bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of a brush‐sampling fecal immunochemical test for hemoglobin with a sensitive guaiac‐based fecal occult blood test in detection of colorectal neoplasia
Authors: Smith Alicia , Young Graeme P. , Cole Stephen R , Bampton Peter 
Published: September 2006

AbstractBACKGROUND.Fecal immunochemical tests (FIT) are an advanced fecal occult blood test (FOBT) technology that reduces barriers to population screening by simplifying the logistics of stool‐sampling. The current study was conducted to undertake a paired comparison of a sensitive guaiac FOBT (GFOBT; Hemoccult II Sensa, Beckman Coulter, Fullerton, CA) with a brush‐sampling FIT (InSure; Enterix, North Ryde, NSW, Australia), to determine whether this FIT improves detection of significant neoplasia.METHODS.Individuals sampled consecutive stools, at home, with both FIT and GFOBT sampling devices while following dietary restrictions appropriate for GFOBT. Study populations included a screening cohort (n = 2351) and a symptomatic diagnostic group (n = 161). Paired comparison of positivity rates was undertaken in those found to have cancer and/or significant adenoma (high‐grade dysplasia, villous change, ≥10 mm, serrated histology or ≥3 polyps), benign pathology, or no pathology.RESULTS.Combined results for both cohorts showed that the FIT returned a true‐positive result significantly more often in cancer (n = 24; 87.5% vs. 54.2%) and in significant adenomas (n = 61; 42.6% vs. 23.0%). Of all UICC Stage I cancers, the FIT was positive in 12 of 13 compared with 4 of 13 with the GFOBT (P = .002). In analyses of just the screening cohort, the FIT remained significantly better at detecting cancers and significant adenomas; the false‐positive rate for any neoplasia was marginally higher with the FIT than the GFOBT (3.4% vs. 2.5%; 95% CI of difference, 0–1.8%), whereas positive predictive values were 41.9% and 40.4%, respectively.CONCLUSIONS.This brush‐sampling FIT is more sensitive for cancers and significant adenomas than a sensitive GFOBT. As such, it should deliver greater reductions in colorectal cancer mortality and incidence than the GFOBT. Cancer 2006; © 2006 American Cancer Society.
JAK2 (V617F) as an acquired somatic mutation and a secondary genetic event associated with disease progression in familial myeloproliferative disorders
Authors: Rumi Elisa , Passamonti Francesco , Pietra Daniela , Porta Matteo G. Della , Arcaini Luca , Boggi Sabrina , Elena Chiara , Boveri Emanuela , Pascutto Cristiana , Lazzarino Mario , Cazzola Mario 
Published: September 2006

AbstractBACKGROUND.A somatic gain‐of‐function mutation of the Janus kinase 2 (JAK2) gene has been identified in chronic myeloproliferative disorders, which appear to have a sporadic occurrence in most individuals. The authors studied the biologic significance of the JAK2 (V617F) mutation in familial myeloproliferative disorders.METHODS.Twenty pedigrees with familial chronic myeloproliferative disorders were identified through an investigation of family history in 264 patients with sporadic myeloproliferative disorders. A quantitative real‐time polymerase chain reaction (qRT‐PCR)‐based allelic discrimination assay was employed for the detection of the V617F mutation in circulating granulocytes and T lymphocytes. An analysis of X‐chromosome inactivation pattern was performed in female patients.RESULTS.Fourteen families had homogeneous phenotypes, and 6 families had mixed phenotypes. By using a qRT‐PCR‐based allelic discrimination assay, the JAK2 (V617F) mutation was detected in circulating granulocytes from 20 of 31 patients, but the mutation was not detected in T lymphocytes. Granulocyte mutant alleles ranged from 2.1% to 91.5% and, on average, increased with time. Discordant distribution of the JAK2 (V617F) mutation was observed in siblings with polycythemia vera. The proportion of granulocytes that carried the JAK2 (V617F) mutation was lower than the proportion of clonal granulocytes, as determined in an analysis of X‐chromosome inactivation patterns in female patients.CONCLUSIONS.The current findings indicated that the JAK2 (V617F) mutation represents an acquired somatic mutation in patients with familial chronic myeloproliferative disorders and probably occurs as a secondary genetic event in the background of preexisting clonal hematopoiesis. Thus, a genetic predisposition to acquisition of JAK2 (V617F) is inherited in families with myeloproliferative disorders. Cancer 2006. © 2006 American Cancer Society.
Plasticity of synaptopodin and the spine apparatus organelle in the rat fascia dentata following entorhinal cortex lesion
Authors: Deller Thomas , Bas Orth Carlos , Vlachos Andreas , Merten Tobias , Del Turco Domenico , Dehn Doris , Mundel Peter , Frotscher Michael 
Published: September 2006

