bluuebunny/crossref_metadata_2025 huggingface dataset:
Hippocampus‐dependent learning is associated with adult neurogenesis in MRL/MpJ mice
Authors: Thuret Sandrine , Toni Nicolas , Aigner Stefan , Yeo Gene W. , Gage Fred H.
Published: January 2009

The hippocampus is involved in declarative memory and produces new neurons throughout adulthood. Numerous experiments have been aimed at testing the possibility that adult neurogenesis is required for learning and memory. However, progress has been encumbered by the fact that abating adult neurogenesis usually affects other biological processes, confounding the interpretation of such experiments. In an effort to circumvent this problem, we used a reverse approach to test the role of neurogenesis in hippocampus‐dependent learning, exploiting the low levels of adult neurogenesis in the MRL/MpJ strain of mice compared with other mouse strains. We observed that adult MRL/MpJ mice produce 75% fewer new neurons in the dentate gyrus than age‐matched C57BL/6 mice. Learning‐induced synaptic remodeling, spatial learning, and visual recognition learning were reduced in MRL/MpJ mice compared with C57BL/6 mice. When MRL/MpJ mice were allowed unlimited access to running wheels, neurogenesis along with spatial learning and visual recognition learning were increased to levels comparable to those in running C57BL/6 mice. Together, these results suggest that adult neurogenesis is correlated with spatial learning and visual recognition learning, possibly by modulating morphological plasticity in the dentate gyrus. © 2009 Wiley‐Liss, Inc.
Hippocampal synapsin isoform levels are linked to spatial memory enhancement by SGS742
Authors: John Julius Paul Pradeep , Sunyer Berta , Höger Harald , Pollak Arnold , Lubec Gert
Published: January 2009

Synapsins are essential proteins for synaptic plasticity and there is no information available for their role in cognitive enhancement (CE) of spatial memory formation. It was therefore the aim of the study to link individual synapsin proteins and their isoforms to spatial memory formation enhanced by SGS742 in the mouse. Extracted hippocampal proteins from a cognitive study treating OF1 mice with the cognitive enhancer SGS742 and tested in the Morris water maze, were run on two‐dimensional gel electrophoresis. Subsequently, protein spots were unambiguously identified by qQ‐TOF mass spectrometry. Quantification of proteins from four groups (NaCl‐treated mice, SGS742‐treated mice, SGS742‐treated yoked controls, and NaCl‐treated yoked controls) was carried out according to an in‐gel stable isotope labeling method. A total of 17 protein spots representing synapsin isoforms were identified and quantified. Using quantification of individual synapsin isoforms showed that these can be clearly assigned to CE by the GABAB antagonist SGS742. Quantitative determination of individual synapsin isoform showed an increase in SGS742‐treated mice (mean ± SD) of ratios between light and heavy stable isotope labeled synapsin protein (SGS742 vs. controls: 2.19 ± 0.41 for synapsin Ia, and 1.41 ± 0.81 for synapsin IIa). Synapsins Ib and IIb were not linked to CE. The NaCl‐treated controls and the use of yoked controls that were ruling out swimming‐ and stress‐mediated changes of synapsins, unequivocally allow to propose a role for synapsins Ia and IIa in the mechanism of CE of spatial memory formation. © 2009 Wiley‐Liss, Inc.
Down‐regulation of the expression of RB18A/MED1, a cofactor of transcription, triggers strong tumorigenic phenotype of human melanoma cells
Authors: Ndong Jean de La Croix , Jean Didier , Rousselet Nathalie , Frade Raymond
Published: January 2009

The RB18A/MED1 human gene, also named TRAP220, DRIP205 and PBP, encodes for a single 205 kDa component, which interacts with nuclear receptors and transcription factors. RB18A/MED1 chromosome localization on locus 17q12‐q21.1 suggests its involvement in human cancers. We herein analyzed RB18A/MED1 expression in human melanoma cell lines. We found that RB18A/MED1 is either highly or weakly expressed in melanoma cells, depending on their respectively non or highly‐tumorigenic phenotype. We therefore investigated the possible existence of a relationship between the RB18A/MED1 expression level and melanoma cell phenotype. For this purpose, we down‐regulated RB18A/MED1 expression by transfecting melanoma cells with a RB18A/MED1 small interfering RNA (siRNA), specific to the 3′‐untranslated region of native RB18A/MED1 RNA, already demonstrated to inhibit specifically RB18A/MED1 protein expression. A nonspecific (scrambled) siRNA was used as control. This RB18A/MED1 siRNA did not modify the expression of cathepsin L forms or lamin A/C, nor the secretion of procathepsin L and MMP2 in transfected cells. Analysis using a microarray membrane with 113 cancer‐related genes, western blot and specific tests, demonstrated that RB18A/MED1 knockdown significantly inhibits tissue inhibitor of metalloproteinase‐3 expression, and increases uPAR expression, two genes well known to be involved in melanoma cell invasion, through modifications of the tumor microenvironment. Indeed, RB18A/MED1 knockdown in melanoma cells in vitro increased their invasive properties, without modification of cell proliferation. Furthermore, RB18A/MED1 knockdown in vivo switched melanoma phenotype from non to strongly‐tumorigenic in nude mice. Our data thus demonstrated for the first time that a decrease of RB18A/MED1 expression in human melanoma cells increases their tumorigenic phenotype. © 2009 Wiley‐Liss, Inc.
Molecular markers predict outcome in squamous cell carcinoma of the head and neck after concomitant cisplatin‐based chemoradiation
Authors: van den Broek Guido B. , Wildeman Maarten , Rasch Coen R.N. , Armstrong Nicola , Schuuring Ed , Begg Adrian C. , Looijenga Leendert H.J. , Scheper Rik , van der Wal Jacqueline E. , Menkema Lorian , van Diest Paul J. , Balm Alfons J.M. , van Velthuysen Marie‐Louise F. , van den Brekel Michiel W.M.
Published: January 2009

