole Clive K. , Schwabl Hubert
Published: December 2005

Male song complexity is a sexually selected trait found in many songbirds,including strains of the domestic canary. Studies on several species have shown that male song can affect the hormonal state of females and may also influence concentrations of maternal hormones in the yolk of their eggs. In this study, we show that the level of circulating androgens and oestrogens of female canaries, as measured in faeces, varies with the quality of male song to which they are exposed. The female-perceived quality of male canary song depends on the production of special `sexy syllables' to which females respond with more sexual displays. Using playback of synthetic song we show that females hearing songs with sexy syllables have higher levels of faecal testosterone than control females hearing songs without them. However, unlike previous studies on the canary, we found no evidence that such females laid eggs with more testosterone (or other steroids) in their egg yolks. We discuss these results in relation to the evolution of male signalling and maternal investment strategies.
Composition, morphology and mechanics of hagfish slime
Authors: Fudge Douglas S. , Levy Nimrod , Chiu Scott , Gosline John M.
Published: December 2005

Hagfish slime consists of mucins and protein threads that are released from slime glands and mix with seawater to produce an ephemeral material with intriguing physical properties. We recently characterized the mechanics of the slime's fibrous component, and here we report the first mechanical properties of the mucin component and the slime as a whole. Our results suggest that hagfishes can produce remarkable quantities of the slime because it is almost three orders of magnitude more dilute than typical mucus secretions. Mechanical experiments using whole slime produced in vitrodemonstrate that the slime threads dominate the slime's material properties and impart elasticity. Mucins impart viscosity at the strain rates tested and are important for rapid deployment of the slime. We also found that slime threads are tapered at both ends, which suggested to us that hagfish slime might best be modeled as a discontinuous fibre-reinforced composite. Our measurements demonstrate that the mucins are not capable of providing shear linkage between threads, but this is not necessary because the threads are long enough to span an entire slime mass. Our findings suggest that hagfish slime consists mainly of bulk seawater entrained between mucin-coated threads,and in this way functions more like a fine sieve than coherent mucus. These results are consistent with the hypothesis that the slime has evolved as a defense against gill-breathing predators.
Constraints on starting and stopping: behavior compensates for reduced pectoral fin area during braking of the bluegill sunfishLepomis macrochirus
Authors: Higham Timothy E. , Malas Brett , Jayne Bruce C. , Lauder George V.
Published: December 2005

Many natural animal movements involve accelerating from a standstill and then stopping. Obstacles in natural environments often limit the straight-line distance available for movement, and decreased braking ability theoretically can limit speed for short distances. Consequently, braking ability can be important for avoiding collisions with obstacles and exploiting resources effectively in complex environments. A presumed morphological correlate of improved braking performance in fish is increased pectoral fin area, because most fish protract these structures as they decelerate. However, the kinematics and modulation of velocity during starting and stopping are poorly understood for most species of fish as well as most species of animals. Thus,for bluegill sunfish Lepomis macrochirus with complete and partially ablated pectoral fins (35% original fin area), we analyzed high speed video recordings (200 images s–1) of predatory attacks with a start and stop in a short, standardized distance (40 cm). We quantified body displacement, velocity, acceleration, deceleration and several fin angle variables during each feeding. Unexpectedly, several variables including maximum velocity and maximum deceleration (grand means 72 cm s–1 and –512 cm s–2, respectively) did not change significantly with reduced pectoral fin area. The average values of braking movements of the median and caudal fins did increase with decreased pectoral fin area but lacked statistically significant differences. The primary mechanism of attaining similar braking performance with decreased area of the pectoral fins was that they were protracted significantly more (mean difference=42°) and with a significantly faster average velocity of protraction. Thus, pectoral fin area appears unlikely to be the primary constraint on braking performance for this particular task.
Derivatization in Mass Spectrometry—6. Formation of Mixed Derivatives of Polyfunctional Compounds
Authors: Zaikin Vladimir G. , Halket John M.
Published: December 2005

