bluuebunny/crossref_metadata_2025 huggingface dataset:
LMP4 regulates Tbx5 protein subcellular localization and activity
Authors: Camarata Troy , Bimber Benjamin , Kulisz Andre , Chew Teng-Leong , Yeung Jennifer , Simon Hans-Georg
Published: August 2006

The limb- and heart-specific Tbx5 transcription factor coexpresses with and directly binds to the novel PDZ-LIM domain protein, LMP4. LMP4 is distributed in the cytoplasm associated with the actin cytoskeleton. In the presence of LMP4, Tbx5 shuttles dynamically between the nucleus and cytoplasm and, in a complex with LMP4, localizes to actin filaments. Nuclear and cytoplasmic Tbx5 distribution in developing chicken wings suggests the functional significance of the LMP4–Tbx5 interaction. In primary epicardial cells, we demonstrate that Tbx5 protein subcellular relocalization can be stimulated by external signals that induce cell differentiation. To test whether the relocalization from nuclear to cytoplasmic sites interferes with downstream gene expression, we used limb-specific Fgf10 and heart-specific Anf promoter-luciferase reporters and demonstrate that LMP4 acts as a repressor of Tbx5 activity. These studies reveal a previously unknown mechanism for Tbx transcription factor regulation in vertebrate limb and heart development and provide a better understanding of the molecular basis of hand/heart birth defects associated with Tbx5 mutations.
Supervillin modulation of focal adhesions involving TRIP6/ZRP-1
Authors: Takizawa Norio , Smith Tara C. , Nebl Thomas , Crowley Jessica L. , Palmieri Stephen J. , Lifshitz Lawrence M. , Ehrhardt Anka G. , Hoffman Laura M. , Beckerle Mary C. , Luna Elizabeth J.
Published: August 2006

Cell–substrate contacts, called focal adhesions (FAs), are dynamic in rapidly moving cells. We show that supervillin (SV)—a peripheral membrane protein that binds myosin II and F-actin in such cells—negatively regulates stress fibers, FAs, and cell–substrate adhesion. The major FA regulatory sequence within SV (SV342-571) binds to the LIM domains of two proteins in the zyxin family, thyroid receptor–interacting protein 6 (TRIP6) and lipoma-preferred partner (LPP), but not to zyxin itself. SV and TRIP6 colocalize within large FAs, where TRIP6 may help recruit SV. RNAi-mediated decreases in either protein increase cell adhesion to fibronectin. TRIP6 partially rescues SV effects on stress fibers and FAs, apparently by mislocating SV away from FAs. Thus, SV interactions with TRIP6 at FAs promote loss of FA structure and function. SV and TRIP6 binding partners suggest several specific mechanisms through which the SV–TRIP6 interaction may regulate FA maturation and/or disassembly.
mBet3p is required for homotypic COPII vesicle tethering in mammalian cells
Authors: Yu Sidney , Satoh Ayano , Pypaert Marc , Mullen Karl , Hay Jesse C. , Ferro-Novick Susan
Published: August 2006

TRAPPI is a large complex that mediates the tethering of COPII vesicles to the Golgi (heterotypic tethering) in the yeast Saccharomyces cerevisiae. In mammalian cells, COPII vesicles derived from the transitional endoplasmic reticulum (tER) do not tether directly to the Golgi, instead, they appear to tether to each other (homotypic tethering) to form vesicular tubular clusters (VTCs). We show that mammalian Bet3p (mBet3p), which is the most highly conserved TRAPP subunit, resides on the tER and adjacent VTCs. The inactivation of mBet3p results in the accumulation of cargo in membranes that colocalize with the COPII coat. Furthermore, using an assay that reconstitutes VTC biogenesis in vitro, we demonstrate that mBet3p is required for the tethering and fusion of COPII vesicles to each other. Consistent with the proposal that mBet3p is required for VTC biogenesis, we find that ERGIC-53 (VTC marker) and Golgi architecture are disrupted in siRNA-treated mBet3p-depleted cells. These findings imply that the TRAPPI complex is essential for VTC biogenesis.
Control of Single Channel Conductance in the Outer Vestibule of the Kv2.1 Potassium Channel
Authors: Trapani Josef G. , Andalib Payam , Consiglio Joseph F. , Korn Stephen J.
Published: August 2006

Current magnitude in Kv2.1 potassium channels is modulated by external [K+]. In contrast to behavior expected from the change in electrochemical driving force, outward current through Kv2.1 channels becomes larger when extracellular [K+] is increased within the physiological range. The mechanism that underlies this unusual property involves the opening of Kv2.1 channels into one of two different outer vestibule conformations, which are defined by their sensitivity to TEA. Channels that open into a TEA-sensitive conformation generate larger macroscopic currents, whereas channels that open into a TEA-insensitive conformation generate smaller macroscopic currents. At higher [K+], more channels open into the TEA-sensitive conformation. In this manuscript, we examined the mechanism by which the conformational change produced a change in current magnitude. We started by testing the simplest hypothesis: that each pharmacologically defined channel conformation produces a different single channel conductance, one smaller and one larger, and that the [K+]-dependent change in current magnitude reflects the [K+]-dependent change in the percentage of channels that open into each of the two conformations. Using single channel and macroscopic recordings, as well as hidden Markov modeling, we were able to quantitatively account for [K+]-dependent regulation of macroscopic current with this model. Combined with previously published work, these results support a model whereby an outer vestibule lysine interferes with K+ flux through the channel, and that the [K+]-dependent change in orientation of this lysine alters single channel conductance by changing the level of this interference. Moreover, these results provide an experimental example of single channel conductance being modulated at the outer end of the conduction pathway by a mechanism that involves channel activation into open states with different outer vestibule conformations.
Indices of Surgical Site Infection Risk and Prediction of Other Adverse Outcomes During Hospitalization
Authors: Delgado-Rodríguez Miguel , Palma Silvia , Gómez-Ortega Antonio , Martínez-Gallego Gabriel , Medina-Cuadros Marcelino
Published: August 2006

Objective.