Not all patients with squamous cell carcinomas of the head and neck (HNSCC) benefit from concurrent cisplatin‐based chemoradiation, but reliable predictive markers for outcome after chemoradiation are scarce. We have investigated potential prognostic biomarkers for outcome in a large group of patients. Ninety‐one tumor biopsies taken from consecutive HNSCC patients were evaluated for protein expression on a tissue microarray. Using immunohistochemistry, 18 biomarkers, involved in various cellular pathways were investigated. Univariable and multivariable proportional hazard analyses were performed to investigate associations between each individual marker and outcome. In addition, the global test was used to test all variables simultaneously and selected combinations of markers for an overall association with local control. Univariable analysis showed statistically significant increased relative risks of RB, P16 and MRP2 for local control and MDR1 and HIF‐1α for overall survival. MRP2, MDR1 and P16 levels were positively associated with outcome whereas RB and HIF‐1α had a negative relationship. Using Goeman's global testing no combination of markers was identified that was associated with local control. Grouping the markers according to their function revealed an association between a combination of 3 markers (P16, P21 and P27) and outcome (p= 0.05) was found. In the multivariable analysis, MRP2 and RB remained significant independent predictive markers for local control. This study describes the prognostic value of biomarkers for the outcome in patients uniformly treated with concurrent chemoradiation. MRP2 and RB were found to be associated with outcome in patients treated with concurrent chemoradiation. © 2009 Wiley‐Liss, Inc.
Retracted: Overexpression of Aurora A by loss of CHFR gene expression increases the growth and survival of HTLV‐1‐infected T cells through enhanced NF‐κB activity
Authors: Tomita Mariko , Toyota Minoru , Ishikawa Chie , Nakazato Tetsuro , Okudaira Taeko , Matsuda Takehiro , Uchihara Jun‐Nosuke , Taira Naoya , Ohshiro Kazuiku , Senba Masachika , Tanaka Yuetsu , Ohshima Koichi , Saya Hideyuki , Tokino Takashi , Mori Naoki
Published: January 2009

Retraction: The following article has been retracted through agreement between the first author and several coauthors, the journal Editor‐in‐Chief, Peter Lichter, and John Wiley & Sons, Ltd.: Tomita M, Toyota M, Ishikawa C, Nakazato T, Okudaira T, Matsuda T, Uchihara JN, Taira N, Ohshiro K, Senba M, Tanaka Y, Ohshima K, Saya H, Tokino T, Mori N. (2009). Overexpression of Aurora A by loss of CHFR gene expression increases the growth and survival of HTLV‐1‐infected T cells through enhanced NF‐kappaB activity. Int J Cancer; 124 (June (11)): 2607‐2615, published online on 12 JAN 2009 in Wiley Online Library (www.onlinelibrary.wiley.com). After an investigation the retraction has been agreed due to inappropriate duplication of images and overlap with other published work.
The prognostic value of cytology and fluorescence in situ hybridization in the follow‐up of nonmuscle‐invasive bladder cancer after intravesical Bacillus Calmette‐Guérin therapy
Authors: Savic Spasenija , Zlobec Inti , Thalmann George N. , Engeler Daniel , Schmauss Martina , Lehmann Kurt , Mattarelli Gianfranco , Eichenberger Tobias , Dalquen Peter , Spieler Peter , Schoenegg René , Gasser Thomas C. , Sulser Tullio , Forster Thomas , Zellweger Tobias , Casella Roberto , Bubendorf Lukas
Published: January 2009

Molecular markers reliably predicting failure or success of Bacillus Calmette‐Guérin (BCG) in the treatment of nonmuscle‐invasive urothelial bladder cancer (NMIBC) are lacking. The aim of our study was to evaluate the value of cytology and chromosomal aberrations detected by fluorescence in situ hybridization (FISH) in predicting failure to BCG therapy. Sixty‐eight patients with NMIBC were prospectively recruited. Bladder washings collected before and after BCG instillation were analyzed by conventional cytology and by multitarget FISH assay (UroVysion®, Abbott/Vysis, Des Plaines, IL) for aberrations of chromosomes 3, 7, 17 and 9p21. Persistent and recurrent bladder cancers were defined as positive events during follow‐up. Twenty‐six of 68 (38%) NMIBC failed to BCG. Both positive post‐BCG cytology and positive post‐BCG FISH were significantly associated with failure of BCG (hazard ratio (HR)= 5.1 and HR= 5.6, respectively; p < 0.001 each) when compared to those with negative results. In the subgroup of nondefinitive cytology (all except those with unequivocally positive cytology), FISH was superior to cytology as a marker of relapse (HR= 6.2 and 1.4, respectively). Cytology and FISH in post‐BCG bladder washings are highly interrelated and a positive result predicts failure to BCG therapy in patients with NMIBC equally well. FISH is most useful in the diagnostically less certain cytology categories but does not provide additional information in clearly malignant cytology. © 2009 UICC
Ascochlorin activates p53 in a manner distinct from DNA damaging agents
Authors: Jeong Ji‐Hak , Nakajima Hiroo , Magae Junji , Furukawa Chiharu , Taki Keiko , Otsuka Kensuke , Tomita Masanori , Lee In‐Seon , Kim Cheorl‐Ho , Chang Hyeun‐Wook , Min Kwan‐Sik , Park Kwang‐Kyun , Park Kwan‐Kyu , Chang Young‐Chae
Published: January 2009

