bluuebunny/crossref_metadata_2025 huggingface dataset:
A systematic review of intravenous gamma globulin for therapy of acute myocarditis
Authors: Robinson Joan L , Hartling Lisa , Crumley Ellen , Vandermeer Ben , Klassen Terry P
Published: June 2005

Background

Intravenous gamma globulin (IVGG) is commonly used in the management of acute myocarditis. The objective of this study was to systematically review the literature evaluating this practice.

Methods

We conducted a comprehensive search (electronic databases, trials registries, conference proceedings, reference lists, contact with authors) to identify studies evaluating the use of IVGG in adults and children with a clinical or histologically proven diagnosis of myocarditis of possible viral etiology and symptoms of less than six months duration. Two reviewers independently screened the searches, applied inclusion criteria, and graded the evidence.

Results

Results were described qualitatively; data were not pooled because only one randomized controlled trial (RCT) with 62 patients was identified. The RCT showed no benefit with respect to cardiac function, functional outcome, or event-free survival. A small, uncontrolled trial (n = 10) showed significant improvement in LVEF from a mean of 24% to 41% 12 months after IVGG in nine survivors. A retrospective cohort study of pediatric patients showed improvement in cardiac function and a trend towards improved survival in patients receiving IVGG (n = 21) versus historic controls (n = 25). Ten case reports and two case series (total n = 21) described improvement in cardiac function after administration of IVGG; two case reports showed no benefit of IVGG. One case of hemolytic anemia was attributed to IVGG.

Conclusion

There is insufficient data from methodologically strong studies to recommend routine use of IVGG for acute myocarditis. Future randomized studies that take into account the etiology of acute myocarditis will be required to determine the efficacy of IVGG.
Prevalence of von Hippel-Lindau gene mutations in sporadic renal cell carcinoma: results from the Netherlands cohort study
Authors: van Houwelingen Kjeld P , van Dijk Boukje AC , Hulsbergen-van de Kaa Christina A , Schouten Leo J , Gorissen Hanneke JM , Schalken Jack A , van den Brandt Piet A , Oosterwijk Egbert
Published: June 2005

Background

Biallelic von Hippel-Lindau (VHL) gene defects, a rate-limiting event in the carcinogenesis, occur in approximately 75% of sporadic clear-cell Renal Cell Carcinoma (RCC). We studied the VHL mutation status in a large population-based case group.

Methods

Cases were identified within the Netherlands cohort study on diet and cancer, which includes 120,852 men and women. After 11.3 years of follow-up, 337 incident cases with histologically confirmed epithelial cancers were identified. DNA was isolated from paraffin material collected from 51 pathology laboratories and revised by one pathologist, leaving material from 235 cases. VHL mutational status was assessed by SSCP followed by direct sequencing, after testing SSCP as a screening tool in a subsample.

Results

The number of mutations was significantly higher for clear-cell RCC compared to other histological types. We observed 131 mutations in 114 out of 187 patients (61%) with clear-cell RCC. The majority of mutations were truncating mutations (47%). The mean tumor size was 72.7 mm for mutated tumors compared to 65.3 mm for wildtype tumors (p = 0.06). No statistically significant differences were observed for nuclear grade, TNM distribution or stage. In other histological types, we observed 8 mutations in 7 out of 48 patients (15%), 1 mutation in 1 of 6 oncocytoma, 3 mutations in 2 of 7 chromophobe RCC, 2 mutations in 2 of 30 papillary RCC, no mutations in 1 collecting duct carcinoma and 2 mutations in 2 of 4 unclassified RCC.

Conclusion

VHL mutations were detected in 61% of sporadic clear-cell RCC. VHL mutated and wildtype clear-cell RCC did not differ with respect to most parameters.
Alterations of prolyl endopeptidase activity in the plasma of children with autistic spectrum disorders
Authors: Momeni Naghi , Nordström Berit M , Horstmann Vibeke , Avarseji Hassan , Sivberg Bengt V
Published: June 2005

Background

Prolyl Endopeptidase (PEP, EC 3.4.21.26), a cytosolic endopeptidase, hydrolyses peptide bonds on the carboxyl side of proline residue in proteins with a relatively small molecular weight. It has been shown that altered PEP activity is associated with various psychological diseases such as schizophrenia, mania and depression. Autistic Spectrum Disorders (ASD) are neuropsychiatric and behavioural syndromes affecting social behaviours and communication development. They are classified as developmental disorders. The aim of this study was to examine the hypothesis that PEP activity is also associated with ASDs.

Methods

Fluorometric assay was used to measure PEP activity in EDTA plasma in children with ASD (n = 18) aged 4–12 years (mean ± SD: 7.9 ± 2.5). These results were then compared to PEP activity in a control group of non-ASD children (n = 15) aged 2–10 years (mean ± SD: 6.4 ± 2.2).

Results

An alteration in PEP activity was found in the children with ASD compared to the control group. There was much greater variation of PEP activity in the group of ASD children when compared to the controls (SD= 39.9 and SD 9.6, respectively). This variation was significant (p < 0.0005), although the mean level of PEP activity in the group of ASD children was slightly higher than in the control group (124.4 and 134.1, respectively).

