bluuebunny/crossref_metadata_2025 huggingface dataset:
Oxidative stress promotes autophagic cell death in human neuroblastoma cells with ectopic transfer of mitochondrial PPP2R2B (Bβ2)
Authors: Cheng Wan-Ting , Guo Zhi-Xuan , Lin Chia-An , Lin Ming-Yi , Tung Li-Chu , Fang Kang
Published: December 2009

Background

The multifunctional protein phosphatase 2A (PP2A) is a heterotrimeric serine/threonine protein phosphatase composed of a scaffolding, catalytic and regulatory subunits. By modifying various downstream signal transducers, the aberrant expression of the brain-targeted regulatory subunit PPP2R2B is associated with the onset of a panel of neuronal disorders. The alternatively splicing of PPP2R2B encodes two regulatory subunit isoforms that determine cellular distribution of the neuron-specific holoenzyme to mitochondria (Bβ2) and cytoplasm (Bβ1), respectively.

Results

Human neuroblastoma cells were transfected with PPP2R2B constructs encoding the complete sequences of Bβ2 and Bβ1, respectively. The colonies with antibiotic resistance were selected as stable cell lines. Both ectopic Bβ1 and Bβ2 clones exhibited characteristics of autophagy. To test how cells respond to reactive oxygen species generators, the cells were treated with either hydrogen peroxide or t-butyl hydroperoxide and Bβ2 clones induced cell death. Suppression of autophagy using either RNA interference of the essential autophagy gene or pharmacological inhibitor rescued cell death caused by oxidative stress.

Conclusions

Cells with ectopically expressed mitochondria-targeted regulatory subunit PPP2R2B of the holoenzyme PP2A were shown predisposed to autophagy and oxidative stress induced cell death that is related to apoptosis. The results promised a model for studying the mechanism and function of aberrant PPP2R2B expression in neuronal cells. The work provided a new target for understanding and prevention of neuropathogenesis.
Model system for the analysis of cell surface expression of human ABCA1
Authors: Kasza Ildikó , Hegyi Zoltán , Szabó Katalin , Andrikovics Hajnalka , Német Katalin , Váradi András , Sarkadi Balázs , Homolya László
Published: December 2009

Background

The ABCA1 protein plays a pivotal role in reverse cholesterol transport, by mediating the generation of HDL particles and removing cellular cholesterol. Both the proper expression of ABCA1 in the plasma membrane and the internalization along with apoA-I are required for function. Therefore, we developed a model system to investigate the effect of clinically relevant drugs on the cell surface appearance of ABCA1.

Results

By retroviral transduction system, we established stable mammalian cell lines expressing functional and non-functional ABCA1 variants, tagged with an extracellular hemagglutinin epitope. After characterization of the expression, proper localization and function of different ABCA1 variants, we followed quantitatively their cell surface expression by immunofluorescent staining, using flow cytometry. As expected, we found increased cell surface expression of ABCA1 after treatment with a calpain inhibitor, and observed a strong decrease in plasma membrane ABCA1 expression upon treatment with a trans-Golgi transport inhibitor, Brefeldin A. We tested cholesterol level lowering drugs and other potential inhibitors of ABCA1. Here we demonstrate that ezetimibe affects ABCA1 cell surface expression only in the case of a functional ABCA1.

Conclusions

Our model system allows a quantitative detection of cell surface expression of ABCA1, screening of substrates or specific inhibitors, and investigating transport regulation.
Genome-wide profiling of Populus small RNAs
Authors: Klevebring Daniel , Street Nathaniel R , Fahlgren Noah , Kasschau Kristin D , Carrington James C , Lundeberg Joakim , Jansson Stefan
Published: December 2009

Background

Short RNAs, and in particular microRNAs, are important regulators of gene expression both within defined regulatory pathways and at the epigenetic scale. We investigated the short RNA (sRNA) population (18-24 nt) of the transcriptome of green leaves from the sequenced Populus trichocarpa using a concatenation strategy in combination with 454 sequencing.

Results

The most abundant size class of sRNAs were 24 nt. Long Terminal Repeats were particularly associated with 24 nt sRNAs. Additionally, some repetitive elements were associated with 22 nt sRNAs. We identified an sRNA hot-spot on chromosome 19, overlapping a region containing both the proposed sex-determining locus and a major cluster of NBS-LRR genes. A number of phased siRNA loci were identified, a subset of which are predicted to target PPR and NBS-LRR disease resistance genes, classes of genes that have been significantly expanded in Populus. Additional loci enriched for sRNA production were identified and characterised. We identified 15 novel predicted microRNAs (miRNAs), including miRNA*sequences, and identified a novel locus that may encode a dual miRNA or a miRNA and short interfering RNAs (siRNAs).

Conclusions

The short RNA population of P. trichocarpa is at least as complex as that of Arabidopsis thaliana. We provide a first genome-wide view of short RNA production for P. trichocarpa and identify new, non-conserved miRNAs.
The commonly-used DNA probe for diffusely-adherent Escherichia coli cross-reacts with a subset of enteroaggregative E. coli
Authors: Snelling Anna M , Macfarlane-Smith Louissa R , Fletcher Jonathan N , Okeke Iruka N
Published: December 2009

Background

The roles of diffusely-adherent Escherichia coli (DAEC) and enteroaggregative E. coli (EAEC) in disease are not well understood, in part because of the limitations of diagnostic tests for each of these categories of diarrhoea-causing E. coli. A HEp-2 adherence assay is the Gold Standard for detecting both EAEC and DAEC but DNA probes with limited sensitivity are also employed.

