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Background Single nucleotide polymorphisms (SNPs) are the foundation of powerful complex trait and pharmacogenomic analyses. The availability of large SNP databases, however, has emphasized a need for inexpensive SNP genotyping methods of commensurate simplicity, robustness, and scalability. We describe a solution-based, microtiter plate method for SNP genotyping of human genomic DNA. The method is based upon allele discrimination by ligation of open circle probes followed by rolling circle amplification of the signal using fluorescent primers. Only the probe with a 3' base complementary to the SNP is circularized by ligation. Results SNP scoring by ligation was optimized to a 100,000 fold discrimination against probe mismatched to the SNP. The assay was used to genotype 10 SNPs from a set of 192 genomic DNA samples in a high-throughput format. Assay directly from genomic DNA eliminates the need to preamplify the target as done for many other genotyping methods. The sensitivity of the assay was demonstrated by genotyping from 1 ng of genomic DNA. We demonstrate that the assay can detect a single molecule of the circularized probe. Conclusions Compatibility with homogeneous formats and the ability to assay small amounts of genomic DNA meets the exacting requirements of automated, high-throughput SNP scoring.

In September 1999, the Cryogenic Dark Matter Search (CDMS) completed a nine month run with the first generation detectors (0.5 kg of germanium BLIPs sensing thermal phonons and ionization). The data acquired in this run significantly decrease the upper limits on WIMP interaction in germanium by a factor 2.5 to 10 depending on the mass region and begin to probe supersymmetry. The results are however strongly inconsistent with the claims of the DAMA experiment.

The Cryogenic Dark Matter Search (CDMS) employs Ge and Si detectors to search for weakly interacting massive particles (WIMPs) via their elastic-scattering interactions with nuclei while discriminating against interactions of background particles. CDMS data, accounting for the neutron background, give limits on the spin-independent WIMP-nucleon elastic-scattering cross section that exclude unexplored parameter space above 10 GeV c-2 WIMP mass and. at > 75% CL. the entire 3σ allowed region for the WIMP signal reported by the DAMA experiment.

PCOS is a common disorder of unknown etiology. Studies of first-degree relatives of women diagnosed with PCOS suggest familial clustering of the disease. A prospective study of first-degree female relatives of women with PCOS conducted by NCPIR found that 46% of ascertainable sisters of women with PCOS were hyperandrogenemic. NCPIR has conducted linkage and association studies using affected sibling-pair analysis and the transmission/disequilibrium test to explore candidate PCOS genes. These studies point a finger at a region 1 MB centromeric to the insulin receptor gene on chromosome 19.

Understanding how viral components collaborate to convert the human immunodeficiency virus type 1 genome from single-stranded RNA into double-stranded DNA is critical to the understanding of viral replication. Not only must the correct reactions be carried out, but unwanted side reactions must be avoided. After minus-strand strong stop DNA (−sssDNA) synthesis, degradation of the RNA template by the RNase H domain of reverse transcriptase (RT) produces single-stranded DNA that has the potential to self-prime at the imperfectly base-paired TAR hairpin, making continued DNA synthesis impossible. Although nucleocapsid protein (NC) interferes with −sssDNA self-priming in reverse transcription reactions in vitro, NC alone did not prevent self-priming of a synthetic −sssDNA oligomer. NC did not influence DNA bending and therefore cannot inhibit self-priming at hairpins by directly blocking hairpin formation. Using DNA oligomers as a model for genomic RNA fragments, we found that a 17-base DNA fragment annealed to the 3′ end of the −sssDNA prevented self-priming in the presence of NC. This implies that to avoid self-priming, an RNA-DNA hybrid that is more thermodynamically stable than the hairpin must remain within the hairpin region. This suggests that NC prevents self-priming by generating or stabilizing a thermodynamically favored RNA-DNA heteroduplex instead of the kinetically favored TAR hairpin. In support of this idea, sequence changes that increased base pairing in the DNA TAR hairpin resulted in an increase in self-priming in vitro. We present a model describing the role of NC-dependent inhibition of self-priming in first-strand transfer.