Conclusion

Our preliminary finding suggests a role for PEP enzyme in the pathophysiology of autism but further research should be conducted to establish its role in the aetiology of psychiatric and neurological disorders, including autism and related spectrum disorders.
Rapid assessment of injection practices in Cambodia, 2002
Authors: Vong Sirenda , Perz Joseph F , Sok Srun , Som Seiharath , Goldstein Susan , Hutin Yvan , Tulloch James
Published: June 2005

Background

Injection overuse and unsafe injection practices facilitate transmission of bloodborne pathogens such as hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV). Anecdotal reports of unsafe and unnecessary therapeutic injections and the high prevalence of HBV (8.0%), HCV (6.5%), and HIV (2.6%) infection in Cambodia have raised concern over injection safety. To estimate the magnitude and patterns of such practices, a rapid assessment of injection practices was conducted.

Methods

We surveyed a random sample of the general population in Takeo Province and convenience samples of prescribers and injection providers in Takeo Province and Phnom Penh city regarding injection-related knowledge, attitudes, and practices. Injection providers were observed administering injections. Data were collected using standardized methods adapted from the World Health Organization safe injection assessment guidelines.

Results

Among the general population sample (n = 500), the overall injection rate was 5.9 injections per person-year, with 40% of participants reporting receipt of ≥ 1 injection during the previous 6 months. Therapeutic injections, intravenous infusions, and immunizations accounted for 74%, 16% and 10% of injections, respectively. The majority (>85%) of injections were received in the private sector. All participants who recalled their last injection reported the injection was administered with a newly opened disposable syringe and needle. Prescribers (n = 60) reported that 47% of the total prescriptions they wrote included a therapeutic injection or infusion. Among injection providers (n = 60), 58% recapped the syringe after use and 13% did not dispose of the used needle and syringe appropriately. Over half (53%) of the providers reported a needlestick injury during the previous 12 months. Ninety percent of prescribers and injection providers were aware HBV, HCV, and HIV were transmitted through unsafe injection practices. Knowledge of HIV transmission through "dirty" syringes among the general population was also high (95%).

Conclusion

Our data suggest that Cambodia has one of the world's highest rates of overall injection usage, despite general awareness of associated infection risks. Although there was little evidence of reuse of needles and syringes, support is needed for interventions to address injection overuse, healthcare worker safety and appropriate waste disposal.
Which young physicians are satisfied with their work? A prospective nationwide study in Norway
Authors: Finset Kristine Benedictow , Gude Tore , Hem Erlend , Tyssen Reidar , Ekeberg Oivind , Vaglum Per
Published: June 2005

Background

Few studies have investigated personality and medical school variables in regard to job satisfaction after graduation. It is of great importance to investigate these factors because this information may be used in the recruitment/admittance process to medical schools, and possibly to improve medical education.

Methods

We conducted a nationwide prospective 10-year follow-up study of medical students at all medical schools in Norway. They were approached three times during their medical training: at very beginning (T1), in the middle (T2), in the last year of medical school (T3), and then four years after graduation (T4). There were 210 participants who responded on all four occasions. Job satisfaction was measured with the Job Satisfaction Scale, which was used as the outcome variable. In addition to conducting multiple regression analysis for the total sample, we also conducted similar analyses separately for men and women.

Results

Among the demographic and personality variables, 'having a father who is a physician' and 'interpersonal functioning (being withdrawn)' were significantly associated with job satisfaction at T4. Among the medical school variables, 'well-being with peers', 'identification with the doctor's role at the end of curriculum', 'perceived medical school stress', and 'perceived clinical skills' were significantly associated with job satisfaction. In the multiple regression analysis only 'father as a physician' and 'perceived clinical skills' yielded an independent influence on the outcome variable in separate analyses within sub-groups of male and female students, 'perceived clinical skills' differentiated among woman only, while 'well-being with peers' differentiated only among men.

Conclusion

The main finding of this study is that the young physicians who are the most satisfied in their work are those whose fathers are physicians and those who have a high level of perceived clinical skills at the end of medical school. There are also differences in regard to predictors of job satisfaction among men and women. These findings indicate that medical schools should invest substantial effort in clinical skills training, and this seems to be especially important among female students.
A GFP-based genetic screen reveals mutations that disrupt the architecture of the zebrafish retinotectal projection
Authors: Xiao Tong , Roeser Tobias , Staub Wendy , Baier Herwig
Published: June 2005

The retinotectal projection is a premier model system for the investigation of molecular mechanisms that underlie axon pathfinding and map formation. Other important features, such as the laminar targeting of retinal axons, the control of axon fasciculation and the intrinsic organization of the tectal neuropil, have been less accessible to investigation. In order to visualize these processes in vivo, we generated a transgenic zebrafish line expressing membrane-targeted GFP under control of the brn3c promoter/enhancer. The GFP reporter labels a distinct subset of retinal ganglion cells (RGCs),which project mainly into one of the four retinorecipient layers of the tectum and into a small subset of the extratectal arborization fields. In this transgenic line, we carried out an ENU-mutagenesis screen by scoring live zebrafish larvae for anatomical phenotypes. Thirteen recessive mutations in 12 genes were discovered. In one mutant, ddl, the majority of RGCs fail to differentiate. Three of the mutations, vrt, late and tard, delay the orderly ingrowth of retinal axons into the tectum. Two alleles of drg disrupt the layer-specific targeting of retinal axons. Three genes, fuzz, beyo and brek, are required for confinement of the tectal neuropil. Fasciculation within the optic tract and adhesion within the tectal neuropil are regulated by vrt, coma, bluk, clew and blin. The mutated genes are predicted to encode molecules essential for building the intricate neural architecture of the visual system.
Essential role of protein kinase Bγ (PKBγ/Akt3) in postnatal brain development but not in glucose homeostasis
Authors: Tschopp Oliver , Yang Zhong-Zhou , Brodbeck Daniela , Dummler Bettina A. , Hemmings-Mieszczak Maja , Watanabe Takashi , Michaelis Thomas , Frahm Jens , Hemmings Brian A.
Published: June 2005

