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At least 1,423 records · Page 79Linked to original sources
PubMed2006

Role of the simultaneous sequential strategy for failed acute sinus restoration after modified left maze procedure for persistent atrial fibrillation with concomitant mitral surgery.

BACKGROUND: We assessed whether the simultaneous sequential strategy could (1) achieve additional sinus restoration for those patients who were not in sinus rhythm while coming off bypass after modified left maze procedure and (2) attain the same long-term success rates as the bi-atrial maze procedure in patients with persistent atrial fibrillation (AF) and mitral valve disease. MATERIALS AND METHODS: Twenty-seven consecutive patients - ten men and 17 women with a mean age of 52 +/- 13 years, all with persistent AF and mitral valve disease - underwent the modified maze procedure with the simultaneous sequential strategy. In the first phase, the modified left atrial maze operation was carried out with concomitant valvular surgery; the right side maze operation was subsequently carried out as a second phase of the sequential strategy only if AF re-appeared following the spontaneous restoration of heart beats during the operation. RESULTS: Twenty patients (74.1%) underwent the left atrial maze procedure only, and seven patients (25.9%) required the subsequent right atrial maze procedure as part of the sequential strategy. At a mean follow-up of 15.1 +/- 7.7 months, six of the 27 patients (22.2%) who underwent additional right atrial maze procedure had restored sinus rhythm. At a mean follow-up of 17.8 +/- 7.3 months, 24 of the 27 patients (88.9%) had restored sinus rhythm and 22 patients (81.5%) had restored bi-atrial transport function (right atrial filling fraction: 40.8 +/- 11.7%; left atrial filling fraction: 22.9 +/- 8.1%) after application of the sequential strategy. CONCLUSIONS: Compared with modified left atrial maze procedure, the application of the simultaneous sequential strategy successfully restored sinus rhythm in an additional 22.2% of patients with persistent AF. The overall sinus conversion rate of 88.9% was comparable with that of the standard bi-atrial maze procedure.

Adult↗
PubMed2004

A novel genetic modifier of p53, mop1, results in embryonic lethality.

The heterogeneity that occurs in the tumor spectrum and latency in Li-Fraumeni syndrome (LFS) patients with inherited mutations in p53 suggest risk modifiers at loci other than the major gene. We developed a mouse model to investigate these risk modifiers. Inbred CE/J mice, which succumb to multiple types of tumors similar to those found in LFS, were crossed with the p53-null 129/Sv (129-Trp53(tm1Tyj)) mouse. In this cross, we uncovered evidence for a genetic modifier of p53, mop1, based on an unexpected mix of genotypes in the F2 progeny from Mendelian expectations. A model in which a recessive CE/J allele in combination with p53 heterozygosity or homozygosity results in lethality most closely fits the data. Using simple-sequence length polymorphism analysis of the entire genome, we identified a putative chromosomal region for this modifier of p53 on mouse chromosome 11 centromeric to p53.

Animals↗
PubMed2005

Genetic modifiers affecting severity of epilepsy caused by mutation of sodium channel Scn2a.

Mutations in the voltage-gated sodium channels SCN 1 A and SCN 2 A are responsible for several types of human epilepsy. Variable expressivity among family members is a common feature of these inherited epilepsies, suggesting that genetic modifiers may influence the clinical manifestation of epilepsy. The transgenic mouse model Scn 2 a(Q 54) has an epilepsy phenotype as a result of a mutation in Scn 2 a that slows channel inactivation. The mice display progressive epilepsy that begins with short-duration partial seizures that appear to originate in the hippocampus. The partial seizures become more frequent and of longer duration with age and often induce secondary generalized seizures. Clinical severity of the Scn 2 a(Q 54) phenotype is influenced by genetic background. Congenic C57BL/6J.Q 54 mice exhibit decreased incidence of spontaneous seizures, delayed seizure onset, and longer survival in comparison with [C57BL/6J x SJL/J]F(1).Q 54 mice. This observation indicates that strain SJL/J carries dominant modifier alleles at one or more loci that determine the severity of the epilepsy phenotype. Genome-wide interval mapping in an N(2) backcross revealed two modifier loci on Chromosomes 11 and 19 that influence the clinical severity of of this sodium channel-induced epilepsy. Modifier genes affecting clinical severity in the Scn 2 a(Q 54) mouse model may contribute to the variable expressivity seen in epilepsy patients with sodium channel mutations.