Results

We demonstrate that the daaC probe, conventionally used to detect DAEC, cross-reacts with a subset of strains belonging to the EAEC category. The cross hybridization is due to 84% identity, at the nucleotide level, between the daaC locus and the aggregative adherence fimbriae II cluster gene, aafC, present in some EAEC strains. Because aaf-positive EAEC show a better association with diarrhoea than other EAEC, this specific cross-hybridization may have contributed to an over-estimation of the association of daaC with disease in some studies. We have developed a discriminatory PCR-RFLP protocol to delineate EAEC strains detected by the daaC probe in molecular epidemiological studies.

Conclusions

A PCR-RFLP protocol described herein can be used to identify aaf-positive EAEC and daaC-positive DAEC and to delineate these two types of diarrhoeagenic E. coli, which both react with the daaC probe. This should help to improve current understanding and future investigations of DAEC and EAEC epidemiology.
Growth in glucose-based medium and exposure to subinhibitory concentrations of imipenem induce biofilm formation in a multidrug-resistant clinical isolate of Acinetobacter baumannii
Authors: Nucleo Elisabetta , Steffanoni Laura , Fugazza Giulia , Migliavacca Roberta , Giacobone Ernesto , Navarra Antonella , Pagani Laura , Landini Paolo
Published: December 2009

Background

Acinetobacter baumannii is emerging as an important nosocomial pathogen. Multidrug resistance, as well as ability to withstand environmental stresses, makes eradication of A. baumannii difficult, particularly from hospital settings.

Results

Over a six-year period, 73 isolates of A. baumannii were collected from infected patients in two hospitals in Italy. While 69 out of the 73 isolates displayed identical multidrug antibiotic resistance pattern, they were susceptible to carbapenems. Genetic profiles of these 69 isolates, determined by Pulsed Field Gel Electrophoresis (PFGE), indicated that they were genetically related and could be clustered in a specific clone, called SMAL. We tested the ability of the SMAL clone to form biofilm, an important determinant for bacterial colonization of the human host and for persistence in the hospital environment. Biofilm formation by A. baumannii SMAL, measured as surface adhesion to polystyrene, is strongly affected by growth conditions, being impaired in rich growth media such as LB, while being favoured in glucose-based medium. Surface adhesion in glucose-based media is inhibited by treatment with cellulase, suggesting that it depends on production of cellulose or of a chemically related extracellular polysaccharide. Exposure of A. baumannii SMAL to subinhibitory concentrations of imipenem resulted in biofilm stimulation and increased production of iron uptake proteins. Growth in iron-supplemented medium also stimulated surface adhesion, thus suggesting that increased intracellular iron concentrations might act as an environmental signal for biofilm formation in A. baumannii SMAL.

Conclusions

Our results indicate that exposure to subinhibitory concentrations of imipenem can stimulate biofilm formation and induce iron uptake in a pathogenic strain of A. baumannii, with potential implications on antibiotic susceptibility and ability to persist in the human host.
Intrinsic chemoresistance to gemcitabine is associated with constitutive and laminin-induced phosphorylation of FAK in pancreatic cancer cell lines
Authors: Huanwen Wu , Zhiyong Liang , Xiaohua Shi , Xinyu Ren , Kai Wang , Tonghua Liu
Published: December 2009

Background

One of the major reasons for poor prognosis of pancreatic cancer is its high resistance to currently available chemotherapeutic agents. In recent years, focal adhesion kinase (FAK), a central molecule in extracellular matrix (ECM)/integrin-mediated signaling, has been thought to be a key determinant of chemoresistance in cancer cells. In this study, we aimed to determine the roles of FAK phosphorylation in the intrinsic chemoresistance of pancreatic cancer cell lines.

Results

Our results showed that, the level of constitutive phosphorylation of FAK at Tyr397 correlated with the extent of intrinsic resistance to Gemcitabine (Gem) in four pancreatic cancer cell lines. Moreover, in Panc-1 cells, which had high expression of pFAK, specific inhibition of constitutive FAK phosphorylation by either RNAi or FRNK overexpression decreased the phosphorylation of Akt, reduced the levels of survivin expression and Bad phosphorylation at Ser136 and increased Gem-induced cytotoxicity and apoptosis. However, in AsPC-1 cells with a low level of pFAK, neither FAK RNAi nor FRNK overexpression affected Gem-induced cell apoptosis. We further found that laminin (LN) induced FAK and Akt phosphorylation in a time-dependent manner, increased the levels of survivin and pBad (pS136) and decreased Gem-induced cytotoxicity and apoptosis in AsPC-1 cells; Specific inhibition of LN-induced FAK phosphorylation by either FAK RNAi or FRNK overexpression suppressed the effects of LN on AsPC-1 cells. Moreover, inhibition of constitutive FAK phosphorylation in Panc-1 cells and LN-induced FAK phosphorylation in AsPC-1 cells by a novel and more specific FAK phosphorylation inhibitor PF-573,228 showed similar results with those of FAK phosphorylation inhibition by FAK RNAi or FRNK overexpression.

Conclusions

In conclusion, our research demonstrates for the first time that both constitutive and LN-induced FAK phosphorylation contribute to increased intrinsic chemoresistance to Gem in pancreatic cancer cell lines and these effects are partly due to the regulation of Akt and Bad phosphorylation and survivin expression. Development of selective FAK phosphorylation inhibitors may be a promising way to enhance chemosensitivity in pancreatic cancer.
Effect of filtration on morphine and particle content of injections prepared from slow-release oral morphine tablets
Authors: McLean Stuart , Bruno Raimondo , Brandon Susan , de Graaff Barbara
Published: December 2009

Background

Injections of mixtures prepared from crushed tablets contain insoluble particles which can cause embolisms and other complications. Although many particles can be removed by filtration, many injecting drug users do not filter due to availability, cost or performance of filters, and also due to concerns that some of the dose will be lost.

