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PubMed2005

Interactions between xenoestrogens and ketoconazole on hepatic CYP1A and CYP3A, in juvenile Atlantic cod (Gadus morhua).

BACKGROUND: Xenoestrogens and antifungal azoles probably share a common route of metabolism, through hepatic cytochrome P450 (CYP) enzymes. Chemical interactions with metabolic pathways may affect clearance of both xenobiotics and endobiotics. This study was carried out to identify possible chemical interactions by those substances on CYP1A and CYP3A, in Atlantic cod liver. We investigated effects of two xenoestrogens (nonylphenol and ethynylestradiol) and of the model imidazole ketoconazole, alone and in combination. RESULTS: Treatment with ketoconazole resulted in 60% increase in CYP1A-mediated ethoxyresorufin-O-deethylase (EROD) activity. Treatment with nonylphenol resulted in 40% reduction of CYP1A activity. Combined exposure to ketoconazole and nonylphenol resulted in 70% induction of CYP1A activities and 93% increase in CYP1A protein levels. Ketoconazole and nonylphenol alone or in combination had no effect on CYP3A expression, as analyzed by western blots. However, 2-dimensional (2D) gel electrophoresis revealed the presence of two CYP3A-immunoreactive proteins, with a more basic isoform induced by ketoconazole. Treatment with ketoconazole and nonylphenol alone resulted in 54% and 35% reduction of the CYP3A-mediated benzyloxy-4-[trifluoromethyl]-coumarin-O-debenzyloxylase (BFCOD) activity. Combined exposure of ketoconazole and nonylphenol resulted in 98% decrease in CYP3A activity. This decrease was greater than the additive effect of each compound alone. In vitro studies revealed that ketoconazole was a potent non-competitive inhibitor of both CYP1A and CYP3A activities and that nonylphenol selectively non-competitively inhibited CYP1A activity. Treatment with ethynylestradiol resulted in 46% decrease in CYP3A activity and 22% decrease in protein expression in vivo. In vitro inhibition studies in liver microsomes showed that ethynylestradiol acted as a non-competitive inhibitor of CYP1A activity and as an uncompetitive inhibitor of CYP3A activity. CONCLUSIONS: Ketoconazole, nonylphenol and ethynylestradiol all interacted with CYP1A and CYP3A activities and protein expression in Atlantic cod. However, mechanisms of interactions on CYP1A and CYP3A differ between theses substances and combined exposure had different effects than exposure to single compounds. Thus, CYP1A and CYP3A mediated clearance may be impaired in situations of mixed exposure to those types of compounds.

Journal Article↗
PubMed2005

The reduction of endplate fractures during balloon vertebroplasty: a detailed radiological analysis of the treatment of burst fractures using pedicle screws, balloon vertebroplasty, and calcium phosphate cement.

STUDY DESIGN: In a human cadaveric burst fracture model, the amount of endplate fracture reduction after posterior instrumentation and balloon vertebroplasty was investigated quantitatively. OBJECTIVES: To assess, in a burst fracture model, the vertebral body and adjacent disc heights, in parallel sagittal planes with 3-dimensional (3D) rotational x-ray imaging, at various phases during pedicle screw fixation and subsequent balloon vertebroplasty. SUMMARY OF BACKGROUND DATA: In recent human cadaveric thoracolumbar fracture studies, it was found that vertebral body height could be restored significantly with inflatable bone tamps. However, limited quantitative data exist on the amount of fracture reduction that can be achieved and how much of the reduction will be lost after deflation and removal of the bone tamps before the cement is injected. METHODS: Twenty burst fractures were created and balloon vertebroplasty with calcium phosphate cement was performed after pedicle screw instrumentation. A 3D dataset was obtained during the following phases: intact, fractured, after reduction and stabilization with pedicle screws, after inflation of the balloons, after deflation and removal of the balloons, after injection of the cement. The fractured vertebral body and adjacent disc heights were measured from five reconstructed sagittal images and compared for the six phases of the procedure. Furthermore, the difference between the vertebral body height centrally and peripherally was calculated. RESULTS: The mean vertebral body height at the thoracic level was Tintact = 19.5 +/- 2.2 mm, Tfractured = 14.6 +/- 3.8 mm, Treduction = 17.3 +/- 2.2 mm, Tinflation = 20.1 +/- 2.0 mm, Tdeflation = 18.0 +/- 2.0 mm, and Tcement = 17.8 +/- 1.8 mm. The overall change in vertebral body height between these phases was significant (P < 0.001). At the lumbar level the mean vertebral body height was Tintact = 23.2 +/- 3.8 mm, Tfractured = 14.7 +/- 3.0 mm, Treduction = 18.4 +/- 2.5 mm, Tinflation = 23.2 +/- 3.5 mm, Tdeflation = 19.3 +/- 2.3 mm, and Tcement = 20.2 +/- 2.8 mm. The overall change in MCVBH between these phases was also significant (P < 0.001). The increase in vertebral body height resulted in a decrease of the adjacent disc height. No difference was found for the amount of endplate reduction in the center or at the periphery. No leakage of cement was detected in the spinal canal. CONCLUSIONS: Reduction of endplate fractures, both in the center and at the periphery, seems feasible and safe with combined fracture reduction and balloon vertebroplasty. The endplate fracture reduction that was gained by inflation of the bone tamps could not be maintained after deflation.

