bluuebunny/crossref_metadata_2025 huggingface dataset:
Epidermal Growth Factor Stimulates Human Trophoblast Cell Migration through Rho A and Rho C Activation
Authors: Han Jian , Li Li , Hu Jiongyu , Yu Lili , Zheng Yingru , Guo Jianxin , Zheng Xiuhui , Yi Ping , Zhou Yuanguo
Published: February 2010

This study investigated the roles of Rho protein in epidermal growth factor (EGF)-induced trophoblast cell migration and its mechanism. Using choriocarcinoma cell lines JEG-3 and JAR and first-trimester human chorionic villus explant cultures on matrigel, we examined EGF-mediated stimulation of trophoblast migration. EGF is shown to have a dose-dependent effect on trophoblast migration. A low concentration of EGF (1 ng/ml) has a stimulatory effect on cell migration, whereas high concentrations of EGF (100 ng/ml) shows an inhibitory effect. EGF (1 ng/ml) activates RhoA and RhoC, but not RhoB, through elevated protein levels and activity. EGF-induced migration was shown to be inhibited by either cell-permeable C3 exoenzyme transferase or selective RhoA or RhoC small interfering RNAs. The inhibition was not mitigated by the addition of EGF, suggesting that RhoA and RhoC play an important role in trophoblast migration and are obligatory for EGF action. Treatment of JEG-3 and JAR cells with RhoA small interfering RNA induced F-actin cytoskeleton disruption and cell shrinkage, which is consistent with the effect of C3 exoenzyme transferase, and this action was not mitigated by EGF treatment. RhoC small interfering RNA had no apparent effect on the F-actin arrangement, suggesting that RhoA but not RhoC takes part in the EGF-induced migration through F-actin rearrangement. These results indicate that RhoA and RhoC play more important roles than RhoB in EGF-mediated migration of trophoblast cells, and RhoA but not RhoC regulates this migration through F-actin cytoskeleton reorganization.
The Trithorax group protein Ash2l and Saf-A are recruited to the inactive X chromosome at the onset of stable X inactivation
Authors: Pullirsch Dieter , Härtel Renate , Kishimoto Hiroyuki , Leeb Martin , Steiner Günter , Wutz Anton
Published: February 2010

Mammals compensate X chromosome gene dosage between the sexes by silencing of one of the two female X chromosomes. X inactivation is initiated in the early embryo and requires the non-coding Xist RNA, which encompasses the inactive X chromosome (Xi) and triggers its silencing. In differentiated cells, several factors including the histone variant macroH2A and the scaffold attachment factor SAF-A are recruited to the Xi and maintain its repression. Consequently, in female somatic cells the Xi remains stably silenced independently of Xist. Here, we identify the Trithorax group protein Ash2l as a novel component of the Xi. Ash2l is recruited by Xist concomitantly with Saf-A and macroH2A at the transition to Xi maintenance. Recruitment of these factors characterizes a developmental transition point for the chromatin composition of the Xi. Surprisingly, expression of a mutant Xist RNA that does not cause gene repression can trigger recruitment of Ash2l, Saf-A and macroH2A to the X chromosome, and can cause chromosome-wide histone H4 hypoacetylation. This suggests that a chromatin configuration is established on non-genic chromatin on the Xi by Xist to provide a repressive compartment that could be used for maintaining gene silencing. Gene silencing is mechanistically separable from the formation of this repressive compartment and, thus, requires additional pathways. This observation highlights a crucial role for spatial organization of chromatin changes in the maintenance of X inactivation.
A conditional mutation in Arabidopsis thaliana separase induces chromosome non-disjunction, aberrant morphogenesis and cyclin B1;1 stability
Authors: Wu Shuang , Scheible Wolf-Rüdiger , Schindelasch Dana , Van Den Daele Hilde , De Veylder Lieven , Baskin Tobias I.
Published: February 2010

The caspase family protease, separase, is required at anaphase onset to cleave the cohesin complex, which joins sister chromatids. However, among eukaryotes, separases have acquired novel functions. Here, we show that Arabidopsis thaliana radially swollen 4 (rsw4), a temperature-sensitive mutant isolated previously on the basis of root swelling, harbors a mutation in At4g22970, the A. thaliana separase. Loss of separase function in rsw4 at the restrictive temperature is indicated by the widespread failure of replicated chromosomes to disjoin. Surprisingly, rsw4 has neither pronounced cell cycle arrest nor anomalous spindle formation, which occur in other eukaryotes upon loss of separase activity. However, rsw4 roots have disorganized cortical microtubules and accumulate the mitosis-specific cyclin, cyclin B1;1, excessive levels of which have been associated with altered microtubules and morphology. Cyclin B1;1 also accumulates in certain backgrounds in response to DNA damage, but we find no evidence for aberrant responses to DNA damage in rsw4. Our characterization of rsw4 leads us to hypothesize that plant separase, in addition to cleaving cohesin, regulates cyclin B1;1, with profound ramifications for morphogenesis.
The HMX/NKX homeodomain protein MLS-2 specifies the identity of the AWC sensory neuron type via regulation of the ceh-36 Otx gene in C. elegans
Authors: Kim Kyuhyung , Kim Rinho , Sengupta Piali
Published: February 2010