Alleles↗
PubMed2006

Diabetic modifier QTLs identified in F2 intercrosses between Akita and A/J mice.

To identify novel genetic modifiers of type 2 diabetes (T2D), we performed quantitative trait loci (QTL) analysis on F(2) progeny of hypoinsulinemic diabetic Akita mice, heterozygous for the Ins2 gene Cys96Tyr mutation, and nondiabetic A/J mice. We generated 625 heterozygous (F(2)-Hetero) and 338 wild-type (F(2)-Wild) mice with regard to the Ins2 mutation in F(2) intercross progeny. We measured quantitative traits, including plasma glucose and insulin concentrations during the intraperitoneal glucose tolerance test (IPGTT), and body weight (BW). We observed three significant QTLs in hypoinsulinemic hyperglycemic male F(2)-Hetero mice, designated Dbm1, Dbm3, and Dbm4 on Chromosomes 6, 14, and 15, respectively. They showed linkage to plasma glucose concentrations, with significant maximum logarithm of odds (LOD) scores of 4.12, 4.17, and 6.17, respectively, all exceeding threshold values by permutation tests. In normoinsulinemic normoglycemic male F(2)-Wild mice, Dbm1 on Chromosome 6 showed linkage to both plasma insulin concentrations and BW, and Dbm2 on Chromosome 11 showed linkage to plasma glucose concentrations only, with LOD scores of 4.52 and 6.32, and 5.78, respectively. Based on these results, we concluded that Dbm1, Dbm2, Dbm3, and Dbm4 represent four major modifier QTLs specifically affecting T2D-related traits and that these diabetic modifier QTLs are conditional on the heterozygous Ins2 gene mutation and sex to exert their modifier functions. Identification of the genes responsible for these QTLs would provide new drug development targets for human T2D.

Animals↗
PubMed2000

ROSA26 mice carry a modifier of Min-induced mammary and intestinal tumor development.

B6.129S7-Gtrosa26 (B6.R26) mice carry a LacZ-neoR insertion on Chromosome (Chr) 6, made by promoter trapping with 129 ES cells. Female C57BL/6J ApcMin/+ (B6Min/+) mice are highly susceptible to intestinal tumors and to the induction of mammary tumors after treatment with ethylnitrosourea (ENU). However, B6.R26/+ Min/+ females develop fewer mammary and intestinal tumors after ENU treatment than do B6 Min/+ mice. B6.R26/+ mice from two independently derived congenic lines show this modifier effect. Each of these congenic lines carries approximately 20 cM of 129-derived DNA flanking the insertion, raising the possibility that the resistance is due to a linked modifier locus. To further map the modifier locus, we have generated several lines of mice carrying different regions of the congenic interval. We have found that resistance to mammary and intestinal tumors in ENU-treated Min/+ mice maps to a minimum 4-cM interval that includes the ROSA26 LacZ-neoR insertion. Therefore, the resistance to tumor development is due to either the ROSA26 insertion or a very tightly linked modifier locus.

Animals↗
PubMed2000

A modifier screen of ectopic Krüppel activity identifies autosomal Drosophila chromosomal sites and genes required for normal eye development.

Irregular facets (If) is a dominant gain-of-function allele of the Drosophila segmentation gene Krüppel (Kr) that interferes with eye development. In a search for genes that interact with Kr activity, we recently performed a systematic genetic screen to identify dominant enhancers and suppressors of the If eye phenotype that are located on the third chromosome. Here we describe locations and candidate genes of the second chromosome that act as dominant modifiers of ectopic Kr activity during eye development. The collection of more than 40 modifiers of Kr activity located on the second and third chromosomes, from which a total of 16 genes were identified, includes genes encoding transcription factors and components of signal transduction pathways that may regulate or be regulated by Kr activity. We also identified genes coding for more general cellular factors that could interfere with the intracellular transport or the half-life of the Kr protein. The data demonstrate that the If mutation provides a means to screen the Drosophila genome for functional components of developmental pathways that depend on or can be modified by Kr activity. Owing to the bias of the screening system applied, these modifier genes will be expressed and are likely to be required during Drosophila wild-type eye development.