Ascochlorin, a prenylphenol antitumor antibiotic, profoundly increases the expression of endogenous p53 by increasing protein stability in the human osteosarcoma cells and human colon cancer cells. Ascochlorin also increases DNA binding activity to the p53 consensus sequence in nuclear extract and enhances transcription of p53 downstream targets. Ascochlorin specifically induces p53 phosphorylation at ser 392 without affecting ser 15 or 20, whereas DNA damaging agents typically phosphorylate these serines. Moreover, ascochlorin does not induce phosphorylation of ATM and CHK1, an established substrate of ATR that is activated by genotoxins, nor does it increase DNA strand break, as confirmed by comet assay. The structure‐activity relationship suggests that p53 activation by ascochlorin is related to inhibition of mitochondrial respiration, which is further supported by the observation that respiratory inhibitors activate p53 in a manner similar to ascochlorin. These results suggest that ascochlorin, through the inhibition of mitochondrial respiration, activates p53 through a mechanism distinct from genotoxins. © 2009 UICC
A cohort study of cancer among sarcoidosis patients
Authors: Boffetta Paolo , Rabkin Charles S. , Gridley Gloria
Published: January 2009

Sarcoidosis is a chronic inflammatory condition that may increase the risk of cancer, but limited information is available on occurrence of cancer in these patients. We compared the incidence of cancer among 2,013 White and 3,755 Black male patients admitted to Veterans hospitals in the United States during 1969–1996 with a diagnosis of sarcoidosis, with that of 2,792,503 White and 662,204 Black nonsarcoidosis patients admitted to the same hospitals. Patients suffering from pulmonary and autoimmune diseases were excluded from the study, as was the first year of follow‐up after first admission for sarcoidosis. A total of 241 malignant neoplasms were diagnosed in sarcoidosis patients [relative risk 0.99, 95% confidence interval (CI) 0.87–1.13]. The risks of rectal cancer (relative risk 2.12; 95% CI 1.27–3.52), colon cancer (relative risk 1.55; 95% CI 0.99–2.43) and kidney cancer (relative risk 1.84; 95% CI 1.02–3.33) were increased in sarcoidosis patients when compared with other Veterans hospital patients, whereas the risk of lung cancer was decreased (relative risk 0.60; 95% CI 0.42–0.85). The risk of kidney cancer remained elevated 10 years after first admission. Results were generally consistent among ethnic groups, although the increased risk of colon and kidney cancer was observed only in White patients. These results provide further evidence for an increased risk of specific cancers in patients with sarcoidosis, but do not support any additional increase in overall cancer risk. © 2009 Wiley‐Liss, Inc.
Modulation of pentose phosphate pathway during cell cycle progression in human colon adenocarcinoma cell line HT29
Authors: Vizán Pedro , Alcarraz‐Vizán Gema , Díaz‐Moralli Santiago , Solovjeva Olga N. , Frederiks Wilma M. , Cascante Marta
Published: January 2009

Cell cycle regulation is dependent on multiple cellular and molecular events. Cell proliferation requires metabolic sources for the duplication of DNA and cell size. However, nucleotide reservoirs are not sufficient to support cell duplication and, therefore, biosynthetic pathways should be upregulated during cell cycle. Here, we reveal that glucose‐6‐phosphate dehydrogenase (G6PDH) and transketolase (TKT), the 2 key enzymes of oxidative and nonoxidative branches of the pentose phosphate pathway (PPP), respectively, which is necessary for nucleotide synthesis, are enhanced during cell cycle progression of the human colon cancer cell line HT29. These enhanced enzyme activities coincide with an increased ratio of pentose monophosphate to hexose monophosphate pool during late G1 and S phase, suggesting a potential role for pentose phosphates in proliferating signaling. Isotopomeric analysis distribution of nucleotide ribose synthesized from 1,2‐13C2‐glucose confirms the activation of the PPP during late G1 and S phase and reveals specific upregulation of the oxidative branch. Our data sustain the idea of a critical oxidative and nonoxidative balance in cancer cells, which is consistent with a late G1 metabolic check point. The distinctive modulation of these enzymes during cell cycle progression may represent a new strategy to inhibit proliferation in anticancer treatments. © 2009 UICC
Transforming growth factor‐β induces CD44 cleavage that promotes migration of MDA‐MB‐435s cells through the up‐regulation of membrane type 1‐matrix metalloproteinase
Authors: Kuo Yi‐Chih , Su Cheng‐Hsi , Liu Chin‐Yi , Chen Tien‐Hua , Chen Chie‐Pein , Wang Hwai‐Shi
Published: January 2009

CD44, a transmembrane receptor for hyaluronic acid, is implicated in various adhesion‐dependent cellular processes, including cell migration, tumor cell metastasis and invasion. Recent studies demonstrated that CD44 expressed in cancer cells can be proteolytically cleaved at the ectodomain by membrane type 1‐matrix metalloproteinase (MT1‐MMP) to form soluble CD44 and that CD44 cleavage plays a critical role in cancer cell migration. Here, we show that transforming growth factor‐β (TGF‐β), a multifunctional cytokine involved in cell proliferation, differentiation, migration and pathological processes, induces MT1‐MMP expression in MDA‐MB‐435s cells. TGF‐β‐induced MT1‐MMP expression was blocked by the specific extracellular regulated kinase‐1/2 (ERK1/2) inhibitor PD98059 and the specific phosphoinositide 3‐OH kinase (PI3K) inhibitor LY294002. In addition, treatment with SP600125, an inhibitor for c‐Jun NH2‐terminal kinase (JNK), resulted in a significant inhibition of MT1‐MMP production. These data suggest that ERK1/2, PI3K, and JNK likely play a role in TGF‐β‐induced MT1‐MMP expression. Interestingly, treatment of MDA‐MB‐435s cells with TGF‐β resulted in a colocalization of MT1‐MMP and CD44 in the cell membrane and in an increased level of soluble CD44. Using an electric cell‐substrate impedance sensing cell‐electrode system, we demonstrated that TGF‐β treatment promotes MDA‐MB‐435s cell migration, involving MT1‐MMP‐mediated CD44 cleavage. MT1‐MMP siRNA transfection‐inhibited TGF‐β‐induced cancer cell transendothelial migration. Thus, this study contributes to our understanding of molecular mechanisms that play a critical role in tumor cell invasion and metastasis. © 2009 Wiley‐Liss, Inc.
The SAAP force field: Development of the single amino acid potentials for 20 proteinogenic amino acids and Monte Carlo molecular simulation for short peptides
Authors: Iwaoka Michio , Kimura Naoki , Yosida Daisuke , Minezaki Toshiya
Published: January 2009