The differentiated features of postmitotic neurons are dictated by the expression of specific transcription factors. The mechanisms by which the precise spatiotemporal expression patterns of these factors are regulated are poorly understood. In C. elegans, the ceh-36 Otx homeobox gene is expressed in the AWC sensory neurons throughout postembryonic development, and regulates terminal differentiation of this neuronal subtype. Here, we show that the HMX/NKX homeodomain protein MLS-2 regulates ceh-36 expression specifically in the AWC neurons. Consequently, the AWC neurons fail to express neuron type-specific characteristics in mls-2 mutants. mls-2 is expressed transiently in postmitotic AWC neurons, and directly initiates ceh-36 expression. CEH-36 subsequently interacts with a distinct site in its cis-regulatory sequences to maintain its own expression, and also directly regulates the expression of AWC-specific terminal differentiation genes. We also show that MLS-2 acts in additional neuron types to regulate their development and differentiation. Our analysis describes a transcription factor cascade that defines the unique postmitotic characteristics of a sensory neuron subtype, and provides insights into the spatiotemporal regulatory mechanisms that generate functional diversity in the sensory nervous system.
Perfusion with cAMP analogue affects pheromone-sensitive trichoid sensilla of the hawkmothManduca sextain a time-dependent manner
Authors: Flecke Christian , Nolte Andreas , Stengl Monika
Published: February 2010

Octopamine causes time-dependent disadaptation of pheromone-sensitive olfactory receptor neurons (ORNs) of Manduca sexta. Because the majority of insect octopamine receptors are positively coupled to adenylyl cyclases we examined whether cyclic adenosine monophosphate (cAMP) mimics octopamine-dependent modulation of pheromone transduction in a time-dependent manner. Long-term tip recordings of single trichoid sensilla of Manduca sexta were performed during three zeitgeber times (ZTs, ZT 0=lights on), while stimulating the sensilla with two doses of the main pheromone component bombykal in a non-adapting protocol. The membrane-permeable cAMP analogue 8bcAMP increased the normalized sensillar potential amplitude in a time- and bombykal dose-dependent way. At the higher bombykal dose only, the applied 8bcAMP antagonized an endogenous decrease in the mean sensillar potential amplitude at ZT 1-4 and ZT 8-11 when ORNs were adapted but not at ZT 22-1, when ORNs were sensitized. In contrast to octopamine, 8bcAMP did not consistently affect the initial pheromone-dependent action potential frequency, the phasic/tonic response pattern, or the time-dependent shift to lower mean action potential frequencies at ZT 8-11. Furthermore, 8bcAMP increased the spontaneous action potential frequency time dependently, but differently from octopamine. In conclusion, our results show that cAMP only partly mimics the octopamine-dependent disadaptation of olfactory receptor neurons during photophase, apparently due to another missing octopamine-dependent synergistic factor such as defined intracellular calcium levels.
Effect of acute and chronic hypoxia on the swimming performance, metabolic capacity and cardiac function of Atlantic cod (Gadus morhua)
Authors: Petersen L. H. , Gamperl A. K.
Published: February 2010

Low water oxygen content (hypoxia) is a common feature of many freshwater and marine environments. However, we have a poor understanding of the degree to which diminished cardiac function contributes to the reduction in fish swimming performance concomitant with acute exposure to hypoxia, or how fish cardiorespiratory physiology is altered by, or adapts to, chronic hypoxia. Thus, we acclimated adult Atlantic cod (Gadus morhua) to either ~8-9 kPa O2 (40-45% air saturation) or ~21 kPa O2 (100% air saturation; normoxia) for 6-12 weeks at 10°C, and subsequently measured metabolic variables [routine oxygen consumption (), maximum , metabolic scope] and cardiac function (cardiac output, ; heart rate, fH; and stroke volume, VS) in these fish during critical swimming speed (Ucrit) tests performed at both levels of water oxygenation. Although surgery (flow probe implantation) reduced the Ucrit of normoxia-acclimated cod by 14% (from 1.74 to 1.50 BL s−1) under normoxic conditions, exposure to acute hypoxia lowered the Ucrit of both groups (surgery and non-surgery) by ~30% (to 1.23 and 1.02 BL s−1, respectively). This reduction in swimming performance was associated with large decreases in maximum and metabolic scope (≥50%), and maximum fH and (by 16 and 22%), but not VS. Long-term acclimation to hypoxia resulted in a significant elevation in normoxic metabolic rate as compared with normoxia-acclimated fish (by 27%), but did not influence normoxic or hypoxic values for Ucrit, maximum or metabolic scope. This was surprising given that resting and maximum values for were significantly lower in hypoxia-acclimated cod at both levels of oxygenation, because of lower values for VS. However, hypoxia-acclimated cod were able to consume more oxygen for a given cardiac output. These results provide important insights into how fish cardiorespiratory physiology is impacted by short-term and prolonged exposure to hypoxia, and further highlight the tremendous capacity of the fish cardiorespiratory system to deal with environmental challenges.
Venom kinematics during prey capture inConus: the biomechanics of a rapid injection system
Authors: Salisbury S. Michael , Martin Gary G. , Kier William M. , Schulz Joseph R.
Published: February 2010