AbstractSynaptopodin is an actin‐associated molecule essential for the formation of a spine apparatus in telencephalic spines. To study whether synaptopodin and the spine apparatus organelle are regulated under conditions of lesion‐induced plasticity, synaptopodin and the spine apparatus were analyzed in granule cells of the rat fascia dentata following entorhinal denervation. Confocal microscopy was employed to quantify layer‐specific changes in synaptopodin‐immunoreactive puncta densities. Electron microscopy was used to quantify layer‐specific changes in spine apparatus organelles. Within the denervated middle and outer molecular layers, the layers of deafferentation‐induced spine loss, synaptogenesis, and spinogenesis, the density of synaptopodin puncta and the number of spine apparatuses decreased by 4 days postlesion and slowly recovered in parallel with spinogenesis by 180 days postlesion. Within the nondenervated inner molecular layer, the zone without deafferentation‐induced spine loss, a rapid loss of synaptopodin puncta and spine apparatuses was also observed. In this layer, spine apparatus densities recovered by 14 days postlesion, in parallel with plastic remodeling at the synaptic level and the postlesional recovery of granule cell activity. These data demonstrate layer‐specific changes in the distribution of synaptopodin and the spine apparatus organelle following partial denervation of granule cells: in the layer of spine loss, spine apparatus densities follow spine densities; in the layer of spine maintenance, however, spine apparatus densities appear to be regulated by other signals. J. Comp. Neurol. 499:471–484, 2006. © 2006 Wiley‐Liss, Inc.
Structural isoforms of the circadian neuropeptide PDF expressed in the optic lobes of the cricket Gryllus bimaculatus: Immunocytochemical evidence from specific monoclonal antibodies
Authors: Honda Takeshi , Matsushima Ayami , Sumida Kazunori , Chuman Yoshiro , Sakaguchi Kazuyasu , Onoue Hitoshi , Meinertzhagen Ian A. , Shimohigashi Yasuyuki , Shimohigashi Miki 
Published: September 2006

AbstractPigment‐dispersing factor (PDF) is an 18‐mer peptide that acts as a principal neurotransmitter of the insect circadian clock. Our previous study, utilizing anti‐Uca β‐PDH polyclonal antibody (pAb) to immunolabel the optic lobe of the cricket Gryllus bimaculatus, suggested the existence of an alternative PDF‐like peptide in the outer cells of the first neuropile, or lamina (La), which were much less immunoreactive than the inner cells of the second neuropile, the medulla (Me). To obtain structural information about such a PDF‐like peptide, we prepared 10 anti‐Gryllus PDF monoclonal (mAb) and pAb antibodies and analyzed their detailed epitope specificities. The PDFMe and PDFLa inner cells and their axonal projections were clearly immunoreactive to all these antibodies, revealing the widespread immunocytochemical organization of the PDF system in the optic lobe, as seen previously with anti‐Uca β‐PDH pAb and anti‐Gryllus PDF mAb, the epitope structures of which were also clarified in this study. The lamina outer cells, which we found lacked a target pdf mRNA, displayed specific immunoreactivities, indicating that the cells contain a distinct PDF‐like peptide possessing both N‐ and C‐terminal structures. These cells were not immunolabeled by some other monoclonal antibodies, however, implying that the PDFLa outer cells have a PDF isoform peptide devoid of Asn at positions 6 and 16. This isoform was also identified in a varicose arborization in the lamina. These results suggest not only the structure of the peptide, but also the possibility of additional functions of this novel PDF isoform. J. Comp. Neurol. 499:404–421, 2006. © 2006 Wiley‐Liss, Inc.
Distinct and gradient distributions of connexin26 and connexin30 in the cochlear sensory epithelium of guinea pigs
Authors: Zhao Hong‐Bo , Yu Ning 
Published: September 2006