Molecular simulation by using force field parameters has been widely applied in the fields of peptide and protein research for various purposes. We recently proposed a new all‐atom protein force field, called the SAAP force field, which utilizes single amino acid potentials (SAAPs) as the fundamental elements. In this article, whole sets of the SAAP force field parameters in vacuo, in ether, and in water have been developed by ab initio calculation for all 20 proteinogenic amino acids and applied to Monte Carlo molecular simulation for two short peptides. The side‐chain separation approximation method was employed to obtain the SAAP parameters for the amino acids with a long side chain. Monte Carlo simulation for Met‐enkephalin (CHO‐Tyr‐Gly‐Gly‐Phe‐Met‐NH2) by using the SAAP force field revealed that the conformation in vacuo is mainly controlled by strong electrostatic interactions between the amino acid residues, while the SAAPs and the interamino acid Lennard‐Jones potentials are predominant in water. In ether, the conformation would be determined by the combination of the three components. On the other hand, the SAAP simulation for chignolin (H‐Gly‐Tyr‐Asp‐Pro‐Glu‐Thr‐Gly‐Thr‐Trp‐Gly‐OH) reasonably reproduced a native‐like β‐hairpin structure in water although the C‐terminal and side‐chain conformations were different from the native ones. It was suggested that the SAAP force field is a useful tool for analyzing conformations of polypeptides in terms of intrinsic conformational propensities of the single amino acid units. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2009
A new hybrid DFT approach to electronic excitation and first hyperpolarizabilities of transition metal complexes
Authors: Lin Jian , Wu Kechen , Zhang Mingxin
Published: January 2009

The electronic excitations and the static first hyperpolarizability of three typical transition metal (TM) aromatic carbonyl complexes, two tungsten pentacarbonyl derivatives (W(CO)5L, L = Py and PyCHO) and one chromium tricarbonyl arene derivative (Cr(CO)3Bz, Bz = benzene), have been theoretically studied by a variety of density functional methods. The assessments reveal that most of the conventional DFT methods including local density approximation, generalized gradient approximation (GGA), and the various kinds of hybrid exchange‐correlation (xc) methods present the first hyperpolarizabilities of these TM‐containing molecules with large deviations from the experiments. A one‐parameter hybrid xc functional is introduced by using the Perdew‐Wang 1991 gradient‐corrected correlation functional (PW91) and the Barone's‐modified PW91 exchange functional (mPW). The ratio between the exact and the density functional exchange is determined to be 0.40 by the adiabatic connection method. The application of the new hybrid functional to the three organometallic carbonyl molecules results in the satisfactory agreement between the calculated first hyperpolarizabilities and the experimental ones. The second‐order nonlinear optical properties of the three organometallic complexes are addressed to the metal‐to‐ligand charge transfers, and the extended π‐delocalization ligands benefit the enhancement of the first hyperpolarizability. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2009
Laboratory diagnosis of syphilis with automated immunoassays
Authors: Marangoni Antonella , Moroni Alessandra , Accardo Silvia , Cevenini Roberto
Published: January 2009

The serological detection of specific antibodies to Treponema pallidum is of particular importance in the diagnosis of syphilis. The purpose of this study was to evaluate diagnostic performances of automated immunoassays in comparison with T. pallidum hemagglutination test (TPHA) and Western Blot (WB). The retrospective study was performed with different panels of sera: 244 clinical and serological characterized syphilitic sera and 203 potentially interfering samples. All the sera were tested by Enzygnost Syphilis, ARCHITECT Syphilis TP, TPHA, and homemade WB. The diagnostic performances of the two assays were very similar: both Enzygnost Syphilis and ARCHITECT Syphilis TP performed with a sensitivity of 99.2%, whereas the specificity was 98.5 and 98.4%, respectively. Considering the suitability for automation, both immunoassays may represent a good choice as a screening test. However, the use of a confirmatory test, such as TPHA or WB, remains a must in order to avoid false‐positive results. J. Clin. Lab. Anal. 23:1–6, 2009. © 2009 Wiley‐Liss, Inc.
First production of fluorescent anti‐ribonucleoproteins conjugate for diagnostic of rabies in Brazil
Authors: Medeiros Caporale Graciane Maria , Rodrigues da Silva Andréa de Cássia , Peixoto Zélia Maria Pinheiro , Chaves Luciana Botelho , Carrieri Maria Luiza , Vassão Ruth Camargo
Published: January 2009

The laboratory tests recommended by the World Health Organization for detection of rabies virus and evaluation of specific antibodies are performed with fluorescent antibodies against the virus, the ribonucleoproteins (RNPs), or by monoclonal antibodies. In this study, we purified the rabies virus RNPs for the production of a conjugate presenting sensibility and specificity compatible with commercial reagents. The method employed for the purification of RNPs was ultracentrifugation in cesium chloride gradient, the obtained product being used for immunizing rabbits, from which the hyperimmune sera were collected. The serum used for conjugate production was the one presenting the highest titer (1/2,560) when tested by indirect immunofluorescence. The antibodies were purified by anion exchange chromatography (QAE‐Sephadex A‐50),conjugated to fluorescein isothiocyanateand separated by gel filtration (Sephadex G‐50). The resulting conjugate presented titers of 1/400 and 1/500 when assayed by direct immunofluorescence (DIF) and simplified fluorescence inhibition microtest, respectively. Sensibility and specificity tests were performed by DIF in 100 central nervous system samples of different animal species, presenting 100% matches when compared with the commercial reagent used as standard, independent of the conservation state of the samples. The quality reached by our conjugate will enable the standardization of this reagent for use by the laboratories performing diagnosis of rabies in Brazil, contributing to the intensification of the epidemiological vigilance and research on this disease. J. Clin. Lab. Anal. 23:7–13, 2009. © 2009 Wiley‐Liss, Inc.
Prognostic value of combination of heart‐type fatty acid‐binding protein and ischemia‐modified albumin in patients with acute coronary syndromes and normal troponin T values
Authors: Liyan Cui , Jie Zhang , Xiaozhou Hu
Published: January 2009