The incidence of malignant melanoma is increasing. Because of increased awareness, early recognition of malignant melanoma has become more common. In 1997, a new staging system for cutaneous melanoma was proposed, with reclassification of thin melanoma1 mm, with and without ulceration. This report evaluates the pathologic and clinical features of thin melanomas influencing recurrence and survival from a tertiary cancer center in an attempt to correlate findings with the proposed staging system. A review of the Roswell Park Cancer Institute tumor registry identified 352 patients with thin cutaneous melanomas (1.0 mm) seen during an 18-year period ending August 30, 1998. Overall survival was 93 and 87 per cent at 5 and 10 years, respectively. Disease-free survival was 94 and 93 per cent at 5 and 10 years, respectively. Local recurrence occurred in 3 per cent of patients, regional recurrence in 3 per cent, and metastatic disease in 3 per cent, for an overall recurrence of 7 per cent, with a median follow-up of 118 months. Only the presence of ulceration was a significant prognostic factor for recurrence by both univariate and multivariate analysis. Failure rates (any recurrence) by Clark levels I, II, and III/IV were 3, 5, and 10 per cent, respectively (P = 0.14). Failure rates by tumor thickness (mm), for 0.0-0.24, 0.25-0.49, 0.50-0.74, and 0.75-0.99 were 3, 4, 7, and 10 per cent, respectively (P = 0.49). Ten-year disease-free survival for ulceration versus no ulceration was 40 and 94 per cent, respectively (P0.0001). We conclude that thin cutaneous melanoma carries an excellent prognosis with appropriate treatment. Our findings support inclusion of ulceration in a new staging system. Lesions 0.76 to 0.99 mm and Clark level III and IV may warrant close observation as a separate subgroup.

These investigations were undertaken to compare and contrast the roles of phosducin and phosducin-like protein in the retina.Phosducin and phosducin-like protein were compared in an in vitro assay measuring their inhibition of transducin binding to light-activated rhodopsin. The two proteins were localized within the retina by immunoblot analyses and immunocytochemistry using affinity-purified antibodies with high specificity for each of the two homologs. The sensitivity of phosducin-like protein to phosphorylation was probed using in vitro protein kinase reactions.Phosducin and phosducin-like protein were found to have similar, though not identical, transducin inhibiting activity in vitro. These two proteins were found to be localized dissimilarly within the retina, with spatial overlap limited to the inner segments of the photoreceptors. Phosducin is found exclusively in photoreceptor cells, including the synaptic and nuclear layers, while phosducin-like protein is found throughout the inner retinal layers, most abundantly in the bipolar cells of the inner nuclear layer. Phosducin-like protein is not efficiently phosphorylated by the protein kinases tested, indicating that its regulation differs from that of phosducin.It appears that phosducin and phosducin-like protein play distinct roles in the retina. While phosducin is likely to be important in feedback regulation of the visual signal, such as in light adaptation, phosducin-like protein probably has little if any function in the phototransduction cascade. Phosducin-like protein may have a role in regulating the processing of visual signals by the neural cells of the inner retina.

The Cryogenic Dark Matter Search (CDMS) employs low-temperature Ge and Si detectors to search for Weakly Interacting Massive Particles (WIMPs) via their elastic-scattering interactions with nuclei while discriminating against interactions of background particles. For recoil energies above 10 keV, events due to background photons are rejected with >99.9% efficiency, and surface events are rejected with >95% efficiency. The estimate of the background due to neutrons is based primarily on the observation of multiple-scatter events that should all be neutrons. Data selection is determined primarily by examining calibration data and vetoed events. Resulting efficiencies should be accurate to about 10%. Results of CDMS data from 1998 and 1999 with a relaxed fiducial-volume cut (resulting in 15.8 kg-days exposure on Ge) are consistent with an earlier analysis with a more restrictive fiducial-volume cut. Twenty-three WIMP candidate events are observed, but these events are consistent with a background from neutrons in all ways tested. Resulting limits on the spin-independent WIMP-nucleon elastic-scattering cross-section exclude unexplored parameter space for WIMPs with masses between 10-70 GeV c^{-2}. These limits border, but do not exclude, parameter space allowed by supersymmetry models and accelerator constraints. Results are compatible with some regions reported as allowed at 3-sigma by the annual-modulation measurement of the DAMA collaboration. However, under the assumptions of standard WIMP interactions and a standard halo, the results are incompatible with the DAMA most likely value at >99.9% CL, and are incompatible with the model-independent annual-modulation signal of DAMA at 99.99% CL in the asymptotic limit., 40 pages, 49 figures (4 in color), submitted to Phys. Rev. D; v.2:clarified conclusions, added content and references based on referee's and readers' comments; v.3: clarified introductory sections, added figure based on referee's comments

This chapter describes the application of the Storage-Treatment (S-T) Block of the EPA Storm Water Management Model (SWMM) to design and/or analyze extended-de…