The review describes chemical transformations of multi-functional compounds (amino acids and peptides, amino alcohols, amino thiols, hydroxy acids, oxo acids, oxo alcohols, compounds containing simultaneously three or more different groups etc.) by using step-wise or one-step modification or protection of functional groups. Some chemical aspects of mixed derivatization performed for improving the physical–chemical properties and mass spectral characteristics are discussed. Application of mixed derivatization to qualitative and quantitative analysis of various multifunctional compounds mainly in biological fluids and other matrices by gas chromatography/mass spectrometry in electron ionization, chemical ionization, negative-ion chemical ionization and selected ion monitoring modes is considered.
Conformation Effects on the Dissociation of Ionized Polymers
Authors: Casey Janeen , Alhazmi Abdulrahman , Mayer Paul M.
Published: December 2005

The collision-induced dissociation (CID) of a series of oligomer chain lengths of four polymer samples have been quantified according to their observed total relative fragment ion abundances. The CID mass spectra of oligomers of ionized poly(methyl methacrylate) (PMMA), with three different types of end groups and polystyrene (PS), were obtained at fixed center-of-mass collision energies and collision numbers. For the PMMA polymers, the total fragment ion abundance increases with increasing chain length, consistent with an increase in internal energy deposition with the size of the ion. A discontinuity in the increase in total fragment ion abundance appears to correspond with a change in conformation of the polymer ions from linear (at short chain lengths) to cyclic (at long chain lengths). Ionized PS does not exhibit this change in conformation as all chain lengths show compact structures and accordingly the total fragment ion abundance does not change with increasing chain length.
A stemmed acetabular component in the management of severe acetabular deficiency
Authors: Badhe N. P. , Howard P. W.
Published: December 2005

We evaluated the use of a stemmed acetabular component in the treatment of severe acetabular deficiency during revision and complex primary total hip arthroplasty.

There were 31 hips of which 24 were revisions (20 for aseptic loosening, four for infection) and the remainder were complex primary arthroplasties. At a mean follow-up of 10.7 years (6 to 12.8), no component had been revised for aseptic loosening; one patient had undergone a revision of the polyethylene liner for wear. There was one failure because of infection. At the latest follow-up, the cumulative survival rate for aseptic loosening, with revision being the end-point, was 100%; for radiographic loosening it was 92% and for infection and radiographic loosening it was 88%. These results justify the continued use of this stemmed component for the reconstruction of severe acetabular deficiency.
Long-term results of rotational acetabular osteotomy for dysplasia of the hip in adult ambulatory patients with cerebral palsy
Authors: Nagoya S. , Nagao M. , Takada J. , Kaya M. , Iwasaki T. , Yamashita T.
Published: December 2005

We performed rotational acetabular osteotomy in order to treat dysplasia of the hip in five ambulatory adults with cerebral palsy. There was one man and four women, with a mean age of 21 years (16 to 27) who were followed up for a mean of 12 years and two months.

The mean Sharp angle improved from 52° to 43°, the mean acetabular index from 30.2° to 2.8°, the mean centre-edge angle from −5.6° to 29.2°, and the mean acetabular head index from 49.2 to 88.2. There was no progression of joint degeneration and relief from pain was maintained.

Our results suggest that rotational acetabular osteotomy is a valuable option for the treatment of acetabular dysplasia in adults with cerebral palsy who have incapacitating pain in the hip.
Interleukin-6 and Nitric Oxide Synthase Expression in the Vasopressin and Corticotrophin-releasing Factor Systems of the Rat Hypothalamus
Authors: González-Hernández Tomás , Afonso-Oramas Domingo , Cruz-Muros Ignacio , Barroso-Chinea Pedro , Abreu Pedro , Pérez-Delgado María del Mar , Rancel-Torres Nélida , González María del Carmen
Published: December 2005

Nitric oxide synthase (NOS) and interleukin-6 (IL-6) are constitutively expressed in hypothalamic cells. However, phenotypic and functional aspects of these cells remain unknown. We have studied the expression pattern of these two molecules in hypothalamic cells expressing corticotropin-releasing factor (CRF) and arginin-vasopressin (AVP), two major regulatory peptides in the hypothalamus-pituitary system, using immunofluorescence, intracerebroventricular injection of colchicine, and the study in parallel of the labeling pattern of axons in the median eminence. Within AVP cells, we distinguished two different populations: large, intensely stained AVP cells coexpressing IL-6; and large, intensely stained AVP cells coexpressing IL-6 and NOS. Within the CRF cells, we distinguished three different populations: large, intensely stained CRF cells immunonegative for AVP, NOS, and IL-6; large cells weakly stained for CRF and AVP, immunopositive for NOS and immunonegative for IL-6; and small cells intensely stained for CRF and AVP and immunonegative for IL-6 and NOS. In addition, we also found AVP cells containing IL-6 in the suprachiasmatic nucleus. These results suggest that neuronal NOS and IL-6 may be involved in different modulatory processes in hypophysiotropic and non-hypophysiotropic cells.
Three Mutations (p.Q36H, p.G418fsX482, and g.IVS19-2A>C) in the Dual Oxidase 2 Gene Responsible for Congenital Goiter and Iodide Organification Defect
Authors: Varela Viviana , Rivolta Carina M , Esperante Sebastián A , Gruñeiro-Papendieck Laura , Chiesa Ana , Targovnik Héctor M
Published: December 2005