Methods

Injection solutions were prepared from slow-release morphine tablets (MS Contin®) replicating methods used by injecting drug users. Contaminating particles were counted by microscopy and morphine content analysed by liquid chromatography before and after filtration.

Results

Unfiltered tablet extracts contained tens of millions of particles with a range in sizes from < 5 μm to > 400 μm. Cigarette filters removed most of the larger particles (> 50 μm) but the smaller particles remained. Commercial syringe filters (0.45 and 0.22 μm) produced a dramatic reduction in particles but tended to block unless used after a cigarette filter. Morphine was retained by all filters but could be recovered by following the filtration with one or two 1 ml washes. The combined use of a cigarette filter then 0.22 μm filter, with rinses, enabled recovery of 90% of the extracted morphine in a solution which was essentially free of tablet-derived particles.

Conclusions

Apart from overdose and addiction itself, the harmful consequences of injecting morphine tablets come from the insoluble particles from the tablets and microbial contamination. These harmful components can be substantially reduced by passing the injection through a sterilizing (0.22 μm) filter. To prevent the filter from blocking, a preliminary coarse filter (such as a cigarette filter) should be used first. The filters retain some of the dose, but this can be recovered by following filtration with one or two rinses with 1 ml water. Although filtration can reduce the non-pharmacological harmful consequences of injecting tablets, this remains an unsafe practice due to skin and environmental contamination by particles and microorganisms, and the risks of blood-borne infections from sharing injecting equipment.
Correlation between expression of p53, p21/WAF1, and MDM2 proteins and their prognostic significance in primary hepatocellular carcinoma
Authors: Zhang Mei-Fang , Zhang Zhi-Yi , Fu Jia , Yang Yu-Feng , Yun Jing-Ping
Published: December 2009

Background

Tumor Protein p53 (p53), cyclin-dependent kinase inhibitor 1A (p21/WAF1), and murine double minute 2 (MDM2) participate in the regulation of cell growth. Altered expression of these gene products has been found in malignant tumors and has been associated with poor prognosis. Our aim was to investigate the expression of the 3 proteins in hepatocellular carcinoma (HCC) and their prognostic significance.

Methods

We examined p53, p21/WAF1, and MDM2 expression in 181 pairs of HCC tissues and the adjacent hepatic tissues by performing immunohistochemistry and examined the expression of the 3 proteins in 7 pairs of HCC tissues and the adjacent hepatic tissues by using western blot analysis.

Results

The expression of p53, p21/WAF1, and MDM2 in the HCC tissues was significantly higher than those in the adjacent hepatic tissues (P < 0.05). A statistical correlation was observed between p53 and p21/WAF1 expression in HCC tissues (R = 0.195, P = 0.008). A statistical correlation was observed between expression of p53 and p21/WAF1 (R = 0.380, P = 0.000), p53 and MDM2 (R = 0.299, P = 0.000), p21/WAF1 and MDM2 (R = 0.285, P = 0.000) in 181 liver tissues adjacent to the tumor. Patients with a low pathologic grade HCC (I+II) had a higher tendency to express p53 on tumor cells than the patients with high pathologic grade HCC (III+IV) (P = 0.007). Survival analysis showed that positive p21/WAF1 expression or/and negative MDM2 expression in HCC was a predictor of better survival of patients after tumor resection (P < 0.05).

Conclusions

The proteins p53, p21/WAF1, and MDM2 were overexpressed in all the HCC cases in this study, and p53 and p21/WAF1 overexpression were positively correlated. The expression of p21/WAF1 and MDM2 can be considered as 2 useful indicators for predicting the prognosis of HCC.
Acute hepatitis in three patients with systemic juvenile idiopathic arthritis taking interleukin-1 receptor antagonist
Authors: Canna Scott , Frankovich Jennifer , Higgins Gloria , Narkewicz Michael R , Nash S Russell , Hollister J Roger , Soep Jennifer B , Dragone Leonard L
Published: December 2009

Purpose

We investigated the etiology of acute hepatitis in three children with systemic Juvenile Idiopathic Arthritis (sJIA) taking Interleukin-1 receptor antagonist (IL1RA).

Methods

Laboratory and clinical data for three children with sJIA diagnosed at ages 13 months to 8 years who developed acute hepatitis during treatment with IL1RA were reviewed for evidence of sJIA flare, infection, macrophage activation syndrome (MAS), malignancy, and drug reaction.

Results

In all patients, hepatitis persisted despite cessation of known hepatotoxic drugs and in absence of known infectious triggers, until discontinuation of IL1RA. Liver biopsies had mixed inflammatory infiltrates with associated hepatocellular injury suggestive of an exogenous trigger. At the time of hepatitis, laboratory data and liver biopsies were not characteristic of MAS. In two patients, transaminitis resolved within one week of discontinuing IL1RA, the third improved dramatically in one month.

