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A robust morphological algorithm for automatic radiation field extraction and correlation of portal images.

Accurate extraction of the radiation field boundary from portal images is required in selective histogram equalization techniques for contrast enhancement of portal images, and and is required in portal-simulator image correlation. Local edge detectors are so noise sensitive that delineation of the field from anatomy edges and noise is very difficult. Optimization in edge localization and noise suppression can be achieved with optimal edge detectors, but the required calculation is very time consuming. A portal-image segmentation algorithm based on morphological edge detection and morphological segmentation techniques is presented. Relying on two predefined criteria that are nonsensitive to variation of portal-image types, the algorithm can automatically search the optimal threshold value, which is sensitive to the variation of the type and quality of portal images. With two stages of the searching procedure, the algorithm can accommodate a large variation of single and double exposure portal images obtained from different therapy machines. Results of our morphological edge detector are also compared to that of an optimal edge detector. Portal-simulator image correlation using the inertia moments of the radiation field mask is investigated, and compared to correlation obtained using inertia moments of the radiation field contour. It is shown that the mask method is less sensitive to small variations and distortions in the field shape, resulting in a more accurate correlation. This would substantially simplify the task of treatment verifications.

Algorithms↗

High-order cumulant-based blind deconvolution of Raman spectra.

The spectra recorded by a dispersion spectrophotometer are usually distorted by the response function of the instrument. To improve the resolving power, double or triple cascade spectrophotometers with narrow slits have been employed, but the total flux of the radiation available decreases accordingly, resulting in a low signal-to-noise ratio and a longer measuring time. The actual spectra can be restored approximately by mathematically removing the effects of the measuring instruments. Based on the Shalvi-Weinstein criterion, a (6, 2)-order normalized cumulant-based blind deconvolution algorithm for Raman spectral data is proposed. The actual spectral data and the unit-impulse response of the measuring instruments can be estimated simultaneously. By conducting experiments on real Raman spectra of some organic compounds, it is shown that this algorithm has a robust performance and fast convergence behavior and can improve the resolving power and correct the relative intensity distortion considerably.

Algorithms↗

An economic evaluation of the Irbesartan in Diabetic Nephropathy Trial (IDNT) in a UK setting.

There are substantial healthcare costs associated with the provision of renal replacement therapy. Patients with diabetes mellitus are the largest and fastest growing group developing end-stage renal disease (ESRD) in the United Kingdom (UK). Treatment leading to a slowing of progression to ESRD in diabetic patients could lead to considerable cost savings. Using treatment-specific probabilities derived from the Irbesartan in Diabetic Nephropathy Trial (IDNT), the cost effectiveness of treating patients with hypertension, type II diabetes and nephropathy with irbesartan, amlodipine or control was calculated using a Markov model. UK-specific ESRD-related data were retrieved from published sources to reflect local management practices, ESRD outcomes and costs. Mean 10-year costs and changes in life expectancy due to ESRD delayed or avoided were calculated. Future costs and clinical benefits were discounted at 6.0 and 1.5% per annum and extensive sensitivity analyses were performed. Delay in the onset of ESRD with irbesartan led to cost savings of pound sterling 5125 and pound sterling 2919/patient and improvements in projected discounted life expectancy of 0.07 and 0.21 years over 10 years vs amlodipine and control, respectively. The costs of treatment of ESRD were the main contributor to the total costs. The cost of trial medications had only a minor impact. These results were robust in a wide range of plausible assumptions. Given that the IDNT efficacy results could be translated to a UK setting, treating patients with hypertension, type II diabetes and overt nephropathy with irbesartan was cost saving over a 10-year period compared to amlodipine and control.

Amlodipine↗

Aberrant neuronal-glial differentiation in Taylor-type focal cortical dysplasia (type IIA/B).