Animals↗
PubMed1998

Genetically modified fibroblasts induce angiogenesis in the rat epigastric island flap.

METHODS: Gene therapy was tested for inducing functional angiogenesis in the superficial rat epigastric island flap to allow earlier pedicle division. Autologous rat fibroblasts were grown, harvested, cultured and retrovirally transfected to produce platelet-derived growth factor AA (PDGF-AA), an angiogenetically active protein. Stable gene expression was monitored by PDGF-AA enzyme-linked immunosorbent assay (ELISA). One hundred and eighty animals were divided into three groups (I-III) and a bilateral flap created in each animal. In all experiments, the right-sided flap was subjected to experimental treatment and the left-sided flap served as control (1ml saline 0.9%). During flap elevation, group I received 5X10(6) GMFB (genetically modified fibroblasts) plus 1 ml Dulbecco's modified Eagle's medium. Group II was treated with 5x10(6) NMFB (non-modified fibroblasts) plus 1 ml medium and group III received 1 ml medium only. The flaps were sutured back and the vascular pedicle was bilaterally ligated and divided in each of ten animals during the following 6 days. After 7 days, the flaps were harvested, the amount of necrosis measured and histologically examined. RESULTS: The GMFB produced up to 560 times more PDGF-AA than the NMFB, measured by ELISA. The GMFB-treated flaps tolerated surgical division of the vascular pedicle significantly earlier than groups II and III. Histologically, fibroblasts persisted in all flaps of groups I and II, without major inflammatory reaction. In all GMFB-treated flaps, massive angiogenesis could be demonstrated. CONCLUSION: By means of retroviral gene transfer, autologous rat fibroblasts can be genetically modified for stable expression of the PDGF-A gene to produce high amounts of PDGF-AA, which is angiogenetically active. After injection into the panniculus carnosus, these cells induce functional angiogenesis to permit earlier division of the vascular pedicle in this flap model.

Animals↗
PubMed2006

Genetic analysis of candidate genes modifying the age-at-onset in Huntington's disease.

The expansion of a polymorphic CAG repeat in the HD gene encoding huntingtin has been identified as the major cause of Huntington's disease (HD) and determines 42-73% of the variance in the age-at-onset of the disease. Polymorphisms in huntingtin interacting or associated genes are thought to modify the course of the disease. To identify genetic modifiers influencing the age at disease onset, we searched for polymorphic markers in the GRIK2, TBP, BDNF, HIP1 and ZDHHC17 genes and analysed seven of them by association studies in 980 independent European HD patients. Screening for unknown sequence variations we found besides several silent variations three polymorphisms in the ZDHHC17 gene. These and polymorphisms in the GRIK2, TBP and BDNF genes were analysed with respect to their association with the HD age-at-onset. Although some of the factors have been defined as genetic modifier factors in previous studies, none of the genes encoding GRIK2, TBP, BDNF and ZDHHC17 could be identified as a genetic modifier for HD.

Acyltransferases↗
PubMed2001

A modified laryngeal mask in the endoscopic management of an esophageal tumor.