To assess which adverse postsurgical outcomes are best predicted by the Study on the Efficacy of Nosocomial Infection Control (SENIC) index and the National Nosocomial Infection Surveillance system (NNIS) index.

Design.

Prospective cohort study.

Setting.

The service of general surgery at a tertiary care hospital.

Patients.

A consecutive series of patients hospitalized for more than 1 day (n = 2,989).

Results.

The outcome best predicted by the SENIC and NNIS indices was assessed by estimating the area under the receiver operating characteristic (ROC) curve. The areas under the ROC curves for nosocomial infection and in-hospital death were higher for the NNIS index than they were for the SENIC index (P<.05). The NNIS index predicted in-hospital death better than it predicted surgical site infection (area under the ROC curve ± SE, 0.836 ± 0.022 vs 0.689 ± 0.017; P = .001).

Conclusions.

The NNIS index is superior to the SENIC index for all adverse postsurgical outcomes. Its ability to predict in-hospital mortality is clearly better than its ability to predict surgical site infection.
Impact of Postdischarge Surveillance on Surgical Site Infection Rates for Several Surgical Procedures Results From the Nosocomial Surveillance Network in The Netherlands
Authors: Manniën Judith , Wille Jan C. , Snoeren Ruud L. M. M. , van den Hof Susan
Published: August 2006

Objective.

To compare the number of surgical site infections (SSIs) registered after hospital discharge with respect to various surgical procedures and to identify the procedures for which postdischarge surveillance (PDS) is most important.

Design.

Prospective SSI surveillance with voluntary PDS. Recommended methods for PDS in the Dutch national nosocomial surveillance network are addition of a special registration card to the outpatient medical record, on which the surgeon notes clinical symptoms and whether a patient developed an SSI according to the definitions; an alternative method is examination of the outpatient medical record.

Setting.

Hospitals participating in the Dutch national nosocomial surveillance network between 1996 and 2004.

Results.

We collected data on 131,798 surgical procedures performed in 64 of the 98 Dutch hospitals. PDS was performed according to one of the recommended methods for 31,134 operations (24%) and according to another active method for 32,589 operations (25%), and passive PDS was performed for 68,075 operations (52%). Relatively more SSIs were recorded after discharge for cases in which PDS was performed according to a recommended method (43%), compared with cases in which another active PDS method was used (30%) and cases in which passive PDS was used (25%). The highest rate of SSI after discharge was found for appendectomy (79% of operations), followed by knee prosthesis surgery (64%), mastectomy (61%), femoropopliteal or femorotibial bypass (53%), and abdominal hysterectomy (53%).

Conclusions.

For certain surgical procedures, most SSIs develop after discharge. SSI rates will be underestimated if no PDS is performed. We believe we have found a feasible and sensitive method for PDS that, if patients routinely return to the hospital for a postdischarge follow-up visit, might be suitable for use internationally.
Adherence to American Society of Health-System Pharmacists Surgical Antibiotic Prophylaxis Guidelines in Iran
Authors: Askarian Mehrdad , Moraweji Ali Reza , Mirkhani Hossein , Namazi Soha , Weed Harrison
Published: August 2006

We assessed the appropriateness of surgical antibiotic prophylaxis in 6 teaching hospitals in Shiraz, Iran, using the American Society of Health-System Pharmacists guideline as a reference. We reviewed the medical records of 1,000 patients who underwent 1 of 9 different surgical procedures (1 procedure per patient). The proportion of procedures in which there was compliance with all guideline recommendations was 0.3%. The most common mistakes were overuse and misuse of antibiotics.
Gentamicin-Susceptible or Gentamicin-Resistant Methicillin-ResistantStaphylococcus aureusA Case-Case Study
Authors: Robert Jérôme , Bismuth Roland , Lemaitre Nadine , Jarlier Vincent
Published: August 2006

Gentamicin-susceptible methicillin-resistantStaphylococcus aureus(GS-MRSA) strains are replacing gentamicin-resistant MRSA (GR-MRSA) strains. We showed that exposure to other patients harboring GS-MRSA was the major risk factor for GS-MRSA acquisition, and that antimicrobial exposure, especially to β-lactams, was strongly associated with acquisition of GR-MRSA but not GS-MRSA. Gentamicin use was significantly associated with GR-MRSA acquisition in univariate analysis only.
Knowledge, Attitude, and Practices Regarding Contact Precautions Among Iranian Physicians
Authors: Askarian Mehrdad , Shiraly Ramin , Aramesh Kiarash , McLaws Mary-Louise
Published: August 2006

Objective.

To assess the knowledge, attitudes, and practices of Iranian physicians regarding contact isolation precautions.

Design.

Data were collected between May and November 2002 using a cross-sectional survey design.

Setting.

Teaching hospitals in Shiraz, Iran.

Participants.

A total of 155 physicians: 78 attending clinicians and 77 resident physician surgeons or internists.

Results.