Protein kinase B is implicated in many crucial cellular processes, such as metabolism, apoptosis and cell proliferation. In contrast to Pkbα and Pkbβ-deficient mice, Pkbγ-/- mice are viable, show no growth retardation and display normal glucose metabolism. However, in adult Pkbγmutant mice, brain size and weight are dramatically reduced by about 25%. In vivo magnetic resonance imaging confirmed the reduction of Pkbγ-/- brain volumes with a proportionally smaller ventricular system. Examination of the major brain structures revealed no anatomical malformations except for a pronounced thinning of white matter fibre connections in the corpus callosum. The reduction in brain weight of Pkbγ-/- mice is caused, at least partially, by a significant reduction in both cell size and cell number. Our results provide novel insights into the physiological role of PKBγ and suggest a crucial role in postnatal brain development.
FGF signals specifically regulate the structure of hair shaft medulla via IGF-binding protein 5
Authors: Schlake Thomas
Published: June 2005

Reciprocal interactions between the dermal papilla and the hair matrix control proliferation and differentiation in the mature hair follicle. Analysis of expression suggests an important role for FGF7 and FGF10, as well as their cognate receptor FGFR2-IIIb, in these processes. Transgenic mice that express a soluble dominant-negative version of this receptor in differentiating hair keratinocytes were generated to interfere with endogenous FGF signalling. Transgenic mice develop abnormally thin but otherwise normal hairs, characterised by single columns of medulla cells in all hair types. All structural defects and the accompanying changes of global gene expression patterns are restricted to the hair medulla. Forced transgenic expression of IGF-binding protein 5, whose expression level is elevated upon suppression of FGFR2-IIIb-mediated signalling largely phenocopies the defect of dnFgfr2-IIIb-expressing hairs. Thus, the results identify Igfbp5-mediated FGFR2-IIIb signals as a key regulator of the genetic program that controls the structure of the hair shaft medulla.
Analysis ofMsx1;Msx2double mutants reveals multiple roles for Msx genes in limb development
Authors: Lallemand Yvan , Nicola Marie-Anne , Ramos Casto , Bach Antoine , Cloment Cécile Saint , Robert Benoît
Published: June 2005

The homeobox-containing genes Msx1 and Msx2 are highly expressed in the limb field from the earliest stages of limb formation and,subsequently, in both the apical ectodermal ridge and underlying mesenchyme. However, mice homozygous for a null mutation in either Msx1 or Msx2 do not display abnormalities in limb development. By contrast, Msx1; Msx2 double mutants exhibit a severe limb phenotype. Our analysis indicates that these genes play a role in crucial processes during limb morphogenesis along all three axes. Double mutant limbs are shorter and lack anterior skeletal elements (radius/tibia, thumb/hallux). Gene expression analysis confirms that there is no formation of regions with anterior identity. This correlates with the absence of dorsoventral boundary specification in the anterior ectoderm, which precludes apical ectodermal ridge formation anteriorly. As a result, anterior mesenchyme is not maintained, leading to oligodactyly. Paradoxically, polydactyly is also frequent and appears to be associated with extended Fgf activity in the apical ectodermal ridge, which is maintained up to 14.5 dpc. This results in a major outgrowth of the mesenchyme anteriorly, which nevertheless maintains a posterior identity, and leads to formation of extra digits. These defects are interpreted in the context of an impairment of Bmp signalling.
Gsh2 is required for the repression of Ngn1 and specification of dorsal interneuron fate in the spinal cord
Authors: Kriks Sonja , Lanuza Guillermo M. , Mizuguchi Rumiko , Nakafuku Masato , Goulding Martyn
Published: June 2005

The molecular programs that specify progenitors in the dorsal spinal cord remain poorly defined. The homeodomain transcription factor Gsh2 is expressed in the progenitors of three dorsal interneuron subtypes, dI3, dI4 and dI5 neurons, whereas Gsh1 is only expressed in dI4 and dI5 progenitors. Mice lacking Gsh2 exhibit a selective loss of dI3 interneurons that is accompanied by an expansion of the dI2 progenitor domain. In Gsh2mutant embryos, expression of the proneural bHLH protein Mash1 is downregulated in dI3 neural progenitors, with Mash1 mutants exhibiting a concordant reduction in dI3 neurons. Conversely, overexpression of Gsh2 and Mash1 leads to the ectopic production of dI3 neurons and a concomitant repression of Ngn1 expression. Our results provide evidence that genetic interactions involving repression of Ngn1 by Gsh2promote the differentiation of dI3 neurons from class A progenitors.
Mouse intraflagellar transport proteins regulate both the activator and repressor functions of Gli transcription factors
Authors: Liu Aimin , Wang Baolin , Niswander Lee A.
Published: June 2005

