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PubMed2005

Improvement on the visualization of cytoskeletal structures of protozoan parasites using high-resolution field emission scanning electron microscopy (FESEM).

The association of high resolution field emission scanning electron microscopy (FESEM), with a more efficient system of secondary electron (SE) collection and in-lens specimen position, provided a great improvement in the specimen's topographical contrast and in the generation of high-resolution images. In addition, images obtained with the use of the high-resolution backscattered electrons (BSE) detector provided a powerful tool for immunocytochemical analysis of biological material. In this work, we show the contribution of the FESEM to the detailed description of cytoskeletal structures of the protozoan parasites Herpetomonas megaseliae, Trypanosoma brucei and Giardia lamblia. High-resolution images of detergent extracted H. megaseliae and T. brucei showed the profile of the cortical microtubules, also known as sub-pellicular microtubules (SPMT), and protein bridges cross-linking them. Also, it was possible to visualize fine details of the filaments that form the lattice-like structure of the paraflagellar rod (PFR) and its connection with the axoneme. In G. lamblia, it was possible to observe the intricate structure of the adhesive disk, funis (a microtubular array) and other cytoskeletal structures poorly described previously. Since most of the stable cytoskeletal structures of this protozoan rely on tubulin, we used the BSE images to accurately map immunolabeled tubulin in its cytoskeleton. Our results suggest that the observation of detergent extracted parasites using FESEM associated to backscattered analysis of immunolabeled specimens represents a new approach for the study of parasite cytoskeletal elements and their protein associations.

Animals↗

PubMed2005

Dynamic kinetic resolution: alternative approach in optimizing S-ibuprofen production.

A lipase catalysed enantioselective hydrolysis process under in situ racemization of the remaining (R)-ibuprofen ester substrate with sodium hydroxide as the catalyst was developed for the production of S-ibuprofen from (R,S)-ibuprofen ester in isooctane. Detailed investigations on parameters study indicated that 0.5 M NaOH, addition of 20% (v/v) co-solvent (dimethyl sulphoxide), operating temperature of 45 degrees C, and 40 mmol/L substrate gave 86% conversion and 99.4% optical purity of S-ibuprofen in dynamic kinetic resolution. Meanwhile, in common enzymatic kinetic resolution process, only 42% conversion of the racemate and 93% enantiomeric excess of the product was obtained which are of lower values as compared to dynamic kinetic resolution. The S-ibuprofen produced during each process was evaluated and approximately 50% increment in concentration of S-acid product was produced when dynamic kinetic resolution was applied into the process.

Combinatorial Chemistry Techniques↗

PubMed2005

Cine magnetic resonance imaging vs high-resolution ultrasonography for detection of adhesions after laparoscopic and open incisional hernia repair: a matched pair pilot analysis.

BACKGROUND: Intraabdominal adhesions represent nonspecific complications before or after laparoscopic or open incisional hernia repair. The objective of this matched control pilot study was to display long-term adhesions noninvasively by applying functional cine magnetic resonance (MR) imaging, as compared with applying high-resolution ultrasonography (US). METHODS: The study group, composed of 17 consecutive patients (12 men and 5 women; mean age, 52 years), underwent laparoscopic intraperitoneal onlay mesh repair using expanded polytetrafluoroethylene (ePTFE) mesh. Their mean body mass index was 30 kg/m(2), and the size of the hernia was 95 cm(2). Another group, matched for age, gender, and type of hernia, was subjected to open abdominal wall repair using the preperitoneal sublay technique with a large-pore, low-weight polypropylene mesh. For cine MR imaging (1.5 T), section-by-section dynamic depiction of induced visceral slide throughout the entire abdomen was achieved by applying transverse or sagittal true fast imaging with steady-state precession sequences. The location and type of adhesions were compared with high-resolution ultrasonography using nine segments of the abdominal map. RESULTS: The patients subjected to laparoscopic and open incisional hernia repair were examined 16 and 28 months after surgery. The findings showed functional cine MR imaging as superior to high-resolution ultrasonography for assessing the amount of intraabdominal adhesions (n = 53 vs n = 3; p < 0.01). Most frequently, adhesions were seen between small bowel loops and the abdominal wall (n = 22), followed by bowel-to-bowel adhesions (n = 19; p < 0.05). However, adhesions between small bowel loops and the abdominal wall occurred more frequently after open mesh repair (p < 0.05). Furthermore, a strong correlation was observed between patient complaints and findings with cine MR imaging (p < 0.05). Maximum pain correlated significantly with the region of the most distinctive adhesions (p < 0.05). CONCLUSIONS: Functional cine MR imaging represents a reliable noninvasive technique for detecting long-term adhesions after open and laparoscopic incisional hernia repair. The study results suggest that this approach has distinct advantages over high-resolution ultrasonography.