The mean scores for knowledge and attitude were acceptable, with 71% of physicians scoring the maximum for knowledge and 65% achieving the maximum scores for attitude, whereas the mean score for practice was low, with only 26% achieving the maximum score. A good level of knowledge be associated with a good attitude (odds ratio [OR], 68.4 [95% confidence interval {CI}, 20.0-285.6]; P< .001), good practices were associated with good knowledge (OR, 22.5 [95% CI, 7.1-91.3]; P< .001), and a good attitude was associated with good practice (OR, 20.0 [95% CI, 5.7-105.2]; P<.001).

Conclusion.

Although strong associations were found among knowledge, attitude, and practice, the level of compliance with precautions was not nearly as high as it should be.
Effects of Oestrogen on Cognition: What Have We Learned From Basic Research?
Authors: Daniel J. M.
Published: August 2006

The results of clinical and basic research conducted over the past two decades have implicated a role for oestrogen in modulating cognitive function. This review focuses on what the results of research using female rodent models have revealed about the effects of oestrogen on mammalian cognition. Increased levels of oestrogen are associated with increased dendritic spine and synapse density in the hippocampus, a brain area important for learning and memory. However, the role of oestrogen in the modulation of performance on tasks of learning and memory is complex because it exerts enhancing effects on some tasks and impairing effects on others. Hypotheses have been offered to explain these varied actions, including differentiating the effects of oestrogen on cognitive processes required to complete tasks and analysing the influence of fluctuating levels of oestrogen on the strategies selected by animals to solve tasks. It is proposed that, when these hypotheses are viewed together and within the context of oestrogen action in the hippocampus and potentially other brain areas, a framework for understanding the varied effects of oestrogen on cognition emerges. The hippocampal‐dependent memory system supports the flexible expression of memories and the hippocampal‐independent memory system supports development of individual representations. Because of the effects exerted by oestrogen on the structure and function of the hippocampus, it would be expected to enhance performance across a variety of tasks that require hippocampal‐dependent flexible expression of memories and would not enhance performance on tasks that involve hippocampal‐independent individual representations. This review offers a theoretical model by which the divergent results of studies assessing the role of oestrogen on cognition can be reconciled and suggests that effects of oestrogen on cognition are best understood within the framework of oestrogen action in the brain and the role of those brain areas affected by oestrogen in the mediation of learning and memory.
Dynamics of root growth stimulation in Nicotiana tabacum in increasing light intensity
Authors: NAGEL KERSTIN A. , SCHURR ULRICH , WALTER ACHIM
Published: August 2006

Light intensity is crucial for plant growth. In this study, the hypothesis was tested whether a sudden increase in light intensity leads to an immediate increase of root growth. Seedlings of Nicotiana tabacum grown in agar‐filled Petri dishes were subjected to light intensities of 60 and 300 µmol m−2 s−1, respectively. Seedling biomass, sucrose, glucose and fructose concentration as well as primary root growth increased significantly with light intensity. The dynamics of the increase in root growth were analysed here in more detail. In transition experiments from low to high light intensities, root growth increased by a factor of four within 4 d, reaching the steady‐state level measured in plants that were cultivated in high‐light conditions. The distribution of relative elemental growth rates along the root growth zone retained a constant shape throughout this transition. During the first three hours after light increase, strong growth fluctuations were repeatedly observed with the velocity of the root tip cycling in a sinusoidal pattern between 120 and 180 µm h−1. These dynamic patterns are discussed in the context of hydraulic and photosynthetic acclimation to the altered conditions. Experiments with externally applied sucrose and with transgenic plants having reduced capacities for sucrose synthesis indicated clearly that increasing light intensity rapidly enhanced root growth by elevating sucrose export from shoot to root.
Silicon deposition in the root reduces sodium uptake in rice (Oryza sativa L.) seedlings by reducing bypass flow
Authors: GONG H. J. , RANDALL D. P. , FLOWERS T. J.
Published: August 2006

Sodium chloride reduces the growth of rice seedlings, which accumulate excessive concentrations of sodium and chloride ions in their leaves. In this paper, we describe how silicon decreases transpirational bypass flow and ion concentrations in the xylem sap in rice (Oryza sativa L.) seedlings growing under NaCl stress. Salt (50 mM NaCl) reduced the growth of shoots and roots: adding silicate (3 mM) to the saline culture solution improved the growth of the shoots, but not roots. The improvement of shoot growth in the presence of silicate was correlated with reduced sodium concentration in the shoot. The net transport rate of Na from the root to shoot (expressed per unit of root mass) was also decreased by added silicate. There was, however, no effect of silicate on the net transport of potassium. Furthermore, in salt‐stressed plants, silicate did not decrease the transpiration, and even increased it in seedlings pre‐treated with silicate for 7 d prior to salt treatment, indicating that the reduction of sodium uptake by silicate was not simply through a reduction in volume flow from root to shoot. Experiments using trisodium‐8‐hydroxy‐1,3,6‐pyrenetrisulphonic acid (PTS), an apoplastic tracer, showed that silicate dramatically decreased transpirational bypass flow in rice (from about 4.2 to 0.8%), while the apparent sodium concentration in the xylem, which was estimated indirectly from the flux data, decreased from 6.2 to 2.8 mM. Direct measurements of the concentration of sodium in xylem sap sampled using Philaenus spumarius confirmed that the apparent reduction was not a consequence of sodium recycling. X‐ray microanalysis showed that silicon was deposited in the outer part of the root and in the endodermis, being more obvious in the latter than in the former. The results suggest that silicon deposition in the exodermis and endodermis reduced sodium uptake in rice (Oryza sativa L.) seedlings under NaCl stress through a reduction in apoplastic transport across the root.
The Occupational Performance History Interview in community mental health case management: Consumer and occupational therapist perspectives
Authors: Ennals Priscilla , Fossey Ellie
Published: August 2006

Background/aim:  This study aimed to explore the use of the Occupational Performance History Interview from the perspectives of mental health consumers and occupational therapist case managers in community mental health settings.