Estrogen-inducible genes contain an enhancer called the estrogen response element (ERE), a double-stranded inverted repeat. The estrogen receptor (ER) is generally thought to bind to the double-stranded ERE. However, some reports provide evidence that an ER homodimer can bind a single strand of the ERE and suggest that single-stranded ERE binding is the preferred binding mode for ER. Since these two models describe quite different mechanisms of receptor action, we have attempted to reconcile the observations. Analyzing DNA structure by nuclease sensitivity, we found that two identical molecules of a single strand of DNA containing the ERE sequence can partially anneal in an antiparallel manner. Bimolecular annealing produces double-stranded inverted repeats, with adjacent unannealed tails. The amount of annealing correlates exactly with the ability of ER to bind bimolecular EREs. Either strand of an ERE could anneal to itself in a way that would bind ER. We conclude that ER binds only the annealed double-stranded ERE both in vitro and in vivo.

Because of its role in cell motility and growth regulation, gelsolin, an actin-binding protein, has been considered a tumor suppressor and a potential prognostic marker in some neoplasias, such as breast and bladder cancer. Little is known about its immunoexpression in prostatic adenocarcinoma (PCA).Formalin-fixed, paraffin-embedded tissues of 72 prostatectomy specimens with adenocarcinoma and 8 nonneoplastic prostates from autopsies were stained with a gelsolin monoclonal antibody using the Avidin-biotin-peroxidase complex (ABC) method after microwave antigen retrieval. Immunoreactivity was evaluated in PCA, prostatic intraepithelial neoplasia (PIN), benign prostatic hyperplasia (BPH), and nonproliferative glandular tissue and stroma. The results were statistically analyzed.Consistent gelsolin immunoreactivity was seen in prostatic stromal cells, smooth muscle, endothelia, and nerves. Variable gelsolin expression was seen in 20-100% (average (A) = 65.5%) of glandular cells in nonproliferative tissue (N = 75); 0-50% (A = 9.7%) in BPH (N = 59); 0-80% (A = 8.9%) in PIN (N = 61); and 0-90% (A = 9.3%) in PCA (N = 71). The level of gelsolin expression in nonproliferative prostatic tissue was similar between prostates with PCA (A = 63.4%) and nonneoplastic prostates (A = 67.5%). The level of gelsolin expression did not correlate with age, tumor size, Gleason score, or stage.Gelsolin is decreased in PCA, PIN, and BPH in comparison to nonproliferative tissue. The role of this downregulation in the development of PCA is not clear. The similar reduction seen in PIN and BPH suggests that this event takes place indiscriminately in hyperplasia and early tumorigenesis in the prostate, which might limit its prognostic significance in PCA.

We developed a high-pressure high-resolution probe for NMR experiments on biological systems which enabled us to study the phase behaviour of model biomembranes by 1D and 2D NMR experiments at temperatures up to 65°C and at pressures up to 3000 bar.

There are technical difficulties in resecting soft tissue sarcomas extending to or crossing a joint. The objective of this study was to determine the rate of amputation and local recurrence rate for these sarcomas and compare them with the respective rates for overall extremity sarcomas.Retrospective review of 78 patients with sarcoma near a joint compared with 215 patients with extremity sarcomas accrued during the same period, 1977-1994. Of these 78 patients, 64 were in the lower and 14 in the upper extremity. Most common histologic subtypes were malignant fibrous histiocytoma (15/78, 19%), synovial sarcoma (11/78, 14%), liposarcoma (11/78, 14%), and leiomyosarcoma (10/78, 13%). The surgical treatment consisted of local excision in 10 (13%), wide excision in 56 (72%), and amputation in 12 (15%). Adjuvant radiation was given to 26 patients.Local recurrence was noted in 20% (16/78) patients. The incidence of local recurrence in the surgery alone group (n = 52) was 15% (8/52) and in the surgery plus adjuvant radiation group (n = 26) it was 31% (8/26); P = 0.11. Of the 16 patients with local recurrence, 9 (56%) required amputation. The 5-year and 10-year survival rates for the entire group of patients were 68% and 60% respectively. On multivariate analysis survival varied according to grade (P = 0.05) and tumor size (P = 0.02).Amputation was finally required in 27% (21/78) for local control of the disease. The local recurrence rate was 20%. These rates appear to be somewhat higher than those reported in our overall extremity sarcoma series and those in most modern series of overall extremity sarcomas, but the 5- and 10-year survival rates are similar to those of the latter.