Focal cortical dysplasia (FCD) type IIA/B (Taylor type) is a malformation of cortical development characterized by laminar disorganization and dysplastic neurons. FCD IIA and FCD IIB denote subtypes in which balloon cells are absent or present, respectively. The etiology of FCD IIA/B is unknown, but previous studies suggest that its pathogenesis may involve aberrant, mixed neuronal-glial differentiation. To investigate whether aberrant differentiation is a consistent phenotype in FCD IIA/B, we studied a panel of neuronal and glial marker antigens in a series of 15 FCD IIB cases, and 2 FCD IIA cases. Double-labeling immunofluorescence and confocal imaging revealed that different combinations of neuronal and glial antigens were co-expressed by individual cells in all cases of FCD IIA/B, but not in control cases of epilepsy due to other causes. Co-expression of neuronal and glial markers was most common in balloon cells, but was also observed in dysplastic neurons. The relative expression of neuronal and glial antigens varied over a broad range. Microtubule-associated protein 1B, an immature neuronal marker, was more frequently co-expressed with glial antigens than were mature neuronal markers, such as neuronal nuclear antigen. Our results indicate that aberrant neuronal-glial differentiation is a consistent and robust phenotype in FCD IIA/B, and support the hypothesis that developmental defects of neuronal and glial fate specification play an important role in its pathogenesis.

Adolescent↗

Population pharmacokinetics of allopurinol in full-term neonates with perinatal asphyxia.

In newborn infants, allopurinol is being tested as a free radical scavenger to prevent brain damage caused by reperfusion and oxygenation after perinatal hypoxia and ischemia (birth asphyxia). To develop rational dosing schemes for future studies, knowledge of the pharmacokinetics in this patient group is essential. In the present study, a population pharmacokinetic model was designed and validated for allopurinol in this specific patient group. One-compartment and 2-compartment models were fitted to plasma concentration time data of 24 newborns entered in 2 clinical trials using nonlinear mixed effects modeling. A bootstrap procedure was performed to check the robustness of the model. The data were best described using a 1-compartment model with linear elimination. Estimated pharmacokinetic parameters were volume of the central compartment (V, 0.79 L/kg) and total body clearance (CL, 0.078 L/h/kg), with 42% and 60% interindividual variability, respectively. The median values for these parameters of 1000 bootstrap replicates were very similar (95% confidence intervals were 0.67 to 0.96 and 0.054 to 0.10 for V and CL, respectively), indicating the robustness of the model. A population pharmacokinetic model has been designed and validated which adequately describes the data of 2 clinical studies in critically ill newborn infants. The model will be used to design dosing strategies for future evaluation of the benefits of allopurinol in these patients.

Allopurinol↗

Reactive astrocytes in neonate brain upregulate intermediate filament gene expression in response to axonal injury.

We have examined the injury response of astrocytes in the immature hamster brain in this study, focusing on alterations in the expression of glial fibrillary acidic protein (GFAP) and vimentin. In the adult CNS these two type III intermediate filament (IF) proteins have been shown to undergo robust increases in expression in response to axonal injury. Since injury to the immature CNS reportedly elicits less glial scar formation than adult brain injury, we examined the possibility that immature astrocytes respond differently than adult astrocytes to CNS injury with respect to IF gene expression. In situ hybridization using a 35S-labeled cDNA GFAP probe was done on brainstem sections obtained 2, 7 and 14 days after unilateral transection of the corticospinal tract in P8 hamster pups. The results indicated that substantial increases in GFAP mRNA were associated with the degenerating portion of the corticospinal tract by 2 days after axotomy and that the levels remained elevated for at least 14 days. Double-label immunofluorescence studies of this material suggested that GFAP as well as vimentin protein levels were also increased in many astrocytes in and around the degenerating corticospinal tract 2-14 days after axotomy. Most of the reactive astrocytes in the degenerating regions exhibited increases in GFAP and vimentin immunostaining but some vimentin-negative GFAP-positive reactive astrocytes were also observed, particularly in regions surrounding the actual degenerative zones.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Hydroxyethylcellulose as an effective polymer network for DNA analysis in uncoated glass microchips: optimization and application to mutation detection via heteroduplex analysis.