Background: Iodide organification defects are associated with mutations in the dual oxidase 2 (DUOX2) gene and are characterized by a positive perchlorate discharge test. These mutations produce a congenital goitrous hypothyroidism, usually transmitted in an autosomal recessive mode.

Methods: We studied the complete coding sequence of the human DUOX2 gene by single-strand conformational polymorphism (SSCP) analysis of DNA from 17 unrelated patients with iodide organification defects. Samples showing an aberrant pattern were directly sequenced. All mutations were validated by SSCP analysis. Finally, the effect of a splicing mutation was studied by construction of minigenes.

Results: Genomic DNA sequencing revealed 3 novel mutations [c.108G>C (p.Q36H), c.1253delG (p.G418fsX482), and g.IVS19-2A>C] and 1 previously reported mutation [c.2895-2898delGTTC (p.S965fsX994)] in 2 families with 1 (family 1) and 2 (family 2) affected members. This implies the inheritance of 2 compound heterozygous mutations, p.Q36H and p.S965fsX994 in family 1 and p.G418fsX482 and g.IVS19-2A>C in family 2. The c.1253delG mutation was associated with a c.1254C>A transversion. In vitro transcription analysis showed that exon 20 is skipped entirely when the g.IVS19-2A>C mutation is present. The wild-type glutamine residue at position 36 is strictly conserved.

Conclusions: Two previously unknown compound heterozygous mutations in the DUOX2 gene, p.Q36H/p.S965fsX994 and p.G418fsX482/g.IVS19-2A>C, are responsible for iodide organification defects in 2 unrelated families. Identification of the molecular basis of this disorder might be helpful for understanding the pathophysiology of this congenital hypothyroidism.
Upregulation of Forebrain NMDA NR2B Receptors Contributes to Behavioral Sensitization after Inflammation
Authors: Wu Long-Jun , Toyoda Hiroki , Zhao Ming-Gao , Lee Yong-Seok , Tang Jianrong , Ko Shanelle W. , Jia Yong Heng , Shum Fanny W. F. , Zerbinatti Celina V. , Bu Guojun , Wei Feng , Xu Tian-Le , Muglia Louis J. , Chen Zhou-Feng , Auberson Yves P. , Kaang Bong-Kiun , Zhuo Min
Published: November 2005

Transgenic overexpression of NMDA NR2B receptors in forebrain regions increased behavioral responses to persistent inflammatory pain. However, it is not known whether inflammation leads to the upregulation of NR2B receptors in these regions. Here, we show that peripheral inflammation increased the expression of NMDA NR2B receptors and NR2B receptor-mediated synaptic currents in the anterior cingulate cortex (ACC). In freely moving mice, the increase in NR2B receptors after inflammation contributed to enhanced NMDA receptor-mediated responses in the ACC. Inhibition of NR2B receptors in the ACC selectively reduced behavioral sensitization related to inflammation. Our results demonstrate that the upregulation of NR2B receptors in the ACC contributes to behavioral sensitization caused by inflammation.
The Purine Synthesis Gene Prat2 Is Required for Drosophila Metamorphosis, as Revealed by Inverted-Repeat-Mediated RNA Interference
Authors: Ji Yingbiao , Clark Denise V
Published: December 2005