Recent studies have suggested that heart‐type fatty acid‐binding protein (H‐FABP) may detect ongoing myocardial damage involved in the progression of acute coronary syndromes (ACS). This study was prospectively designed to examine whether the combination of H‐FABP, a marker for ongoing myocardial damage, and ischemia‐modified albumin (IMA), a marker for myocardial ischemia, would effectively diagnose patients with ACS. H‐FABP values above 1.5 µg/l can be correctly measured via an ELISA and 6 µg/l is the currently used cut‐off value (1–3). We measured serum H‐FABP and IMA of 108 patients on admission within 12 hr after onset of chestpain and normal troponin T. serum samplesfrom ACS group (n=82) had decreased capacity of ACB [64 (61–67) U/ml] compared with non‐ACS ischemic chest pain group (n=26) samples [75 (71–78) U/ml] (P<0.05). The combination of IMA and H‐FABP usually had better sensitivity [96.3% (92.2–100%)] (P<0.05) and accuracy [92.6 (87.7–97.5%)] (P<0.05) than when individually used. Thus, the combination of H‐FABP and IMA measurements after initiation of chest pain may be highly effective for risk stratification in patients with ACS and normal cardiac troponin T. J. Clin. Lab. Anal. 23:14–18, 2009. © 2009 Wiley‐Liss, Inc.
Comparison of PCR methods for detecting fetal RhDin maternal plasma
Authors: Atamaniuk Johanna , Stuhlmeier Karl M. , Karimi Alireza , Mueller Mathias M.
Published: January 2009

Background: Aim of this study was to establish the method yielding the highest sensitivity routinely used to determine fetal RhD type and gender from maternal cell‐free plasma DNA in different periods of gestation.

Methods: Plasma DNA concentrations were measured from 46 pregnant women in different gestational periods and tested for RhD using three different PCR methods on exon 7: Thermal Cycler, Taqman method on LightCycler, and melting curve analysis on LightCycler. In addition, fetal gender was determined by PCR. Cell‐free plasma DNA was measured in 100 healthy volunteers as a reference group.

Results: The mean value of cell‐free plasma DNA in the reference group was 10.9 pg/µL mean, (standard deviation (SD): 3.66) in 50 healthy women and 12.7 pg/µL (SD: 8.2) in 50 healthy men. In the firsttrimester of pregnancy cell‐free plasmaDNA was 14.9 pg/µL mean, (SD: 4.2), in the second trimester 15.4 pg/µL mean, (SD: 4.96), and the maximum was achieved in the third trimester of pregnancy 15.6 pg/µl mean, (SD: 6.49). TaqMan probes had the same accuracy, when compared with Thermal Cycler technology (46 samples, 6 failures). Using real‐time PCR with melting curve analysis 12 of 17 samples were correctly tested. Gender determination was correctly in 41 of 46 samples.

Conclusion: RhD determinations with TaqMan and Thermal Cycler technology are useful methods for fetal RhD prediction. To increase the accuracy of RhD determination it is necessary to test on other exons in addition. J. Clin. Lab. Anal. 23:24–28, 2009. © 2009 Wiley‐Liss, Inc.
Effect of anticoagulants on the plasma hyaluronidase activities
Authors: Sharma R. , Mahadeswaraswamy Y.H. , Harish Kumar K. , Devaraja S. , Kemparaju K. , Vishwanath B.S. , Girish K.S.
Published: January 2009

Mammalian hyaluronidases (HAases) are an endo‐β‐N‐acetyl‐hexosaminidases that degrade hyaluronan (HA) and have been implicated in diverse pathophysiological functions. Several pathological conditions, such as diabetes, monoclonal gammapathy, and bladder and prostate tumors, report the distorted plasma HAase activity. However, the plasma HAase (hHyal‐1) activity has been presumed to change with the circulating HA level and serves as an early marker for several diseases. It has been generally practised to use the anticoagulants such as tri‐sodium citrate/di‐sodium EDTA/heparin for the preparation of plasma for both biochemical and clinical analyses. In the present investigation, theeffect of anticoagulants on plasma HAaseactivity was evaluated and compared with the serum HAase activity that is devoid of anticoagulants as no study provides information in this regard. The results suggested that the plasma HAase activity in the presence of the recommended concentration of EDTA was highly comparable/similar to that of the serum HAase activity. In contrast, citrated or heparinized plasma recorded a significantly reduced level of activity than that of the serum HAase activity. In conclusion, our results suggested that the EDTA‐treated plasma samples are a better choice compared with heparin and citrated samples to assess the HAase activity. J. Clin. Lab. Anal. 23:29–33, 2009. © 2009 Wiley‐Liss, Inc.
Magnesium and calcium in exhaled breath condensate of children with asthma and gastroesophageal reflux disease
Authors: Dodig Slavica , Vlašić Željka , Čepelak Ivana , Zrinski Topić Renata , Turkalj Mirjana , Nogalo Boro
Published: January 2009

Background: Magnesium and calcium physiologic functions are closely related. Magnesium is primarily an intracellular cation, the action of which also involves maintenance of cellular ionic balance, while influencing calcium homeostasis by blocking calcium channels. The aim of this study was to compare the concentrations of magnesium and calcium in exhaled breath condensate (EBC) of children with asthma and gastroesophageal reflux disease (GERD).

