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Brie A. Stotler Ronit Reich-Slotky Joseph Schwartz William B. Inabnet James Lee Fann Wu Phyllis Della-Latta Jeffrey S. Jhang 《Cell and tissue banking》2011,12(2):111-116
Cryopreservation of parathyroid tissue (PT) provides patients undergoing parathyroidectomy with an option for delayed autologous
heterotopic parathyroid transplantation. A standard protocol for quality monitoring of PT has not been established. This article
describes a method for detecting the presence of bacterial contamination in PT tissue intended for autologous transplantation.
PT was received in the tissue bank, processed under aseptic conditions, and placed into cryopreservation medium. Sterility
testing was performed at 2 time points prior to cryopreservation. From January 2005 to October 2008, 47 PT samples were cryopreserved.
The following bacteria were isolated from 11 PT specimens: Staphylococcus epidermidis, Staphylococcus capitis subspecies ureolyticus, Staphylococcus lugdunensis, Bacillus pumilus, and corynebacteria (diphtheroids). 23% of PTs were contaminated at the time of collection, predominantly with indigenous bacteria. Quality monitoring using
this protocol is a useful tool to identify tissues contaminated with bacteria. 相似文献
33.
M.-C. Fann A.H. Davies A. Varadhachary T. Kuroda C. Sevier T. Tsuchiya P.C. Maloney 《The Journal of membrane biology》1998,164(2):187-195
Three lines of evidence indicate that arginine-46 (R46) and arginine-275 (R275) are essential to the function of UhpT, the
Pi-linked antiport protein of Escherichia coli. A role for arginine was initially suggested by the sensitivity of UhpT to inhibition by 2,3-butanedione, an arginine-directed
probe. Since the presence of substrate protected against this inhibition, this work further suggested that arginine(s) may
lie at or near the UhpT active site. In other work, each UhpT arginine was examined individually by using site-directed mutagenesis
to generate a cysteine or a lysine derivative. With two exceptions (R46, R275), all arginines could be replaced by either
cysteine (10 of 14 residues) or lysine (12 of 14) without loss of function, implicating R46 and R275 as essential to UhpT
function. This idea was strengthened by examining a multiple alignment of the eleven known UhpT-related proteins (≥30% identity).
That alignment showed R46 and R275 were two of the only three arginines strongly conserved in this group of proteins. Considered
together, these different approaches lead us to conclude that UhpT and its relatives have only two arginine residues (R46,
R275) whose presence is essential to function. Prior biochemical work had placed R275 at the external entrance to the translocation
pathway, and a symmetry argument emerging from the multiple alignment suggests a similar position for R46. Accordingly, by
virtue of their locations at the entrance to this pathway, we speculate that R46 and R275 function in establishing substrate
specificity.
Received: 29 January 1998/Revised: 13 April 1998 相似文献
34.
Endophenotypes such as behavior disorders have been increasingly adopted in genetic studies for complex traits. For efficient gene mapping, it is essential that an endophenotype is associated with the disease of interest and is inheritable or co-segregating within families. In this study, we proposed a strategy to construct endophenotypes to analyze the Genetic Analysis Workshop 14 simulated dataset. Initially, generalized estimating equation models were employed to identify phenotypes that were correlated to the disease (affected status) in combination with the family structures in data. Endophenotypes were then constructed with consideration of heterogeneity as functions of the identified phenotypes. Genome scans on the constructed endophenotypes were carried out using family-based association analysis. For comparison, genome scans were also performed with the original affected status. The family-based association analysis using the endophenotypes correctly identified the same susceptible gene in about 80 of the 100 replicates. 相似文献
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Yu-Li Liu Sheng-Chang Wang Hai-Gwo Hwu Cathy Shen-Jang Fann Ueng-Cheng Yang Wei-Chih Yang Pei-Chun Hsu Chien-Ching Chang Chun-Chiang Wen Jyy-Jih Tsai-Wu Tzung-Jeng Hwang Ming H. Hsieh Chen-Chung Liu Yi-Ling Chien Chiu-Ping Fang Stephen V. Faraone Ming T. Tsuang Wei J. Chen Chih-Min Liu 《PloS one》2016,11(3)