Methods:  This qualitative study, based on naturalistic inquiry, involved 10 participants: four occupational therapists who interviewed six mental health consumers, using the Occupational Performance History Interview. All participants participated in follow‐up interviews or in focus groups to gain an understanding of their perspectives of this experience. Transcribed data were analysed using the constant comparative method.

Results:  Consumer participants valued telling their occupational stories. Occupational therapist participants described the interview as supporting them to be more occupationally focused within their case management roles.

Conclusion:  The findings suggest that the Occupational Performance History Interview can potentially enhance client‐centred and occupationally focused practice in community mental health case management.
Note on Longevity and Aggregate Savings
Authors: Sheshinski Eytan
Published: August 2006

In a recent paper in this journal, Bloom, Canning and Graham (2003) model the effect of changes in longevity on individual savings. They proceed to present empirical findings about the relation between longevity and aggregate savings. There is a missing link between their empirics and theory: the changes in the populations age density distribution due to increased longevity. This note provides such an aggregation analysis within a simple model with uncertain survival, endogenous life‐cycle consumption and retirement age. It is shown that, with continuous annuitization, an increase in expected longevity raises aggregate steady‐state savings. The magnitude of this effect depends on the economy's age‐specific distribution and on the elasticity of optimum retirement to changes in longevity.
Conflict and the Social Contract*
Authors: Bester Helmut , Wärneryd Karl
Published: August 2006

We consider social contracts for resolving conflicts between two agents who are uncertain about each other's fighting potential. Applications include international conflict, litigation and elections. Even though only a peaceful agreement avoids a loss of resources, if this loss is small enough, then any contract must assign a positive probability of conflict. We show how the likelihood of conflict outbreak depends on the distribution of power between the agents and their information about each other.
Career Concerns, Monetary Incentives and Job Design*
Authors: Kaarbøe Oddvar M. , Olsen Trond E.
Published: August 2006

We study optimal incentive contracts when commitments are limited, and agents have multiple tasks and career concerns. The agent's career concerns are determined by the outside market. We show that the principal might want to give the strongest explicit incentives to agents far from retirement to account for the fact that career concerns might induce behavior in conflict with the principal's preferences. Furthermore, we show that maximized welfare might be decreasing in the strength of career concerns, that optimal incentives can be positively correlated with various measures of uncertainty, and that career incentives have strong implications for optimal job design.
Novel C59T leader peptide mutation in the MPZ gene associated with late‐onset, axonal, sensorimotor polyneuropathy
Authors: Finsterer J. , Miltenberger G. , Rauschka H. , Janecke A.
Published: August 2006

The objective of this study was to report a novel exon‐1 mutation in the myelin protein zero (MPZ) gene, resulting in axonal Charcot–Marie–Tooth neuropathy with recurrent hyper‐CK‐emia. In a 64‐year‐old woman slowly progressive distal lower limb weakness, muscle cramps in the lower limb muscles, and stocking‐type numbness had developed from the age of 61. Neurologic examination revealed discrete hip flexor weakness, weakness for foot extension, diffuse wasting of the distal lower limb muscles, reduced patella tendon reflexes, and absent Achilles tendon reflexes. There was recurrently elevated creatine kinase with a maximum of 607 U/l (n, <145 U/l). Stimulation of the peroneal and tibial nerves did not evoke a muscular response. Electromyography was neurogenic. Biopsy of the right sural nerve showed diffuse axonal degeneration and loss of axons of all diameters. Muscle biopsy showed increased fiber‐size variability, angulated fibers, internalized nuclei, accumulations of nuclei, grouped atrophic muscle fibers, and fiber splitting. Molecular genetic analysis by PCR and direct nucleotide sequencing revealed the heterozygous C59T exon‐1 MPZ gene mutation, resulting in the amino acid exchange S20F of the MPZ signal protein domain (leader peptide). The novel C59T mutation in the leader peptide of the MPZ gene is pathogenic and manifests as severe, late‐onset, axonal, symmetric sensorimotor polyneuropathy (CMT2) and hyper‐CK‐emia.
Acute autonomic dysfunction contralateral to acute strokes: a prospective study of 100 consecutive cases
Authors: Diserens K. , Vuadens P. , Michel P. , Reichhart M. , Herrmann F. R. , Arnold P. , Bogousslavsky J. , Ghika J.
Published: August 2006