Cone snails use an extensile, tubular proboscis as a conduit to deliver a potent cocktail of bioactive venom peptides into their prey. Previous studies have focused mainly on understanding the venom's role in prey capture but successful prey capture requires both rapid physiological and biomechanical mechanisms. Conus catus, a fish-hunting species, uses a high-speed hydraulic mechanism to inject its hollow, spear-like radular tooth into prey. We take an integrated approach to investigating the biomechanics of this process by coupling kinematic studies with morphological analyses. Taking advantage of the opaque venom and translucent proboscis of a mollusc-hunting juvenile cone snail, Conus pennaceus, we have determined that a high-speed prey capture mechanism is not unique to cone species that hunt fish prey. Two morphological structures were found to play crucial roles in this process. A constriction of the lumen near the tip of the proboscis, composed of tall epithelial cells densely packed with microfilaments, impedes forward movement of the radular tooth prior to its propulsion. Proximal to the constriction, a muscular sphincter was found to regulate venom flow and pressurization in the proboscis. In C. pennaceus, the rapid appearance and flushing of venom within the proboscis during prey capture suggests a mechanism involving the delivery of a discrete quantity of venom. The interplay between these elements provides a unique and effective biomechanical injection system for the fast-acting cone snail venom peptides.
Insect form vision as one potential shaping force of spider web decoration design
Authors: Cheng R.-C. , Yang E.-C. , Lin C.-P. , Herberstein M. E. , Tso I.-M.
Published: February 2010

Properties of prey sensory systems are important factors shaping the design of signals generated by organisms exploiting them. In this study we assessed how prey sensory preference affected the exploiter signal design by investigating the evolutionary relationship and relative attractiveness of linear and cruciate form web decorations built by Argiope spiders. Because insects have an innate preference for bilaterally symmetrical patterns, we hypothesized that cruciate form decorations were evolved from linear form due to their higher visual attractiveness to insects. We first reconstructed a molecular phylogeny of the Asian members of the genus Argiope using mitochondrial markers to infer the evolutionary relationship of two decoration forms. Results of ancestral character state reconstruction showed that the linear form was ancestral and the cruciate form derived. To evaluate the luring effectiveness of two decoration forms, we performed field experiments in which the number and orientation of decoration bands were manipulated. Decoration bands arranged in a cruciate form were significantly more attractive to insects than those arranged in a linear form, no matter whether they were composed of silks or dummies. Moreover, dummy decoration bands arranged in a cruciate form attracted significantly more insects than those arranged in a vertical/horizontal form. Such results suggest that pollinator insects' innate preference for certain bilateral or radial symmetrical patterns might be one of the driving forces shaping the arrangement pattern of spider web decorations.
Mounting a specific immune response increases energy expenditure of the subterranean rodentCtenomys talarum(tuco-tuco): implications for intraspecific and interspecific variation in immunological traits
Authors: Cutrera A. P. , Zenuto R. R. , Luna F. , Antenucci C. D.
Published: February 2010

It was recently hypothesised that specific induced defences, which require substantial time and resources and are mostly beneficial against repeated infections, are more likely to be favoured in ‘slow-living-pace’ species. Therefore, understanding how different types of immune defences might vary with life history requires knowledge of the costs and benefits of defence components. Studies that have explored the energetic costs of immunity in vertebrates have done so with a focus primarily on birds and less so on mammals, particularly surface-dwelling rodents. In this study, we evaluated whether an experimental induction of the immune system with a non-pathogenic antigen elevates the energetic expenditure of a subterranean rodent: Ctenomys talarum (tuco-tucos). In both seasons studied, a significant increase in oxygen consumption was verified in immune-challenged tuco-tucos injected with sheep red blood cells (SRBC) compared with control animals. The increase in oxygen consumption 10 days after the exposure to SRBC was lower for female tuco-tucos monitored in the breeding season compared with females in the non-breeding season. Interestingly, antibody titres of female tuco-tucos did not decrease during the breeding season. Our results add new insight into the role of other factors such as basal metabolic rate or degree of parasite exposure besides ‘pace of life’ in modulating the interspecific immunological variation observed in natural populations of mammals.
The influence of tendon compliance on muscle power output and efficiency during cyclic contractions
Authors: Lichtwark G. A. , Barclay C. J.
Published: February 2010

Muscle power output and efficiency during cyclical contractions are influenced by the timing and duration of stimulation of the muscle and the interaction of the muscle with its mechanical environment. It has been suggested that tendon compliance may reduce the energy required for power production from the muscle by reducing the required shortening of the muscle fibres. Theoretically this may allow the muscle to maintain both high power output and efficiency during cyclical contraction; however, this has yet to be demonstrated experimentally. To investigate how tendon compliance might act to increase muscle power output and/or efficiency, we attached artificial tendons of varying compliance to muscle fibre bundles in vitro and measured power output and mechanical efficiency during stretch—shorten cycles (2 Hz) with a range of stretch amplitudes and stimulation patterns. The results showed that peak power, average power output and efficiency (none of which can have direct contributions from the compliant tendon) all increased with increasing tendon compliance, presumably due to the tendon acting to minimise muscle energy use by allowing the muscle fibres to shorten at optimal speeds. Matching highly compliant tendons with a sufficiently large amplitude length change and appropriate stimulation pattern significantly increased the net muscle efficiency compared with stiff tendons acting at the same frequency. The maximum efficiency for compliant tendons was also similar to the highest value measured under constant velocity and force conditions, which suggests that tendon compliance can maximise muscle efficiency in the conditions tested here. These results provide experimental evidence that during constrained cyclical contractions, muscle power and efficiency can be enhanced with compliant tendons.
Dual control of cytosolic metals by lysosomal transporters in lobster hepatopancreas
Authors: Sterling K. M. , Roggenbeck B. , Ahearn G. A.
Published: February 2010