The nature of the sieving matrix for DNA fragment separation is of immense importance in capillary and microchip electrophoresis. The chemical nature of the surface of the capillary or microchannel wall is equally as important, particularly with DNA electrophoresis where a substantial electroosmotic flow (EOF) may be detrimental to the separation. Although DNA analysis has been carried out successfully in both coated and uncoated capillaries, analysis of unpurified polymerase chain reaction products has been carried out primarily with covalently coated surfaces, especially with microchip electrophoresis. In this report, double-stranded (ds) DNA fragment analysis using hydroxyethylcellulose (HEC) buffered in 1xTris-borate-EDTA is demonstrated both in uncoated capillaries and in microchips. EOF was suppressed 20% in the presence of 1.5% HEC, and the effectiveness of HEC as a polymer for dsDNA fragment analysis was dependent on the pH, with pH 8.6 being optimal. Using separation efficiency (number of theoretical plates) and resolution to gauge the effectiveness of a variety of polymers for the capillary separation of dsDNA fragments in the size range 60-587bp, HEC was found to be comparable in performance to polydimethylacrylamide (PDMA), and superior to linear polyacrylamide and polyethylene oxide for DNA analysis. With respect to longevity and robust performance, HEC could be used effectively in an uncoated capillary for more than 40 runs and for more than 90 runs (without replenishing the polymer) in an uncoated microchip. Application of the optimized HEC conditions is demonstrated through its ability to facilitate heteroduplex analysis.

Buffers↗

Gradual development of the interferon-gamma response of swine to porcine reproductive and respiratory syndrome virus infection or vaccination.

Infection of swine with virulent porcine reproductive and respiratory syndrome (PRRS) virus induced a rapid, robust antibody response that comprised predominantly nonneutralizing antibodies and waned after approximately 3 months. In contrast, the initial onset of virus-specific interferon (IFN)-gamma-secreting cells (SC) in the pig lymphocyte population remained at a fairly low level during this period and then increased gradually in frequency, plateauing at 6 months postinfection. A similar polarization of the host humoral and cellular immune responses was also observed in pigs immunized with a PRRS-modified live virus (MLV) vaccine. Even coadministration of an adjuvant that enhanced the immune response to a pseudorabies (PR) MLV vaccine failed to alter the induction of PRRS virus-specific IFN-gamma SC (comprising predominantly CD4/CD8 alpha double positive memory T cells with a minority being typical CD4(-)/CD8 alpha beta(+) T cells) and the generation of neutralizing antibodies. Moreover, unlike inactivated PR virus, nonviable PRRS virus did not elicit virus-neutralizing antibody production. Presumably, an intrinsic property of this pathogen delays the development of the host IFN-gamma response and preferentially stimulates the synthesis of antibodies incapable of neutralization.

Animals↗

Evaluation of a 3D reconstruction algorithm for multi-slice PET scanners.

A fully 3D reconstruction algorithm based on filtered backprojection was evaluated for the reconstruction of data obtained with multi-slice positron emission tomography (PET) scanners which have had the septa removed. This algorithm uses forward-projection through the reconstructed images of a 2D subset of the data to complete the 3D dataset thus satisfying the condition of shift invariance. This is followed by 3D filtered backprojection. Axial sampling was doubled by combining adjacent polar angles, thus improving reconstructed axial resolution. The algorithm was tested using real and simulated datasets and gave high quality reconstructions without artifacts over a wide range of imaging conditions. Events are placed accurately throughout the imaging volume as determined by measurements with a MRI/PET registration phantom. The forward-projection step leads to degradation in image resolution due to insufficient axial and transaxial sampling. This effect is amplified if multiple iterations of the algorithm are used, with little decrease in image noise. Changing the filter employed in the initial 2D reconstruction can be used to alter the noise and resolution characteristics of the 3D images. This algorithm has proved very robust at reconstructing 3D PET data and is relatively fast. Those small problems which exist can be attributed to detector sampling problems, especially in the axial direction, which is a consequence of the geometry of these scanners, which are designed primarily for 2D data acquisition.

Algorithms↗

Evaluating alleged drinking after driving--the hip-flask defence. Part 1. Double blood samples and urine-to-blood alcohol relationship.