Aged↗
PubMed1993

[Transesophageal echocardiography study with dynamic spontaneous contrast of the left atrial appendage, its relationship with the anatomy and thrombus formation].

OBJECTIVE: Transesophageal two-dimensional echocardiographic study of anatomical characteristics of the left atrial appendage and its relation to spontaneous dynamic echocardiographic contrast. DESIGN: Outpatients undergoing a prospective two-dimensional transesophageal echocardiographic study. SETTING: Consecutive outpatients studied at the Echocardiographic Laboratory of Gregorio Marañon General Hospital, Madrid. MATERIAL AND METHODS: In each patient at the level of the left atrial appendage we calculated the following transesophageal echocardiographic parameters: end-systolic and end-diastolic maximal longitudinal and transversal diameters, total systolic and diastolic areas, percentage of systolic fractional shortening, presence of left atrial appendage thrombus and spontaneous dynamic echo-contrast. MAIN RESULTS: Left atrial appendage spontaneous dynamic contrast was observed in 48% of the total population. In the group of patients with left atrial spontaneous echo-contrast we observed larger longitudinal systolic (44 +/- 14 mm vs 28 +/- 13 mm, p = 0.01) and diastolic (52 +/- 16 mm vs 38 +/- 12 mm, p = 0.005) diameters, larger transversal systolic (25 +/- 10 mm vs 19 +/- 6 mm, p = 0.03) and diastolic (28 +/- 8 mm vs 25 +/- 9 mm, p = NS) diameters and also larger systolic (601 +/- 204 mm2 vs 337 +/- 110 mm2, p < 0.0001) and diastolic (715 +/- 230 mm2 vs 507 +/- 184 mm2, p = 0.001) areas, compared to the group without this dynamic echocardiographic phenomena. Left atrial appendage percentage of fractional shortening was considerably reduced in patients with spontaneous dynamic echo-contrast (15 +/- 14% vs 39 +/- 18%, p = 0.001) and related to local thrombus formation (13% vs 1%, p < 0.001). CONCLUSIONS: Left atrial spontaneous dynamic echo-contrast is more common in patients with enlarged left atrial appendage systo-diastolic diameters and areas. In this group of patients the presence of left atrial spontaneous echo-contrast is related to a significant reduction in left atrial appendage contractile function and thrombus formation. Parameter analysis of left atrial appendage anatomy by two-dimensional transesophageal echocardiography may have clinical relevance in the assessment of patients with high risk for left atria thromboembolic phenomena.

Diastole↗
PubMed1991

A perspective on recent trends for scoliosis correction.