Subjects and methods: EBC was collected from 66 children aged 7–14 years (23 children with acute asthma, 17 children with GERD, and 26 healthy children).Determination of magnesium and calciumconcentrations was preceded by optimization and validation for low concentrations.

Results: No difference was recorded for either magnesium or calcium concentration between study groups. However, the magnesium to calcium ratio was statistically significantly lower in both GERD and asthma children as compared with control group.

Conclusion: Study results showed the magnesium to calcium ratio to be a statistically significantly better indicator of certain pathologic changes than absolute concentration of either ion. J. Clin. Lab. Anal. 23:34–39, 2009. © 2009 Wiley‐Liss, Inc.
A rapid enzyme‐linked immunosorbent assay with two modes of detection for measuring cytokine concentration
Authors: Miwa Keishi , Shibayama Naoko , Moriguchi Takeshi , Goto Junko , Yanagisawa Masahiko , Yamazaki Yoshiaki , Jung Giman , Matsuda Kenichi
Published: January 2009

Interleukin (IL)‐6 and IL‐8 were measured in 101 serum samples collected from eight intensive‐care unit patients using a polystyrene‐based stick enzyme‐linked immunosorbent assay (STICKELISA) system. This system consisted of an immobilized‐antibody ELISA stick and a noncontact spectrophotometer. Cytokine concentration was detected by two ways: first, rapidly and semi‐quantitatively by naked‐eye observation of the color change and second, quantitatively using the spectrophotometer for accurate concentration determination. The spectrophotometric assay enabled the quantitation of as little as 100 pg/mL cytokine and took only 45 min to complete. There was a good agreement between the STICKELISA observations and data ob‐tained using a plate ELISA system. The agreement between STICKELISA naked‐eye observation and plate ELISA determination was 94 and 85% for IL‐6 and IL‐8, respectively. The correlation coefficients between the STICKELISA spectrophotometric determination and plate ELISA determination were 0.88 and 0.91 for IL‐6 and IL‐8, respectively, in a 0.1–5 ng/mL cytokine concentration range. These results demonstrate that the STICKELISA system is a simple, rapid, and quantitative method for bedside cytokine measurement in critical‐care settings. J. Clin. Lab. Anal. 23:40–44, 2009. © 2009 Wiley‐Liss, Inc.
Establishment of a cut‐point value of serum TNF‐α levels in the metabolic syndrome
Authors: Gurrola‐Díaz Carmen M. , Sánchez‐Enriquez Sergio , Oregon‐Romero Edith , García‐López Pedro M. , Garzón de la Mora Pedro , Bastidas‐Ramírez Blanca E. , González‐Hita Mercedes , Muñoz‐Valle José F.
Published: January 2009

Cardiovascular diseases and type 2 diabetes are the major causes of mortality in Mexico. Metabolic syndrome (MS) is a cluster of factors that increase the risk to develop such diseases. Previous studies have shown that MS is associated with high tumor necrosis factor (TNF‐α) levels. In fact, TNF‐α has been proposed to be a useful marker for clinical diagnosis of inflammation at an early stage. Therefore, we analyzed TNF‐α concentrations in Mexican individuals with or without MS and related these levels to the associated MS components. Clinical, anthropometric, and biochemical data were analyzed in 41 healthy and 39 MS individuals. Individuals were similarly grouped by age and gender.The serum TNF‐α levels measured bya highly sensitive enzyme‐linked immunosorbent assay (ELISA) kit were increased significantly in MS subjects compared with healthy individuals (P<0.001). The assay showed 78.1% sensitivity and 61.5% specificity with a cut‐point level of 1.36 pg/mL. TNF‐α levels higher than the cut‐point value were correlated with insulin resistance indices. These findings support the hypothesis that serum TNF‐α concentration could be a useful marker for early MS diagnosis. Nevertheless, we suggest the establishment of specific cut‐point values in each studied population to evaluate potential clinical applications. J. Clin. Lab. Anal. 23:51–56, 2009. © 2009 Wiley‐Liss, Inc.
Cellular immune response profile in patients with American tegumentary leishmaniasis prior and post chemotherapy treatment
Authors: Reis Luiza C. , Brito Maria Edilenza F. , Souza Marina A. , Medeiros Angela C.R. , Silva Claudio J. , Luna Carlos F. , Pereira Valéria R.A.
Published: January 2009

In this study, we have the objective of evaluating the lymphoproliferative response and determining interferon (IFN)‐γ and interleukin (IL)‐10 cytokine production in the peripheral blood mononuclear cells (PBMC) of patients with American tegumentary leishmaniasis prior and post 12 months of chemotherapy treatment with meglumine antimoniate compared with the PBMC of noninfected donors. Lymphoproliferation, such as cytokine production, was evaluated through in vitro stimulus with the soluble antigenic fraction from Leishmania (Viannia) braziliensis promastigotes (1.25 µg/ml) and Concanavalin A (2.5 µg/ml). Patients showed a significant lymphoproliferative response prior and post treatment compared with the control group. Similar result, prior to chemotherapy treatment, was observed in IFN‐γ and IL‐10 production when patients were compared with the control group. After chemotherapy treatment, PBMC lymphoproliferative response of the patients revealed an increase, whereas patients have shown a decrease in IFN‐γ levels and an increase in IL‐10, although without statistical difference. These results may indicate that the patients produced a specific cellular response to the soluble antigenic fraction suggesting that besides Th1 and Th2 dichotomy, immunological regulation mechanisms with the participation of memory T cells and regulatory T cells could be present in the clinical evolution of these patients. This understanding will allow the study and identification of new L. (V.) braziliensis molecules potentially candidates to vaccines. J. Clin. Lab. Anal. 23:63–69, 2009. © 2009 Wiley‐Liss, Inc.
Using viscosity‐time plots of Escherichia coli cells undergoing chemical lysis to measure the impact of physiological changes occurring during batch cell growth
Authors: Kong Simyee , Day Andrew F. , O'Kennedy Ronan D. , Shamlou Parviz A. , Titchener‐Hooker Nigel J.
Published: January 2009