Two infants with chromosome 22q11 deletion syndrome were noted to have symmetrically enlarged Sylvian fissures on cranial MRI. We compared the size of the Sylvian fissures in neuroimaging studies from 17 other subjects with del 22q11 to age-matched disease controls. The mean anterior interopercular distance was used as an index of Sylvian fissure enlargement. Symmetric enlargement of the Sylvian fissures was present in 10 of 17 subjects with del 22q11. The age-incidence pattern, as well as follow-up scans in 2 patients, suggests delayed growth of the opercular region in these patients. Subjects with del 22q11 consistently had disproportionate enlargement of the left Sylvian fissure compared to the right. This observation suggests that a gene (or genes) in the deleted region affects the development of the left and right perisylvian cortex in different ways. Abnormal development of the operculum may explain some of the neurodevelopmental features that are common among individuals with 22q11 deletion syndrome.

Patients with squamous cell carcinoma (SCC) of the skin may exhibit locally advanced or metastatic disease and present a challenging management problem.A retrospective review of 40 patients with advanced SCC of the trunk or extremity managed at Roswell Park Cancer Institute from 1982 through 1992 was performed to identify clinical and pathologic factors that influenced outcome.There were 27 males and 13 females with a median age of 61 years. Median follow-up was 24 months. Surgical resection to control the primary tumor was often extensive. Amputation was required in nine patients, hemipelvectomy in three patients, and hemicorporectomy in one patient. Median survival was 28 months, and 5-year survival was 43%. Univariate analysis identified stage (P = 0.04), size (P = 0.0001), type of surgical procedure (P = 0.009), and margins of resection (P = 0.005) as having prognostic significance. On multivariate analysis, stage (P = 0.04) and size (P = 0.02) were found to be significant.Optimum treatment for advanced SCC of the trunk and extremity involves surgical resection with uninvolved margins. The role of elective node dissection remains undefined. Investigation is needed to define the role of neoadjuvant therapy that may improve functional and cosmetic results.

This paper focuses on performance variations associated with lamp geometry and distribution in portable table luminaires. If correctly retrofit with compact fluorescent lamps (CFLs), these high use fixtures produce significant energy savings, but if misused, these products could instead generate consumer dissatisfaction with CFLs. It is the authors assertion that the lumen distribution of the light source within the luminaires plays a critical role in total light output, fixture efficiency and efficacy, and, perhaps most importantly, perceived brightness. The authors studied nearly 30 different integral (screw-based) CFLs available on the market today in search of a lamp, or group of lamps, which work best in portable table luminaires. The findings conclusively indicate that horizontally oriented CFLs outperform all other types of CFLs in nearly every aspect.

To (a) introduce arterial tonometry, which may offer a greater opportunity to detect alterations of the cardiovascular system with spinal manipulative therapy, (b) discuss the limitations of standard blood pressure measurements and (c) discuss factors that govern the contour of arterial pressure pulses.Information was obtained from peer-reviewed medical and chiropractic journal articles and medical textbooks written in English (mainly from 1980 to 1995).Arterial tonometry is a noninvasive method to continuously measure the contour of arterial pressure pulses. Pulses can be simultaneously measured from several arterial sites because vessel occlusion is not required. Arterial tonometry supplements standard blood pressure measurements that measure only the systolic and diastolic pressures by permitting the investigation of additional parameters of cardiovascular functions. Changes in large artery function can be inferred through alterations of pulse contour, wave velocity and harmonic transmission ratios. Analysis of the beat-to-beat fluctuations of arterial pressure and heart rate can be used to determine whether SMT alters the autonomic nervous system. Reductions of arterial pressure in central vessels may not be apparent from standard blood pressure measurements at peripheral vessels. The contour of a pressure pulse depends on the magnitude and timing between incident and reflected pressure waves from the upper and lower body, body shape and size, the duration and pattern of ventricular systole, and dispersion and attenuation. Therefore, arterial tonometry may improve the ability to detect alterations of the cardiovascular system in chiropractic research.

New diagnostic, modelling and plant capability on the Mega Ampère Spherical Tokamak (MAST) have delivered important results in key areas for ITER/DEMO and the upcoming MAST Upgrade, a step towards future ST devices on the path to fusion currently under procurement. Micro-stability analysis of the pedestal highlights the potential roles of micro-tearing modes and kinetic ballooning modes for the pedestal formation. Mitigation of edge localized modes (ELM) using resonant magnetic perturbation has been demonstrated for toroidal mode numbers n = 3, 4, 6 with an ELM frequency increase by up to a factor of 9, compatible with pellet fuelling. The peak heat flux of mitigated and natural ELMs follows the same linear trend with ELM energy loss and the first ELM-resolved Ti measurements in the divertor region are shown. Measurements of flow shear and turbulence dynamics during L-H transitions show filaments erupting from the plasma edge whilst the full flow shear is still present. Off-axis neutral beam injection helps to strongly reduce the redistribution of fast-ions due to fishbone modes when compared to on-axis injection. Low-k ion-scale turbulence has been measured in L-mode and compared to global gyro-kinetic simulations. A statistical analysis of principal turbulence time scales shows them to be of comparable magnitude and reasonably correlated with turbulence decorrelation time. Te inside the island of a neoclassical tearing mode allow the analysis of the island evolution without assuming specific models for the heat flux. Other results include the discrepancy of the current profile evolution during the current ramp-up with solutions of the poloidal field diffusion equation, studies of the anomalous Doppler resonance compressional Alfvén eigenmodes, disruption mitigation studies and modelling of the new divertor design for MAST Upgrade. The novel 3D electron Bernstein synthetic imaging shows promising first data sensitive to the edge current profile and flows. © 2013 IAEA, Vienna.