The basic principles for scoliosis surgery learned during the Harrington era are still valid today. Experience has confirmed the need for careful selection of the vertebrae to be instrumented, the value of anterior release for rigid curves in imparting convertibility of the deformity, and the importance of careful fusion techniques. During the last decade, further development has occurred because of an increased knowledge of the biomechanical needs for the internally instrumented spine and a three-dimensional appreciation of the scoliotic curve. Biomechanical advances have centered on an understanding of the load-sharing properties afforded by the multiple spinal purchase sites (segmental spinal instrumentation) and the value of two-rod systems linked by couplers. These advances have provided an increased stiffness of the instrumental spine, a reduction in correction loss, improved fatigue properties of the implant, and fewer pseudarthroses. The most important advance of the last decade is an improved awareness of the three-dimensional approach to the scoliotic deformity with the need to preserve or improve sagittal contours. In particular, the importance of the loss of normal thoracic kyphosis or lumbar lordosis has been emphasized. These conceptual gains have led to the development of many new instrument systems to correct deformity. Each is associated with advantages, problems, and risks that must be understood to make intelligent choices for treatment.

Biomechanical Phenomena↗
PubMed2005

Structural insight into the interactions of SoxV, SoxW and SoxS in the process of transport of reductants during sulfur oxidation by the novel global sulfur oxidation reaction cycle.

Microbial redox reactions involving inorganic sulfur compounds, mainly the sulfur anions, are one of the vital reactions responsible for the environmental sulfur balance. These reactions are mediated by phylogenetically diverse prokaryotes, some of which also take part in the extraction of metal ions from their sulfur containing ores. These sulfur oxidizers oxidize inorganic sulfur compounds like sulfide, thiosulfate etc. to produce reductants that are used for carbon dioxide fixation or in respiratory electron transfer chains. The sulfur oxidizing gene cluster (sox) of alpha-Proteobacteria comprises of at least 15 genes, forming two transcriptional units, viz., soxSR and soxVWXYZABCDEFGH. SoxV is known to be a CcdA homolog involved in the transport of reductants from cytoplasm to periplasm. SoxW and SoxS are periplasmic thioredoxins, which (SoxW) interact with SoxV and thereby help in the redox reactions. We have employed homology modeling to construct the three-dimensional structures of the SoxV, SoxW and SoxS proteins from Rhodovulum sulfidophilum. With the help of docking and molecular dynamics simulations we have identified the amino acid residues of these proteins involved in the interaction. The probable biochemical mechanism of the transport of reductants through the interactions of these proteins has also been investigated. Our study provides a rational basis to interpret the molecular mechanism of the biochemistry of sulfur anion oxidation reactions by these ecologically important organisms.

Alphaproteobacteria↗
PubMed1997

Modeling the mechanical behavior of vertebral trabecular bone: effects of age-related changes in microstructure.

Age-related reductions in the thickness and number of trabeculae in vertebral trabecular bone have been documented by several workers, yet the relative effects of these changes on mechanical properties are not known. We developed a two-dimensional model of human vertebral trabecular bone and investigated its mechanical behavior using finite element analysis. The stress-strain behavior, failure mode, and strain distributions predicted using the model were consistent with those observed for vertebral trabecular bone under compressive loading. Random reductions in the number of trabeculae reduced the modulus and strength of the models two to five times more than uniform reductions in the thickness of trabeculae that caused the same loss of bone volume. For example, randomly removing longitudinal trabeculae to achieve a reduction in density of 10% reduced the strength by approximately 70%, whereas removing the same amount of bone by uniformly reducing the thickness of the longitudinal trabeculae only reduced the strength by approximately 20%. For a simulation of aged bone, in which the thickness and number of trabeculae were reduced concurrently, the strength was 23% of its intact ("young") value. When the bone mass of the aged model was restored to its intact level by increasing the thickness but not the number of trabeculae, the strength increased by 60%, but was still only 37% of its intact value. These combined findings, based on a two-dimensional, idealized model of vertebral trabecular bone, illustrate the importance of maintaining trabecular number and suggest that it may not be possible to restore bone strength following a period of advanced bone loss if a substantial number of trabeculae have been resorbed. Thus, until treatments exist that can increase trabecular number, the most effective treatment strategy is to prevent the degradation of bone strength by maintaining the number of trabeculae at a healthy level.

Aged↗
PubMed1997

Bilateral reductions in hippocampal volume in adults with epilepsy and a history of febrile seizures.