Complex painful reflex syndrome is sometimes described in the chronic phase of stroke. Acute autonomic dysfunction (AAD), which is occasionally present in cases of acute stroke, has not been studied prospectively. The aim of the study was to investigate AAD on the hemibody contralateral to the lesion in the acute phase of stroke. One hundred consecutive patients (median age ± interquartile range, 74 ± 21; range 19–93; 51 women: 80 ± 17 and 49 men: 70 ± 17 years) in the acute phase of stroke were studied prospectively. Changes in skin temperature or coloration, diaphoresis, pain, or edema were noted in the first 3 days post‐stroke. Associations between AAD and topography (cortical pre‐ and/or post‐central, insular, corona radiata, basal ganglia, internal capsule, thalamus, and brainstem), age, gender, ischemic or hemorrhagic etiology, or the presence of sensorimotor deficits or ataxia were examined using the chi‐squared or Fisher's exact test and logistic regression analysis. AAD was found in 71% of the patients and showed a significant positive association with the presence of a lesion in the post‐central cortex (P = 0.037), internal capsule (P = 0.005), basal ganglia (P = 0.002), or insula (P = 0.011) and a negative association with the presence of a lesion in the brainstem (P = 0.004). Multivariate logistic regression analysis including all studied topographic variables showed that only brainstem lesions were significantly associated with a decreased risk of developing AAD (odds ratio = 0.08, 95% confidence interval: 0.01–0.69, P = 0.022). AAD was not associated with age, gender, the ischemic or hemorrhagic nature of the lesion, the side of lesion, hypertonic or hypotonic paresis, or hyperreflexia or hyporeflexia. AAD was found in association with sensory deficits (P = 0.001) and contralateral hyperkinesia (P = 0.004). Acute AAD is significantly more likely to occur in the presence of hemispheric lesions involving sensory pathways from the cortex to the internal capsule and insula and is significantly less prevalent in the presence of brainstem lesions.
Changes in expression of the δ subunit of the GABAAreceptor and in receptor function induced by progesterone exposure and withdrawal
Authors: Mostallino Maria Cristina , Mura Maria Luisa , Maciocco Elisabetta , Murru Luca , Sanna Enrico , Biggio Giovanni
Published: August 2006

Type A receptors for GABA (GABAAreceptors) that contain the δ subunit are located predominantly at extrasynaptic sites and are implicated in modulation of neuronal excitability through tonic inhibition. We have examined the effects of chronic exposure to and subsequent withdrawal of progesterone or the progesterone metabolite 3α‐hydroxy‐5α‐pregnan‐20‐one (3α,5α‐THPROG) on expression of the δ subunit of GABAAreceptors and on receptor function in cultured rat hippocampal neurons. Progesterone treatment for 1 day increased the amounts of both δ subunit mRNA and protein, whereas such treatment for 6 days induced marked decreases in the abundance of both the mRNA and protein. Subsequent progesterone withdrawal up‐regulated expression of the δ subunit, which was significantly increased at 9–12 h after withdrawal. These effects of progesterone were mimicked by 3α,5α‐THPROG and blocked by the 5α‐reductase inhibitor finasteride. They were also accompanied by parallel changes in the function of GABAAreceptors in hippocampal neurons. These results show that chronic exposure to and withdrawal of progesterone induce differential effects on both expression of the δ subunit of GABAAreceptors and receptor function that are mediated by 3α,5α‐THPROG. They are consistent with the notion that this progesterone metabolite plays a key physiological role in modulation of GABAergic synapses.
Characterization of the localization and function of NECAP 1 in neurons
Authors: Murshid Ayesha , Srivastava Archana , Kumar Rohit , Presley John F.
Published: August 2006

NECAPs (adaptin ear‐binding clathrin‐associated protein) are a new family of clathrin accessory proteins identified through a proteomic analysis of clathrin‐coated vesicles (CCVs) from the brain. One member of this family, NECAP 1, is found primarily in tissues from the central nervous system and has been shown to be complexed tightly with a substantial portion of adaptor protein‐2 (AP‐2) in brain extracts. However, the function and intracellular location of this protein is unknown. In this study, we find that endogenous and epitope‐tagged NECAP 1 co‐localizes well with clathrin and AP‐2 in punctate structures, many of which also contain the presynaptic markers synaptophysin, synaptotagmin or synaptic vesicle protein 2 (SV2). NECAP 1 was also detected by western blot in synaptic vesicle preparations. Overexpression of a truncation mutant of NECAP 1 (BC‐NECAP 1) in neurons inhibited transferrin endocytosis but not epidermal growth factor (EGF) endocytosis, and this inhibition was dependent on an AP‐2‐binding WVQF motif. Moreover, overexpression of BC‐NECAP 1 results in inhibition of synaptotagmin endocytosis both in unstimulated neurons and in neurons stimulated with potassium chloride. This inhibition was abrogated by truncation of the WVQF domain. We conclude from these observations that NECAP 1 plays a role in clathrin‐mediated neuronal endocytosis, including a role in presynaptic endocytosis.
Up‐regulation of heat shock protein HSP 20 in the hippocampus as an early response to hypoxia of the newborn
Authors: David Jean‐Claude , Boelens Wilbert C. , Grongnet Jean‐François
Published: August 2006

Hypoxia is an important challenge for newborn mammals. Stress generated at the brain level under low oxygenation conditions results in up‐regulation of heat shock proteins (HSPs) and other stress proteins. The aim of the present work was to determine the effect of hypoxia in the newborn on some newly described small molecular weight HSPs (HSP 20 and B8) in the hippocampus, cortex and cerebellum of newborn piglets. These effects will be compared with those of other closely related proteins such as αB crystallin, HSP 27, heme oxygenase (HO)‐1, HO‐2, cyclooxygenase (COX)‐1 and COX‐2. The piglets were submitted to hypoxia (5% O2; 95% N2) over either 1 or 4 h, with recovery periods ranging from 0 to 68 h. Western blot analysis showed that HSP 20 was rapidly induced only in the hippocampus, long before hypoxia‐inducible transcription factor HIF‐1α, while HSP 27 was rapidly induced in the cortex and cerebellum. Vascular epithelial growth factor was increased simultaneously in the three regions. Moreover, an increase in the expression of, respectively, HO‐1 and COX‐2 was observed later, but at the same time, in the three regions tested. It appears that HSP 20 can be an early marker of hypoxia in the hippocampus. The other small HSPs or stress proteins display different temporal patterns of up‐regulation (HSP 27 and HO‐1, COX‐2) or do not show changes in their expressions (αB crystallin, HSP B8, HO‐2 and COX‐1).
Mitigating scoring errors in microsatellite data from wild populations
Authors: DEWOODY JENNIFER , NASON JOHN D. , HIPKINS VALERIE D.
Published: August 2006