Conclusions

Although sJIA symptoms improved significantly on IL1RA, it appeared that IL1RA contributed to the development of acute hepatitis. Hepatitis possibly occurred as a result of an altered immune response to a typical childhood infection while on IL1RA. Alternatively, hepatitis could have represented an atypical presentation of MAS in patients with sJIA taking IL1RA. Further investigation is warranted to determine how anti-IL1 therapies alter immune responsiveness to exogenous triggers in patients with immune dysfunction such as sJIA. Our patients suggest that close monitoring for hepatic and other toxicities is indicated when treating with IL1RA.
Spatiotemporal regulation of the cough motor pattern
Authors: Wang Cheng , Saha Sourish , Rose Melanie J , Davenport Paul W , Bolser Donald C
Published: December 2009

The purpose of this study was to identify the spatiotemporal determinants of the cough motor pattern. We speculated that the spatial and temporal characteristics of the cough motor pattern would be regulated separately. Electromyograms (EMG) of abdominal muscles (ABD, rectus abdominis or transversus abdominis), and parasternal muscles (PS) were recorded in anesthetized cats. Repetitive coughing was produced by mechanical stimulation of the lumen of the intrathoracic trachea. Cough inspiratory (CTI) and expiratory (CTE) durations were obtained from the PS EMG. The ABD EMG burst was confined to the early part of CTEand was followed by a quiescent period of varying duration. As such, CTEwas divided into two segments with CTE1defined as the duration of the ABD EMG burst and CTE2defined as the period of little or no EMG activity in the ABD EMG. Total cough cycle duration (CTTOT) was strongly correlated with CTE2(r2>0.8), weakly correlated with CTI(r2<0.3), and not correlated with CTE1(r2<0.2). There was no significant relationship between CTIand CTE1or CTE2. The magnitudes of inspiratory and expiratory motor drive during cough were only weakly correlated with each other (r2<0.36) and were not correlated with the duration of any phase of cough. The results support: a) separate regulation of CTIand CTE, b) two distinct subphases of CTE(CTE1and CTE2), c) the duration of CTE2is a primary determinant of CTTOT, and d) separate regulation of the magnitude and temporal features of the cough motor pattern.
Superior sagittal sinus thrombosis presenting as a continuous headache: a case report and review of the literature
Authors: Gupta Rishi K , Jamjoom Aimun AB , Devkota Upendra P
Published: December 2009

Cerebral venous sinus thrombosis is a rare but dangerous condition, occurring with an incidence of 3-4 cases/million/year. Cerebral venous sinus thrombosis presents a diagnostic challenge due to its varied presentation patterns. We report a case of a 42 year old Nepali man diagnosed with cerebral venous sinus thrombosis after presenting with a week long history of continuous headache. He improved rapidly following prompt anticoagulation. Despite thorough investigation no discernable underlying cause could be found. Our report highlights the value of prompt diagnosis of cerebral venous sinus thrombosis through neuroimaging and the importance of immediate anticoagulation as part of patient management.
HES 130/0.4 impairs haemostasis and stimulates pro-inflammatory blood platelet function
Authors: Sossdorf Maik , Marx Sascha , Schaarschmidt Barbara , Otto Gordon P , Claus Ralf A , Reinhart Konrad , Hartog Christiane S , Lösche Wolfgang
Published: December 2009

Introduction

Hydroxyethyl starch (HES) solutions are widely used for volume replacement therapy but are also known to compromise coagulation, impair renal function and increase long-term mortality. To test the hypotheses that HES 130/0.4 has fewer adverse effects than HES 200/0.5 and exerts anti-inflammatory properties, we compared the effects of HES 130/0.4, HES 200/0.5 and saline on in vitro haemostasis and pro-inflammatory platelet function.

Methods

Whole blood samples from healthy volunteers were mixed with 6% HES 130/0.4, 10% HES 200/0.5, or normal saline to achieve a final haemodilution rate of 10% or 40%. Haemostatic capacity was characterised by thromboelastography (ROTEM) and measurement for FXIIIa activity. Platelet activation and pro-inflammatory platelet functions were characterised by flow cytometry measuring the platelet activation marker CD62P and binding of fibrinogen to platelets as well as the formation of heterotypic platelet-leukocyte conjugates.

Results

Compared with saline, HES 130/0.4 dose-dependently impaired formation and firmness of the fibrin clot but did not affect the fibrin crosslinking activity of FXIIIa. At 40% but not at 10% haemodilution rate, HES 200/0.5 also increased platelet fibrinogen binding and both HES solutions increased expression of CD62P, the main receptor for platelet-leukocyte adhesion. HES 130/0.4 but not HES 200/0.5 increased formation of platelet-neutrophil conjugates and, to a lesser degree, platelet-monocyte conjugates.

Conclusions

Our data demonstrate that HES 130/0.4 has similar adverse effects as HES 200/0.5. In particular, both types of HES impair coagulation capacity and stimulate, rather than attenuate, pro-inflammatory platelet function.
Characterizing the admixed African ancestry of African Americans
Authors: Zakharia Fouad , Basu Analabha , Absher Devin , Assimes Themistocles L , Go Alan S , Hlatky Mark A , Iribarren Carlos , Knowles Joshua W , Li Jun , Narasimhan Balasubramanian , Sidney Steven , Southwick Audrey , Myers Richard M , Quertermous Thomas , Risch Neil , Tang Hua
Published: December 2009

Background

Accurate, high-throughput genotyping allows the fine characterization of genetic ancestry. Here we applied recently developed statistical and computational techniques to the question of African ancestry in African Americans by using data on more than 450,000 single-nucleotide polymorphisms (SNPs) genotyped in 94 Africans of diverse geographic origins included in the HGDP, as well as 136 African Americans and 38 European Americans participating in the Atherosclerotic Disease Vascular Function and Genetic Epidemiology (ADVANCE) study. To focus on African ancestry, we reduced the data to include only those genotypes in each African American determined statistically to be African in origin.