OBJECTIVES: To examine the degree and frequency of reductions in hippocampal volume in patients with temporal lobe epilepsy with and without a history of febrile seizures. METHODS: In vivo measures of hippocampal volume were computed from three dimensional gradient echo (FLASH) images in 44 patients undergoing comprehensive evaluations for epilepsy surgery. Twenty one patients (48%) reported a history of febrile seizures. The volumes from these patients were compared with those from 23 patients without a history of febrile seizures and 34 healthy controls. RESULTS: The febrile seizure group had significant reductions in volume, both ipsilateral (30% decrease) and contralateral (15% decrease), to the EEG seizure focus. Twelve of 18 patients with febrile seizures exhibited clinically significant ipsilateral volume reductions, defined as volumes falling 2 SD below the mean obtained from the control sample. Only four of 19 patients without febrile seizures exhibited this degree of reduction. No significant correlations were found between seizure variables (for example, duration of epilepsy, seizure frequency) and ipsilateral reductions in volume. However, a significant inverse correlation (r=-0.45, P<0.05) between seizure frequency and the volume of the hippocampus contralateral to the seizure focus was found in the febrile seizure group. CONCLUSION: These results suggest that a history of febrile seizures is associated with the finding of a smaller hippocampus on the side ipsilateral to the subsequent temporal lobe focus whereas chronic factors seem to be be related to pathology contralateral to the seizure focus.

Adolescent↗
PubMed2003

Usefulness of coronary flow reserve over regional wall motion when added to dual-imaging dipyridamole echocardiography.

Vasodilator stress echocardiography allows semi-simultaneous imaging of left anterior descending (LAD) coronary flow and regional wall function. To assess the relative (and additive?) value of regional flow and function for noninvasive identification of angiographically assessed LAD disease in patients with chest pain syndrome, we studied 230 consecutive in-hospital patients (134 men, aged 63.5 +/- 11 years) with chest pain syndrome and normal regional and global left ventricular function. All patients underwent stress echocardiography with dipyridamole (up to 0.84 mg/kg over 10 minutes), including wall motion analysis by 2-dimensional echocardiography and coronary flow reserve (CFR) evaluation of the LAD artery by Doppler, with or without contrast injection. A new regional wall motion abnormality in >or=2 contiguous segments was required for 2-dimensional echocardiographic positivity. CFR was evaluated as the ratio of dipyridamole to peak diastolic coronary blood flow velocity at rest. All patients underwent coronary angiography within 60 days; a quantitatively assessed diameter reduction >50% of the LAD artery was considered significant. Of the 230 patients, 70 had LAD disease. A regional wall motion abnormality in LAD territory was present in 52 patients, and reduced CFR (<1.9) in 62 patients. Sensitivity for detecting LAD disease was 74% for 2-dimensional echocardiography (95% confidence interval [CI] 64% to 84%) and 81% for CFR <1.9 (95% CI 72% to 90%); specificity was 91% (95% CI 87% to 96%) for 2-dimensional echocardiography and 84% for CFR (95% CI 79% to 90%). Accuracy was 86% for 2-dimensional echocardiography (95% CI 82% to 91%) and 83.5% for CFR (95% CI 79% to 88%). When 2-dimensional echocardiography and CFR criteria were considered, sensitivity increased to 93% (95% CI 87% to 99%), with 80.6% specificity (95% CI 74.5% to 86.7%). CFR was assessed during vasodilator stress echocardiography. Its diagnostic accuracy for detecting LAD disease was comparable to regional wall motion abnormalities. However, the data for flow and function can be complementary in terms of predicting underlying angiographic anatomy, because abnormal wall motion can include coronary artery disease, and negative CFR can exclude it.

Chest Pain↗
PubMed1994

Disparate structural effects on left and right ventricles by angiotensin-converting enzyme inhibitors and calcium antagonists in essential hypertension.

Reduced left ventricular (LV) mass and wall thicknesses with angiotensin-converting enzyme (ACE) inhibitors and calcium antagonists occur in essential hypertension. Experimental studies have shown that reduction in LV mass is related to hemodynamic as well as nonhemodynamic factors that may influence LV and also right ventricular (RV) structure. Sixty-eight patients with essential hypertension were studied echocardiographically 4 to 8 weeks after antihypertensive therapy was initiated. LV dimensions and wall thicknesses, as well as RV free wall thickness, were measured using 2-dimensionally guided techniques. Patients were divided in 2 groups: those receiving either ACE inhibitors or calcium antagonists. This short-term therapeutic period produced similar hemodynamic changes and reductions in LV mass in both groups. In these 2 groups of patients, we found opposite effects in RV free wall thickness. Thus, RV wall thickness increased from 0.44 +/- 0.02 to 0.56 +/- 0.02 cm (p < 0.01) after therapy in the calcium antagonist group, whereas no change was found in the ACE inhibitor group (0.36 +/- 0.04 vs 0.34 +/- 0.04). These structural changes suggest that ACE inhibitors and calcium antagonists produce dissimilar effects on cardiac cell growth and growth reversal, which may be mediated through different nonhemodynamic effects after short-term therapy.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed2005