Microsatellite data are widely used to test ecological and evolutionary hypotheses in wild populations. In this paper, we consider three typical sources of scoring errors capable of biasing biological conclusions: stuttering, large‐allele dropout and null alleles. We describe methods to detect errors and propose conventions to mitigate scoring errors and report error rates in studies of wild populations. Finally, we discuss potential bias in ecological or evolutionary conclusions based on data sets containing these scoring errors.
Microsatellite DNA markers for population genetic studies in the dinoflagellate Karenia brevis
Authors: RENSHAW MARK A. , SOLTYSIAK KELLY , ARREOLA DANIEL , LORET PASCAL , PATTON JOHN C. , GOLD JOHN R. , CAMPBELL LISA
Published: August 2006

Nine nuclear‐encoded microsatellites from an enriched genomic DNA library of the HAB (harmful algal bloom) dinoflagellate Karenia brevis were isolated and characterized. The microsatellites include five perfect (three dinucleotide and two trinucleotide) and four imperfect (two dinucleotide and two trinucleotide) repeat motifs. Gene (haplotype) diversity ranged from 0.153 to 0.750 among a sample of 13 isolates; the number of alleles among the isolates ranged from two to six and pairwise tests of genotypic disequilibria were nonsignificant. The microsatellites developed in this study will provide insight into the genetic diversity of this HAB species and tools that may prove useful in predicting source populations and physiological parameters of individual K. brevis blooms.
Identification and characterization of simple sequence repeat (SSR) markers derived in silico from Brassica oleracea genome shotgun sequences
Authors: BURGESS BENJAMIN , MOUNTFORD HAYLEY , HOPKINS CLARE J. , LOVE CHRISTOPHER , LING ALISON E. , SPANGENBERG GERMAN C. , EDWARDS DAVID , BATLEY JACQUELINE
Published: August 2006

The availability of sequence data derived from shotgun sequencing programs enables mining for simple sequence repeats (SSRs), providing useful genetic markers for crop improvement. This study presents the development and characterization of 40 SSR markers from Brassica oleracea shotgun sequence and their cross‐amplification across Brassica species. The markers show reliable amplification, genome specificity and considerable polymorphism, demonstrating the utility of SSRs for genetic analysis of commercial Brassica germplasm.
Y chromosome microsatellite markers identified from the tammar wallaby (Macropus eugenii) and their amplification in three other macropod species
Authors: MACDONALD ANNA J. , SANKOVIC NATASHA , SARRE STEPHEN D. , FITZSIMMONS NANCY N. , WAKEFIELD MATTHEW J. , GRAVES JENNIFER A. MARSHALL , ZENGER KYALL R.
Published: August 2006

Microsatellites were identified from three fully sequenced Y chromosome‐specific bacterial artificial chromosome clones from the tammar wallaby, Macropus eugenii. Ten microsatellites were genotyped in male tammar wallabies. Four loci were polymorphic with between two and six alleles per locus. Eleven different haplotypes were identified from 22 male tammar wallabies. No amplifications were obtained from female samples. Each microsatellite was also shown to amplify reliably in at least one other macropod species. These markers may therefore prove useful as some of the first male‐specific genetic markers for marsupials, with potential application to studies of male‐biased dispersal and mating systems.
Nest‐site fidelity and breeding success in White StorkCiconia ciconia
Authors: VERGARA PABLO , AGUIRRE JOSÉ I. , FARGALLO JUAN A. , DÁVILA JOSÉ A.
Published: August 2006

Nest‐site fidelity is a life‐history trait of birds that may produce benefits in terms of fitness. We studied the relationship between breeding success and nest fidelity in the White StorkCiconia ciconia. We also studied how other factors such as age, sex, habitat, colony size and productivity in previous breeding attempts might interfere with this relationship. Our results showed that pairs with higher fidelity rates also have lower failure rates, and that breeding failure and productivity in the previous season influenced the frequency of nest change in the following season. In addition, a curvilinear relationship was found between age and nest fidelity. These results suggest that age is a major factor related to nest fidelity and therefore individual experience could explain this behaviour in the White Stork. Changing the nest involves a reproductive cost for which nest fidelity can be considered as an adaptive strategy to increase fitness.
How robust are Palearctic migrants to habitat loss and degradation in the Sahel?
Authors: WILSON JARED M. , CRESSWELL WILL
Published: August 2006

The Sahel region of West Africa is an important wintering ground for Palearctic migrants. It is, however, undergoing significant anthropogenic degradation that is likely to reduce its value in this regard. Despite there being evidence that the populations of several Palearctic species may be determined by conditions on their Sahelian wintering grounds, there are almost no baseline data on their habitat use there and so the effects that habitat change might have on their future density and distribution cannot be determined. This study measured which habitat variables were associated with the presence of Common Whitethroat Sylvia communis, Lesser Whitethroat S. curruca and Subalpine Warbler S. cantillans by carrying out point counts and habitat measurements at 1861 points at 16 sites in northern Nigeria during the winters of 2001 and 2002. Common Whitethroats were most common at intermediate tree heights and densities of Salvadora persica (with this effect strongest in the pre‐migratory period), and increased as Balanites aegyptiaca density increased initially, although this effect levelled off at higher Balanites densities. Subalpine Warblers were most common at intermediate tree heights and Balanites densities, increased as Acacia and Salvadora density increased, decreased with tree density and were least common at intermediate shrub densities. Lesser Whitethroats were most common at intermediate tree heights, Acacia and Salvadora densities, increased with Balanites density, and decreased with tree density and diversity. The shallow slopes of the significant relationships found between Palearctic warbler species and habitat parameters suggest that the group may be robust to habitat degradation, with densities remaining relatively constant across a wide range of habitat types. Common Whitethroats, in particular, appear to be able to survive in extremely degraded habitats, yet may be vulnerable to the disappearance of Salvadora, the fruit of which may allow pre‐migratory fattening.
Dinoflagellate, Euglenid, or Cercomonad? The Ultrastructure and Molecular Phylogenetic Position ofProtaspis grandisn. sp.
Authors: HOPPENRATH MONA , LEANDER BRIAN S.
Published: August 2006