PRAT (phosphoribosylamidotransferase; E.C. 2.4.2.14) catalyzes the first reaction in de novo purine nucleotide biosynthesis. In Drosophila melanogaster, the Prat and Prat2 genes are both highly conserved with PRAT sequences from prokaryotes and eukaryotes. However, Prat2 organization and expression during development is different from Prat. We used RNA interference (RNAi) to knock down expression of both Prat and Prat2 to investigate their functions. Using the GAL4–UAS system, Prat RNAi driven by Act5c–GAL4 or tubP–GAL4 causes variable pupal lethality (48–100%) and ∼50% female sterility, depending on the transgenic strains and drivers used. This observation agrees with the phenotype previously observed for Prat EMS-induced mutations. Prat2 RNAi driven by Act5C–GAL4 or tubP–GAL4 also results in variable pupal lethality (61–93%) with the different transgenic strains, showing that Prat2 is essential for fly development. However, Prat2 RNAi-induced arrested pupae have a head eversion defect reminiscent of the “cryptocephal” phenotype, whereas Prat RNAi-induced arrested pupae die later as pharate adults. We conclude that Prat2 is required during the prepupal stage while Prat is more important for the pupal stage. In addition, Prat and Prat2 double RNAi results in more severe pupal lethal phenotypes, suggesting that Prat and Prat2 have partially additive functions during Drosophila metamorphosis.
gon-14 Functions With Class B and Class C Synthetic Multivulva Genes to Control Larval Growth in Caenorhabditis elegans
Authors: Chesney Michael A , Kidd Ambrose R , Kimble Judith
Published: December 2005

Previous work showed that C. elegans gon-14 is required for gonadogenesis. Here we report that gon-14 encodes a protein with similarity to LIN-15B, a class B synMuv protein. An extensive region of GON-14 contains blocks of sequence similarity to transposases of the hAT superfamily, but key residues are not conserved, suggesting a distant relationship. GON-14 also contains a putative THAP DNA-binding domain. A rescuing gon-14∷GON-14∷VENUS reporter is broadly expressed during development and localizes to the nucleus. Strong loss-of-function and predicted null gon-14 alleles have pleiotropic defects, including multivulval (Muv) defects and temperature-sensitive larval arrest. Although the gon-14 Muv defect is not enhanced by synMuv mutations, gon-14 interacts genetically with class B and class C synMuv genes, including lin-35/Rb, let-418/Mi-2β, and trr-1/TRRAP. The gon-14; synMuv double mutants arrest as larvae when grown under conditions supporting development to adulthood for the respective single mutants. The gon-14 larval arrest is suppressed by loss of mes-2/E(Z), mes-6/ESC, or mes-4, which encodes a SET domain protein. Additionally, gon-14 affects expression of pgl-1 and lag-2, two genes regulated by the synMuv genes. We suggest that gon-14 functions with class B and class C synMuv genes to promote larval growth, in part by antagonizing MES-2,3,6/ESC-E(z) and MES-4.
The Effects of Dominance, Regular Inbreeding and Sampling Design on QST, an Estimator of Population Differentiation for Quantitative Traits
Authors: Goudet Jérôme , Büchi Lucie
Published: December 2005

To test whether quantitative traits are under directional or homogenizing selection, it is common practice to compare population differentiation estimates at molecular markers (FST) and quantitative traits (QST). If the trait is neutral and its determinism is additive, then theory predicts that QST = FST, while QST > FST is predicted under directional selection for different local optima, and QST < FST is predicted under homogenizing selection. However, nonadditive effects can alter these predictions. Here, we investigate the influence of dominance on the relation between QST and FST for neutral traits. Using analytical results and computer simulations, we show that dominance generally deflates QST relative to FST. Under inbreeding, the effect of dominance vanishes, and we show that for selfing species, a better estimate of QST is obtained from selfed families than from half-sib families. We also compare several sampling designs and find that it is always best to sample many populations (>20) with few families (five) rather than few populations with many families. Provided that estimates of QST are derived from individuals originating from many populations, we conclude that the pattern QST > FST, and hence the inference of directional selection for different local optima, is robust to the effect of nonadditive gene actions.
Mesenchymal Stem Cells Spontaneously Express Neural Proteins in Culture and Are Neurogenic after Transplantation
Authors: Deng Jie , Petersen Bryon E. , Steindler Dennis A. , Jorgensen Marda L. , Laywell Eric D.
Published: December 2005