AbstractConnexin26 (Cx26) and Cx30 are predominant isoforms of gap junction channels in the cochlea and play a critical role in hearing. In this study, the cellular distributions of Cx26 and Cx30 in the cochlear sensory epithelium of guinea pigs were examined by immunofluorescent staining and confocal microscopy in whole mounts of the cochlear sensory epithelium and dissociated cell preparations. The expression of Cx26 and Cx30 demonstrated a longitudinal gradient distribution in the epithelium and was reduced threefold from the cochlear apex to base. The reduction was more pronounced in the Deiters cells and pillar cells than in the Hensen cells. Cx26 was expressed in all types of supporting cells, but little Cx30 labeling was seen in the Hensen cells. Cx26 expression in the Hensen cells was concentrated mainly in the second and third rows, forming a distinct band along the sensory epithelium at its outer region. In the dissociated Deiters cells and pillar cells, Cx30 showed dense labeling at the cell bodies and processes in the reticular lamina. Cx26 labeling largely overlapped that of Cx30 in these regions. Cx26 and Cx30 were also coexpressed in the gap junctional plaques between Claudius cells. Neither Cx26 nor Cx30 labeling was seen in the hair cells and spiral ganglion neurons. These observations demonstrate that Cx26 and Cx30 have a longitudinal gradient distribution and distinct cellular expression in the auditory sensory epithelium. This further supports our previous reports that Cx26 and Cx30 can solely and concertedly perform different functions in the cochlea. J. Comp. Neurol. 499:506–518, 2006. © 2006 Wiley‐Liss, Inc.
Immunocytochemical analysis of GABA‐positive and calretinin‐positive horizontal cells in the tiger salamander retina
Authors: Zhang Jian , Zhang Ai‐Jun , Wu Samuel M. 
Published: September 2006

AbstractBy using immunocytochemical techniques, we demonstrate that there are two distinct, nonoverlapping populations of horizontal cells (HCs) in the tiger salamander retina: GABA‐positive cells account for about 72% and GABA‐negative (calretinin‐positive) cells account for 28% of the total HC somas. The calretinin‐positive HCs have relatively sparse and thick dendrites: soma diameter of 19.72 ± 0.29 μm, and soma density of 140 ± 13 cells/mm2, morphological features very much like the A‐type HCs described in the accompanying article. The GABA‐positive HCs have thinner dendritic and coarse axon‐terminal‐like processes of higher density: soma diameter of 18 ± 0.18 μm, and soma density of 364 ± 18 cells/mm2, features that very much resemble the B‐type HCs and B‐type HC axon terminals in the accompanying article. By using double and triple immunostaining techniques we found that only 18% of the non‐GABAergic HC dendritic clusters contact rods, whereas the remaining 82% of the dendritic clusters contact cones. This is consistent with the physiological finding in the accompanying article that the A‐type HCs are cone‐dominated. On the other hand, 32% of GABAergic HC dendrites contact rod pedicles and 68% contact cone pedicles, consistent with the physiological finding that B‐type HCs and B‐type HC axon terminals receive mixed rod/cone inputs. Detailed confocal microscope analysis shows that 4% rods, 6% principal double cones/single cones, and 100% accessory double cones contact calretinin‐positive HCs, and 79% rods, 100% principal double cones, 14% accessory double cones, and 82% single cones contact GABAergic HCs. These results suggest that GABAergic and non‐GABAergic HC input/output synapses differ and they may mediate different functional pathways in the outer retina. J. Comp. Neurol. 499:432–441, 2006. © 2006 Wiley‐Liss, Inc.
Flow cytometry for assessment of the efficacy of siRNA
Authors: Ho Hung‐Yao , Cheng Mei‐Ling , Wang Yi‐Hui , Chiu Daniel Tsun‐Yee 
Published: September 2006

AbstractBackground:Small interfering RNA (siRNA) has emerged as a powerful tool to study the loss‐of‐function phenotype by specifically silencing a target gene. The success of gene silencing depends on the choice of appropriate target sequence, and requires a rapid and sensitive assay for quantifying siRNA efficiency. Conventional assays include Western blotting and reverse transcription coupled with polymerase chain reaction. However, these methods are somewhat inaccurate owing to the variation in transfection efficiency. To avoid this, we have developed a flow cytometric method to provide a quantitative analysis of siRNA efficacy.Methods:We constructed a novel vector pHyper1G, which can express enhanced green fluorescent protein (EGFP). It contains a H1 promoter, which can drive expression of short hairpin RNA (shRNA) directed against a target gene. The target gene was cloned into pDsRed1‐N1. The resulting construct can express a fusion protein between target protein and DsRed. These vectors were co‐transfected into 293T cells. The transfected cells were analyzed by flow cytometry. The percentage of EGFP+, DsRed+cells and the change in mean fluorescence intensity (MFI) of DsRed channel indicate changes in expression of target gene in a cell population, and hence the efficacy of the corresponding shRNA. In addition, the cells transfected with pHyper1G derivative were sorted, and analyzed for the activity of target gene.Results:We designed an oligonucleotide duplex encoding shRNA against glucose‐6‐phosphate dehydrogenase (G6PD) gene, and cloned this into pHyper1G. The resulting vector pHyper1G101 effectively knocked down the expression of G6PD‐DsRed from pDsRed301, as shown by significant reduction in both the percentage of EGFP+, DsRed+cells, and MFI. Changes in these parameters were consistent with decreases in protein level and activity of G6PD. Moreover, albeit at low suboptimal transfection efficiency, cells transfected with pHyper1G101 alone were successfully sorted for those expressing shRNA, and their G6PD activity was found to be suppressed.Conclusion:Flow cytometric analysis provides a reliable assessment of the efficiency of siRNA in a cell population. This method can be easily automated and used for screening of appropriate shRNAs against certain genes. Moreover, using the present system, we can deliberately sort for and analyze those cells expressing shRNA. This can be applied to the hard‐to‐transfect cell types, and greatly facilitates the analyses of gene silencing in these cells. © 2006 International Society for Analytical Cytology
High‐throughput fluorescent screening of transgenic animals: Phenotyping and haplotyping
Authors: Hellrung Daniel J. , Rossi Gabriela , Link Charles J. 
Published: September 2006