Structural and kinetic basis for substrate selectivity in Populus tremuloides sinapyl alcohol dehydrogenase.

We describe the three-dimensional structure of sinapyl alcohol dehydrogenase (SAD) from Populus tremuloides (aspen), a member of the NADP(H)-dependent dehydrogenase family that catalyzes the last reductive step in the formation of monolignols. The active site topology revealed by the crystal structure substantiates kinetic results indicating that SAD maintains highest specificity for the substrate sinapaldehyde. We also report substantial substrate inhibition kinetics for the SAD-catalyzed reduction of hydroxycinnamaldehydes. Although SAD and classical cinnamyl alcohol dehydrogenases (CADs) catalyze the same reaction and share some sequence identity, the active site topology of SAD is strikingly different from that predicted for classical CADs. Kinetic analyses of wild-type SAD and several active site mutants demonstrate the complexity of defining determinants of substrate specificity in these enzymes. These results, along with a phylogenetic analysis, support the inclusion of SAD in a plant alcohol dehydrogenase subfamily that includes cinnamaldehyde and benzaldehyde dehydrogenases. We used the SAD three-dimensional structure to model several of these SAD-like enzymes, and although their active site topologies largely mirror that of SAD, we describe a correlation between substrate specificity and amino acid substitution patterns in their active sites. The SAD structure thus provides a framework for understanding substrate specificity in this family of enzymes and for engineering new enzyme specificities.

Acrolein↗
PubMed1999

Human ceruloplasmin. Intramolecular electron transfer kinetics and equilibration.

Pulse radiolytic reduction of disulfide bridges in ceruloplasmin yielding RSSR(-) radicals induces a cascade of intramolecular electron transfer (ET) processes. Based on the three-dimensional structure of ceruloplasmin identification of individual kinetically active disulfide groups and type 1 (T1) copper centers, the following is proposed. The first T1 copper(II) ion to be reduced in ceruloplasmin is the blue copper center of domain 6 (T1A) by ET from RSSR(-) of domain 5. The rate constant is 28 +/- 2 s(-1) at 279 K and pH 7.0. T1A is in close covalent contact with the type 3 copper pair and indeed electron equilibration between T1A and the trinuclear copper center in the domain 1-6 interface takes place with a rate constant of 2.9 +/- 0.6 s(-1). The equilibrium constant is 0.17. Following reduction of T1A Cu(II), another ET process takes place between RSSR(-) and T1B copper(II) of domain 4 with a rate constant of 3.9 +/- 0.8. No reoxidation of T1B Cu(I) could be resolved. It appears that the third T1 center (T1C of domain 2) is not participating in intramolecular ET, as it seems to be in a reduced state in the resting enzyme.

Ceruloplasmin↗
PubMed1999

The CXXC motif: crystal structure of an active-site variant of Escherichia coli thioredoxin.

The 2.2 A crystalline structure of an oxidized active-site variant of Escherichia coli thioredoxin (Trx) has been solved. Trx is a 12 kDa enzyme which catalyzes the oxidation of dithiols and the reduction and isomerization of disulfides in other proteins. Its active site contains the common structural motif CXXC. Protein-disulfide isomerase (PDI), a 57 kDa homolog of Trx, contains four Trx-like domains. The three-dimensional structure of PDI is unknown. PDI-deficient Saccharomyces cerevisiae are inviable. An active-site variant of Trx which complements PDI-deficient yeast has the active-site sequence Cys32-Val33-Trp34-Cys35 (CVWC). The reduction potential of oxidized CVWC Trx (E degrees ' = -0.230 V) is altered significantly from that of the wild-type enzyme (E degrees ' = -0.270 V). However, the structure of the oxidized CVWC enzyme is almost identical to that of wild-type Trx. The addition of valine and tryptophan in the active site is likely to increase the reduction potential, largely by decreasing the pK(a) of the Cys32 thiol in the reduced enzyme. Unlike in wild-type Trx, significant protein-protein contacts occur in the crystal. Protein molecules related by a crystallographic twofold axis form a dimer in the crystal. The dimer forms as an extension of the twisted mixed beta-sheet which composes the backbone of each Trx structure.