Cerebral blood flow plays a central role in maintaining homeostasis in the brain, and its dysfunction leads to pathological conditions such as stroke. Moreover, understanding the dynamics of blood flow is central to the interpretation of data from imaging modalities—such as intrinsic optical signaling and functional magnetic resonance imaging—that rely on changes in cerebral blood flow and oxygen level to infer changes in the underlying neural activity. Recent advances in imaging techniques have allowed detailed studies of blood flow in vivo at high spatial and temporal resolutions. We discuss techniques to accurately measure cerebral blood flow at the level of individual blood vessels using two-photon laser-scanning microscopy. By directing the scanning laser along a user-defined path, it is possible to measure red blood cell (RBC) velocity and vessel diameter across multiple vessels simultaneously. The combination of these measurements permits accurate assessment of total flux with sufficient time resolution to measure fast modulations in flux, such as those caused by heartbeat, as well as slower signals caused by vasomotion and hemodynamic responses to stimulus. Here, we discuss general techniques for animal preparation and measurement of blood flow with two-photon microscopy. We incorporate extensions to existing methods to accurately acquire flux data simultaneously across multiple vessels in a single trial. Central to these measurements is the ability to generate scan paths that smoothly connect user-defined lines of interest while maintaining high accuracy of the scan path.

The cerebral vascular system services the constant demand for energy during neuronal activity in the brain. Attempts to delineate the logic of neurovascular coupling have been greatly aided by the advent of two-photon laser scanning microscopy to image both blood flow and the activity of individual cells below the surface of the brain. Here we provide a technical guide to imaging cerebral blood flow in rodents. We describe in detail the surgical procedures required to generate cranial windows for optical access to the cortex of both rats and mice and the use of two-photon microscopy to accurately measure blood flow in individual cortical vessels concurrent with local cellular activity. We further provide examples on how these techniques can be applied to the study of local blood flow regulation and vascular pathologies such as small-scale stroke.

Quantitative DNase I footprinting assays were employed to simultaneously measure the amount of estrogen receptor (ER) bound to each site in constructs containing multiple estrogen response elements (EREs). These assays revealed identical, high affinity ER-ERE binding, K d of approximately 0.25 nM, for estradiol-liganded ER (E 2 -ER), 4-hydroxytamoxifen liganded ER (4-OHT-ER), tamoxifen aziridine liganded ER (TAz-ER), and unliganded dimeric ER, for each ERE in constructs containing up to four tandem EREs. Increasing concentrations of ER resulted in the same pattern of occupancy for each ERE, whether or not the site was located near other EREs. Similarly, the presence or absence of E 2 , 4-OHT, or TAz ligand did not change ER-ERE interaction. Since activated ER-ERE binding affinity is identical, whether ER is liganded or unliganded, ligand cannot regulate ER-ERE binding affinity. These results support the hypothesis that ligand-dependent conformational changes primarily determine how ER interacts with components of the transcription initiation complex that mediate gene transactivation. In addition, footprint assays revealed that, following ER binding, an AT-rich site adjacent to the ERE becomes hypersensitive to DNase I digestion. This sequence may be easily or intrinsically bent, assisting in recruiting ER to ERE sites.