Results

From cluster analysis, we found that all the African Americans are admixed in their African components of ancestry, with the majority contributions being from West and West-Central Africa, and only modest variation in these African-ancestry proportions among individuals. Furthermore, by principal components analysis, we found little evidence of genetic structure within the African component of ancestry in African Americans.

Conclusions

These results are consistent with historic mating patterns among African Americans that are largely uncorrelated to African ancestral origins, and they cast doubt on the general utility of mtDNA or Y-chromosome markers alone to delineate the full African ancestry of African Americans. Our results also indicate that the genetic architecture of African Americans is distinct from that of Africans, and that the greatest source of potential genetic stratification bias in case-control studies of African Americans derives from the proportion of European ancestry.
Interaction with surrounding normal epithelial cells influences signalling pathways and behaviour of Src-transformed cells
Authors: Kajita Mihoko , Hogan Catherine , Harris Andrew R. , Dupre-Crochet Sophie , Itasaki Nobue , Kawakami Koichi , Charras Guillaume , Tada Masazumi , Fujita Yasuyuki
Published: December 2009

At the initial stage of carcinogenesis, transformation occurs in a single cell within an epithelial sheet. However, it remains unknown what happens at the boundary between normal and transformed cells. Using Madin-Darby canine kidney (MDCK) cells transformed with temperature-sensitive v-Src, we have examined the interface between normal and Src-transformed epithelial cells. We show that Src-transformed cells are apically extruded when surrounded by normal cells, but not when Src cells alone are cultured, suggesting that apical extrusion occurs in a cell-context-dependent manner. We also observe apical extrusion of Src-transformed cells in the enveloping layer of zebrafish gastrula embryos. When Src-transformed MDCK cells are surrounded by normal MDCK cells, myosin-II and focal adhesion kinase (FAK) are activated in Src cells, which further activate downstream mitogen-activated protein kinase (MAPK). Importantly, activation of these signalling pathways depends on the presence of surrounding normal cells and plays a crucial role in apical extrusion of Src cells. Collectively, these results indicate that interaction with surrounding normal epithelial cells influences the signalling pathways and behaviour of Src-transformed cells.
Fixative Composition Alters Distributions of Immunoreactivity for Glutaminase and Two Markers of Nociceptive Neurons, Nav 1.8 and TRPV1, in the Rat Dorsal Root Ganglion
Authors: Hoffman E. Matthew , Schechter Ruben , Miller Kenneth E.
Published: December 2009

Most, if not all, dorsal root ganglion (DRG) neurons use the neurotransmitter glutamate. There are, however, conflicting reports of the percentages of DRG neurons that express glutaminase (GLS), the enzyme that synthesizes glutamate, ranging from 30% to 100% of DRG neurons. Defining DRG neuron populations by the expression of proteins like GLS, which indicates function, is routinely accomplished with immunolabeling techniques. Proper characterization of DRG neuron populations relies on accurate detection of such antigens. It is known intuitively that fixation can alter immunoreactivity (IR). In this study, we compared the effects of five formaldehyde concentrations between 0.25% and 4.0% (w/v) and five picric acid concentrations between 0.0% and 0.8% (w/v) on the IR of GLS, the voltage-gated sodium channel 1.8 (Nav 1.8), and the capsaicin receptor TRPV1. We also compared the effects of five incubation time lengths from 2 to 192 hr, in primary anti-serum on IR. Lowering formaldehyde concentration elevated IR for all three antigens, while raising picric acid concentration increased Nav 1.8 and TRPV1 IR. Increasing IR improved detection sensitivity, which led to higher percentages of labeled DRG neurons. By selecting fixation conditions that optimized IR, we found that all DRG neurons express GLS, 69% of neurons express Nav 1.8, and 77% of neurons express TRPV1, indicating that some previous studies may have underestimated the percentages of DRG neurons expressing these proteins. This manuscript contains online supplemental material at http://www.jhc.org . Please visit this article online to view these materials.
The Effect of Averaging Time on Oximetry Values in the Premature Infant
Authors: Ahmed Sally Jasmine Maia , Rich Wade , Finer Neil Norman
Published: December 2009

BACKGROUND: The effect of a pulse oximeter's averaging time on time spent by premature infants in established saturation ranges, as well as on desaturation events, has not previously been reported.

OBJECTIVE: Our goal for this pilot study was to evaluate the effect of 2 distinct averaging times on the amount of time spent outside target oxygen saturation (SPo2) as well as the types and durations of desaturation events.

METHODS: Neonates <32 weeks' gestation requiring respiratory support were included. Each infant was studied with 2 simultaneous pulse oximeters: 1 with a short averaging time (2 seconds) and 1 with a longer averaging time (16 seconds). Time spent within different saturation ranges and the number, duration, and severity of desaturation events for each averaging time were compared by using Student's 2-tailed t test, and a P value of <.05 was considered significant.

RESULTS: The number of desaturations was greater with the 2-second averaging time; however, it did not reach significance when only desaturations of clinically significant duration were included (22.3 vs 19.4 significant events were detected by 2- and 16-second averaging time; P = .1). There was a trend for the longer averaging time to underestimate brief desaturation lasting <30 seconds (16 vs 41). Longer averaging time also tended to overestimate events of long duration that lasted >300 seconds (40 vs 20 long events). Longer averaging time tended to underestimate desaturation events of greater severity when SPo2 was <70% (P = .01). Averaging time did not affect the time spent within various target SPo2 ranges (P > .05).