Adult↗

PubMed2000

Feasibility of optical coherence tomography for high-resolution imaging of human gastrointestinal tract malignancies.

Optical coherence tomography (OCT) is a new imaging technology which can perform high-resolution, cross-sectional imaging of the internal microstructure of biological tissues. OCT is analogous to ultrasound, except that it measures the intensity of back-reflected infrared light rather than sound waves. OCT performs two- and three-dimensional imaging of tissue microstructure in situ and in real time. It can achieve image resolutions approaching the cellular level over approximately the same imaging depths as a conventional biopsy. In this article we examine the feasibility of OCT for high-resolution imaging of gastrointestinal malignancies with ex-vivo imaging of normal and pathologic microstructures. Tissue, both normal and neoplastic, was obtained from patients undergoing surgical resection after an initial diagnosis of a gastrointestinal malignancy. The tissue samples were imaged prior to fixation using a laboratory OCT system. The OCT system consists of a fiber optic-based Michelson interferometer, a commercially available amplified superluminscent light source, and a computer for data acquisition. The images were subsequently compared with histological cross-sections corresponding to the imaged areas. The stratified squamous epithelium of the normal esophagus was clearly visible in the OCT images and contrasted to the disorganized and non-uniform nature of the mucosal layers of Barrett's esophagus and squamous carcinoma. The columnar epithelial morphology as well as other mucosal structures in normal colon were distinctly visible using OCT. In contrast, disorganization of the normal mucosal layers and ulcerative lesions were identified in tissues from ulcerative colitis and adenocarcinoma of the colon. The ability of OCT to image tissue microstructure at high resolutions makes it a potentially powerful technology for minimally invasive assessment of the gastrointestinal tract and the evaluation of early neoplastic changes.

Adenocarcinoma↗

PubMed2002

Three high-resolution crystal structures of cadmium-substituted carboxypeptidase A provide insight into the enzymatic function.

Three high-resolution crystal structures of Cd(II)-substituted carboxypeptidase A (CPA) have been determined by X-ray diffraction from crystals prepared in three different buffer systems to assess the effect of pH and ionic strength on the Cd(II) coordination geometry. All crystallize in the space group P2(1) with identical cell dimensions. Cd-CPA(7.5): Cd(II)-substituted CPA prepared at pH 7.5 with [Cl(-)]=2 mM determined to 1.70 A resolution ( R=17.4% and R(free)=19.8%); Cd-CPA(5.5): Cd(II)-substituted CPA prepared at pH 5.5 with [Cl(-)]=2 mM to 2.00 A resolution ( R=16.1% and R(free)=18.6%); Cd-CPA(7.5)-Cl: Cd(II)-substituted CPA prepared at pH 7.5 with [Cl(-)]=250 mM to 1.76 A resolution ( R=16.7% and R(free)=17.8%). No noticeable structural changes were observed between the three structures. Two water molecules coordinate to Cd(II), in contrast to the single water molecule coordinating to Zn(II) in the Zn-CPA structure. No binding sites for anions could be identified, even in the structure with a high concentration of chloride ions. It is suggested that the anion inhibition is due to weak outer-sphere association of Cl(-) ions at several binding sites, shielding the strong positive charge distribution at the surface of the protein near the active site. Based on structural data and a sequence alignment of 18 non-redundant carboxypeptidases, a more elaborate version of the earlier reaction model is proposed that also addresses the transport of water to and from the active site. Conserved residues whose function was not addressed previously delineate the proposed pathways used in the transport of water during catalysis.

Amino Acid Sequence↗

PubMed2005

Is preoperative high-resolution magnetic resonance imaging accurate in predicting neurovascular compression in patients with trigeminal neuralgia? A single-blind study.