Reports of neural transdifferentiation of mesenchymal stem cells (MSCs) suggest the possibility that these cells may serve as a source for stem cell–based regenerative medicine to treat neurological disorders. However, some recent studies controvert previous reports of MSC neurogenecity. In the current study, we evaluate the neural differentiation potential of mouse bone marrow–derived MSCs. Surprisingly, we found that MSCs spontaneously express certain neuronal phenotype markers in culture, in the absence of specialized induction reagents. A previously published neural induction protocol that elevates cytoplasmic cyclic AMP does not upregulate neuron-specific protein expression significantly in MSCs but does significantly increase expression of the astrocyte-specific glial fibrillary acidic protein. Finally, when grafted into the lateral ventricles of neonatal mouse brain, MSCs migrate extensively and differentiate into olfactory bulb granule cells and periventricular astrocytes, without evidence of cell fusion. These results indicate that MSCs may be “primed” toward a neural fate by the constitutive expression of neuronal antigens and that they seem to respond with an appropriate neural pattern of differentiation when exposed to the environment of the developing brain.
Flt3-Ligand–Mobilized Peripheral Blood, but Not Flt3-Ligand–Expanded Bone Marrow, Facilitating Cells Promote Establishment of Chimerism and Tolerance
Authors: Huang Yiming , Kucia Magda , Rezzoug Francine , Ratajczak Janina , Tanner Michael K. , Ratajczak Mariusz Z. , Schanie Carrie L. , Xu Hong , Fugier-Vivier Isabelle , Ildstad Suzanne T.
Published: December 2005

Facilitating cells (CD8+/TCR−) (FCs) enhance engraftment of limiting numbers of hematopoietic stem cells (HSCs). The primary component of FCs is precursor-plasmacytoid dendritic cells (p-preDCs), a tolerogenic cell expanded by Flt3-ligand (FL). In this study, we evaluated the function and composition of FL-expanded FCs. FL treatment resulted in a significant increase of FCs in bone marrow (BM) and peripheral blood (PB). When FL-expanded FCs were transplanted with c-Kit+/Sca-1+/Lin− (KSL) cells into allogeneic recipients, BM-FCs exhibited significantly impaired function whereas PB-FCs were potently functional. A significant upregulation of P-selectin expression and downregulation of VCAM-1 (vascular cell adhesion molecule 1) were present on FL-expanded PB-FCs compared with FL BM-FCs. Stromal cell–derived factor-1 (SDF-1), and CXCR4 transcripts were significantly increased in FL PB-FCs and decreased in FL BM-FCs. Supernatant from FL PB-FCs primed HSC migration to SDF-1, confirming production of the protein product. The FL PB-FCs contained a predominance of p-preDCs and natural killer (NK)–FCs, and NK-FCs were lacking in FL BM-FCs. The impaired function for BM-FCs was restored within 5 days after cessation of treatment. Taken together, these data suggest that FCs may enhance HSC homing and migration via the SDF-1/CXCR4 axis and adhesion molecule modulation. These findings may have implications in development of strategies for retaining function of ex vivo manipulated FCs and HSCs.
Genomic actions of the androgen receptor are required for normal male sexual differentiation in a mouse model
Authors: Notini A J , Davey R A , McManus J F , Bate K L , Zajac J D
Published: December 2005

Androgens mediate their effects in target cells via the androgen receptor (AR), which acts predominantly as a ligand-dependent transcription factor. In addition, androgens induce rapid activation of second messenger signal transduction cascades, and this is thought to occur via non-genomic mechanisms. We have used the Cre/loxP system to generate an AR knockout (ARKO) mouse targeting exon 3, which encodes the second zinc finger of the DNA-binding domain. To generate universal ARKO mice, floxed AR mice were mated with CMV-Cre mice, which express Cre recombinase ubiquitously. Deletion of the floxed allele in our mice does not disrupt the reading frame, and has been designed so that the mutant AR can bind ligand but not target genes. ARKO males displayed a complete androgen insensitivity phenotype, with female external genitalia and a reduction in body weight compared with wild-type males (P < 0.001). Testes of ARKO males were smaller than control males (P < 0.0001) and were located intra-abdominally. We have demonstrated that genotypically XY mice lacking the second zinc finger of the AR have a female phenotype, and we conclude that the genomic actions of the AR (mediated by DNA binding) are indispensable for normal male sexual differentiation.
Wave overtopping at vertical and steep seawalls
Authors: Allsop W. , Bruce T. , Pearson J. , Besley P.
Published: November 2005