AbstractBackground:Methods for genotyping transgenic animals currently consist of extracting genomic DNA from blood or tissue followed by PCR or Southern blot analysis. These methods when used to screen large numbers of animals can be time consuming and expensive. Therefore, we developed a novel method that allows high‐throughput screening of phenotypic changes on leukocytes, resulting from the transgenic genotype. This technique allows investigators to quickly screen a large number of animals without the need to extract DNA from each one. Moreover, since blood is collected for the initial screening, putative homozygotes can be confirmed by conventional methods using the same blood samples.Methods:We collected blood from wild‐type αgal positive and αgal knockout mice and probed for the presence of Galα(1→3)Gal (αgal) epitopes. Also, alloantigen specific antibodies were used to determine the haplotype of our outbred mouse colony in order to develop an inbred line.Results:αgal epitopes were detected in wild‐type but not αgal knock‐out samples. To validate these results, PCR was used to demonstrate the native αgal gene in wild‐type and the pGKneo construct in αgal knock‐out mice. Furthermore, haplotypes were determined and mice divided for backcrosses.Conclusions:This screening method is useful for both preliminary screening of transgenic mice and the development of an inbred mouse colony by rapid determination of MHC I haplotype. Here, we demonstrate the use of this technique and show how it can be a valuable tool, saving time and resources in both investigator effort and animal husbandry. © 2006 International Society for Analytical Cytology
Flow cytometric method for the assessment of the minimal inhibitory concentrations of antibacterial agents to Mycoplasma agalactiae
Authors: Assunção Patrícia , Antunes Nuno T. , Rosales Ruben S. , de la Fe Christian , Poveda Carlos , Poveda José B. , Davey Hazel M. 
Published: September 2006

AbstractIn this study, flow cytometry was evaluated for the determination of the minimal inhibitory concentrations (MIC) of seven antibacterial agents (enrofloxacin, ciprofloxacin, gentamicin, streptomycin, chloramphenicol, oxytetracycline, and tylosin) on Mycoplasma (M.) agalactiae. Flow cytometry was able to detect M. agalactiae inhibition from 6 h postincubation, although it seems that definitive MIC values determined by flow cytometry were only possible at 12‐h postincubation. However, the results obtained by the traditional method were only obtained at 24 h, when a visible change in the medium had occurred. At 24 h, both methods gave the same result for six antibacterial agents (enrofloxacin, ciprofloxacin, gentamicin, streptomycin, chloramphenicol, and oxytetracycline); whereas flow cytometry gave slightly higher MIC for tylosin. This was attributed to the fact that the M. agalactiae growth that had occurred in the tubes containing tylosin was not enough to visibly change the color of the medium. Futhermore, flow cytometry detected that inhibitory concentrations of oxytetracycline, chloramphenicol, and tylosin as judged at 24 h were not able to inhibit the M. agalactiae growth after 48 h. MIC values of enrofloxacin and ciprofloxacin were sufficient only to maintain the total counts per milliliter throughout the time matched samples, whereas higher concentrations of theses antibacterial agents reduced the total counts per milliliter over the course of the experiment. The main advantage of the flow cytometric method is that MIC results for M. agalactiae can be obtained in a shorter time than is possible with the traditional method. The method presented makes identification of resistant populations of M. agalactiae possible and, unlike the traditional method, allows the effect of each antibacterial agent to be determined in real‐time at the single‐cell level. © 2006 International Society for Analytical Cytology
Single and three‐color flow cytometry assay for intracellular zinc ion availability in human lymphocytes with Zinpyr‐1 and double immunofluorescence: Relationship with metallothioneins
Authors: Malavolta Marco , Costarelli Laura , Giacconi Robertina , Muti Elisa , Bernardini Gianni , Tesei Silvia , Cipriano Catia , Mocchegiani Eugenio 
Published: September 2006