Intraflagellar transport (IFT) is an active event in which cargo is transported along microtubules by motor proteins such as kinesin and dynein. IFT proteins are required for the formation and maintenance of flagella and cilia. We have previously shown that mouse mutants for two IFT proteins, IFT88 and IFT172, as well as Kif3a, a subunit of mouse kinesin 2, exhibit ventral spinal cord patterning defects that appear to result from reduced hedgehog(Hh) signaling. Although genetic epistasis experiments place IFT proteins downstream of the Hh receptor and upstream of the Gli transcription factors,the mechanism by which IFT regulates Gli function is unknown.

The developing limb provides an excellent system to study Hh signaling, in particular as it allows a biological and molecular readout of both Gli activator and repressor function. Here we report that homozygous mutants for flexo (Fxo), a hypomorphic allele of mouse IFT88 generated in our ENU mutagenesis screen, exhibit polydactyly in all four limbs. Molecular analysis indicates that expression domains of multiple posteriorly restricted genes are expanded anteriorly in the mutant limbs, similar to loss of Gli3 transcriptional repressor function. Sonic hedgehog (Shh) expression is normal, yet Ptch1 and Gli1, two known targets of Hh signaling, are greatly reduced, consistent with loss of Shh signaling. Expression of Gli3 and Hand2 in the mutant limb indicates that the limb prepattern is abnormal. In addition, we show that partial loss-of-function mutations in another mouse IFT gene, Ift52(Ngd5), result in similar phenotypes and abnormal Hh signaling as Fxo, indicating a general requirement for IFT proteins in Hh signaling and patterning of multiple organs. Analysis of Ift88 and Shh double mutants indicates that, in mouse, IFT proteins are required for both Gli activator and repressor functions, and Gli proteins are insensitive to Hh ligand in the absence of IFT proteins. Finally, our biochemical studies demonstrate that IFT proteins are required for proteolytic processing of Gli3 in mouse embryos. In summary, our results indicate that IFT function is crucial in the control of both the positive and negative transcriptional activities of Gli proteins, and essential for Hh ligand-induced signaling cascade.
Do monarch butterflies use polarized skylight for migratory orientation?
Authors: Stalleicken Julia , Mukhida Maya , Labhart Thomas , Wehner Rüdiger , Frost Barrie , Mouritsen Henrik
Published: June 2005

To test if migratory monarch butterflies use polarized light patterns as part of their time-compensated sun compass, we recorded their virtual flight paths in a flight simulator while the butterflies were exposed to patches of naturally polarized blue sky, artificial polarizers or a sunny sky. In addition, we tested butterflies with and without the polarized light detectors of their compound eye being occluded. The monarchs' orientation responses suggested that the butterflies did not use the polarized light patterns as a compass cue, nor did they exhibit a specific alignment response towards the axis of polarized light. When given direct view of the sun, migratory monarchs with their polarized light detectors painted out were still able to use their time-compensated compass: non-clockshifted butterflies, with their dorsal rim area occluded, oriented in their typical south–southwesterly migratory direction. Furthermore, they shifted their flight course clockwise by the predicted ∼90° after being advance clockshifted 6 h. We conclude that in migratory monarch butterflies, polarized light cues are not necessary for a time-compensated celestial compass to work and that the azimuthal position of the sun disc and/or the associated light-intensity and spectral gradients seem to be the migrants' major compass cue.
Specificity of the fluorescein transport process in Malpighian tubules of the cricketAcheta domesticus
Authors: Neufeld Douglas S. G. , Kauffman Ross , Kurtz Zachary
Published: June 2005

We demonstrate the presence of an efficient, multispecific transport system for excretion of organic anions in the Malpighian tubules of the cricket Acheta domesticus using fluorescein (FL) as a model substrate. Malpighian tubules rapidly accumulated FL via a high affinity process(Km=7.75 μmol l–1); uptake was completely eliminated by the prototypical organic anion transport inhibitor probenecid (1 mmol l–1), but not by p-aminohippuric acid (3 mmol l–1). FL uptake was inhibited by monocarboxylic acids at a high concentration (3 mmol l–1), and inhibition was more effective with an increase in the carbon chain of the monocarboxylic acid (37% inhibition by 5-carbon valeric acid, and 89% inhibition by 7-carbon caprylic acid). Likewise, tests using a series of aliphatic glutathione conjugates indicated that only the compound with the longest side-chain(decyl-glutathione) significantly inhibited FL uptake (81% inhibition). FL uptake was inhibited by a number of xenobiotics, including a plant alkaloid(quinine), herbicides (2,4-dichlorophenoxyacetic acid and 4-(2,4-dichlorophenoxy)-butyric acid), and the insecticide metabolites malathion monocarboxylic acid (MMA) and 3-phenoxybenzoic acid (PBA),suggesting that this transport system plays an active role in excretion of xenobiotics from Acheta by Malpighian tubules. HPLC quantification of MMA and PBA accumulation into Malpighian tubules verified that MMA accumulation was via a mediated transport process, but suggested that PBA accumulation was by nonspecific binding. The presence of a transport system in Malpighian tubules that handles at least one pesticide metabolite(MMA) suggests that transport processes could be a mechanism conferring resistance to xenobiotic exposure in insects.
Patterns of red muscle strain/activation and body kinematics during steady swimming in a lamnid shark, the shortfin mako (Isurus oxyrinchus)
Authors: Donley Jeanine M. , Shadwick Robert E. , Sepulveda Chugey A. , Konstantinidis Peter , Gemballa Sven
Published: June 2005