High-resolution magnetic resonance imaging (HR-MRI) using three-dimensional fast imaging employing steady-state acquisition (3D-FIESTA) and double-dose contrast-enhanced three-dimensional fast spoiled gradient echo (3D-FSPGR) sequences is considered to be a useful tool in detecting neurovascular compression in patients with trigeminal neuralgia. The purpose of this study was to analyze the accuracy and preoperative diagnostic value of these high-resolution imaging techniques in patients with trigeminal neuralgia, in a single-blind study. The preoperative MRI images of 21 consecutive patients were matched to one neuroradiologist, who was blind as to which side exhibited the symptoms. The images and post-processing multiplanar reconstructions were compared with the video-documented operative observations. HR-MRI using only 3D-FSPGR sequences demonstrated neurovascular compression in accordance with the intraoperative finding in 11 patients (52.4%). In the subgroup where, additionally, 3D-FIESTA sequences were available, neurovascular compression was in accordance with the intraoperative finding in 71.4% (n = 7). High-resolution magnetic resonance imaging using double-dose contrast-enhanced 3D-FSPGR and 3D-FIESTA sequences is currently not sufficient enough to make an accurate prediction of neurovascular compression in a single-blind setting. These 3D imaging techniques currently provide only limited information, and one should consider their use carefully when identifying patients with trigeminal neuralgia from operation until image quality is improved by superior image resolution that can accurately discriminate vessels surrounding the trigeminal root entry zone.

Adult↗

PubMed2004

Cervical cancer tissue characterized by high-resolution magic angle spinning MR spectroscopy.

OBJECTIVE: In recent years, high-resolution magic angle spinning (HR MAS) has provided the opportunity to explore detailed biochemical composition of intact tissue. Previous studies of intact cervical biopsies with high-resolution magnetic resonance spectroscopy (MRS) have correlated well with histopathology. Lactate level in cervical cancer tissue has been found to correlate to metastatic spread. The purpose of this study was to explore the potential of the HR MAS technique as a tool for chemical characterization of cervical cancer tissue. MATERIALS AND METHODS: Tissue samples from the cervix were collected after hysterectomy from patients with cervical cancer (n=8) and patients with nonmalignant disease (n=8). The tissue specimens were analyzed using HR MAS MR spectroscopy combined with principal component analysis (PCA). RESULTS: The resulting spectra showed resolution comparable with high-resolution MR spectra of extracts. Multivariate analysis confirmed that MAS spectra classified according to patient diagnosis. CONCLUSION: Malignant tissue of the cervix differed from nonmalignant tissue with regard to higher levels of cholines and amino acid residues and lower levels of glucose.

Adenocarcinoma↗

PubMed2006

Prevalence of perianal intraepithelial neoplasia in HIV-infected patients referred for high-resolution anoscopy.

PURPOSE: This study was designed to describe perianal disease in a cohort of HIV-infected patients referred for high-resolution anoscopy. METHODS: A retrospective chart review was performed on 52 HIV-infected patients referred for high-resolution anoscopy from 2001 to 2005. All patients underwent anal canal and perianal high-resolution anoscopy in the office with biopsy of suspicious areas. Patients with high-grade intraepithelial perianal lesions underwent multiple biopsies under general anesthesia in the operating room to rule out malignancy. RESULTS: Of the 52 patients, 19 (37 percent) had perianal abnormalities noted on high-resolution anoscopy and underwent punch biopsy. The mean duration of known HIV infection in these 19 patients (15 males) was 10.6 years, with 17 on highly active antiretroviral therapy for the last 3-month period. Mean CD4 count was 371 cells/microl. Office perianal biopsies diagnosed two patients with invasive squamous-cell carcinoma and nine with high-grade squamous intraepithelial lesion. Seven of the nine patients with perianal high-grade squamous intraepithelial lesion on office biopsy were submitted to multiple biopsies under general anesthesia. One of these seven had an occult perianal squamous-cell carcinoma. CONCLUSIONS: Perianal disease was common in this group of HIV-infected patients; 11 patients (21 percent of total) were diagnosed with squamous-cell carcinoma or high-grade squamous intraepithelial lesion. Because only 19 patients had clinically suspicious perianal lesions biopsied, this may be an underestimate. Our data suggest that anal canal neoplasia often is accompanied by perianal disease and illustrates the need for biopsy of any suspicious perianal lesions.