AbstractBackground:The amount of available intracellular zinc is pivotal to regulate many cellular processes, including oxidative stress response and apoptotic mechanisms. Therefore it is not surprising that zinc homeostasis and dyshomeostasis is involved in many physiological and pathological states, respectively. Cell permeable zinc probes allow intracellular applications with microscopy technology, but flow cytometry (FC) applications have been scarcely explored, albeit they can be suited to study zinc homeostasis in different cell types, including rare cells.Methods:We describe a FC method able to estimate intracellular zinc ion availability and the intracellular capability to activate a zinc signal after treatment with an NO‐donor (AcOM‐DEA/NO) in human PBMCs, using the fluorescent zinc‐specific probe, Zinpyr‐1 (ZP1), alone or in association with CD4‐PE and CD8‐Cychrome mAb.Results:This method was able to detect an increase/decrease of intracellular zinc available in human fresh cultured PBMC and in immune subsets using AcOM‐DEA/NO or TPEN, respectively. ZP1 mean fluorescence on gated histograms was sensitive to the amount of zinc added in the culture medium and significantly correlated to metallothioneins and total intracellular zinc.Conclusions:FC applications using ZP1 may be a fast and useful tool to study zinc homeostasis in immune cells. © 2006 International Society for Analytical Cytology
Dynamic quantification of the tyrosine phosphorylation of the sperm surface proteins during capacitation
Authors: Piehler Erik , Petrunkina Anna M. , Ekhlasi‐Hundrieser Mahnaz , Töpfer‐Petersen Edda 
Published: September 2006

AbstractBackground:Spermatozoa acquire active fertilizing competence only after deposition in the female tract and subsequent capacitation. Recent studies on the cellular location of major sperm phosphoproteins suggest that capacitation is associated with tyrosine phosphorylation of proteins exposed on the sperm surface. However, these changes have not yet been quantified objectively. A calcium influx seems to be required for the completion of tyrosine phosphorylation in some species; however, the exact temporal coordination between these processes is still poorly understood.Methods:Flow cytometry was used to quantify the degree of phosphorylation of the sperm surface proteins by probing with fluorescein isothiocyanate‐conjugated anti‐phosphotyrosine (pY) antibody raised in mouse. Dynamic changes in other sperm parameters (calcium influx, membrane integrity, and spontaneous acrosome reaction) were assessed to analyze their temporal coordination.Results:The changes in specific phosphotyrosine (pY) fluorescence signal detected in live, nonpermeabilized boar cell suspensions were biphasic during incubation under capacitating conditions. After 120 min of incubation, the degree of pY fluorescence increased threefold, indicating the changes in proteins exposed on sperm surface. At the same time there was a gradual increase in cytosolic calcium ion levels with the maximal rate at 60 min of incubation. This rate slowed immediately before the onset of the massive rise in tyrosine phosphorylation and decreased by 90% after its completion. The integrity of plasma and acrosome membranes decreased only slowly, illustrating that the changes observed were not due to the process of spontaneous acrosome reaction.Conclusions:These data provide quantitative evidence for the appearance of tyrosine‐phosphorylated proteins on the surface of live boar spermatozoa during capacitation. An exact temporal coordination exists between cytosolic calcium ion content and protein tyrosine phosphorylation under these conditions. This novel approach has the advantage of making possible a precise quantification and kinetic comparison of molecular processes in different cell subpopulations. © 2006 International Society for Analytical Cytology
Ad hocnetwork routing and security: A review
Authors: Zhang Congzhe , Zhou MengChu , Yu Ming 
Published: September 2006

AbstractAd hocnetwork is a collection of wireless mobile nodes dynamically forming a local area network without the use of any existing network infrastructure or centralized administration. One problem in the design of such network is its wireless vulnerabilities and limited physical security. This paper considers the security attributes and goals ofad hocnetwork, reviews the latest developments of current routing and security mechanissm including encryption, protocols, user authentication, and physical security. It also explores the new approaches trying to solve current problems and points out the future research directions. These methods will be helpful in the military and other security‐sensitive fields. Copyright © 2006 John Wiley & Sons, Ltd.