This two-part article examines the strengths and weaknesses of various ways of investigating claims of drinking alcohol after driving, commonly known as the hip-flask or glove-compartment defence. In many countries the onus of proof in hip-flask cases rests on the prosecution. With good co-operation from the police and timely sampling of body fluids, such as blood and urine for forensic analysis of ethanol, useful evidence can be mustered to support or challenge the truthfulness of alleged drinking after driving. The person's blood-alcohol concentration (BAC) can be compared with values expected on the basis of the amount of alcohol consumed after driving, according to theoretical Widmark calculations. The actual BAC measured is then adjusted for the additional amount of alcohol consumed in the after-drink. Double blood samples, that is, taking two specimens of venous blood about 30-60 minutes apart and looking at the magnitude and direction of change in BAC provides little or no more information than a single blood specimen. However, the relationship between alcohol in blood and urine is very useful in hip-flask cases whereby the concentration expected in the primary urine is compared with the concentration in the bladder urine voided. The concentration of alcohol determined in a second urine sample collected 30-60 min later gives supporting evidence in hip-flask cases. A graphical method, which entails plotting ethanol concentrations in blood and urine as a function of time provides a robust and practical way to investigate hip-flask defences. In the second part of the review, congener analysis is presented, which entails comparing the concentrations of n-propanol, isobutanol and occasionally other congeners in the alcoholic beverage allegedly consumed after driving with the volatiles present in the suspect's blood and urine determined by headspace gas chromatography.

Alcohol Drinking↗

Restoration of bone mass in the severely osteopenic senescent rat.

Studies in humans and rats suggest that age impairs the ability to form bone. This impairment may be due to a depletion or deficit in osteoprogenitor stem cells. Such a deficit would be expected to reduce the ability of the skeleton to respond to therapy designed to restore lost bone. This study evaluated whether severely osteopenic senescent rats are capable of responding to a potent anabolic factor in bone, prostaglandin E2 (PGE). Growing female Sprague Dawley rats were ovariectomized at 3 months and aged until the start of treatment at 23 months. Rats were treated daily with PGE (3 mg/kg sc) or vehicle for 56 days. Tibiae were harvested for bone histomorphometry and femora were obtained for mRNA analysis of bone matrix proteins. The cancellous bone area was fivefold greater in PGE-treated rats than in vehicle-treated controls and not different from age-matched ovary-intact rats. PGE approximately doubled the bone-forming surface and the mineral apposition rate and increased the bone formation rate fourfold. The increased cancellous bone area in PGE-treated rats was primarily due to an increase in osteoblasts over osteoclasts. One hundred percent of the endocortical surface and 72 +/- 9% of the periosteal surface of cortical bone was undergoing mineralization in PGE-treated rats, whereas no mineratization was evident in vehicle-treated rats. An architectural analysis of cancellous bone indicates that trabecular number and thickness were increased and separation decreased in the treated rats. Imaging by microcomputed tomography further revealed that with PGE treatment, trabeculae in the medial plane of the proximal tibial metaphysis were more robust and continuous with the endocortical surface. PGE also significantly induced message levels for the prepro-alpha (I) subunit of type I collagen (collagen), osteonectin, and osteocalcin. In summary, bone mass can be restored to severely osteopenic senescent rats, suggesting that aging does not necessarily diminish the capacity of the skeleton to form bone.

Aging↗

Relationship of glia to GnRH axonal outgrowth from third ventricular grafts in hpg hosts.

The homozygous mutant hypogonadal (hpg) mouse lacks a functional gene for the neuropeptide gonadotropin releasing hormone (GnRH). The consequence of this defect is an infantile reproductive tract in adulthood. This condition can be reversed by the implantation of normal fetal preoptic area tissue that contains GnRH neurons. Reversal is always preceded by the outgrowth of GnRH axons into the host target tissue, the median eminence, by a stereotyped pathway. In the current experiments we investigated the cellular nature of the path taken by early emerging GnRH axons focusing on their relationship with astrocytic components and with the specialized ependymal population of this area, the tanycytes. In control tissue glial fibrillary acid protein (GFAP) immunoreactivity was confined to the exterior of cerebral blood vessels and glial limitans. Both GFAP and vimentin, another intermediate filament protein, marked the specialized ependymal cells of this region, the tanycytes. There was a robust reactive astrocytic response to the injury of transplantation in both the donor and host tissue within 5 days of implantation and the reactive astrocytes persisted for 60 days. These cells were GFAP-positive and were present in many areas of the host along the cannula tract and not confined to the area of GnRH axonal outgrowth. Vimentin, another intermediate filament, marked only the specialized ependymal cells of this region, the tanycytes, in both control and grafted tissue. Despite the profound reactive gliosis, GnRH axons were shown to exit the implant as early as 5 days after grafting suggesting that the gliotic process did not constitute a barrier to this phenomenon. At the light microscopic level, double label immunocytochemical studies did not reveal any specific association between GFAP or vimentin-positive cellular processes and these pioneer GnRH fibers. However, since normal GnRH axons had been reported to travel in tanycytic channels through the medial basal hypothalamus we reinvestigated the pattern of early emerging GnRH axons at the ultrastructural level. With this higher resolution, GnRH axons were found adjacent to glial elements along their entire traverse from the graft-host interface, through the host basal hypothalamus to their termination on the hypophysial portal capillaries. At the interface, GnRH-positive axons appeared to exit via glial channels similar to those described in other developing and regenerating systems. In the host, GnRH immunoreactive axonal profiles were surrounded by glial processes though the latter could not be further defined as tanycytic or astroglial. Other, immunonegative, axons were frequently seen in axonal bundles or fascicles and not necessarily in contact with glia.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Intravenous levosimendan treatment is cost-effective compared with dobutamine in severe low-output heart failure: an analysis based on the international LIDO trial.