Wave overtopping is the critical response of most sea defence structures and one of the more important responses for many coast defences around the UK and other developed shorelines. Sea defences in rural areas are commonly provided by embankment seawalls. Steep or vertical seawalls are more commonly used in urban areas to protect against erosion, flooding and local overtopping hazards, and to protect the base of eroding cliffs in urban or rural areas. Vertically faced breakwaters have been common around many European countries, and caisson-type breakwaters are heavily used in Japan. For simple slopes or embankments, overtopping performance can be predicted by simple monotonic empirical formulae. Overtopping of vertical or slightly battered walls is however rather more complicated, with substantial differences in overtopping volumes and velocities depending on the form of wave interaction at or close to the wall. The present paper draws together results from a number of UK and European research projects over the last 10 years. Their results improve and validate prediction methods for wave overtopping discharges and velocities for steep battered, composite and vertical seawalls/breakwaters. The methods presented herein support, extend and qualify guidance given in the UK Environment Agency overtopping manual. The present paper shows how the use of these methods can now explain why overtopping of some seawalls/breakwaters can be greatest at mid-water level, rather than at the highest water levels.
Individual and Contextual Determinants of Domestic Violence in North India
Authors: Koenig Michael A. , Stephenson Rob , Ahmed Saifuddin , Jejeebhoy Shireen J. , Campbell Jacquelyn
Published: November 2005

Objectives. We examined individual- and community-level influences on domestic violence in Uttar Pradesh, North India.

Methods. Multilevel modeling was used to explore domestic violence outcomes among a sample of 4520 married men.

Results. Recent physical and sexual domestic violence was associated with the individual-level variables of childlessness, economic pressure, and intergenerational transmission of violence. A community environment of violent crime was associated with elevated risks of both physical and sexual violence. Community-level norms concerning wife beating were significantly related only to physical violence.

Conclusions. Important similarities as well as differences were evident in risk factors for physical and sexual domestic violence. Higher socioeconomic status was found to be protective against physical but not sexual violence. Our results provide additional support for the importance of contextual factors in shaping women’s risks of physical and sexual violence.
Inequities in Mental Health Care After Health Care System Reform in Chile
Authors: Araya Ricardo , Rojas Graciela , Fritsch Rosemarie , Frank Richard , Lewis Glyn
Published: November 2005

Objectives. We compared differences in mental health needs and provision of mental health services among residents of Santiago, Chile, with private and public health insurance coverage.

Methods. We conducted a cross-sectional survey of a random sample of adults. Presence of mental disorders and use of health care services were assessed via structured interviews. Individuals were classified as having public, private, or no health insurance coverage.

Results. Among individuals with mental disorders, only 20% (95% confidence interval [CI]=16%, 24%) had consulted a professional about these problems. A clear mismatch was found between need and provision of services. Participants with public insurance coverage exhibited the highest prevalence of mental disorders but the lowest rates of consultation; participants with private coverage exhibited exactly the opposite pattern. After adjustment for age, income, and severity of symptoms, private insurance coverage (odds ratio [OR]=2.72; 95% CI=1.6, 4.6) and higher disability level (OR=1.27, 95% CI=1.1, 1.5) were the only factors associated with increased frequency of mental health consultation.

Conclusions. The health reforms that have encouraged the growth of the private health sector in Chile also have increased risk segmentation within the health system, accentuating inequalities in health care provision.
Junking Good Science: Undoing Daubert v Merrill Dow Through Cross-Examination and Argument
Authors: Givelber Daniel , Strickler Lori
Published: November 2005

For more than 40 years, the tobacco industry prevailed in lawsuits brought by injured smokers, despite overwhelming epidemiological evidence that smoking caused lung cancer. Tobacco lawyers were able to create doubt about causation. They sought to persuade jurors that “everybody knew” smoking was harmful but “nobody knows” what causes cancer by recreating in court the scientific debate resolved by the 1964 Surgeon General’s Report.

The particularistic structure of jury trials combined with the law’s mechanistic view of causation enables a defendant to contest virtually any claim concerning disease causation. Despite judicial efforts to eliminate “junk science” from lawsuits, a well-financed defendant may succeed in persuading jurors of the epidemiological equivalent of the proposition that the earth is flat.