The dynamics of steady swimming were examined in the shortfin mako(Isurus oxyrinchus), a member of the cartilaginous fish family Lamnidae, a family known for their morphological adaptations for high-performance locomotion and their similarity in hydromechanical design to tunas. Patterns of red muscle (RM) strain (i.e. relative length change) and activation were quantified at two axial positions (∼0.4 and 0.6L,where L is total body length), using sonomicrometry and electromyography (EMG), and correlated with simultaneous measurements of dorsal midline kinematics during steady swimming (∼0.5–1 Ls–1). RM strain varied longitudinally with strain amplitudes ranging from 5.5±1.1% (s.e.m.) in the anterior to 8.7±0.9% in the posterior. We found no significant longitudinal variation in patterns of RM activation, with mean onset of activation occurring at 83–84° (90° is peak length) and offset at 200–210° at both body positions. Likewise, duty cycles were similar:35.5±1.0% in the anterior and 32.2±1.6% in the posterior. Comparison of the timing of waves of dorsal midline curvature and predicted strain relative to measured RM strain revealed a phase shift between RM shortening and local body bending. Furthermore, when the body is bent passively, RM shortens synchronously with the surrounding white muscle (WM)and skin, as expected. During active swimming, peaks in RM strain were delayed relative to peaks in WM strain by a mean of ∼10% of the tailbeat cycle,with one individual as high as ∼17% in the anterior and nearly 50% in the posterior. The longitudinal consistency in the EMG/strain phase relationship in the mako is similar to that in the leopard shark, suggesting a consistent trend among sharks using different locomotor modes. However, unlike in the leopard shark, RM shortening in the mako is physically uncoupled from deformation of the surrounding body during steady swimming, a characteristic shared between the mako and tunas.
Beta-adrenergic control of plasma glucose and free fatty acid levels in the air-breathing African catfishClarias gariepinusBurchell 1822
Authors: van Heeswijk Johannes C. F. , Vianen Gerjanne J. , van den Thillart Guido E. E. J. M. , Zaagsma Johan
Published: June 2005

In several water-breathing fish species, β-adrenergic receptor stimulation by noradrenaline leads to a decrease in plasma free fatty acid(FFA) levels, as opposed to an increase in air-breathing mammals. We hypothesised that this change in adrenergic control is related to the mode of breathing. Therefore, cannulated air-breathing African catfish were infused for 90 min with noradrenaline or with the nonselective β-agonist,isoprenaline. To identify the receptor type involved, a bolus of either a selective β1-antagonist (atenolol) or a selectiveβ 2-antagonist (ICI 118,551) was injected 15 min prior to the isoprenaline infusion. Both noradrenaline and isoprenaline led to an expected rise in glucose concentration. Isoprenaline combined with both theβ 1- and β2-antagonist led to higher glucose concentrations than isoprenaline alone. This could indicate the presence of a stimulatory β-adrenoceptor different from β1 andβ 2-adrenoceptors; these two receptors thus seemed to mediate a reduction in plasma glucose concentration. Both noradrenaline and isoprenaline led to a significant decrease in FFA concentration. Whereas theβ 1-antagonist had no effect, the β2-antagonist reduced the decrease in FFA concentration, indicating the involvement ofβ 2-adrenoceptors. It is concluded that the air-breathing African catfish reflects water-breathing fish in the adrenergic control of plasma FFA and glucose levels.
Determination of the exact copy numbers of particular mRNAs in a single cell by quantitative real-time RT-PCR
Authors: Wagatsuma Akiko , Sadamoto Hisayo , Kitahashi Takashi , Lukowiak Ken , Urano Akihisa , Ito Etsuro
Published: June 2005