BACKGROUND: Levosimendan, a novel calcium sensitiser, improves cardiac performance and symptoms without increasing oxygen consumption, and decreases the mortality of patients with low-output heart failure. AIMS: To estimate the cost-effectiveness of intravenous treatment with levosimendan compared with dobutamine in patients with severe low-output heart failure. METHODS: This economic evaluation was based on a European clinical trial (LIDO), in which 203 patients with severe heart failure randomly received a 24 h infusion with either levosimendan or dobutamine. Survival and resource utilisation data were collected for 6 months; survival was extrapolated assuming a mean additional lifetime of 3 years based on data from the Cooperative North Scandinavian Enalapril Survival Study trial. Costs were based on study drug usage and hospitalisation in the 6-month follow-up. A sensitivity analysis on dosage of drug and duration of survival was performed. RESULTS: The mean survival over 6 months was 157+/-52 days in the levosimendan group and 139+/-64 days in the dobutamine group (P<0.01). When extrapolated up to 3 years, the gain in life expectancy was estimated at 0.35 years (discounted at 3%). Levosimendan increased the mean cost per patient by 1108, which was entirely due to the cost of the study drug. The incremental cost per life-year saved (LYS) was 3205 at the European level; in the individual countries the cost per LYS ranged between 3091 and 3331. The result was robust in the sensitivity analysis. CONCLUSIONS: Although the patients in the levosimendan group were alive for more days and thus at risk of hospitalisation for longer, there was no increase in hospitalisation or hospitalisation costs with levosimendan treatment. The cost per LYS using levosimendan compares favourably with other cost-effectiveness analyses in cardiology.

Aged↗

Coded pulse excitation for ultrasonic strain imaging.

Decorrelation strain noise can be significantly reduced in low echo-signal-to-noise (eSNR) conditions using coded excitation. Large time-bandwidth-product (>30) pulses are transmitted into tissue mimicking phantoms with 2.5-mm diameter inclusions that mimic the elastic properties of breast lesions. We observed a 5-10 dB improvement in eSNR that led to a doubling of the depth of focus for strain images with no reduction of spatial resolution. In high eSNR conditions, coded excitation permits the use of higher carrier frequencies and shorter correlation windows to improve the attainable spatial resolution for strain relative to that obtained with conventional short pulses. This paper summarizes comparative studies of strain imaging in noise-limited conditions obtained by short pulses and four common aperiodic codes (chirp, Barker, suboptimal, and Golay) as a function of attenuation, eSNR and applied strain. Imaging performance is quantified using SNR for displacement (SNRd), local modulation transfer function (LMTF), and contrast-to-noise ratio for strain (CNRepsilon). We found that chirp and Golay codes are the most robust for imaging soft tissue deformation using matched filter decoding. Their superior performance is obtained by balancing the need for low-range lobes, large eSNR improvement, and short-code duration.

Animals↗

Low TRBP levels support an innate human immunodeficiency virus type 1 resistance in astrocytes by enhancing the PKR antiviral response.