ABSTRACT.Protaspisis an enigmatic genus of marine phagotrophic biflagellates that have been tentatively classified with several different groups of eukaryotes, including dinoflagellates, euglenids, and cercomonads. This uncertainty led us to investigate the phylogenetic position ofProtaspis grandisn. sp. with ultrastructural and small subunit (SSU) rDNA sequence data. Our results demonstrated that the cells were dorsoventrally flattened, shaped like elongated ovals with parallel lateral sides, 32.5–55.0 μm long and 20.0–35.0 μm wide. Moreover, two heterodynamic flagella emerged through funnels that were positioned subapically, each within a depression and separated by a distinctive protrusion. A complex multilayered wall surrounded the cell. Like dinoflagellates and euglenids, the nucleus contained permanently condensed chromosomes and a large nucleolus throughout the cell cycle. Pseudopodia containing numerous mitochondria with tubular cristae emerged from a ventral furrow through a longitudinal slit that was positioned posterior to the protrusion and flagellar apparatus. Batteries of extrusomes were present within the cytoplasm and had ejection sites through pores in the cell wall. The SSU rDNA phylogeny demonstrated a very close relationship between the benthicP. grandisn. sp. and the planktonicCryothecomonas longipes. These ultrastructural and molecular phylogenetic data forProtaspisindicated that the current taxonomy ofProtaspisandCrythecomonasis in need of re‐evaluation. The composition and identity ofProtaspisis reviewed and suggestions for future taxonomic changes are presented. Problems within the genusCryothecomonasare highlighted as well, and the missing data needed to resolve ambiguities between the two genera are clarified.
Merkel cell carcinoma: evaluation of KIT (CD117) expression and failure to demonstrate activating mutations in the C‐KIT proto‐oncogene – implications for treatment with imatinib mesylate
Authors: Swick Brian L. , Ravdel Larisa , Fitzpatrick James E. , Robinson William A.
Published: August 2006

Background:  Merkel cell carcinomas (MCCs) express the tyrosine kinase receptor KIT. However, it is not known if MCCs have activating mutations in KIT that would make them responsive to treatment with imatinib mesylate. The purpose of this article is to describe the KIT immunohistological staining pattern (CD117) of MCCs and analyze those MCCs for mutations in areas of KIT where mutations are found in gastrointestinal stromal cell tumors.

Methods:  We evaluated KIT immunostaining in nine MCCs from nine patients. In addition, we extracted DNA from the same MCCs, performed PCR amplification of C‐KIT exons 9, 11, 13, and 17, and sequenced those gene products for mutations.

Results:  Eight of nine (88.8%) MCCs expressed KIT. No mutations were found.

Conclusions:  The majority of MCCs express KIT but do not contain activating mutations in exons 9, 11, 13, or 17 of KIT. Imatinib mesylate is unlikely to provide effective therapy in MCC unless activating mutations in other areas of KIT or activating mutations in other related genes can be detected.
Dual porphyrias revisited
Authors: Poblete‐Gutiérrez Pamela , Badeloe Sadhanna , Wiederholt Tonio , Merk Hans F. , Frank Jorge
Published: August 2006

Abstract:  The porphyrias are clinically and genetically heterogeneous metabolic diseases, which predominantly result from a hereditary dysfunction in the pathway of haeme biosynthesis. Currently, at least eight different forms of porphyrias can be differentiated, all of them characterized by a specific enzyme deficiency that is either inherited in an autosomal‐dominant fashion, autosomal recessively or, in the case of porphyria cutanea tarda, might also be acquired. All genes encoding these enzymes have been cloned and several mutations underlying the different types of porphyrias have been reported. Traditionally, the diagnosis of porphyria is made on the basis of clinical symptoms, characteristic biochemical findings and enzyme assays. In some porphyria patients and families, however, these diagnostic tools can reveal simultaneous findings compatible with two different forms of porphyria, a phenomenon referred to as dual porphyria. Here, we give an overview on what is currently known about these peculiar variants of porphyria and suggest that, whenever feasible, molecular genetic analysis should complement the analytical techniques used to characterize patients and families in which a double enzymatic deficiency within the haeme biosynthetic pathway is assumed.
Botanical antioxidants in the prevention of photocarcinogenesis and photoaging
Authors: Afaq Farrukh , Mukhtar Hasan
Published: August 2006