Adult↗

PubMed2006

A high resolution approach to estimating time-frequency spectra and their amplitudes.

A high resolution approach to estimating time-frequency spectra (TFS) and associated amplitudes via the use of variable frequency complex demodulation (VFCDM) is presented. This is a two-step procedure in which the previously developed time-varying optimal parameter search (TVOPS) technique is used to obtain TFS, followed by using the VFCDM to obtain even greater TFS resolution and instantaneous amplitudes associated with only the specific frequencies of interest. This combinational use of the TVOPS and the VFCDM is termed the TVOPS-VFCDM. Simulation examples are provided to demonstrate the performance of the TVOPS-VFCDM for high resolution TFS as well as instantaneous amplitude estimation. The simulation results show that the TVOPS-VFCDM approach provides the highest resolution and most accurate amplitude estimates when compared to the smoothed pseudo Wigner-Ville, continuous wavelet transform and Hilbert-Huang transform methods. Application of the TVOPS-VFCDM to renal blood flow data indicates some promise of a quantitative approach to understanding the dynamics of renal autoregulatory mechanisms as well as a possible approach to quantitatively discriminating between different strains of rats.

Algorithms↗

PubMed2006

Mapping turbidity in the Charles River, Boston using a high-resolution satellite.

The usability of high-resolution satellite imagery for estimating spatial water quality patterns in urban water bodies is evaluated using turbidity in the lower Charles River, Boston as a case study. Water turbidity was surveyed using a boat-mounted optical sensor (YSI) at 5 m spatial resolution, resulting in about 4,000 data points. The ground data were collected coincidently with a satellite imagery acquisition (IKONOS), which consists of multispectral (R, G, B) reflectance at 1 m resolution. The original correlation between the raw ground and satellite data was poor (R2 = 0.05). Ground data were processed by removing points affected by contamination (e.g., sensor encounters a particle floc), which were identified visually. Also, the ground data were corrected for the memory effect introduced by the sensor's protective casing using an analytical model. Satellite data were processed to remove pixels affected by permanent non-water features (e.g., shoreline). In addition, water pixels within a certain buffer distance from permanent non-water features were removed due to contamination by the adjacency effect. To determine the appropriate buffer distance, a procedure that explicitly considers the distance of pixels to the permanent non-water features was applied. Two automatic methods for removing the effect of temporary non-water features (e.g., boats) were investigated, including (1) creating a water-only mask based on an unsupervised classification and (2) removing (filling) all local maxima in reflectance. After the various processing steps, the correlation between the ground and satellite data was significantly better (R2 = 0.70). The correlation was applied to the satellite image to develop a map of turbidity in the lower Charles River, which reveals large-scale patterns in water clarity. However, the adjacency effect prevented the application of this method to near-shore areas, where high-resolution patterns were expected (e.g., outfall plumes).

Boston↗

PubMed1988

High-resolution two-dimensional electrophoresis of plant proteins.

A technique for the analysis of plant proteins from seed, leaf, root, and coleoptile tissues by high resolution two-dimensional electrophoresis is described. This technique is based primarily on the procedure of P. O'Farrell (1975, J. Biol. Chem. 250, 4007-4021); however, a number of improvements and simplifications have been introduced. We have found that resolution of polypeptides from a range of plant tissues is improved if the concentrations of nonionic detergent and ampholytes used in the isoelectric focusing (IEF) step are increased to 4 and 5% (w/v), respectively. Further increase in the concentrations of these two components results in gels of decreased resolution and low mechanical strength. We have also found that substitution of n-octyl beta-D-glucopyranoside or 3-[(cholamidopropyl)dimethylammonio]-1-propanesulfonate for Triton X-100 or Nonidet-P40 in the IEF dimension significantly increases the resolution of polypeptides in these gels. This technique also allows minor polypeptide differences between closely related cultivars of plants to be identified.

Ampholyte Mixtures↗

PubMed1995

Resolution of experimental pneumothorax in rabbits by oxygen therapy.