This study describes the membrane transport mechanisms used by lobster (Homarus americanus) hepatopancreatic epithelial lysosomes to accumulate and sequester heavy metals from the cytosol, and thereby aid in the regulation of these ions entering the animal from dietary constituents. The present investigation extends previous work describing lysosomal metal uptake by cation exchange with protons and suggests that a second, parallel, lysosomal transport process involving metal—thiol conjugates may work in conjunction with the cation antiporter to control cytoplasmic metal concentrations. Transport of 65Zn2+ by lysosomal membrane vesicles (LMV) incubated in 1 mmol l−1 glutathione (GSH) was not significantly different from metal transport in the absence of the tripeptide. However, preloading LMV with 1 mmol l−1 α-ketoglutarate (AKG), and then incubating in a medium containing 1 mmol l−1 GSH, more than doubled metal uptake, compared with vesicles equilibrated with chloride or possessing an outwardly directed chloride gradient. Kinetic analysis of lysosomal 65Zn2+ influx as a function of zinc concentration, in vesicles containing 1 mmol l−1 AKG and incubated in 1 mmol l−1 GSH, revealed the presence of a sigmoidal, low affinity, high capacity carrier process transporting the metal into the organelle. These data indicated the possible presence of an organic anion exchanger in lobster lysosomal membranes. Western blot analysis of LMV with a rabbit anti-rat OAT1 antibody showed the presence of an orthologous OAT1-like protein (approximate molecular mass of 80 kDa) signal from these membranes. These results, and those published previously, suggest the occurrence of two metal transporters on hepatopancreatic membranes, a high affinity, low capacity cation antiporter and a low affinity, high capacity organic anion exchanger. Together these two systems have the potential to regulate cytoplasmic metals over a wide concentration range.
Tracking the oxidative kinetics of carbohydrates, amino acids and fatty acids in the house sparrow using exhaled 13CO2
Authors: McCue M. D. , Sivan O. , McWilliams S. R. , Pinshow B.
Published: February 2010

Clinicians commonly measure the 13CO2 in exhaled breath samples following administration of a metabolic tracer (breath testing) to diagnose certain infections and metabolic disorders. We believe that breath testing can become a powerful tool to investigate novel questions about the influence of ecological and physiological factors on the oxidative fates of exogenous nutrients. Here we examined several predictions regarding the oxidative kinetics of specific carbohydrates, amino acids and fatty acids in a dietary generalist, the house sparrow (Passer domesticus). After administering postprandial birds with 20 mg of one of seven 13C-labeled tracers, we measured rates of 13CO2 production every 15 min over 2 h. We found that sparrows oxidized exogenous amino acids far more rapidly than carbohydrates or fatty acids, and that different tracers belonging to the same class of physiological fuels had unique oxidative kinetics. Glycine had a mean maximum rate of oxidation (2021 nmol min−1) that was significantly higher than that of leucine (351 nmol min−1), supporting our prediction that nonessential amino acids are oxidized more rapidly than essential amino acids. Exogenous glucose and fructose were oxidized to a similar extent (5.9% of dose), but the time required to reach maximum rates of oxidation was longer for fructose. The maximum rates of oxidation were significantly higher when exogenous glucose was administered as an aqueous solution (122 nmol min−1), rather than as an oil suspension (93 nmol min−1), supporting our prediction that exogenous lipids negatively influence rates of exogenous glucose oxidation. Dietary fatty acids had the lowest maximum rates of oxidation (2-6 nmol min−1), and differed significantly in the extent to which each was oxidized, with 0.73%, 0.63% and 0.21% of palmitic, oleic and stearic acid tracers oxidized, respectively.
A simplified liquid chromatography tandem mass spectrometry assay, using on-line solid-phase extraction, for the quantitation of cortisol in saliva and comparison with a routine DELFIA method
Authors: Owen L J , Haslam S , Adaway J E , Wood P , Glenn C , Keevil B G
Published: February 2010

Background

We have developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for measuring salivary cortisol, which requires only 200 μL sample and no extraction.

Methods

Sample (200 μL) and 25 μL internal standard were added directly to a 96-deep-well plate. Of this, 50 μL was loaded onto a guard cartridge, the cartridge was then washed and the eluate was diverted to waste. The compounds were eluted from the guard cartridge onto the C18 analytical column. Cortisol and deuterated cortisol were monitored using transitions m/z 363.2 > 121.1 and 365.1 > 122.2, respectively.

Results

The method had a lower limit of quantitation of 2 nmol/L. Intra-assay and inter-assay imprecision were better than 9.5%. Comparison with an established dissociation enhanced lanthanide fluorescent immunoassay (DELFIA) gave good agreement for the majority of samples; LC-MS/MS = 1.0065 × DELFIA − 3.7 ( n = 130). The reference range was determined to be 5.8–45.7 nmol/L at 08:00 h and <6.4 nmol/L at 23:00 h ( n = 44).

Conclusions

We have developed a simple, robust assay to measure salivary cortisol using on-line solid-phase extraction to reduce sample clean-up requirements.
Role of miR-34c microRNA in the late steps of spermatogenesis
Authors: Bouhallier Frantz , Allioli Nathalie , Lavial Fabrice , Chalmel Frédéric , Perrard Marie-Hélène , Durand Philippe , Samarut Jacques , Pain Bertrand , Rouault Jean-Pierre
Published: February 2010