CONCLUSIONS: These preliminary results suggest that use of longer averaging time reduces the detection of brief periodic desaturation events and of greater severity. It may also interpret a cluster of shorter events as a single, prolonged episode and, thus, potentially overestimate the frequency of long events. The significance of these findings lies in the potential impact on neurodevelopmental outcomes and growth, which will need additional study.
Improved Care and Growth Outcomes by Using Hybrid Humidified Incubators in Very Preterm Infants
Authors: Kim Sung Mi , Lee Edward Y. , Chen Jie , Ringer Steven Alan
Published: December 2009

OBJECTIVE: To identify changes in temperature, fluid and electrolyte management, growth, and short-term outcome in extremely low birth weight (ELBW) infants nursed in humidified hybrid incubators (HI group) compared with a cohort of patients cared for in nonhumidified conventional incubators (CI group).

METHODS: Body temperature (BT), fluid and electrolyte balance, and growth velocity (GV) were collected retrospectively on 182 ELBW infants. The CI group included ELBW infants cared for with radiant warmers followed by an incubator without humidity. The HI group included ELBW infants cared for in the radiant warmer mode in a Giraffe OmniBed, followed by the incubator mode using high humidity.

RESULTS: The CI group included more multiple births (50.6%) than the HI group (35.8%; P < .05), but there was no difference in demographic characteristics. BT was similar during the first week. The HI group had less fluid intake, urine output, and insensible water loss, less maximum weight loss, and a lower incidence of hypernatremia during the first week than did the CI group (P < .05). The HI group also had a lower frequency of electrolyte sampling and packed red cell transfusion (P < .05), a higher incidence of hyponatremia on postnatal day 1 than the CI group (P < .05), and a higher GV than the CI group (15.2 ± 5.0 vs 13.5 ± 4.8 g/kg per day), especially among those with a birth weight of ≤749 g (P < .01). There was no difference in sepsis, necrotizing enterocolitis, intraventricular hemorrhage, and all bronchopulmonary dysplasia (BPD), but there was a decreased incidence of severe BPD (5.1% [HI] vs 16.4% [CI]; P < .05) and duration of assisted ventilation in the HI group compared with the CI group (15.5 ± 2.1 vs 19.6 ± 2.4 days, respectively; P = .068).

CONCLUSIONS: Use of a humidified hybrid incubator improved care for ELBW infants by making it possible to decrease fluid intake, improve electrolyte balance, and enhance GV without a disturbance of BT compared with conventional care. By adjusting fluid intake when using these devices, benefits may be enhanced and the risk of BPD and severe BPD may be reduced.
Can the OSA-18 Quality-of-Life Questionnaire Detect Obstructive Sleep Apnea in Children?
Authors: Constantin Evelyn , Tewfik Ted L. , Brouillette Robert T.
Published: December 2009

BACKGROUND: Polysomnography is the best tool available for diagnosing obstructive sleep apnea (OSA) in children. However, polysomnography is relatively inaccessible and costly, and studies are needed to evaluate other diagnostic approaches. It has been suggested that the OSA-18 quality-of-life questionnaire (OSA-18) is a useful measure that could replace polysomnography. The purpose of our study was to determine if the OSA-18, is an accurate measure for the detection of moderate-to-severe OSA.

PATIENTS AND METHODS: Children who were referred to our sleep laboratory for evaluation of suspected OSA and who had a nocturnal pulse oximetry study were included in our cross-sectional study. The results of the oximetry study were interpreted by using the McGill oximetry score (MOS). Abnormal scores were consistent with moderate-to-severe OSA. We analyzed demographic and medical data in addition to the OSA-18 results. We estimated sensitivity and negative predictive values for the OSA-18 to detect an abnormal MOS. We also conducted logistic regression analyses with MOS as the dependent variable and the OSA-18 score, age, gender, comorbidities, and race as independent variables.

RESULTS: We studied 334 children (mean age: 4.6 years; 58% male). The OSA-18 had a sensitivity of 40% and a negative predictive value of 73% for detecting an abnormal MOS. While controlling for other variables in the regression model, for each unit increase in the OSA-18 score, the odds of having an abnormal MOS were increased by 2%. For each 1-year increase in age, the odds of having an abnormal MOS were decreased by 17%.

CONCLUSIONS: Among children who are referred to a sleep laboratory, the OSA-18 does not accurately detect which children will have an abnormal MOS and cannot be used to exclude moderate-to-severe OSA. The OSA-18 should not be used in the place of objective testing to identify moderate-to-severe OSA in children.
Prenatal Low-Dose Aspirin and Neurobehavioral Outcomes of Children Born Very Preterm
Authors: Marret Stéphane , Marchand Laetitia , Kaminski Monique , Larroque Béatrice , Arnaud Catherine , Truffert Patrick , Thirez Gérard , Fresson Jeanne , Rozé Jean-Christophe , Ancel Pierre-Yves ,   for the EPIPAGE Study Group
Published: December 2009

OBJECTIVE: Low-dose aspirin (LDA) given during pregnancy may alter brain development in very preterm infants. We report the short- and long-term outcomes of very preterm infants according to LDA treatment.