The treatment of asymptomatic patients with small pneumothoraces (ie, less than 20% by volume) has included observation, tube thoracostomy, and operation. When observation is used, the anticipated expansion of the lung has been estimated to be 1.25% of the lung volume daily. This study was designed to evaluate the use of inhaled oxygen as a method to accelerate the resolution of a pneumothorax in a rabbit model. Experimental pneumothoraces were created in 23 white New Zealand rabbits. Group 1 (9 rabbits) were placed in a cage with room air and group 2 (11 rabbits) were placed in a cage with high oxygen concentration. Three rabbits died before completion of the study. Serial chest roentgenograms were performed until the pneumothoraces resolved. The majority of rabbits treated with oxygen had resolution of their pneumothoraces by 36 hours, whereas the majority of rabbits treated with room air did not show complete resolution before 48 hours. Biopsies showed no evidence of damage secondary to oxygen treatment. Oxygen treatment was found to be significantly better in the early resolution of pneumothoraces when compared with room air. This establishes an alternative treatment for some pneumothoraces that are small and asymptomatic.

Animals↗

PubMed1996

High resolution evoked potential imaging of the cortical dynamics of human working memory.

High resolution evoked potentials (EPs), sampled from 115 channels and spatially sharpened with the finite element deblurring method, were recorded from 8 subjects during working memory (WM) and control tasks. The tasks required matching each stimulus with a preceding stimulus on either verbal or spatial attributes. All stimuli elicited a central P200 potential that was larger in the spatial tasks than in the verbal tasks, and larger in the WM tasks than in the control tasks. Frequent, non-matching stimuli elicited a frontal, positive peak at 305 msec that was larger in the spatial WM task relative to the other tasks. Irrespective of whether subjects attended to verbal or spatial stimulus attributes, non-matching stimuli in the WM tasks also elicited an enhanced P450 potential over the left frontal cortex, followed by a sustained potential over the superior parietal cortex. A posterior P390 potential elicited by infrequent, matching stimuli was smaller in amplitude for both spatial and verbal WM tasks compared to control tasks, as was a central prestimulus CNV. These results indicate that WM is a function of a distributed system with both task-specific and task-independent components. Lesion studies and course temporal resolution functional imaging methods, such as PET and fMRI, tend to paint a fairly static picture of the cortical regions which participate in the performance of WM tasks. In contrast, the fine-grain time resolution provided by imaging brain function with EP methods provides a dynamic picture of subsecond changes in the spatial distribution of WM effects over the course of individual trials, as well as evidence for differences in the activity elicited by matching and non-matching stimuli within sequences of trials. This information about the temporal dynamics of WM provides a critical complement to the fine-grain spatial resolution provided by other imaging modalities.

Acoustic Stimulation↗

PubMed1995

Resolution of the enantiomers of oxamniquine by capillary electrophoresis and high-performance liquid chromatography with cyclodextrins and heparin as chiral selectors.

The methods of separation of the enantiomers of the chiral drug oxamniquine are compared, between HPLC with either cyclodextrins and their related derivatives as chiral selectors in the mobile phase or immobilised in a chiral stationary phase (as Cyclobond I and II) and between capillary zone electrophoresis (CZE) where the cyclodextrins are added to the buffer solution. The HPLC experiments, which included structured method optimisation were largely unsuccessful in resolving the enantiomers, with the exception of when a Chiral-AGP protein stationary phase was introduced into the programme. However although this chiral stationary phase provided baseline resolution of the enantiomers the stability of the method was suspect to small changes in the pH (0.2 units). In contrast the CZE method developed for both cyclodextrins and their derivatives gave good resolution of the enantiomers and method stability (R.S.D. < 1%, n = 10 on precision). The basis of the interaction mechanism between selector and selectand was shown as a 1:2 relationship of cyclodextrin to analyte by NMR. In addition the polysaccharide, heparin was investigated as a chiral additive and excellent resolution of the oxamniquine was achieved with 3 mM heparin in 50 mM sodium dihydrogenphosphate (pH 3.0) as buffer in CZE, which also gave a stable procedure. This method allowed the detection of each of the enantiomers in the presence of the other down to 0.23% (m/m). The overall composition of the heparin material from different sources can however be slightly variable and this can result in small differences in resolution capability.

Capillary Action↗

PubMed1985

The structure of the adenovirus capsid. I. An envelope model of hexon at 6 A resolution.