Spermatogenesis is a cyclic process in which diploid spermatogonia differentiate into haploid spermatozoa. This process is highly regulated, notably at the post-transcriptional level. MicroRNAs (miRNAs), single-stranded noncoding RNA molecules of about 20–25 nucleotides, are implicated in the regulation of many important biological pathways such as proliferation, apoptosis, and differentiation. We wondered whether miRNAs could play a role during spermatogenesis. The miRNA expression repertoire was tested in germ cells, and we present data showing that miR-34c was highly expressed only in these cells. Furthermore, our findings indicate that in male gonads, miR-34c expression is largely p53 independent in contrast to previous results showing a direct link in somatic cells between the miR-34 family and this tumor suppressor protein. In order to identify target genes involved in germinal lineage differentiation, we overexpressed miR-34c in HeLa cells, analyzed the transcriptome of these modified cells, and noticed a shift of the expression profile toward the germinal lineage. Recently, it has been shown that exogenous expression of Ddx4/Vasa in embryonic chicken stem cells (cESC) induces cESC reprogramming toward a germ cell fate. When we simultaneously expressed miR-34c in such cells, we could detect an up-regulation of germ cell-specific genes whereas the expression of other lineage specific markers remained unchanged. These data suggest that miR-34c could play a role by enhancing the germinal phenotype of cells already committed to this lineage.
Immunohistochemical Evaluation of p16INK4A, E-Cadherin, and Cyclin D1 Expression in Melanoma and Spitz Tumors
Authors: George Evan , Polissar Nayak L. , Wick Mark
Published: February 2010

We evaluated the usefulness of immunohistochemical examination for E-cadherin, p16, and cyclin D1 in discriminating melanoma from Spitz tumors. Immunoperoxidase staining was performed on formalin-fixed tissue specimens from 46 Spitz tumors and 42 concurrent melanoma specimens. The percentages of immunoreactive melanocytes in the epidermis and dermis were estimated semiquantitatively. Qualitatively abnormal immunoreactivity patterns were also tabulated. Dermal p16 immunoreactivity was the best quantitative discriminator: decreased nuclear immunoreactivity (<25% of dermal melanocytes) was 3-fold more likely in melanoma than in Spitz tumors (P = .004). Loss of both nuclear and cytoplasmic dermal p16 immunoreactivity was 8-fold more likely in melanoma (P = .01). Qualitative irregularities in the zonal distribution of E-cadherin immunoreactivity were 2-fold higher in melanoma (P = .01), but these were often focal or subtle. There was no statistically significant difference in cyclin D1 immunoreactivity. In atypical Spitz tumors, the dermal p16 immunoreactivity and frequency of qualitative E-cadherin abnormalities were intermediate between those of ordinary Spitz nevi and melanoma. Also, contrasting immunoreactivity patterns were helpful in determining Breslow thickness in specimens containing melanoma and contiguous dermal nevi.
The Usefulness of IgG and IgM Immunostaining of Periportal Inflammatory Cells (Plasma Cells and Lymphocytes) for the Distinction of Autoimmune Hepatitis and Primary Biliary Cirrhosis and Their Staining Pattern in Autoimmune Hepatitis–Primary Biliary Cirrhosis Overlap Syndrome
Authors: Lee Hwajeong , Stapp Robert T. , Ormsby Adrian H. , Shah Veena V.
Published: February 2010

Autoimmune hepatitis (AIH)–primary biliary cirrhosis (PBC) overlap syndrome (OS) is a vaguely defined entity demonstrating features of AIH and PBC. We investigated the usefulness of IgG and IgM immunostaining for the distinction of AIH and PBC and their staining pattern in cases of possible OS. The approximate quantity of IgG+ and IgM+ periportal inflammatory cells in immunohistochemical analysis were compared in cases of AIH, PBC, and OS. AIH cases showed predominant IgG immunostaining of periportal inflammatory cells. A significant number of PBC cases also demonstrated IgG predominance rather than IgM. Six OS cases had IgG predominance, 4 had IgM predominance, and 1 was equivocal. The usefulness of IgG and IgM immunostaining is limited in PBC cases with IgM predominance for excluding AIH. IgG predominance is not specific for AIH. OS does not demonstrate either IgG or IgM predominance (P > .2) and does not help classify OS into either category.
Hormone Receptor and c-ERBB2 Status in Distant Metastatic and Locally Recurrent Breast Cancer
Authors: Idirisinghe Pushpalatha K.A. , Thike Aye Aye , Cheok Poh Yian , Tse Gary Man-Kit , Lui Philip Chi-Wai , Fook-Chong Stephanie , Wong Nan Soon , Tan Puay Hoon
Published: February 2010

Estrogen receptor (ER), progesterone receptor (PR), and c-ERBB2 (HER2/neu) are therapeutically and prognostically important markers in the management of breast carcinoma. They are not always analyzed in distant metastatic and locally recurrent breast cancers. We compared immunohistochemical expression in a series of primary breast carcinomas with their distant metastases (n = 72) and local recurrences (n = 45) and analyzed the impact of any changes on survival. Discordance rates between primary and metastatic and between primary and locally recurrent lesions, respectively, were 18% (13/72) and 13% (6/45) for ER, 42% (30/72) and 33% (15/45) for PR, and 7% (5/72) and 2% (1/45) for c-ERBB2. There was statistically significant discordance between primary and metastatic PR status (P = .017; κ = 0.201). Among locally recurrent tumors, 15 (33%) of 45 revealed discordance for PR (P = .006; κ = 0.366). We observed a trend for shorter survival among women with ER– metastatic and locally recurrent tumors regardless of the primary tumor ER status. Our findings suggest a benefit for routine evaluation of ER, PR, and c-ERBB2 status in distant metastatic and locally recurrent breast cancer for therapeutic and prognostic purposes.
Clinicopathologic Features of Agranulocytosis in the Setting of Levamisole-Tainted Cocaine
Authors: Czuchlewski David R. , Brackney Monica , Ewers Christina , Manna Jonaki , Fekrazad M. Houman , Martinez Afton , Nolte Kurt B. , Hjelle Brian , Rabinowitz Ian , Curtis Brian R. , McFarland Janice G. , Baumbach Joan , Foucar Kathryn
Published: February 2010