Acute human immunodeficiency virus type 1 (HIV-1) replication in astrocytes produces minimal new virus particles due, in part, to inefficient translation of viral structural proteins despite high levels of cytoplasmic viral mRNA. We found that a highly reactive double-stranded (ds) RNA-binding protein kinase (PKR) response in astrocytes underlies this inefficient translation of HIV-1 mRNA. The dsRNA elements made during acute replication of HIV-1 in astrocytes triggers PKR activation and the specific inhibition of HIV-1 protein translation. The heightened PKR response results from relatively low levels of the cellular antagonist of PKR, the TAR RNA binding protein (TRBP). Efficient HIV-1 production was restored in astrocytes by inhibiting the innate PKR response to HIV-1 dsRNA with dominant negative PKR mutants, or PKR knockdown by siRNA gene silencing. Increasing the expression of TRBP in astrocytes restored acute virus production to levels comparable to those observed in permissive cells. Therefore, the robust innate PKR antiviral response in astrocytes results from relatively low levels of TRBP expression and contributes to their restricted infection. Our findings highlight TRBP as a novel cellular target for therapeutic interventions to block productive HIV-1 replication in cells that are fully permissive for HIV-1 infection.

Astrocytes↗

Information hiding technique with double phase encoding.

We propose a technique for information hiding using double phase encoding. The proposed method uses a weighted double phase-encoded hidden image added to a host image referred to as the transmitted image. We develop an analytical presentation for the system performance using the statistical properties of double phase encoding. The peak signal-to-noise-ratio metric is used as a measure for the degradation in the quality of the host image and the recovered hidden image. We test, analytically, the distortion of the hidden image that is due to the host image and the effect of occlusion of the pixels of the transmitted image (that is, the host image containing the hidden image). Moreover, we discuss the effect of using only the real part of the transmitted image to recover the hidden image. Computer simulations are presented to test the system performance against these types of distortion. The simulations illustrate the system ability to recover the hidden image under distortions and the robustness of the hidden image against removal trials.

Journal Article↗

Valproate robustly enhances AP-1 mediated gene expression.

Valproic acid (VPA) is a potent broad spectrum anticonvulsant with demonstrated efficacy in the treatment of Bipolar Affective Disorder, but the biochemical basis for VPA's antimanic or mood-stabilizing actions have not been fully elucidated. It has been demonstrated that VPA, at therapeutically relevant concentrations, increases AP-1 DNA binding activity in cultured cells in vitro. These findings raise the possibility that VPA may produce its mood-stabilizing effects by regulating the expression of subsets of genes via its effects on the AP-1 family of transcription factors. To determine if VPA does, in fact, enhance AP-1 mediated gene expression, the effects of VPA on the expression of a luciferase reporter gene were studied in transiently transfected rat C6 glioma and human SH-SY5Y neuroblastoma cells using the pGL2-control vector. The luciferase gene in the vector is driven by an SV40 promoter which contains well characterized AP-1 sites. VPA produced a greater than doubling of luciferase activity in a time- and concentration-dependent manner in both cell lines. Furthermore, mutations of the AP-1 sites in the SV40 promoter markedly attenuated the VPA-induced increases in luciferase activity. These effects of VPA on AP-1 mediated gene expression are very similar to the effects observed with lithium, and suggest that the temporal regulation of AP-1 mediated gene expression in critical neuronal circuits may play a role in the long-term therapeutic efficacy of these agents.

Animals↗

Inducible systemic RNA silencing in Caenorhabditis elegans.

Introduction of double-stranded RNA (dsRNA) can elicit a gene-specific RNA interference response in a variety of organisms and cell types. In many cases, this response has a systemic character in that silencing of gene expression is observed in cells distal from the site of dsRNA delivery. The molecular mechanisms underlying the mobile nature of RNA silencing are unknown. For example, although cellular entry of dsRNA is possible, cellular exit of dsRNA from normal animal cells has not been directly observed. We provide evidence that transgenic strains of Caenorhabditis elegans transcribing dsRNA from a tissue-specific promoter do not exhibit comprehensive systemic RNA interference phenotypes. In these same animals, modifications of environmental conditions can result in more robust systemic RNA silencing. Additionally, we find that genetic mutations can influence the systemic character of RNA silencing in C. elegans and can separate mechanisms underlying systemic RNA silencing into tissue-specific components. These data suggest that trafficking of RNA silencing signals in C. elegans is regulated by specific physiological and genetic factors.

Animals↗