197 Background: There is a wide variation of compliance with National Comprehensive Cancer Network (NCCN) guidelines for diagnosis and treatment. Overutilization of preoperative testing and underutilization of adjuvant treatments have been documented. The former is associated with higher costs of care; the latter with poor patient outcomes. Prior reports of adherence to NCCN Guidelines contain methodological limitations due to lack of contemporary review and incomplete listing of reasons for non-compliance. This limits the “real time” analysis of breast cancer quality and delays action plans to address quality and cost issues of care. Methods: NCCN Guideline compliance was recorded prospectively by use of electronic synoptic templates for all newly diagnosed breast cancer patients treated at a single institution between January 2010 and December 2011. A retrospective review of the synoptic templates was then conducted. Accuracy of the synoptic auditing method was assessed as well as NCCN compliance and reasons for non-compliance. Results: A total of 312 new breast cancer patients who underwent surgery as initial treatment were identified. Compliance with NCCN Guidelines for preoperative testing, breast surgery and lymph node surgery was 98% (306/312), 99.7% (311/312) and 93% (290/312) respectively. Reasons for non-compliance include patient refusal, comorbidities, advanced age and overutilization of systemic imaging. There was 100% compliance to College of American Pathologist (CAP) molecular profile documentation for all breast cancer cases. There were two discrepancies noted between the prospective template reporting and medical record review. The average time needed to populate data into synoptic templates was less than 2 minutes per patient. Conclusions: Prospective auditing of adherence to NCCN Guidelines with electronic synoptic templates is accurate and time efficient. High compliance rates with NCCN Guidelines were demonstrated. Electronic synoptic NCCN templates potentially facilitate early recognition of quality gaps in compliance and provide a framework for peer performance comparison.

Arterial stiffness in hypertension and heart failure may increase afterload on the left ventricle. Pulse wave velocity and transmission ratio measurements are noninvasive methods to assess arterial stiffness. Since noninvasive pressure pulse recording requires sufficient applied force to distort the vessel wall, we hypothesized that the pulse wave velocity, transmission ratios, and distal pulse amplitudes and contours may be altered by the recording technique. Brachial and radial arterial pressure pulses were recorded simultaneously using a piezoelectric pulse transducer in 14 young, normal, male subjects using 10 brachial artery recording forces (0.35-3.58N, approximately equal to 0.36N increments) applied in a random order and a constant radial force (2.35N). Pulses were Fourier analyzed. One subject was excluded from analysis because of improper transducer positioning over the brachial artery. In 8 subjects, no significant changes occurred in any variable over all brachial recording forces. In the remaining 5 subjects, the measured variables remained constant until brachial artery recording forces exceeded 2.42 +/- 0.03N. The pulse wave velocity (p0.04), transmission ratios (harmonics 2-5, p0.0001), radial pulse amplitude (p0.0003), and relative powers (harmonics 2-5, p0.02) then decreased. In these subjects, brachial artery depths were less than the other subjects (5.9 +/- 0.4 vs. 7.7 +/- 0.4 mm, p0.05). The brachial and radial artery recording forces normally used during clinical measurements by 2 investigators were 1.43 +/- 0.01N (95% confidence intervals (CI) = 1.23N, 1.62N) and 1.88 +/- 0.11N (95% CI = 1.65N, 2.10N), respectively. Therefore, at forces normally used by clinical investigators, the pulse wave velocity, harmonic transmission ratios, and pulse amplitudes and contours obtained at the brachial and radial artery are not significantly influenced by forces applied at the brachial artery. However, these variables may be decreased in subjects with more superficial arteries when higher recording forces are used.

Developmental Toxicity of Inhaled N-Methylformamide in the Rat. Rickard, L. B., Driscoll, C. D., Kennedy, G. L., Jr., Staples, R. E., and Valentine, R. (1995). Fundam. Appl. Toxicol. 28, 167-176. The developmental toxicity of N-methylformamide (MMF), an industrial chemical intermediate used in the production of agrichemicals, was examined in pregnant rats. MMF was administered by nose-only inhalation, 6 hr daily on Days 7-16 of gestation (the day copulation was confirmed was termed Day 1 of gestation, Day 1G) at exposure concentrations of 0, 15, 50, or 150 ppm. Dams were regularly monitored throughout gestation for body weight gain, feed consumption, and clinical signs. Cesarean sections were performed on Day 22G and the offspring were examined. Maternal toxicity was evident in dams exposed to 50 or 150 ppm; one dam exposed to 150 ppm died on Day 14G (considered to be treatment. related) and dams in the 50 and 150 ppm groups exhibited concentration-related clinical findings. Clinical signs of wheezing and rattling were observed both during and after the exposure period. The 150 ppm group also showed significant decreases in weight gain and feed consumption. A significant increase in the mean number of resorptions per litter at the 150 ppm level indicated an embryolethal effect. Developmental toxicity was apparent by a significant decrease in mean fetal body weight and increases in fetal malformations (subcutaneous cysts on the head, microphthalmia, anophthalmia, fused ribs and/or vertebra, and distended brain ventricles) and variations (misaligned and fused sternebrae) due to retarded development at 150 ppm. Significant fetal body weight decreases were also present at 50 ppm. Thus, in this study, the no-observable-adverse-effect level for both dam and fetus was 15 ppm MMF, indicating that for the parameters included in this study, the conceptus is not uniquely sensitive to MMF.