Levamisole is a known contaminant of cocaine and, via this route, has been associated with otherwise unexplained agranulocytosis. Levamisole is currently present in the majority of cocaine samples seized by the US Drug Enforcement Agency. We identified 20 cases of unexplained agranulocytosis in our practice locations of Albuquerque, NM, and Vancouver, Canada. Epidemiologic investigation revealed recent or ongoing cocaine use in 14 cases (70%). Certain morphologic features, including circulating plasmacytoid lymphocytes, increased bone marrow plasma cells, and mild megakaryocytic hyperplasia, were associated with the cocaine-exposed group. Of 5 patients tested, 3 (60%) were HLA-B27+ and showed antineutrophil antibodies, consistent with known associations of levamisole-induced agranulocytosis. One patient, who was positive for cocaine and levamisole by toxicology testing, died of infectious complications. Inadvertent consumption of levamisole via cocaine is a severely underappreciated risk factor for agranulocytosis, and specific laboratory features are suggestive of this etiology.
Conjunctive p16INK4a Testing Significantly Increases Accuracy in Diagnosing High-Grade Cervical Intraepithelial Neoplasia
Authors: Bergeron Christine , Ordi Jaume , Schmidt Dietmar , Trunk Marcus J. , Keller Thomas , Ridder Ruediger
Published: February 2010

The histopathologic interpretation of cervical intraepithelial neoplasia (CIN) is subject to a high level of interobserver variability and a substantial number of false-positive and false-negative results. We assessed the impact of the conjunctive interpretation of p16INK4a-immunostained slides on the accuracy of community-based pathologists in diagnosing high-grade cervical intraepithelial neoplasia (CIN; CIN 2 and CIN 3) in biopsy specimens. Twelve pathologists rendered independent diagnoses on a set of 500 H&E-stained cervical punch and conization specimens. Results were compared with a dichotomized “gold standard” established by consensus of 3 gynecopathology experts. When p16INK4a-immunostained slides were added and conjunctively interpreted with the H&E-stained slides, a significant increase in diagnostic accuracy for the detection of high-grade CIN was observed (P = .0004). Sensitivity for high-grade CIN was increased by 13%, cutting the rate of false-negative results in half. Agreement of community-based pathologists in diagnosing high-grade CIN was significantly improved (mean κ values advanced from 0.566 to 0.749; P < .0001). Reproducibility of p16INK4a stain interpretation was excellent (κ = 0.899). Our results show that conjunctive interpretation of p16INK4a-stained slides could significantly improve the routine interpretation of cervical histopathology.
Efficacy and Safety of Ibuprofen and Acetaminophen in Children and Adults: A Meta-Analysis and Qualitative Review
Authors: Pierce Catherine A , Voss Bryan
Published: February 2010

Objective:

To evaluate the analgesic and antipyretic efficacy and safety of ibuprofen compared to acetaminophen in children and adults.

Data Sources:

Literature searches were performed using PubMed/MEDLINE (through August 2009) and EMBASE (through January 2008) and were restricted to the English language. In PubMed/MEDLINE, search terms used were ibuprofen, acetaminophen, paracetamol, clinical trials, and randomized controlled trials. EMBASE search terms included ibuprofen and acetaminophen, restricted to human and clinical trials.

Study Selection And Data Extraction:

All English-language articles identified from the data sources were reviewed. Multiple review articles were studied for any pertinent references and this yielded additional articles. Only articles that directly compared ibuprofen and acetaminophen were eligible for this review.

Data Synthesis:

Eighty-five studies that directly compared ibuprofen to acetaminophen were identified; 54 contained analgesic efficacy data, 35 contained antipyretic/temperature reduction data, and 66 contained safety data (some articles contained more than 1 type of data). Qualitative review of the literature revealed that, for the most part, ibuprofen was more efficacious than acetaminophen for the treatment of pain and fever in both pediatric and adult populations, and that these 2 drugs were equally safe. Meta-analyses on the subset of randomized clinical trial articles that reported sufficient quantitative information to calculate either an odds ratio (adverse event [AE]) or standardized mean difference (pain and fever) confirmed the qualitative results for adult (standardized mean difference [SMD] 0.69; 95% CI 0.57 to 0.81) and pediatric (SMD 0.28; 95% CI 0.10 to 0.46) pain at 2 hours postdose and pediatric fever (SMD 0.26; 95% CI 0.10 to 0.41) at 4 hours postdose. Conclusions regarding adult fever/temperature reduction could not be made due to a lack of evaluable data. The combined odds ratio for the proportion of adult subjects experiencing at least 1 AE slightly favored ibuprofen; however, the difference was not statistically significant (1.12; 95% CI 1.00 to 1.25). No significant difference between drugs in AE incidence was found for pediatric patients (0.82; 95% CI 0.60 to 1.12).

Conclusions:

Ibuprofen is as or more efficacious than acetaminophen for the treatment of pain and fever in adult and pediatric populations and is equally safe.
Shared perceptions of perceived justice as a predictor of externally rated departmental level academic performance
Authors: Lipponen Jukka , Wisse Barbara
Published: February 2010

The purpose of this study was to investigate if shared perceptions of organizational justice affect externally rated group‐level performance. Previous studies did not consider distributive justice, nor did they control for the possible confounding effects of baseline performance. The present study relates shared perceptions of procedural, interactional, and distributive justice to externally rated departmental level academic performance and takes previous performance into account. We found support for the distributive justice–performance link, partial support for the procedural justice‐performance link, but contrary to our expectations interactional justice was unrelated to performance. All in all, the results seem to support the call for a re‐examination of the role of distributive justice in organizational processes.
Interpersonal problems in alexithymia: A study in three primary care groups
Authors: Vanheule Stijn , Vandenbergen Jan , Verhaeghe Paul , Desmet Mattias
Published: February 2010

Objectives.