Gene expression is differently regulated in every cell even though the cells are included in the same tissue. For this reason, we need to measure the amount of mRNAs in a single cell to understand transcription mechanism better. However, there are no accurate, rapid and appropriate methods to determine the exact copy numbers of particular mRNAs in a single cell. We therefore developed a procedure for isolating a single, identifiable cell and determining the exact copy numbers of mRNAs within it. We first isolated the cerebral giant cell of the pond snail Lymnaea stagnalis as this neuron plays a key role in the process of memory consolidation of a learned behavior brought about by associative learning of feeding behavior. We then determined the copy numbers of mRNAs for the cyclic AMP-responsive element binding proteins (CREBs). These transcription factors play an important role in memory formation across animal species. The protocol uses two techniques in concert with each other: a technique for isolating a single neuron with newly developed micromanipulators coupled to an assay of mRNAs by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). The molecular assay determined the mRNA copy numbers, each of which was compared with a standard curve prepared from cDNA solutions corresponding to the serially diluted solutions of Lymnaea CREB mRNA. The standard curves were linear within a range of 10 to 105 copies, and the intra-assay variation was within 15%. Each neuron removed from the ganglia was punctured to extract the total RNA directly and was used for the assay without further purification. Using this two-step procedure, we found that the mRNA copy number of CREB repressor (CREB2) was 30–240 in a single cerebral giant cell, whereas that of CREB activator (CREB1) was below the detection limits of the assay (<25). These results suggest that the CREB cascade is regulated by an excess amount of CREB2 in the cerebral giant cells. Our procedure is the only quantitative analysis for elucidation of the dynamics of gene transcription in a single cell.
Maximal metabolic rates during voluntary exercise, forced exercise, and cold exposure in house mice selectively bred for high wheel-running
Authors: Rezende Enrico L. , Chappell Mark A. , Gomes Fernando R. , Malisch Jessica L. , Garland Theodore
Published: June 2005

Selective breeding for high wheel-running activity has generated four lines of laboratory house mice (S lines) that run about 170% more than their control counterparts (C lines) on a daily basis, mostly because they run faster. We tested whether maximum aerobic metabolic rates(V̇O2max) have evolved in concert with wheel-running, using 48 females from generation 35. Voluntary activity and metabolic rates were measured on days 5+6 of wheel access (mimicking conditions during selection), using wheels enclosed in metabolic chambers. Following this, V̇O2max was measured twice on a motorized treadmill and twice during cold-exposure in a heliox atmosphere (HeO2). Almost all measurements, except heliox V̇O2max, were significantly repeatable. After accounting for differences in body mass(S<C) and variation in age at testing, S and C did not differ in V̇O2max during forced exercise or in heliox, nor in maximal running speeds on the treadmill. However, running speeds and V̇O2max during voluntary exercise were significantly higher in S lines. Nevertheless, S mice never voluntarily achieved the V̇O2max elicited during their forced treadmill trials, suggesting that aerobic capacity per se is not limiting the evolution of even higher wheel-running speeds in these lines. Our results support the hypothesis that S mice have genetically higher motivation for wheel-running and they demonstrate that behavior can sometimes evolve independently of performance capacities. We also discuss the possible importance of domestication as a confounding factor to extrapolate results from this animal model to natural populations.
Molecular characterization and expression of the UV opsin in bumblebees:three ommatidial subtypes in the retina and a new photoreceptor organ in the lamina
Authors: Spaethe Johannes , Briscoe Adriana D.
Published: June 2005

Ultraviolet-sensitive photoreceptors have been shown to be important for a variety of visual tasks performed by bees, such as orientation, color and polarization vision, yet little is known about their spatial distribution in the compound eye or optic lobe. We cloned and sequenced a UV opsin mRNA transcript from Bombus impatiens head-specific cDNA and, using western blot analysis, detected an eye protein band of ∼41 kDa,corresponding to the predicted molecular mass of the encoded opsin. We then characterized UV opsin expression in the retina, ocelli and brain using immunocytochemistry. In the main retina, we found three different ommatidial types with respect to the number of UV opsin-expressing photoreceptor cells,namely ommatidia containing two, one or no UV opsin-immunoreactive cells. We also observed UV opsin expression in the ocelli. These results indicate that the cloned opsin probably encodes the P350 nm pigment, which was previously characterized by physiological recordings. Surprisingly, in addition to expression in the retina and ocelli, we found opsin expression in different parts of the brain. UV opsin immunoreactivity was detected in the proximal rim of the lamina adjacent to the first optic chiasm, which is where studies in other insects have found expression of proteins involved in the circadian clock, period and cryptochrome. We also found UV opsin immunoreactivity in the core region of the antennal lobe glomeruli and different clusters of perikarya within the protocerebrum, indicating a putative function of these brain regions, together with the lamina organ, in the entrainment of circadian rhythms. In order to test for a possible overlap of clock protein and UV opsin spatial expression, we also examined the expression of the period protein in these regions.
Nitric oxide and the control of catecholamine secretion in rainbow trout Oncorhynchus mykiss
Authors: McNeill B. , Perry S. F.
Published: June 2005

An in situ saline-perfused posterior cardinal vein preparation was used to assess the role of nitric oxide (NO) in the regulation of basal and stimulus-evoked catecholamine secretion from rainbow trout Oncorhynchus mykiss chromaffin cells. Addition of the NO donor, sodium nitroprusside(SNP) to the inflowing perfusate abolished catecholamine secretion during electrical field stimulation, thereby establishing the potential for NO to act as a potent inhibitor of catecholamine release. A possible role for endogenously produced NO was established by demonstrating that stimulus-evoked(depolarizing levels of KCl or electrical field stimulation) catecholamine secretion was markedly stimulated in the presence of the nitric oxide synthase(NOS) inhibitors l-NAME and 7-NI. Although in vitroexperiments demonstrated that catecholamine degradation was enhanced by NO in a dose-dependent manner, the dominant factor contributing to the reduction in catecholamine appearance in the perfusate was specific inhibition of catecholamine secretion. Subsequent experiments were performed to identify the NOS isoform(s) contributing to the inhibition of stimulus-evoked catecholamine secretion. Inducible NOS (iNOS; an enzyme that can be activated in the absence of Ca2+), although present in the vicinity of the chromaffin cells(based on mRNA measurements), does not appear to play a role because stimulus-evoked NO production was eliminated during perfusion with Ca2+-free saline. The potential involvement of endothelial NOS(eNOS) was revealed by showing that hypoxic perfusate evoked NO production and corresponded with an inhibition of stimulus-evoked catecholamine secretion;chemical removal of the endothelium (using saponin) prevented the production of NO during hypoxia. However, because removal of the endothelium did not affect NO production during electrical field stimulation, it would appear that the neuronal form of NOS (nNOS) is the key isoform modulating catecholamine secretion from trout chromaffin cells.
The behavioural transition from straight to curve walking: kinetics of leg movement parameters and the initiation of turning
Authors: Dürr Volker , Ebeling Wiebke
Published: June 2005