The laryngeal mask airway (LMA) can be used for gastroscopy, but its use can result in loss of the seal and/or displacement of the cuff. We describe an LMA that was specifically modified for gastroscopy and report its use in a patient with an esophageal tumor. The modified LMA has (a) a second tube that allows instruments to be directed toward the esophagus and (b) a second cuff mounted on the dorsal surface that increases the efficacy of the seal with the larynx. A 78-year-old man weighing 65 kg presented with a large mediastinal adenocarcinoma that was infiltrating the lateral wall of the thoracic esophagus. An esophagoscopy under anesthesia was planned to debulk the tumor. The modified LMA was inserted easily following induction with propofol. Anesthesia was maintained with propofol and 50% O2 in air and spontaneous ventilation. A lubricated 10.5-mm external diameter gastroscope was inserted into the second tube and passed easily into the esophagus. The tumor was successfully debulked using a polypectomy snare and an argon plasma coagulator. There was no loss of seal or displacement of the cuff, and the patient was stable throughout the procedure. We conclude that gastroscopy is feasible with the modified LMA. The device has a potential application in patients who require ventilatory support during gastroscopy.

Aged↗
PubMed2001

Modified Talairach landmarks.

BACKGROUND: Brain atlas-assisted operations, such as targeting in stereotactic and functional neurosurgery, localisation analysis and metanalysis in human brain mapping, or structure segmentation and labelling in neuroradiology, highly depend on the accurate localisation of the landmarks used for atlas-to-data registration. One of practical and widely used registration methods is the Talairach proportional grid system transformation based on the Talairach landmarks. However, there are several problems associated with the original Talairach landmarks. In the Talairach-Tournoux brain atlas, some Talairach landmarks are not available and locations of others contradict their definitions. When dealing with patient-specific data, the definitions of the Talairach landmarks are not constructive enough or make their identification time consuming. Moreover, there is an inconsistency between the Talairach landmarks and Talairach grid. METHOD: The modified Talairach landmarks, conceptually equivalent to the original Talairach landmarks, are introduced here. They have several advantages and overcome some limitations of the original Talairach landmarks. Three various intercommissural distances are defined: central, internal, and tangential, and the formulas determining their lengths and errors associated are derived. An efficient method for calculating the modified cortical landmarks is proposed. FINDINGS: The internal intercommissural distance is the closest to the original Talairach intercommissural distance. Its relative intercommissural error is only 0.5%, as opposed to the central and tangential intercommissural distances resulting in high (about 10%) relative intercommissural errors. On the other hand, the internal and central intercommissural distances result in a high maximum displacement error at cortex amounting to about 11 mm while the tangential intercommissural distance gives only 1 mm error. The sensitivity of the internal intercommissural distance to the actual location of the intercommissural plane is high reaching above 10%. On the other hand, the sensitivity of the central intercommissural distance is below 0.5%. Each of the Talairach cortical landmarks needs three coordinates for its identification that requires the availability of two two-dimensional projections, the generation and analysis of which is computationally expensive. A modified cortical landmark is identified on a single, one-dimensional projection. INTERPRETATION: The modified Talairach landmarks facilitate the rapid and automated calculation of the Talairach transformation and give more flexibility in their use for specific applications. In stereotactic and functional neurosurgery, the internal intercommissural distance is the most suitable to provide a high accuracy for subcortical structures. In localisation analysis in human brain mapping research, a high accuracy has to be achieved at the cortex and the tangential intercommissural line is superior. Human brain mapping metanalysis may benefit from the use of the central intercommissural distance minimising errors due to the intercommissural plane positioning. In neuroradiology, a high accuracy is required both for subcortical and cortical structures and the tangential intercommissural line should be used.

Anthropometry↗
PubMed2004

Evidence for a modifier of onset age in Huntington disease linked to the HD gene in 4p16.

Huntington disease (HD) is a neurodegenerative disorder caused by the abnormal expansion of CAG repeats in the HD gene on chromosome 4p16.3. A recent genome scan for genetic modifiers of age at onset of motor symptoms (AO) in HD suggests that one modifier may reside in the region close to the HD gene itself. We used data from 535 HD participants of the New England Huntington cohort and the HD MAPS cohort to assess whether AO was influenced by any of the three markers in the 4p16 region: MSX1 (Drosophila homeo box homologue 1, formerly known as homeo box 7, HOX7), Delta2642 (within the HD coding sequence), and BJ56 ( D4S127). Suggestive evidence for an association was seen between MSX1 alleles and AO, after adjustment for normal CAG repeat, expanded repeat, and their product term (model P value 0.079). Of the variance of AO that was not accounted for by HD and normal CAG repeats, 0.8% could be attributed to the MSX1 genotype. Individuals with MSX1 genotype 3/3 tended to have younger AO. No association was found between Delta2642 (P=0.44) and BJ56 (P=0.73) and AO. This study supports previous studies suggesting that there may be a significant genetic modifier for AO in HD in the 4p16 region. Furthermore, the modifier may be present on both HD and normal chromosomes bearing the 3 allele of the MSX1 marker.