The present study investigated the relation between alexithymia and interpersonal problems in a sample of primary care patients with either chronic fatigue syndrome (CFS); a chronic cardiovascular or auto‐immune disease; or a minor medical condition. It was hypothesized that the relation between cold interpersonal functioning would account for the differences in alexithymia scores between the patient groups.

Design and methods.

Participants were 155 primary care patients that were recruited through 52 general practitioners: 52 CFS patients; 52 patients with a chronic cardiovascular or auto‐immune disease; 51 patients with a minor medical condition. Interpersonal problems were assessed by means of the Inventory of Interpersonal Problems and alexithymia was assessed by means of the Toronto Alexithymia Scale.

Results.

CFS patients and patients with a chronic cardiovascular or auto‐immune disease have substantially higher alexithymia scores than patients with a minor medical condition. Alexithymia is positively related to cold and distant interpersonal functioning; negatively related to self‐sacrificing and overly accommodating in relation to others; and positively related to vindictiveness and self‐centredness. The relation between alexithymia and these interpersonal problems accounts for the differences in alexithymia scores between the patient groups.

Conclusions.

Alexithymia and interpersonal problems should be considered together, in terms of one deficient affect regulatory system, and the relation between alexithymia and specific illness conditions is secondary to this. Clinical assessment of patients with problems in naming and discussing affective states should never be isolated from an examination of their interpersonal problems, and vice versa. Mentalization‐based therapy is recommended for patients with problems in naming affective states, interpersonal problems, and concomitant CFS or other alexithymia‐related diseases.
Correlation between glucose and bupivacaine levels in cerebrospinal fluid after spinal anesthesia: glycorrhachia as predictor for duration of sensory block
Authors: Estañ-Capell Nuria , Quiñones-Torrelo Carmen , Sánchez-Morillo Jorge , Estañ-Jago Luis
Published: February 2010

Background: Prediction of the duration of motor block after injection of a local anesthetic into cerebrospinal fluid (CSF) would be a very useful tool in clinical practice. However, previous attempts have not shown conclusive results. In this work, glycorrhachia is demonstrated to be an adequate predictive parameter after spinal anesthesia using 0.5% hyperbaric bupivacaine.

Methods: Two mL of local anesthetic through a continuous spinal catheter was administered to 40 patients. CSF was sampled at different time intervals from the onset of infusion to motor recovery. CSF bupivacaine concentrations were measured using chromatography. An automated analyzer was used for determining glycorrhachia in the same samples.

Results: For all patients, good correlation (r2>0.95, p<0.05) was obtained. From these results, it was possible to develop a general model which establishes the relationship between CSF glucose and bupivacaine concentrations (R2=0.987, p<0.05). Motor block is reached when CSF glucose concentration is about 245 mg/dL (13.5 mmol/L), which corresponds to 35 μg/mL of bupivacaine.

Conclusions: Glycorrhachia measured during surgical intervention in patients undergoing spinal anesthesia with hyperbaric bupivacaine provides a mechanism for predicting the duration of motor block in a rapid and simple manner.

Clin Chem Lab Med 2010;48:523–30.
Pregnancy-associated plasma protein-A as a marker for long-term mortality in patients with peripheral atherosclerosis: inconclusive findings from the Linz Peripheral Arterial Disease (LIPAD) study
Authors: Mueller Thomas , Dieplinger Benjamin , Forstner Thomas , Poelz Werner , Haltmayer Meinhard
Published: February 2010

Background: Pregnancy-associated plasma protein-A (PAPP-A) has been associated with peripheral artery disease (PAD). The aim of this study was to evaluate the utility of PAPP-A as a marker for long-term mortality in patients with atherosclerotic PAD.

Methods: PAPP-A serum concentrations were measured using an enzymatically amplified two-step sandwich-type immunoassay in 487 consecutive patients admitted to a tertiary care hospital with symptomatic PAD. The main outcome measure was all-cause mortality at 5 years.

Results: During follow-up, 114 patients died and 373 survived. The median PAPP-A concentration was higher among decedents compared with survivors (0.96 vs. 0.78 mU/L, p=0.024). The area under the receiver operating characteristic curve for the prediction of 5-year mortality by PAPP-A was 0.57 [95% confidence interval (CI), 0.53–0.61; p=0.026]. Survival probability was not significantly associated with PAPP-A concentrations using Kaplan-Meier curve analysis. However, univariate Cox proportional-hazards regression analysis revealed that PAPP-A was associated with 5-year mortality [risk ratio 1.25; 95% CI, 1.05–1.50; p=0.013 per one standard deviation (SD) increase in log transformed values]. In the multivariate model using a bootstrapping method, the predictive value of PAPP-A remained significant (risk ratio 1.31; 95% CI, 1.01–1.73; p=0.024 per 1 SD increase in log transformed values), even after adjustment for clinical confounders and other biomarkers, such as high-sensitivity C-reactive protein and amino terminal pro-B-type natriuretic peptide.

Conclusions: In this study, PAPP-A was an independent predictor of 5-year all-cause mortality in patients with symptomatic PAD. However, based on the weak association between PAPP-A and outcome in our cohort, we consider PAPP-A measurements to not be useful in clinical practice for prognostic purposes in patients with PAD.