Dibasic esters (DBE) are a mixture of 3 carboxylic acid esters which are used in the paint and coatings industry. In this study, groups of pregnant Crl:CD BR rats were exposed to either 0.16, 0.4, or 1.0 mg DBE/L by inhalation for 6 hr/day from Days 7 through 16 of gestation (day in which copulation plug was detected was designated Day 1G). A control group of chambered pregnant rats was exposed simultaneously to air only. All female rats were euthanized on Day 21G and the fetuses were examined. A suppression of both food consumption and the rate of body weight gain was seen in the 0.4 and 1.0 mg/L groups during the first 6 exposure days. Staining on the fur and perineal area was seen in rats exposed to 1.0 mg/L and liver weight decreases, although not statistically significant, occurred in the 2 high exposure groups. None of the reproductive parameters were altered in any of the groups and no fetal effects were detected. DBE is not a developmental toxin in the rat following inhalation exposures as high as 1.0 mg DBE/L during the period of organogenesis.

Arterial pressure pulses are often measured noninvasively, but the influence of external recording forces has not been well documented. Accordingly, the main goal was to delineate pulse contour and amplitude distortion with progressively compressive forces applied to an external transducer. Ten young normal male subjects were studied while supine. Pulses were recorded at the end of a normal expiration over a range of forces (0.45-4.29, 0.42N increments) applied in a randomized order over first the brachial and then the radial artery. Pulse contours were Fourier analyzed and harmonic powers were normalized to the peak power at the fundamental frequency. Brachial artery pulse amplitudes progressively decreased (p = 0.013), whereas, those at the radial artery peaked at a recording force of 1.79 +/- 0.01N (p0.001) and subsequently decreased parallel to the brachial data with larger forces. No significant pulse contour distortions occurred at either the brachial or radial artery with applied forces of up to 2.18 +/- 0.02 and 2.59 +/- 0.02N, respectively, as indicated by the similarity of the relative power for harmonics 2-7. Radial artery pulses were distorted at and beyond a force of 2.99 +/- 0.01N as indicated by the increased relative power of harmonics 3-7 (p0.05). Therefore, despite significant alterations in pulse amplitude, the relative shape of pulses remains similar over a large range of lower recording forces.

To better understand risk factors associated with early postoperative death or failure, we reviewed our entire experience with 702 consecutive patients who had the modified Fontan operation at the Mayo Clinic between October 1973 and December 1989. The event rate for takedown of repair or death during the initial hospitalization or within 30 days of the operation was 14.8% (successful takedown of the repair, n = 6; death, n = 98). To identify variables associated with early death or Fontan takedown, we analyzed 33 clinical and hemodynamic variables in a univariate and multivariate manner. On the basis of a stepwise logistic discriminant analysis, patients who were younger and operated on before 1980 with a higher preoperative pulmonary artery mean pressure, asplenia, higher intraoperative (after Fontan operation) right atrial pressure, longer aortic crossclamp time, and pulmonary artery ligation were more likely to have the outcome event of interest (p values0.05). A new variable, corrected pulmonary artery pressure (that is, mean preoperative pulmonary artery pressure divided by the ratio of pulmonary to systemic flow if the ratio of pulmonary to systemic flow is greater than 1.0), was significantly associated with the outcome event univariately (p = 0.002), but was no more predictive than the preoperative pulmonary artery mean pressure. Variables less predictive of the outcome event in this analysis included multiple prior operations, polysplenia syndrome, complex anatomy other than asplenia syndrome, and systemic atrioventricular valve regurgitation. These results represent the largest single-institution review of the Fontan operation and suggest that some anatomic and hemodynamic variables previously predictive of poor early outcome have been nullified by current operative methods.