Adolescent↗
PubMed2006

CT angiography in highly calcified arteries: 2D manual vs. modified automated 3D approach to identify coronary stenoses.

BACKGROUND: Two-dimensional axial and manually-oriented reformatted images are traditionally used to analyze coronary data provided by multidetector-row computed tomography angiography (MDCTA). While apparently more accurate in evaluating calcified vessels, 2D methods are time-consuming compared with automated 3D approaches. The purpose of this study was to evaluate the performance of a modified automated 3D approach (using manual vessel isolation and different window and level settings) in a population with high calcium scores who underwent coronary half-millimeter 16-detector-row CT angiography (16 x 0.5-MDCTA). METHODS: ECG-gated 16 x 0.5-MDCTA (16 x 0.5 mm cross-sections, 0.35 x 0.35 x 0.35 mm3 isotropic voxels, 400 ms rotation) was performed after injection of iopamidol (120-ml, 300 mg/ml) in 19 consecutive patients (11 male, 62+/-10 years-old). Native arteries were independently evaluated for >or=50%-stenoses using both manual 2D and modified automated 3D approaches. Stents and bypass grafts were excluded. Conventional coronary angiography was visually analyzed by 2 observers. RESULTS: Median Agatston calcium score was 434. Sensitivities, specificities, positive and negative predictive values for detection of >or=50% coronary stenoses using the 2D and modified 3D approaches were, respectively: 74%/63%, 76%/80%, 45%/34%, and 91%/93% (p=NS for all comparisons). Overall diagnostic accuracies were 75 and 78%, respectively (p=NS). Uninterpretable vessels were, respectively: 37% (77/209) and 35% (73/209) - p=NS. Time to analyze a single study was 160+/-23 and 53+/-11 min, respectively (p<0.01). CONCLUSIONS: This modified automated 3D approach is equivalent to and significantly less time consuming than the traditional manual 2D method for evaluation of >or=50%-stenoses by 16 x 0.5-MDCTA in native coronary arteries of patients with high calcium scores.

Adult↗
PubMed2005

Modified natural cycle using GnRH antagonist can be an optional treatment in poor responders undergoing IVF.

PURPOSE: To investigate the efficacy of gonadotrophin-releasing hormone (GnRH) antagonist supplementation during natural cycles in poor responders undergoing IVF-ET treatment. METHODS: We retrospectively evaluated 540 cycles of 433 suitable patients who were divided by treatment protocol into modified natural, antagonist, and long agonist groups. There were 52 modified natural cycles with GnRH antagonist supplementation, 200 stimulated cycles with GnRH antagonist, and 288 long GnRH agonist cycles. Cycle characteristics and treatment outcomes were compared between the groups. RESULTS: The mean number of oocytes retrieved in the modified natural group was significantly lower than in the stimulated antagonist and long agonist groups (1.4 +/- 0.5 vs. 2.3 +/- 1.1 and 2.5 +/- 1.1, respectively, p < 0.05). The respective implantation and pregnancy rates were 10% and 14.3%, 6.75% and 10.2%, and 7.4% and 10.6%. Cycle outcome and cycle properties were similar. CONCLUSIONS: Modified natural IVF cycle with GnRH antagonist supplementation is a feasible alternative to ovarian stimulation protocols in poor responders.

Adult↗
PubMed2006

Enhanced immunogenicity of the modified GP5 of porcine reproductive and respiratory syndrome virus.