Abstract:  Exposure of the skin to ultraviolet (UV) radiation, particularly its UV‐B component (280–320 nm), from the sun results in erythema, edema, hyperplasia, hyperpigmentation, sunburn cells, immunosuppression, photoaging, and skin cancer. Amongst these various adverse effects of UV‐B radiation, skin cancer and photoaging are of great concern. More recent changes in lifestyle have led to a significant increase in the amount of UV‐B radiation people receive leading to a surge in the incidence of skin cancer and photoaging. As these trends are likely to continue in the foreseeable future, the adverse effect of UV‐B has become a major human health concern. Therefore, development of novel strategies to reduce the occurrence of skin cancer and delay the process of photoaging are highly desirable goals. One approach to reduce their occurrence is through photochemoprevention, which we define as the use of agents capable of ameliorating the adverse effects of UV‐B on the skin. Photochemoprevention via use of botanical antioxidants, present in the common diet of human have gained considerable attention as photochemopreventive agents for human use. Many such agents have also found a place in skin care products. This review will focus on the effects of selected botanical antioxidants in the prevention of photocarcinogenesis and photoaging.
Expression of chitinase‐like proteins in the skin of chronic proliferative dermatitis (cpdm/cpdm) mice
Authors: HogenEsch Harm , Dunham Anisa , Seymour Rosemarie , Renninger Matthew , Sundberg John P.
Published: August 2006

Abstract:  Mammalian chitinase‐like proteins belong to a family of proteins structurally related to chitinases but devoid of enzymatic activity. They have a postulated role in remodeling of extracellular matrix and defense mechanisms against chitin‐containing pathogens. The expression of these proteins is increased in parasitic infections and allergic airway disease, but their expression in dermatitis has not been examined. The mRNA expression of two chitinase 3‐like (Chi3L) proteins, Chi3L3 (Ym1) and Chi3L4 (Ym2), was determined in the skin of normal mice, chronic proliferative dermatitis (cpdm/cpdm) mutant mice and mice with experimentally induced contact hypersensitivity reaction. The localization of Chi3L3 and Chi3L4 proteins in cells was determined by fluorescence microscopy of double‐labeled frozen sections of skin, and confirmed in vitro by stimulation of macrophages and mast cells with cytokines. Quantitative RT‐PCR demonstrated a 976‐fold increase of Chi3l4 mRNA expression and a 24‐fold increase of Chi3l3 mRNA expression in the skin of cpdm/cpdm mice. Their expression was also increased in the ears of mice with 2,4‐dinitrofluorobenzene‐induced contact hypersensitivity, but the increase was greater for Chi3l3 mRNA (51‐fold) than Chi3l4 mRNA (32‐fold). Western blot analysis with an antibody against Chi3L3 and Chi3L4 confirmed the increased amount of these proteins in the skin of cpdm/cpdm mice. Two‐color immunofluorescence identified macrophages, dendritic cells and mast cells as cellular sources of Chi3L3 and Chi3L4 proteins. Eosinophils and neutrophils did not contain detectable concentrations of these proteins. Treatment of macrophages and mast cells in vitro with interleukin‐4 induced expression of Chi3l3 and Chi3l4 mRNA.
Development, validation and application of a chemiluminescent immunoassay for the measurement of circulating chorionic gonadotropin levels in the laboratory macaque
Authors: Lohstroh Pete , Laughlin Lisa , Gee Nancy , Lasley Bill
Published: August 2006

Background  Methods for the measurement of chorionic gonadotropin for pregnancy detection in laboratory macaques chorionic gonadotropin (mCG) have been limited by the paucity of specific reagents. Current assays use reagents developed for human chorionic gonadotropin (hCG) assays and have been limited to radio‐ or enzyme‐immuno assays.

Methods  This report describes an automated assay for mCG using reagents for hCG detection that has been adapted to the automated chemiluminescence platform of the Bayer ACS‐180 autoanalyzer.

Results  This automated assay performs similarly to previous assays in terms of sensitivity and specificity but provides additional speed, reliability, and more easily accessible reagents compared with previous formats.

Conclusions  When implemented broadly, this assay could provide standardization of mCG measurements for both research and colony management purposes.
Change in structure of the N‐terminal region of transthyretin produces change in affinity of transthyretin to T4 and T3
Authors: Prapunpoj Porntip , Leelawatwatana Ladda , Schreiber Gerhard , Richardson Samantha J.
Published: August 2006

The relationship between the structure of the N‐terminal sequence of transthyretin (TTR) and the binding of thyroid hormone was studied. A recombinant human TTR and two derivatives of Crocodylus porosus TTRs, one with the N‐terminal sequence replaced by that of human TTR (human/crocTTR), the other with the N‐terminal segment removed (truncated crocTTR), were synthesized in Pichia pastoris. Subunit mass, native molecular weight, tetramer formation, cross‐reactivity to TTR antibodies and binding to retinol‐binding protein of these recombinant TTRs were similar to TTRs found in nature. Analysis of the binding affinity to thyroid hormones of recombinant human TTR showed a dissociation constant (Kd) for triiodothyronine (T3) of 53.26 ± 3.97 nm and for thyroxine (T4) of 19.73 ± 0.13 nm. These values are similar to those found for TTR purified from human serum, and gave a Kd T3/T4 ratio of 2.70. The affinity for T4 of human/crocTTR (Kd = 22.75 ± 1.89 nm) was higher than those of both human TTR and C. porosus TTR, but the affinity for T3 (Kd = 5.40 ± 0.25 nm) was similar to C. porosus TTR, giving a Kd T3/T4 ratio of 0.24. A similar affinity for both T3 (Kd = 57.78 ± 5.65 nm) and T4 (Kd = 59.72 ± 3.38 nm), with a Kd T3/T4 ratio of 0.97, was observed for truncated crocTTR. The obtained results strongly confirm the hypothesis that the unstructured N‐terminal region of TTR critically influences the specificity and affinity of thyroid hormone binding to TTR.