D-amino acid oxidase (DAO) has been reported to be associated with schizophrenia. This study aimed to search for genetic variants associated with this gene. The genomic regions of all exons, highly conserved regions of introns, and promoters of this gene were sequenced. Potentially meaningful single-nucleotide polymorphisms (SNPs) obtained from direct sequencing were selected for genotyping in 600 controls and 912 patients with schizophrenia and in a replicated sample consisting of 388 patients with schizophrenia. Genetic associations were examined using single-locus and haplotype association analyses. In single-locus analyses, the frequency of the C allele of a novel SNP rs55944529 located at intron 8 was found to be significantly higher in the original large patient sample (p = 0.016). This allele was associated with a higher level of DAO mRNA expression in the Epstein-Barr virus-transformed lymphocytes. The haplotype distribution of a haplotype block composed of rs11114083-rs2070586-rs2070587-rs55944529 across intron 1 and intron 8 was significantly different between the patients and controls and the haplotype frequencies of AAGC were significantly higher in patients, in both the original (corrected p < 0.0001) and replicated samples (corrected p = 0.0003). The CGTC haplotype was specifically associated with the subgroup with deficits in sustained attention and executive function and the AAGC haplotype was associated with the subgroup without such deficits. The DAO gene was a susceptibility gene for schizophrenia and the genomic region between intron 1 and intron 8 may harbor functional genetic variants, which may influence the mRNA expression of DAO and neurocognitive functions in schizophrenia. 相似文献
37.
Hsin‐Liang Chen Jack C.‐C. Hsu Man Hoang Viet Mai Suan Li Chin‐Kun Hu Chia‐Hsun Liu Frederick Y. Luh Silvia S.‐W. Chen Evan S.‐H. Chang Andrew H.‐J. Wang Min‐Feng Hsu Wunshain Fann Rita P.‐Y. Chen 《Proteins》2010,78(14):2973-2983
Kinetic measurement of protein folding is limited by the method used to trigger folding. Traditional methods, such as stopped flow, have a long mixing dead time and cannot be used to monitor fast folding processes. Here, we report a compound, 4‐(bromomethyl)‐6,7‐dimethoxycoumarin, that can be used as a “photolabile cage” to study the early stages of protein folding. The folding process of a protein, RD1, including kinetics, enthalpy, and volume change, was studied by the combined use of a phototriggered caging strategy and time‐resolved photoacoustic calorimetry. The cage caused unfolding of the photolabile protein, and then a pulse UV laser (~10?9 s) was used to break the cage, leaving the protein free to refold and allowing the resolving of two folding events on a nanosecond time scale. This strategy is especially good for monitoring fast folding proteins that cannot be studied by traditional methods. Proteins 2010. © 2010 Wiley‐Liss, Inc. 相似文献
38.
Qiong Guo Jean T Corbett Guihua Yue Yang C Fann Steven Y Qian Kenneth B Tomer Ronald P Mason 《The Journal of biological chemistry》2002,277(8):6104-6110
The redox properties and thiol reactivity of quinones play critical roles in their therapeutic and toxicological properties. The present study was undertaken to investigate the binding activity of ubiquinone 0 (UQ(0)) to human oxyhemoglobin (HbO(2)) using electron spin resonance (ESR). Addition of UQ(0) to HbO(2) resulted in the immediate detection of a five-line ESR spectrum characteristic of the semiquinone radical of UQ(0) (UQ(0)). With time the HbO(2) adduct with UQ(0), which was characterized by a broad immobilized ESR spectrum, was gradually formed. Matrix-assisted laser desorption/ionization time-of-flight mass spectra analysis showed that UQ(0) bound to the beta-chain of HbO(2). Superoxide dismutase dose-dependently suppressed the intensity of the broad spectrum and accelerated its formation. However, N-ethylmaleimide, a thiol-blocking agent, completely eliminated its formation. The nonspecific protease mixture pronase also prevented its formation and resulted in the gradual appearance of a 4-line spectrum from the 5-line spectrum of UQ(0). The structure of the species responsible for the 4-line spectrum was confirmed and identified by the reaction of UQ(0) with reduced glutathione. In human red blood cells, UQ(0) rapidly bound to glutathione but more slowly to HbO(2). These results suggest that UQ(0) reacted with both ferrous heme and the reactive beta-93 cysteinyl residue of HbO(2) to generate its corresponding semiquinone radical. Subsequently UQ(0) bound to the beta-93 cysteinyl residue of HbO(2) to form a covalent-binding adduct responsible for the broad spectrum. 相似文献
39.