The control of locomotion requires the ability to adapt movement sequences to the behavioural context of the animal. In hexapod walking, adaptive behavioural transitions require orchestration of at least 18 leg joints and twice as many muscle groups. Although kinematics of locomotion has been studied in several arthropod species and in a range of different behaviours,almost nothing is known about the transition from one behavioural state to another. Implicitly, most studies on context-dependency assume that all parameters that undergo a change during a behavioural transition do so at the same rate. The present study tests this assumption by analysing the sequence of kinematic events during turning of the stick insect Carausius morosus, and by measuring how the time courses of the changing parameters differ between legs. Turning was triggered reliably at a known instant in time by means of the optomotor response to large-field visual motion. Thus, knowing the start point of the transition, the kinematic parameters that initiate turning could be ranked according to their time constants.

Kinematics of stick insect walking vary considerably among trials and within trials. As a consequence, the behavioural states of straight walking and curve walking are described by the distributions of 13 kinematic parameters per leg and of orientation angles of head and antennae. The transitions between the behavioural states are then characterised by the fraction of the variance within states by which these distributions differ,and by the rate of change of the corresponding time courses. The antennal optomotor response leads that of the locomotor system. Visually elicited turning is shown to be initiated by stance direction changes of both front legs. The transition from straight to curve walking in stick insects follows different time courses for different legs, with time constants of kinematic parameters ranging from 1.7 s to more than 3 s. Therefore, turning is a behavioural transition that involves a characteristic orchestration of events rather than synchronous parallel actions with a single time constant.
The tube feet of sea urchins and sea stars contain functionally different mutable collagenous tissues
Authors: Santos Romana , Haesaerts Delphine , Jangoux Michel , Flammang Patrick
Published: June 2005

Echinoderms possess mutable collagenous tissues (MCTs), which are capable of undergoing rapid changes in their passive mechanical properties mediated by secretions from a specific cell type, the juxtaligamental cell. In this study,the possible presence of MCTs in the tube feet of the echinoid Paracentrotus lividus and the asteroid Marthasterias glacialis was investigated by measuring their extensibility, tensile strength, stiffness and toughness after different treatments known to influence the physiological state of MCTs. Calcium removal reversibly induced a significant plasticization of the tube feet of both species. When exposed to cell-disrupting solutions, the tube foot stem of sea urchins and sea stars showed a significant increase in strength, stiffness and toughness in the absence of calcium. This response, combined with the ultrastructural observation of juxtaligamental-like cells in the connective tissue, confirms that an MCT is present in both echinoid and asteroid tube feet. It was observed, however, that the tube foot stems of P. lividus and M. glacialis are affected differently by exposure to cell-disrupting solutions in the presence of calcium, indicating that their MCTs could be functionally different. In their soft state, MCTs could assist the muscles in tube foot protraction, bending and retraction; in their stiff state, they could play a role in the energy-sparing maintenance of position; for example,during strong attachment to the substratum to resist hydrodynamically generated loads.
The mechanical scaling of coasting in zebrafish (Danio rerio)
Authors: McHenry Matthew J. , Lauder George V.
Published: June 2005

Many fish species span two or three orders of magnitude in length during the growth from larvae to adults, and this change may have dramatic consequences for locomotor performance. We measured how the performance of coasting changes over the life history of zebrafish (Danio rerio) and examined the scaling of mechanics underlying this change. Adult zebrafish coast disproportionately further and faster and maintain their speed for a longer duration than do larvae and juveniles. Measurements of drag on tethered dead fish suggest that adult fish operate in an inertial regime by coasting at relatively high Reynolds numbers (Re>1000), and in vivodrag measurements showed adults to operate with a drag coefficient(Cinert≈0.024) that was consistent with previously published estimates. However, drag scaled differently at lower Revalues than those assumed in previous studies. We found a viscous regime at Re<300, which corresponds to the routine coasting of larvae and juveniles. Despite these changes in hydrodynamics over growth, a mathematical model of coasting mechanics suggests that the disproportionately longer coasting of adults is caused primarily by their large body mass and high speed at the beginning of coasting. We therefore propose that changes in coasting performance with growth are dictated primarily by the scaling of momentum rather than resulting from hydrodynamic changes. These results provide an opportunity for new interpretations of function in the growth and evolution of fish.
Adaptations to an extreme environment: retinal organisation and spectral properties of photoreceptors in Antarctic notothenioid fish
Authors: Pointer Marie A. , Cheng Chi-Hing Christina , Bowmaker James K. , Parry Juliet W. L. , Soto Nelyn , Jeffery Glen , Cowing Jill A. , Hunt David M.
Published: June 2005