Amino Acid Sequence↗
PubMed2007

Studies of a cobalt(III) complex of the MMP inhibitor marimastat: a potential hypoxia-activated prodrug.

We report a potential means of selectively delivering matrix metalloproteinase (MMP) inhibitors to target tumour sites by use of a bioreductively activated Co(III) carrier system. The carrier, comprising a Co(III) complex of the tripodal ligand tris(methylpyridyl)amine (tpa), was investigated with the antimetastatic MMP inhibitor marimastat (mmstH(2)). The X-ray crystal structure of [Co(mmst)(tpa)]ClO(4) x 4H(2)O was determined and two-dimensional NMR revealed the existence of two isomeric forms of the complex in solution. Electrochemical analysis showed that the reduction potential of the complex is suitable for it to be bioreductively activated at hypoxic tumour sites. In vitro assays confirmed the stability of the prodrug in solution prior to reduction and revealed very low cytotoxicity against A2780 cells. In vivo testing in mice showed a higher level of tumour-growth inhibition by the complex than by free marimastat. Both free marimastat and and its Co(III) complex increased metastasis in the model used, with the complex significantly more active.

Amines↗
PubMed2003

Ultimate intrinsic signal-to-noise ratio for parallel MRI: electromagnetic field considerations.

A method is described for establishing an upper bound on the spatial encoding capabilities of coil arrays in parallel MRI. Ultimate intrinsic signal-to-noise ratio (SNR), independent of any particular conductor arrangement, is calculated by expressing arbitrary coil sensitivities in terms of a complete set of basis functions that satisfy Maxwell's equations within the sample and performing parallel imaging reconstructions using these basis functions. The dependence of the ultimate intrinsic SNR on a variety of experimental conditions is explored and a physically intuitive explanation for the observed behavior is provided based on a comparison between the electromagnetic wavelength and the distance between aliasing points. Imaging at high field strength, with correspondingly short wavelength, is shown to offer advantages for parallel imaging beyond those already expected due to the larger available spin polarization. One-dimensional undersampling of k-space yields a steep drop in attainable SNR for more than a 5-fold reduction of scan time, while 2D undersampling permits access to much higher degrees of acceleration. Increased tissue conductivity decreases baseline SNR, but improves parallel imaging performance. A procedure is also provided for generating the optimal coil sensitivity pattern for a given acceleration, which will serve as a useful guide for future coil designs.

Algorithms↗
PubMed1997

Improved 2D time-of-flight angiography using a radial-line k-space acquisition.

For flow imaging applications, radial-line k-space acquisition methods offer advantages over conventional 2DFT methods. Specifically, radial-line acquisition methods mitigate artifacts resulting from pulsatile flow while offering a potential reduction in scan times. In this paper, radial-line and 2DFT acquisitions are compared in a two-dimensional time-of-flight angiography sequence. The twisting radial-line (TwiRL) trajectory, a variant of 2D projection reconstruction, is used to represent the family of radial-line trajectories. In both phantom and in vivo studies, the TwiRL images demonstrate improved vessel depiction including a more uniform signal intensity and better delineation of the vasculature in comparison with images obtained via the 2DFT method.

Arterial Occlusive Diseases↗
PubMed1985

Monoclonal antibodies to human plasma protein X alias complement S-protein.