The ORF5-encoded major envelope glycoprotein (GP5) is one of the key immunogenic proteins of the porcine reproductive and respiratory syndrome virus (PRRSV) and is the leading target for the development of the new generation of vaccines against PRRS. However, weak and tardy neutralizing antibodies have been elicited in several developed experimental vaccines expressing PRRSV GP5. More recent evidence has demonstrated a non-neutralizing decoy epitope upstream of the neutralizing epitope of GP5, which might prevent the development of a strong neutralizing antibody response against PRRSV. In the present study, we modified the ORF5 gene by inserting a Pan DR T-helper cell epitope (PADRE) between the neutralizing epitope and the decoy epitope to minimize or eliminate the decoy effect of the non-neutralizing epitope. The immunogenicity of the modified GP5 was further evaluated using DNA vaccination. The results showed that significantly enhanced neutralizing antibodies were elicited in mice immunized with the DNA construct expressing the modified GP5 compared with the native GP5. Slightly increased levels of GP5-specific ELISA antibodies and T-cell proliferative activities were also observed. These results indicate that the high immunogenicity of the modified GP5 might facilitate the development of improved PRRS vaccines in the future.

Amino Acid Sequence↗
PubMed1992

Estimation of right ventricular volume by modified echocardiographic subtraction method.

To evaluate the accuracy and clinical utility of right ventricular volume estimated by a modified echocardiographic subtraction method versus Krebs' original subtraction method, an experiment was performed on hearts excised from 25 animals (dogs, pigs, and cows) followed by a clinical study of 41 patients with heart disease. Right ventricular volume was measured by subtracting the left ventricular volume from that of the whole heart based on echocardiographic apical two- and four-chamber views by means of the area-length method. In the animal heart study, the coefficient of variation between the right ventricular volume estimated by the modified method and the true volume was +/- 13%. The regression equation was y = 0.94x + 4.15 (r = 0.987, p less than 0.001) and showed good correlation, whereas the right ventricular volume obtained by the original method underestimated the true volume (coefficient of variation = +/- 25%, y = 0.59x + 1.11; r = 0.976, p less than 0.001). In the clinical study, the coefficient of variation between right ventricular volume estimated by the modified echocardiographic method and RV volume estimated by radionuclide ventriculography was +/- 15%. The regression equation was y = 0.80x + 13.3 (r = 0.935, p less than 0.001). This correlation was better than that obtained by the original method (coefficient of variation = +/- 16%), where the regression equation was y = 0.60x + 2.43 (r = 0.888, p less than 0.001). Thus the accuracy of the modified subtraction method was validated, and this method showed a better correlation than the original method both experimentally and clinically.

Adolescent↗
PubMed1993

Comparison of the Asymptomatic Cardiac Ischemia Pilot and modified Asymptomatic Cardiac Ischemia Pilot versus Bruce and Cornell exercise protocols.

The Asymptomatic Cardiac Ischemia Pilot (ACIP) and modified ACIP treadmill exercise protocols were developed to test patients with coronary artery disease and to linearly increase work load between stages. The physiologic changes that occurred with ACIP and modified ACIP were compared to those with the Bruce and Cornell protocols in 28 normal subjects and 16 men with coronary artery disease. The exercise protocols were randomly assigned over 2 days, and gas exchange data were obtained continuously with each test. In normal subjects, the peak heart rate, systolic blood pressure, peak oxygen consumption rate (VO2) and minute ventilation were similar for the 4 protocols tested, with exercise time shortest for the Bruce protocol in comparison with the ACIP, modified ACIP and Cornell protocols (10.2 +/- 3.1 vs 13.4 +/- 4.9, 13.9 +/- 4.5, and 15.0 +/- 4.2 minutes, respectively; p < 0.001). The difference between predicted and observed VO2 was smallest for the ACIP protocol (37.0 +/- 11.0 vs 35.8 +/- 13.5 ml/kg/min) and greatest for the Bruce protocol (41.1 +/- 11.8 vs 36.7 +/- 15.0 ml/kg/min) in normal subjects, as well as in patients with coronary artery disease (ACIP protocol 26.9 +/- 7.1 vs 22.5 +/- 6.7, and Bruce protocol 29.1 +/- 7 vs 22.6 +/- 5.7 ml/kg/min, respectively). The ratio of VO2 to work rate, expressed as a slope, was similar in normal subjects for the 4 protocols tested. However, in patients with coronary artery disease, the slope was 0.84 and 0.83 for the ACIP and modified ACIP protocols, respectively, versus 0.61 and 0.71 for the Bruce and Cornell protocols, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1984