PATIENTS AND METHODS: Data were from the Etude Epidemiologique des Petites Ages Gestationnels (EPIPAGE) cohort study, which included all infants born before 33 weeks of gestation in 9 French regions in 1997. This study was restricted to 656 children who were born to 584 women with an obstetric history of placental vascular disease or with chronic hypertension or renal or autoimmune diseases. The main outcome measures were mortality, cerebral lesions, and outcome at 5 years of age, which were measured by a diagnosis of cerebral palsy; behavioral difficulties, which were assessed with the Strength and Difficulties Questionnaire; and cognitive impairment, which was measured by the mental processing composite scale of the Kaufman Assessment Battery for Children (an IQ-equivalent measure of cognitive ability in 2 dimensions: sequential and simultaneous processing scores).

RESULTS: LDA treatment was administered to 125 of 584 (21%) mothers and was not significantly associated with mortality, cerebral lesions, cerebral palsy, or global cognitive impairment of the children at 5 years of age. The proportion of low simultaneous processing scores (<70) was lower in the group with LDA (7% vs 19% without LDA; P = .04). This association was not significant after adjustment for propensity score, prognostic factors, and social class (adjusted odds ratio [aOR]: 0.59 [95% confidence interval (CI): 0.17–2.06]). LDA treatment was associated with a reduction, at the limit of significance, in total behavioral difficulties (aOR: 0.44 [95% CI: 0.19–1.02]) and hyperactivity (aOR: 0.43 [95% CI: 0.17–1.05]).

CONCLUSIONS: LDA was not associated with adverse neonatal or long-term outcomes. Moreover, the results suggest that LDA may be associated with a reduction in neurobehavioral difficulties. More research is needed to assess the effects of aspirin alone or combined with other neuroprotective agents.
Enzyme-Replacement Therapy in a 5-Month-Old Boy With Attenuated Presymptomatic MPS I: 5-Year Follow-up
Authors: Gabrielli Orazio , Clarke Lorne A. , Bruni Stefano , Coppa Giovanni V.
Published: December 2009

Mucopolysaccharidosis type I (MPS I) is a progressive and multisystemic disease, even in its attenuated Hurler-Scheie and Scheie forms. Clinical trials of enzyme-replacement therapy in MPS I have shown clinical benefit in patients with considerable preexisting disease, but no data exist on the effect of beginning enzyme replacement before the onset of significant clinical signs of disease. Here we present the 5-year follow-up of a boy with attenuated MPS I who had laronidase therapy initiated at the age of 5 months and compare his clinical course to that of his older sister, who began treatment at 5 years of age after she had developed typical signs of MPS I. After 5 years of treatment, the younger sibling has not developed any clinical manifestations of MPS I except for mild corneal clouding. In contrast, although many of the older sibling's clinical features have improved after 5 years of treatment, her dysostosis multiplex, cardiac valve involvement, and corneal clouding, although stabilized, have persisted. We suggest that early treatment of attenuated MPS I may significantly delay or prevent the onset of the major clinical signs, substantially modifying the natural history of the disease.
Annual Summary of Vital Statistics: 2007
Authors: Heron Melonie , Sutton Paul D. , Xu Jiaquan , Ventura Stephanie J. , Strobino Donna M. , Guyer Bernard
Published: December 2009

The number of births in the United States increased between 2006 and 2007 (preliminary estimate of 4 317 119) and is the highest ever recorded. Birth rates increased among all age groups (15 to 44 years); the increase among teenagers is contrary to a long-term pattern of decline during 1991–2005. The total fertility rate increased 1% in 2007 to 2122.5 births per 1000 women. This rate was above replacement level for the second consecutive year.

The proportion of all births to unmarried women increased to 39.7% in 2007, up from 38.5% in 2006, with increases noted for all race and Hispanic-origin groups and within each age group of 15 years and older. In 2007, 31.8% of all births occurred by cesarean delivery, up 2% from 2006. Increases in cesarean delivery were noted for most age groups and for non-Hispanic white, non-Hispanic black, and Hispanic women.

Multiple-birth rates, which rose rapidly over the last several decades, did not increase during 2005–2006. The 2007 preterm birth rate was 12.7%, a decline of 1% from 2006. The low-birth-weight rate also declined in 2007 to 8.2%.

The infant mortality rate was 6.77 infant deaths per 1000 live births in 2007, which is not significantly different from the 2006 rate. Non-Hispanic black infants continued to have much higher rates than non-Hispanic white and Hispanic infants. States in the southeastern United States had the highest infant and fetal mortality rates. The United States continues to rank poorly in international comparisons of infant mortality.

Life expectancy at birth reached a record high of 77.9 years in 2007. Crude death rates for children aged 1 to 19 years decreased by 2.5% between 2006 and 2007. Unintentional injuries and homicide were the first and second leading causes of death, respectively, accounting for 53.7% of all deaths to children and adolescents in 2007.
Specific expression of an oxytocin-enhanced cyan fluorescent protein fusion transgene in the rat hypothalamus and posterior pituitary
Authors: Katoh Akiko , Fujihara Hiroaki , Ohbuchi Toyoaki , Onaka Tatsushi , Young W Scott , Dayanithi Govindan , Yamasaki Yuka , Kawata Mitsuhiro , Suzuki Hitoshi , Otsubo Hiroki , Suzuki Hideaki , Murphy David , Ueta Yoichi
Published: December 2009