Clin Chem Lab Med 2010;48:537–42.
The puzzling uniqueness of the heterotrimeric G15 protein and its potential beyond hematopoiesis
Authors: Giannone Flavia , Malpeli Giorgio , Lisi Veronica , Grasso Silvia , Shukla Priyanka , Ramarli Dunia , Sartoris Silvia , Monsurró Vladia , Krampera Mauro , Amato Eliana , Tridente Giuseppe , Colombatti Marco , Parenti Marco , Innamorati Giulio
Published: February 2010

Heterotrimeric G proteins transduce the signals of the largest family of membrane receptors (G protein-coupled receptors, GPCRs) hence triggering the activation of a wide variety of physiological responses. G15 is a G protein characterized by a number of functional peculiarities that make its signaling exceptional: 1) it can couple a variety of Gs-, Gi/o-, and Gq-linked receptors to phospholipase C activation; 2) relatively to other G proteins, it is poorly affected by β-arrestin-dependent desensitization, the general mechanism that regulates GPCR function and 3) at the protein level, its expression is only detected in highly specific cell types (hematopoietic and epithelial cells). G15 α-subunit displays unique structural and biochemical properties, and is phylogenetically the most recent and divergent component of the Gαq/11 subfamily. All these aspects shed a mysterious light on G15 biological role, which remains substantially elusive. Thus, far, G15 signaling has been analyzed in the context of hematopoiesis. Here, we highlight observations supporting the view that G15 functions may extend further beyond the immune system. In addition, we describe puzzling aspects of G15 signaling that offer a novel perspective in the understanding of its physiological role.
Association of Periodontitis With Rheumatoid Arthritis: A Pilot Study
Authors: Dissick Addie , Redman Robert S. , Jones Miata , Rangan Bavana V. , Reimold Andreas , Griffiths Garth R. , Mikuls Ted R. , Amdur Richard L. , Richards John S. , Kerr Gail S.
Published: February 2010

Background: Similarities exist in the epidemiology and immunopathogenesis of periodontitis and rheumatoid arthritis (RA), but the associations between their respective disease activities and severities are less well documented. We evaluated the prevalence and severity of periodontitis in United States (U.S.) veterans with RA and their relationship to RA disease activity and severity.

Methods: Patients with RA from an outpatient rheumatology clinic were eligible, and patients with osteoarthritis (OA) served as controls. Dentists, masked to the rheumatologic diagnoses, performed periodontal probing and examined dental panoramic radiographs to assess the presence and severity of periodontitis. Associations of periodontitis with RA were examined using multivariate regression, whereas the association of periodontitis with disease‐severity measures in RA was examined using the χ2 test.

Results: Sixty‐nine patients with RA (57 males and 12 females) and 35 patients with OA (30 males and five females) were studied. Moderate to severe periodontitis was more prevalent in patients with RA (51%) than controls (26%) (P = 0.03), an association independent of age, race, smoking, diabetes mellitus, and gender. Patients with RA who were seropositive for rheumatoid factor (RF) were more likely to have moderate to severe periodontitis (59%) than patients who were RF negative (15%) (P = 0.02). Likewise, patients with RA who were positive for the anti–cyclic citrullinated peptide (CCP) antibodies were more likely to have moderate to severe periodontitis (56%) than patients who were anti‐CCP negative (22%) (P = 0.01). There were no associations of periodontitis status with other measures of RA disease activity or severity.

Conclusions: In a cohort of U.S. veterans, periodontitis was more common and severe in patients with RA compared to patients with OA. Although unrelated to disease activity, the presence of periodontitis in patients with RA was associated with seropositivity for RF and the anti‐CCP antibody, which was highly relevant given the associations of these autoantibodies with poor outcomes and disease pathogenesis in RA.
Periodontal Diseases, Preterm Births, and Low Birth Weight: Findings From a Homogeneous Cohort of Women in Madagascar
Authors: Rakoto‐Alson Simone , Tenenbaum Henri , Davideau Jean‐Luc
Published: February 2010

Background: Studies on the possible influence of periodontal disease on preterm birth (PB) and/or low birth weight (LBW) remain inconclusive especially due to the difficulty of assessing the numerous pregnancy risk factors. The aim of this study is to compare periodontal parameters in non‐smoking pregnant women in Madagascar.

Methods: A cohort study with 204 pregnant women (mean age: 25.6 years) was conducted in public prenatal care health clinics in Madagascar. Socioeconomic and obstetric information was obtained. Periodontal parameters, such as periodontal probing depth and clinical attachment loss (AL), were recorded during the second semester of pregnancy.

Results: The mean gestational age was 37.8 weeks. At delivery, 42 newborns (20.6%) were PBs (<37 weeks) and 22 newborns (10.8%) were of LBW (<2,500 g). Nine (4.4%) newborns were PBs and of LBW (PLBW), and 149 newborns (73%) were of normal birth. The mean plaque index and papillary bleeding index were significantly higher (P <0.05 and P <0.001, respectively) in the PB group than in the normal birth group. Periodontitis (at least three sites from different teeth with clinical AL ≥4 mm) was significantly associated with PB (P <0.001), LBW (P <0.001), and PLBW (P <0.01). The rates of periodontitis were considerably higher in PB (78.6%), LBW (77.3%), and PLBW (77.8%) groups than in the full‐term (8.6%), normal weight (16.5%), and normal birth (2.7%) groups.

Conclusion: The strong association among periodontitis, PB, and LBW in this study highlights the need to consider the periodontal status of pregnant women in Madagascar.