Preparation of alkylamine and 125I-radiolabeled derivatives of hyaluronic acid uniquely modified at the reducing end.

Present procedures to obtain radiolabeled hyaluronic acid derivatives are limited to low-specific-activity isotopes and small amounts of material, and often involve multiple points of chemical modification within the polymer. A synthesis has been developed which affords large quantities of a unique, chemically modified derivative of hyaluronic acid containing a single hydroxyphenyl group at the reducing end, which can be radioiodinated to high specific activity. Very little alteration in oligosaccharide structure is expected since only the terminal reducing sugar is modified. Oligosaccharides of hyaluronic acid, which have no free amino groups, were first converted to alkylamine derivatives to allow subsequent reaction with the Bolton-Hunter reagent, N-succinimidyl-3(4-hydroxyphenyl)propionate. Synthesis of the hyaluronate-amine was achieved by (i) reduction of the terminal reducing sugar with sodium borohydride, (ii) controlled sodium periodate oxidation to generate an aldehyde group only at the reduced end, and (iii) coupling this aldehyde to an alpha,omega- alkyldiamine (e.g., 1,6- hexanediamine ) in the presence of sodium cyanoborohydride. Purified hyaluronate-amine oligosaccharides were then reacted with the Bolton-Hunter reagent, and the hydroxyphenyl derivative thus obtained was radioiodinated with Na125I. Specific activities up to 8 X 10(9) cpm/nmol oligosaccharide can be obtained. This approach yields a uniquely modified, highly radioactive probe which will be useful in studies of cellular and extracellular matrix interactions with hyaluronic acid. In addition, the uniquely modified alkylamine derivative of hyaluronic acid has been used to prepare affinity chromatography media and synthetic cell culture surfaces.

Amines↗
PubMed1991

5-Enolpyruvylshikimate-3-phosphate synthase from Escherichia coli--the substrate analogue bromopyruvate inactivates the enzyme by modifying Cys-408 and Lys-411.

In order to identify the essential reactive amino acid residues of 5-enolpyruvylshikimate-3-phosphate synthase, the reaction of the enzyme with its substrate analogue bromopyruvate was investigated. Incubation of the enzyme with bromopyruvate resulted in a time-dependent loss of enzyme activity. The inactivation followed pseudo-first-order and saturation kinetics with a Kinact of 28 microM and a maximum rate constant of 0.31 min-1. The inactivation was prevented by preincubation of the enzyme with the substrates shikimate 3-phosphate, 5-enolpyruvylshikimate 3-phosphate or by the combination of shikimate 3-phosphate plus glyphosate (N-phosphonomethylglycine), an inhibitor of the enzyme. Addition of sodium [3H]borohydride to the reaction mixture had no effect on the rate of inactivation but resulted in the incorporation of 3H label to the modified enzyme. Upon 90% inactivation, approximately 1 mol of bromo[14C]pyruvate was incorporated per mole of enzyme modified in the absence or presence of sodium borohydride. When the enzyme was incubated with bromopyruvate in the presence of sodium [3H]borohydride, approximately 1 mol of 3H label was found to be associated per mole of the modified enzyme. Tryptic digestion of these labeled proteins followed by reverse phase chromatographic separation resulted in the isolation of three radioactive peptides. Analyses of these three peptides indicated that bromopyruvate inactivated the enzyme by modifying Cys-408 and Lys-411, which are conserved in all enzyme sequences studied to date.

3-Phosphoshikimate 1-Carboxyvinyltransferase↗