We have generated rats bearing an oxytocin (OXT)-enhanced cyan fluorescent protein (eCFP) fusion transgene designed from a murine construct previously shown to be faithfully expressed in transgenic mice. In situ hybridisation histochemistry revealed that the Oxt–eCfp fusion gene was expressed in the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) in these rats. The fluorescence emanating from eCFP was observed only in the SON, the PVN, the internal layer of the median eminence and the posterior pituitary (PP). In in vitro preparations, freshly dissociated cells from the SON and axon terminals showed clear eCFP fluorescence. Immunohistochemistry for OXT and arginine vasopressin (AVP) revealed that the eCFP fluorescence co-localises with OXT immunofluorescence, but not with AVP immunofluorescence in the SON and the PVN. Although the expression levels of the Oxt–eCfp fusion gene in the SON and the PVN showed a wide range of variations in transgenic rats, eCFP fluorescence was markedly increased in the SON and the PVN, but decreased in the PP after chronic salt loading. The expression of the Oxt gene was significantly increased in the SON and the PVN after chronic salt loading in both non-transgenic and transgenic rats. Compared with wild-type animals, euhydrated and salt-loaded male and female transgenic rats showed no significant differences in plasma osmolality, sodium concentration and OXT and AVP levels, suggesting that the fusion gene expression did not disturb any physiological processes. These results suggest that our new transgenic rats are a valuable new tool to identify OXT-producing neurones and their terminals.
Absent Smooth Muscle Actin Immunoreactivity of the Small Bowel Muscularis Propria Circular Layer in Association with Chromosome 15q11 Deletion in Megacystis-Microcolon-Intestinal Hypoperistalsis Syndrome
Authors: Szigeti Reka , Chumpitazi Bruno P. , Finegold Milton J. , Ranganathan Sarangarajan , Craigen William J. , Carter Beth A. , Tatevian Nina
Published: December 2009

Megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS; OMIM%249210) is a rare and severe form of congenital intestinal and urinary dysfunction and malformation. Histologic studies suggest that the predominant intestinal manifestation is smooth muscle myopathy. Molecular observations have linked the disease to the neuronal nicotinic acetylcholine receptor (ηAChR), namely the absence of a functional α3 subunit of the ηAChR in patients with MMIHS. We describe a case of MMIHS in association with a de novo deletion of the proximal long arm of chromosome 15 (15q11.2). Histologic evaluation revealed an appropriate light microscopic appearance of both the circular and longitudinal layers of the small bowel muscularis propria. Immunohistochemical staining for smooth muscle actin, however, was selectively absent in the circular layer, demonstrating isolated absence in a unique and previously undescribed pattern. These observations raise the possibility that the proximal long arm of chromosome 15 (15q11) may be of clinical significance in MMIHS.
Evaluation of a Team-Based Learning Tutor Training Workshop on Research and Publication Ethics by Faculty and Staff Participants
Authors: Ju Young-Su
Published: December 2009

A team-based Learning (TBL) tutor training workshop on research and publication ethics was offered to 8 faculty members and 3 staff at Hallym University in 2009. To investigate the effect of the workshop and any attitude changes, a questionnaire survey was performed after the 8-hr course. Questions in four categories-general course content, change in attitudes toward research and publication ethics, the TBL format, and an open-ended question about the course--were included. Participants responded positively to all items on general course content. There was a positive change in attitude on research and publication ethics. Participants also responded positively to six items on team-based learning. The overall positive response to the workshop on research and publication ethics suggested the effectiveness of this kind of TBL tutor training course for university faculty and staff.
Algorithmic Error Correction of Impedance Measuring Sensors
Authors: Starostenko Oleg , Alarcon-Aquino Vicente , Hernandez Wilmar , Sergiyenko Oleg , Tyrsa Vira
Published: December 2009

This paper describes novel design concepts and some advanced techniques proposed for increasing the accuracy of low cost impedance measuring devices without reduction of operational speed. The proposed structural method for algorithmic error correction and iterating correction method provide linearization of transfer functions of the measuring sensor and signal conditioning converter, which contribute the principal additive and relative measurement errors. Some measuring systems have been implemented in order to estimate in practice the performance of the proposed methods. Particularly, a measuring system for analysis of C-V, G-V characteristics has been designed and constructed. It has been tested during technological process control of charge-coupled device CCD manufacturing. The obtained results are discussed in order to define a reasonable range of applied methods, their utility, and performance.
Quantitative Assessment of Bandage Soft Contact Lens Wear Immediately After LASIK
Authors: Orucov Faik , Frucht-Pery Joseph , Raiskup Frederick D. , Strasman Eyal , Landau David , Solomon Abraham
Published: December 2009

Purpose:

To evaluate the efficacy and tolerability of overnight bandage soft contact lenses applied immediately after LASIK.

Methods:

Bandage soft contact lenses were applied immediately following bilateral simultaneous LASIK in 161 consecutive patients. Signs and symptoms associated with contact lens wear were prospectively recorded at 1 hour, 4 hours, and the following morning, using a standardized questionnaire. Contact lenses were removed at 1 hour if patients were symptomatic of intolerance. If no signs of contact lens intolerance appeared at 1 hour, the lenses were removed the following morning. No contact lenses were applied in 61 patients after LASIK.

Results:

One hour after the procedure, contact lenses were removed from 47 patients due to intolerance. In these patients, contact lens removal was associated with a significant decrease of the total score the next morning (0.31±0.55) compared to 1 hour (5.65±1.60,
P
<.0001). A similar decrease of the total score was noted in those patients who tolerated the contact lenses (0.20±0.52 vs 1.30±1.78) and those in whom the contact lenses were not applied (0.76±1.19 vs 3.24±1.93). Mucoid discharge at the lid margins was noted in 52.6% of patients and mild corneal edema was noted in 28.8% of patients with bandage soft contact lenses at 12 to 18 hours.

Conclusions:

Bandage soft contact lens application after LASIK is associated with significant reduction of symptoms during the immediate postoperative period. However, removal of these lenses may be necessary shortly after the procedure.