BACKGROUND: Viscosity–time plots for plasmid‐bearing E. coli cells undergoing alkaline lysis are reported in this study. The plots demonstrate generic features that reflect the progress of fermentation and allow an assessment of the genomic DNA denaturation following cellular release into the alkaline solution. This rheological analysis could offer useful insights to the state of fermentation or the selection of operational specifications and predictions of the performance of subsequent downstream operations.

RESULTS: Studies showed a distinct change in the rheological profile throughout the batch fermentation, with different viscosity versus time profiles for lag, exponential and stationary microbial growth phases. The DNA denaturation time was found to increase with fermentation time from about 120 s after 3 h of fermentation to about 180 s after 7 h of fermentation.

CONCLUSION: The increase of denaturation time was mainly caused by a rise in the genomic content of cells during the exponential growth phase. The viscosity–time profiles were found to provide a good indication of the cellular contents, reflecting the physiological changes occurring during a batch fermentation process. Copyright © 2009 Society of Chemical Industry
In situ SEM indentation experiments: Instruments, methodology, and applications
Authors: Ghisleni Rudy , Rzepiejewska‐Malyska Karolina , Philippe Laetitia , Schwaller Patrick , Michler Johann
Published: January 2009

The purpose of this article is to present the design and capabilities of two in situ scanning electron microscope (SEM) indentation instruments covering a large load range from μN to N. The capabilities and advantages of in situ SEM indentation are illustrated by two applications: indentation of a thin film and a nanowire. All the experiments were performed on electrodeposited cobalt, whose outstanding magnetic properties make it a candidate material for MEMS and NEMS devices. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.
In vivo comparison of 2′‐O‐methyl phosphorothioate and morpholino antisense oligonucleotides for Duchenne muscular dystrophy exon skipping
Authors: Heemskerk Hans A. , de Winter Christa L. , de Kimpe Sjef J. , van Kuik‐Romeijn Petra , Heuvelmans Niki , Platenburg Gerard J. , van Ommen Gert‐Jan B. , van Deutekom Judith C. T. , Aartsma‐Rus Annemieke
Published: January 2009

Background

Antisense‐mediated exon skipping is a putative treatment for Duchenne muscular dystrophy (DMD). Using antisense oligonucleotides (AONs), the disrupted DMD reading frame is restored, allowing generation of partially functional dystrophin and conversion of a severe Duchenne into a milder Becker muscular dystrophy phenotype. In vivo studies are mainly performed using 2′‐O‐methyl phosphorothioate (2OMePS) or morpholino (PMO) AONs. These compounds were never directly compared.

Methods

mdx and humanized (h)DMD mice were injected intramuscularly and intravenously with short versus long 2OMePS and PMO for mouse exon 23 and human exons 44, 45, 46 and 51.

Results

Intramuscular injection showed that increasing the length of 2OMePS AONs enhanced skipping efficiencies of human exon 45, but decreased efficiency for mouse exon 23. Although PMO induced more mouse exon 23 skipping, PMO and 2OMePS were more comparable for human exons. After intravenous administration, exon skipping and novel protein was shown in the heart with both chemistries. Furthermore, PMO showed lower intramuscular concentrations with higher exon 23 skipping levels compared to 2OMePS, which may be due to sequestration in the extracellular matrix. Finally, two mismatches rendered 2OMePS but not PMO AONs nearly ineffective.

Conclusions

The results obtained in the present study indicate that increasing AON length improves skipping efficiency in some but not all cases. It is feasible to induce exon skipping and dystrophin restoration in the heart after injection of 2OMePS and unconjugated PMO. Furthermore, differences in efficiency between PMO and 2OMePS appear to be sequence and not chemistry dependent. Finally, the results indicate that PMOs may be less sequence specific than 2OMePS. Copyright © 2009 John Wiley & Sons, Ltd.
Concordant activity of transgene expression cassettes inserted into E1, E3 and E4 cloning sites in the adenovirus genome
Authors: Pham Linh , Nakamura Takafumi , Gabriela Rosales A. , Carlson Stephanie K. , Bailey Kent R. , Peng Kah‐Whye , Russell Stephen J.
Published: January 2009

Background

Expression cassettes can be inserted at several positions into recombinant adenoviral genomes but the implications of this choice for transgene expression level have not been determined. Knowledge of the relative expression levels of transgenes inserted at different sites in the adenoviral genome is of particular significance for transgene expression monitoring approaches that rely on the concordant expression of a marker transgene inserted elsewhere in the viral genome.

Methods

Three expression cassettes, each comprising a cytomegalovirus promoter driving one of three marker peptides [serum carcinoembryonic antigen (sCEA), beta subunit of human chorionic gonadotropin (βhCG) or human sodium iodide symporter (hNIS)], were inserted into E1, E3 or E4 cloning sites in a recombinant adenoviral vector backbone. High titer stocks of bicistronic adenoviral vectors coding for combinations of marker peptides were prepared. A panel of human cells of various lineages was infected with the vectors and expression ratios of the transgene‐encoded proteins were analysed. Serum levels of the soluble proteins and hepatic uptake of radioactive iodine were also compared in vivo in nude rats after intravenous vector infusion.