The Notothenioid suborder of teleosts comprises a number of species that live below the sea ice of the Antarctic. The presence of `antifreeze'glycoproteins in these fish as an adaptation to freezing temperature has been well documented but little is known about the adaptations of the visual system of these fish to a light environment in which both the quantity and spectral composition of downwelling sunlight has been reduced by passage through ice and snow. In this study, we show that the red/long-wave sensitive (LWS) opsin gene is not present in these fish but a UV-sensitive short-wave sensitive(SWS1) pigment is expressed along with blue-sensitive (SWS2) and green/middle-wave sensitive (Rh2) pigments. The identity and spectral location of maximal absorbance of the SWS1 and Rh2 pigments was confirmed by in vitro expression of the recombinant opsins followed by regeneration with 11-cis retinal. Only the SWS2 pigment showed interspecific variations in peak absorbance. Expression of the Rh2 opsin is localised to double cone receptors in both the central and peripheral retina, whereas SWS2 opsin expression is present only in the peripheral retina. SWS1 cones could not be identified by either microspectrophotometry or in situ hybridisation,presumably reflecting their low number and/or uneven distribution across the retina. A study of photoreceptor organisation in the retina of two species,the shallower dwelling Trematomus hansoni and the deeper dwelling Dissostichus mawsoni, identified a square mosaic in the former, and a row mosaic in the latter species; the row mosaic in Dissostichus mawsoni with less tightly packed cone photoreceptors allows for a higher rod photoreceptor density.
Digestive state influences the heart rate hysteresis and rates of heat exchange in the varanid lizardVaranus rosenbergi
Authors: Clark T. D. , Butler P. J. , Frappell P. B.
Published: June 2005

To maximize the period where body temperature (Tb)exceeds ambient temperature (Ta), many reptiles have been reported to regulate heart rate (fh) and peripheral blood flow so that the rate of heat gain in a warming environment occurs more rapidly than the rate of heat loss in a cooling environment. It may be hypothesized that the rate of cooling, particularly at relatively cool Tbs, would be further reduced during postprandial periods when specific dynamic action (SDA) increases endogenous heat production (i.e. the heat increment of feeding). Furthermore, it may also be hypothesized that the increased perfusion of the gastrointestinal organs that occurs during digestion may limit peripheral blood flow and thus compromise the rate of heating. Finally, if the changes in fh are solely for the purpose of thermoregulation, there should be no associated changes in energy demand and, consequently, no hysteresis in the rate of oxygen consumption(V̇O2). To test these hypotheses, seven individual Varanus rosenbergi were heated and cooled between 19°C and 35°C following at least 8 days fasting and then approximately 25 h after consumption of a meal (mean 10% of fasted body mass). For a given Tb between the range of 19-35°C, fh of fasting lizards was higher during heating than during cooling. Postprandial lizards also displayed a hysteresis in fh, although the magnitude was reduced in comparison with that of fasting lizards as a result of a higher fh during cooling in postprandial animals. Both for fasting and postprandial lizards,there was no hysteresis in V̇O2 at any Tb throughout the range although, as a result of SDA,postprandial animals displayed a significantly higher V̇O2 than fasting animals both during heating and during cooling at Tbs above 24°C. The values of fh during heating at a given Tb were the same for fasting and postprandial animals,which, in combination with a slower rate of heating in postprandial animals,suggests that a prioritization of blood flow to the gastrointestinal organs during digestion is occurring at the expense of higher rates of heating. Additionally, postprandial lizards took longer to cool at Tbs below 23°C, suggesting that the endogenous heat produced during digestion temporarily enhances thermoregulatory ability at lower temperatures, which would presumably assist V. rosenbergiduring cooler periods in the natural environment by augmenting temperature-dependent physiological processes.
Formation of Selective Adsorption Cavity by Chemical Vapor Deposition of Molecular Sieving Silica Overlayer on Alumina using Molecular Template in the Presence of Acetic Acid
Authors: Katada Naonobu , Akazawa Sachiko , Nishiaki Noriko , Yano Yuiko , Yamakita Shohei , Hayashi Kentaro , Niwa Miki
Published: June 2005

Chemical vapor deposition (CVD) of tetramethoxysilane was carried out on alumina after the pre-adsorption of aldehyde as the molecular template, in order to form a molecular-sieving silica overlayer with controlled cavities. The adsorption of a molecule (1-naphthaldehyde) larger than the template was almost completely suppressed when acetic acid was added during CVD, showing a high selectivity based upon the shapes of the controlled cavity and the adsorbed molecule, while the selectivity was low when acetic acid was not used. The infrared (IR) spectrum and nuclear magnetic resonance (NMR) of 29Si showed that the acetic acid enhanced the oligomerization of Si alkoxide via hydrolysis, probably by acid catalysis, resulting in the formation of a dense network of siloxane. Such a dense wall of silica is speculated to determine the shape of the adsorption cavity precisely, and to generate high selectivity.