Hsin-Chou Yang Yu-Jen Liang Yi-Lin Wu Chia-Min Chung Kuang-Mao Chiang Hung-Yun Ho Chih-Tai Ting Tsung-Hsien Lin Sheng-Hsiung Sheu Wei-Chuan Tsai Jyh-Hong Chen Hsin-Bang Leu Wei-Hsian Yin Ting-Yu Chiu Chin-Iuan Chen Cathy S. J. Fann Jer-Yuarn Wu Teng-Nan Lin Shing-Jong Lin Yuan-Tsong Chen Jaw-Wen Chen Wen-Harn Pan 《PloS one》2009,4(5)
Young-onset hypertension has a stronger genetic component than late-onset counterpart; thus, the identification of genes related to its susceptibility is a critical issue for the prevention and management of this disease. We carried out a two-stage association scan to map young-onset hypertension susceptibility genes. The first-stage analysis, a genome-wide association study, analyzed 175 matched case-control pairs; the second-stage analysis, a confirmatory association study, verified the results at the first stage based on a total of 1,008 patients and 1,008 controls. Single-locus association tests, multilocus association tests and pair-wise gene-gene interaction tests were performed to identify young-onset hypertension susceptibility genes. After considering stringent adjustments of multiple testing, gene annotation and single-nucleotide polymorphism (SNP) quality, four SNPs from two SNP triplets with strong association signals (−log10(p)>7) and 13 SNPs from 8 interactive SNP pairs with strong interactive signals (−log10(p)>8) were carefully re-examined. The confirmatory study verified the association for a SNP quartet 219 kb and 495 kb downstream of LOC344371 (a hypothetical gene) and RASGRP3 on chromosome 2p22.3, respectively. The latter has been implicated in the abnormal vascular responsiveness to endothelin-1 and angiotensin II in diabetic-hypertensive rats. Intrinsic synergy involving IMPG1 on chromosome 6q14.2-q15 was also verified. IMPG1 encodes interphotoreceptor matrix proteoglycan 1 which has cation binding capacity. The genes are novel hypertension targets identified in this first genome-wide hypertension association study of the Han Chinese population. 相似文献
40.
Site-directed and second site suppressor mutagenesis identify an intrahelical salt bridge in the eleventh transmembrane segment of UhpT, the sugar phosphate carrier of Escherichia coli. Glucose 6-phosphate (G6P) transport by UhpT is inactivated if cysteine replaces either Asp388 or Lys391 but not if both are replaced. This suggests that Asp388 and Lys391 are involved in an intrahelical salt bridge and that neither is required for normal UhpT function. This interpretation is strengthened by the finding that mutations at Lys391 (K391N, K391Q, and K391T) are recovered as revertants of the inactive D388C variant. Further work shows that although the D388C variant is null for G6P transport, movement of 32Pi by homologous Pi/Pi exchange is unaffected. This raises the possibility that this derivative may have latent function, a possibility confirmed by showing that D388C is a gain-of-function mutation in which phosphoenolpyruvate (PEP) is the preferred substrate. Added study of the Pi/Pi exchange shows that in wild type UhpT this partial reaction is readily blocked by G6P but not PEP. By contrast, in the D388C variant, Pi/Pi exchange is unaffected by G6P but is inhibited by both PEP and 3-phosphoglycerate. These latter substrates are used by PgtP, a related Pi-linked antiporter, which lacks the Asp388-Lys391 salt bridge but has instead an uncompensated arginine at position 391. For this reason, we conclude that in both UhpT and PgtP position 391 can serve as a determinant of substrate selectivity by acting as a receptor for the anionic carboxyl brought into the translocation pathway by PEP. 相似文献