Results

High concordance of expression between the inserted transgenes was observed in all of the bicistronic vectors irrespective of whether the expression cassettes were placed in the E1, E3 or E4 regions. Concordance was maintained across multiple cell lineages. In vivo, in athymic rats, blood and urine levels of βhCG were highly concordant with serum levels of sCEA at all timepoints after intravenous infusion of the bicistronic vectors encoding both of these soluble markers. Hepatic radioiodine uptake was concordant with serum CEA concentration in mice infused with a bicistronic vector expressing CEA and NIS.

Conclusions

The expression level of a given transgene in an adenoviral vector genome can be accurately and quantitatively inferred from the expression of a marker protein encoded by a second transgene inserted elsewhere in the vector genome. Copyright © 2009 John Wiley & Sons, Ltd.
Methicillin‐resistant Staphylococcus aureus bacteremia due to prostatic abscess
Authors: Chao Bo H. , Kidd Jason M. , Dow Alan W.
Published: January 2009

Journal of Hospital Medicine 2009;4:E9–E11. © 2009 Society of Hospital Medicine.
Pericardial effusion presenting with anuric acute renal failure and hepatocellular damage
Authors: Khan Raza , Gessert Charles , Bockhold Stephen
Published: January 2009

A 50‐year‐old male with anuria, creatinine of 5.5 and potassium of 6.5 was referred to our hospital for hemodialysis. Before hemodialysis could be initiated, his blood pressure dropped and liver function tests were found to be increasing rapidly. This prompted us to look for cardiac causes of liver ischemia. An echocardiogram was non‐diagnostic due to the patient's obese body habitus. Pericardial fluid was documented on CT scan. Pericardiocentesis was performed and nearly 1500 ml of bloody pericardial fluid was removed. This resulted in immediate urine output, with 80 ml in the first hour, and an increase in blood pressure. Journal of Hospital Medicine 2009;4:68–70. © 2009 Society of Hospital Medicine.
Perceptions of resident physicians about management of inpatient hyperglycemia in an urban hospital
Authors: Cheekati Vasundhara , Osburne Robert C. , Jameson Kimberly A. , Cook Curtiss B.
Published: January 2009

BACKGROUND:

Information regarding practitioner beliefs about inpatient diabetes care is limited.

OBJECTIVE:

To assess resident physician attitudes about inpatient hyperglycemia and determine perceived barriers to optimal glycemic control in an urban hospital setting.

DESIGN:

A previously developed questionnaire was modified and administered. Residents were asked about the importance of inpatient glucose control, desirable glucose ranges, and problems encountered when managing hyperglycemia.

SETTING:

Urban teaching hospital.

RESULTS:

Of 85 resident physicians, 66 completed the survey (mean age, 31 years; 47% men; 33% in first residency year). Most respondents categorized glucose control as “very important” in critically‐ill and perioperative patients but only “somewhat important” in non‐critically‐ill patients. Most residents said they would target a therapeutic glucose range within the recommended levels. Most residents (88%) also said they felt “very comfortable” or “somewhat comfortable” using subcutaneous insulin therapy, whereas some were “not at all comfortable” with either subcutaneous (11%) or intravenous (18%) administration. In general, respondents were not very familiar with existing institutional policies and preprinted order sets. The most commonly reported barrier to management of inpatient hyperglycemia was lack of knowledge about appropriate insulin regimens and their use. Anxiety about hypoglycemia was only the third most frequent concern.

CONCLUSION:

Most residents acknowledged the importance of good glucose control in hospitalized patients and chose target glucose ranges consistent with existing guidelines. Lack of knowledge about insulin treatment options was the most commonly cited barrier to ideal management. Educational programs should emphasize inpatient treatment strategies for glycemic control. Journal of Hospital Medicine 2009;4:E1–E8. © 2009 Society of Hospital Medicine.
Effects of a subcutaneous insulin protocol, clinical education, and computerized order set on the quality of inpatient management of hyperglycemia: Results of a clinical trial
Authors: Schnipper Jeffrey L. , Ndumele Chima D. , Liang Catherine L. , Pendergrass Merri L.
Published: January 2009

BACKGROUND:

Inpatient hyperglycemia is associated with poor patient outcomes. It is unknown how best to implement glycemic management strategies in the non–intensive care unit (ICU) setting.

OBJECTIVE:

To determine the effects of a multifaceted quality improvement intervention on the management of medical inpatients with diabetes mellitus or hyperglycemia.

DESIGN:

Before‐after trial.

SETTING:

Geographically localized general medical service staffed by physician's assistants (PAs) and hospitalists.

PATIENTS:

Consecutively enrolled patients with type 2 diabetes or inpatient hyperglycemia.

INTERVENTION:

A detailed subcutaneous insulin protocol, an admission order set built into the hospital's computerized order entry system, and case‐based educational workshops and lectures to nurses, physicians, and PAs.

MEASUREMENTS:

Mean percent of glucose readings per patient between 60 and 180 mg/dL; percent patient‐days with hypoglycemia; insulin use patterns; and hospital length of stay.

RESULTS:

The mean percent of readings per patient between 60 and 180 mg/dL was 59% prior to the intervention and 65% afterward (adjusted effect size 9.7%; 95% confidence interval [CI], 0.6%‐18.8%). The percent of patient days with any hypoglycemia was 5.5% preintervention and 6.1% afterward (adjusted odds ratio 1.1; 95% CI, 0.6–2.1). Use of scheduled nutritional insulin increased from 40% to 75% (odds ratio 4.5; 95% CI, 2.0–9.9) and adjusted length of stay decreased by 25% (95% CI, 9%‐44%). Daily insulin adjustment did not improve, nor did glucose control beyond hospital day 3.

CONCLUSIONS:

This multifaceted intervention, which was easy to implement and required minimal resources, was associated with improvements in both insulin ordering practices and glycemic control among non‐ICU medical patients. Journal of Hospital Medicine 2009;4:16–27. © 2009 Society of Hospital Medicine.