The structure of hexon, the major coat protein of adenovirus, has been determined by X-ray crystallography. Electron density maps were obtained with phases based on five heavy-atom derivatives at 2.9 A resolution. The main experimental finding derives from a low resolution electron density map calculated at 6.0 A resolution, but based on phases determined by the multiple isomorphous replacement method at 2.9 A resolution. Hexon consists of three subunits together forming two major components of different morphological symmetry. A triangular top with three towers of density is superimposed on a more bulky pseudo-hexagonal base. The symmetry of the top is in accord with the trimeric nature of hexon, but that of the base derives from the molecular function, which is to provide a densely packed impenetrable protective outer layer for the virion. A close-packed array of hexons forms a planar facet of the icosahedral capsid, with the tops presenting a spiky appearance that is consistent with electron micrographs of the adenovirus capsid. Hexon is hollow, permitting it to occupy a larger volume than normal for the same quantity of protein. The polypeptide chain has been traced in the 2.9 A electron density map for several non-contiguous stretches, allowing C alpha co-ordinates to be measured for 820 out of the 967 amino acid residues. The overall folding pattern confirms the assignment of shape, but the lack of connectivity so far precludes its complete description. The modest amount of alpha-helix and beta-sheet present is in accord with spectroscopic results.

Adenoviridae↗

PubMed1991

High-resolution electron crystallography of light-harvesting chlorophyll a/b-protein complex in three different media.

Large two-dimensional crystals of the light-harvesting chlorophyll a/b-protein complex (LHC-II) from the photosynthetic membrane of pea chloroplasts were grown by a new method from detergent solution. The structure of these crystals was examined by electron crystallography, using three different media to preserve high-resolution detail: vitrified water, glucose and tannin. The crystals diffracted electrons to at least 3.2 A resolution in all three media. R-factors between the three data sets of electron diffraction amplitudes ranged from 6.4% to 14.3%. Fourier difference maps were generated and compared to a projection map of the complex at 3.4 A resolution. No significant differences were found, proving that all three media preserved the native structure of LHC-II at high resolution. The probability of recording high-quality electron diffraction patterns with tannin was 90%. With glucose and water this probability was lower by a factor of 10 to 20, suggesting that tannin may be preferable as a preserving medium for sensitive biological specimens.

Chlorophyll↗

PubMed1994

Resolution of splenic injury after nonoperative management.

Numerous studies have demonstrated success with nonoperative management of splenic injuries in pediatric patients. However, the resolution of the splenic injury has not been previously evaluated. The records of 50 pediatric patients with splenic injuries from blunt trauma treated nonoperatively between 1984 to 1992 were reviewed retrospectively. Abdominal computed tomography (CT) was performed at the time of injury and 6 weeks postinjury in 25 patients. These scans were reviewed and categorized by a modification of a previously reported grading system for parenchymal injury. All patients had healing of the splenic injuries, with complete resolution of the healing process observed at 6 weeks postinjury in 44%. Even those with shattered spleens (n = 6) had consistent improvement in splenic architecture, with resolution of fractures and/or contusions and return of splenic perfusion. Ten (77%) of 13 grade 1 and 2 injuries were completely resolved by the 6-week follow-up examination, whereas only one (8%) of 12 grade 3 to 5 injuries showed radiological resolution of splenic injuries. None of the 25 follow-up CT scans affected clinical decision-making or led to a deviation from the established protocol, which included a 3-month period of reduced activity. All 50 patients did well, without evidence of morbidity, mortality, or complications after return to full activity 3 months postinjury.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

PubMed1991

Effect of x-ray tube motion on dental radiographic resolution.

X-ray tube movement during intraoral radiography is generally discouraged; however, the effects have not been quantified or reported. A portable x-ray tube head was mounted on the horizontal cross-beam of Instron model 1125 universal testing machine. Precise movement from 0 to 25 mm of oscillation was accomplished in increments of 1 mm. Four test objects were imaged during x-, y-, and z-axis movement of the tube head. Pinhole camera and star test pattern images were used to measure focal spot size for each increment of motion. Images of a resolution plate and a section of dry mandible were used to evaluate levels of resolution objectively and subjectively for each increment of motion. Results indicate that the size of the effective focal spot increases predictably with increasing increments of tube motion accompanied by a corresponding loss of resolution. Movement of the tube along the x- and y-axis relative to the plane of the film produced similar losses of resolution and were much greater than loss associated with z-axis movement.

Motion↗