Protein X alias complement S-protein was isolated by dissociation from purified XC5b-9 (fluid-phase terminal C5b-9) complexes with 250 mM deoxycholate and subsequent sucrose density gradient centrifugation and Sephacryl gel chromatography. Polyclonal rabbit and monoclonal mouse antibodies were used to preliminarily characterize the protein in human serum and plasma. In plasma, Protein X yielded a symmetrical immunoprecipitate of alpha 2-mobility in a crossed immunoelectrophoresis assay. However, a second immunoprecipitate of alpha 1-mobility was observed when serum was analysed; this precipitate represented Protein X in complex with antithrombin-III. The co-precipitation of Protein X with serum antithrombin-III was exploited for establishing a simple screening test for unequivocal identification of monoclonal anti-Protein X antibodies. SDS-PAGE immunoblotting with monoclonal antibodies showed that Protein X exhibits pronounced microheterogeneity, migrating as a diffuse moiety of approx. Mr 80-90 000. Additionally, a small amount of polymeric aggregates appear to be present in plasma. Reduction of disulfide bonds led to liberation of a polypeptide of approx. 15 K as discerned by two-dimensional SDS-PAGE immunoblotting. Protein X is not cleaved to lower molecular weight entities during the process of blood coagulation or during formation of fluid-phase terminal complement complexes. The plasma concentrations in healthy adults were in the range of 500-700 micrograms/ml. The availability of methods for isolating Protein X and raising monoclonal antibodies will facilitate further studies on the dual role of this protein in the terminal complement and coagulation cascades.

Animals↗
PubMed2006

Self-assembly of Co-based nanosheets into novel nest-shaped nanostructures: Synthesis and characterization.

We report the first observation of the formation of novel Co-based three-dimensional (3D) self-assembly hollow nanostructures, i.e., nest-shaped nanospheres composed of sheet-like particles, via reduction of cobalt salt with sodium tetrahydroboride in cetyltrimethylammonium bromide (CTAB)-cyclohexane-NH4F aqueous solutions. It was found that the cyclohexane has a significant influence on the formation of the nest-shaped Co-based nanospheres, because when the experiments are carried out in the absence of cyclohexane, only sheet-like particles are formed. NH4F plays also an important role in the formation of the hollow nanostructures because without this salt mainly solid spherical structures, composed of sheet-like particles, instead of nest-shaped structures are obtained. The nanostructures are mainly formed by Co, but also a minor amount (17%) of Co2B is present in the final compounds. The structures are characterized by X-ray diffraction (XRD), energy dispersive spectroscopy (EDS), transmission electron microscopy (TEM), high resolution transmission electron microscopy (HRTEM), and field-emission scanning electron microscopy (FESEM). A possible mechanism for the formation of the novel Co-based nanostructures is proposed.

Journal Article↗
PubMed1998

Effect of oxytocin, prostaglandin F2 alpha, and clenbuterol on uterine dynamics in mares.

The effects of oxytocin, prostaglandin F2 alpha (PGF2 alpha), and clenbuterol on uterine contractility and tone during anestrus and diestrus, and during mobility and postfixation of the embryonic vesicle were studied in 51 pony mares. Contractility was assessed by scoring real-time ultrasound images, and tone was assessed by transrectal digital compression. Scoring was done by an operator who had no knowledge of treatment assignments. In anovulatory mares primed with progesterone for 16 d, oxytocin did not significantly alter contractility but did stimulate an increase in tone, whereas clenbuterol depressed both contractility and tone. The PGF2 alpha given on Days 12, 15, and 18 did not significantly alter uterine contractility in pregnant mares, but it increased contractility on all days in nonpregnant mares. Clenbuterol decreased both tone and contractility when given to pregnant mares on the day of embryonic-vesicle fixation, while it decreased tone but not contractility when given on Day 19. Clenbuterol treatment was associated with dislodgment of the fixed embryo in only 1 of 5 mares. However, on Day 19, clenbuterol treatment was associated with a change in shape of the conceptus when viewed in a cross section of the uterine horn. The conceptus shape became more circular rather than irregular or triangular, as indicated by a significant decrease in the variation in the distances between adjacent walls measured in 4 different directions. Results indicated that: 1) oxytocin increased uterine tone but did not alter contractility in progesterone-primed anestrous mares; 2) on Days 12, 15 and 18, PGF2 alpha increased uterine contractility in nonpregnant mares but not in pregnant mares; 3) clenbuterol decreased both tone and contractility at all reproductive states except for a lack of a decrease in contractility on Day 19 of pregnancy; and 4) reduction in uterine tone from clenbuterol treatment on Day 19 was associated with a change in the two-dimensional shape of the in situ conceptus from irregular to a more circular form.

Animals↗