The synthetic retinobenzoic acid RE-80 was evaluated for its potential as an inductor of tumor cell differentiation and as a chemopreventive agent. A minimally toxic dose of RE-80 in vitro produced morphologic changes typical differentiation in epidermal tumor cell colonies. Indirect immunofluorescence indicated induction of a differentiation-associated keratin of internal stratified epithelia, K13, and inhibition of the differentiation-associated epidermal keratin K1. Cultured cells were skin-grafted to athymic nu/nu mice to evaluate RE-80 effects on early stage malignant progression in vivo. When tumors had grown to 3 to 4 mm in diameter, mice were treated by intraperitoneal injection with RE-80 (67 micrograms, 170 mmol, i.p., two times per week) or vehicle (100 microliters 20% ethanol). Papillomas (benign) and moderately differentiated squamous cell carcinomas were reduced in volume about 4-fold by RE-80 treatment. Larger, poorly differentiated squamous cell carcinomas were unaffected. RE-80 resulted in a lower proportion of proliferative cells (detectable by bromodeoxyuridine incorporation) and a higher proportion of moderately to well differentiated tumors after 40 days of treatment compared with control tumors, which were mainly poorly differentiated squamous cell carcinomas. K13 induction in vitro was correlated with response to retinoid in vivo. Induction of differentiation may be mechanism of the response to RE-80 in vivo since carcinoma cells expressing K13 did not incorporate bromodeoxyuridine and were on a terminal pathway.

The CDMS experiment utilizes Ge and Si detectors operating at 20 mK to search for the Dark Matter of the Universe hypothesized to exist in the form of weakly interacting massive particles (WIMPs). In early 2000, CDMS set the most competitive exclusion limit for scalar-interaction WIMPs in the Stanford Underground Facility (SUF). A new search (CDMS II) is now commencing with several improvements: a deep-site facility in the Soudan mine, Minnesota; and the detector technology has been further improved to aid in the rejection of surface-electron (β) events. A new generation of detectors, sensitive to the initial athermal phonon flux from a particle event, have been in operation for the past year at Stanford's shallow site and are ready for installation at the deep site.

The authors sought to evaluate survival of patients with ovarian cancer treated with second-line intraperitoneal cisplatin-based chemotherapy.From August 1985 to September 1991, 63 patients with recurrent or persistent ovarian cancer after first-line cisplatin-based chemotherapy were on a protocol and treated with cisplatin, cytarabine, with or without bleomycin. Eligibility included Stage III/IV invasive adenocarcinoma of the ovary, documentation of the size of residual disease at reoperation, and prior treatment with first-line intravenous cisplatin-based chemotherapy.The median survival from intraperitoneal chemotherapy was 29 months. For patients who responded to first-line chemotherapy and second-line intraperitoneal chemotherapy, the 5-year survival was 60%, but for patients with response to first-line chemotherapy but no response to intraperitoneal chemotherapy the 5-year survival was only 17%, and no patient survived 5 years who did not have a response to first- or second-line therapy (P0.0001). There was significant improvement in the 2-year survival from initiation of intraperitoneal chemotherapy for patients withor = 5 mm residual tumor in greatest dimension (74%), compared with patients with residual tumor larger than 5 mm and smaller than or equal to 2 cm (38%), and patients with residual tumor larger than 2 cm (0%) (P0.0001).Survival was increased for patients who (1) had a response to first-line intravenous chemotherapy and second-line intraperitoneal chemotherapy and (2) who had residual tumor 5 mm or smaller at the initiation of intraperitoneal therapy. The conclusion that the prolonged survival is secondary to cisplatin-based chemotherapy administered intraperitoneally must be made with caution because survival may have been secondary to the small residual disease at initiation of second-line chemotherapy or the systemic effect of cisplatin.

Between 1975 and 1987, 128 patients with infiltrating breast cancer, categorized as clinical Stage I and II disease, were treated by breast conservation surgery without radiation therapy.After a median disease-free interval of 20 months (range, 8-64 months), 25 of 128 patients had local recurrence, for which salvage mastectomy was performed. The results of modified radial mastectomy as a salvage procedure were analyzed in these 25 patients.After a median disease-free interval of 52 months (range, 8-75 years) after the salvage procedure, 12 patients had chest wall and distant recurrences, whereas 13 patients remained free of disease. The 5-year actuarial disease-free and overall survival rates after the salvage mastectomy were 51% and 65%, respectively.Univariate analyses of factors affecting disease-free survival and overall survival showed that the size of the local recurrence (or = 2 cm) (P = 0.009) and the number of pathologically positive axillary nodes at the time of the salvage procedure (fewer than four nodes) (P = 0.002) were associated with a better prognosis.

In a study for the Federal Highway Administration (FHWA) completed several years ago, my co-workers and I assembled and analyzed pollutant runoff data for high…

We have investigated the possibility that the generation of hydrogen peroxide (H2O2) by spermatozoa plays a physiological role during capacitation. Capacitation is defined as the incubation period required for fertilization in mammals. Capacitation culminates in an exocytotic event, the acrosome reaction (AR). Mammalian sperm generate H2O2 during aerobic incubation and do not contain catalase, the enzyme that promotes scavenging of H2O2. In the present work we show that added catalase inhibited the AR, while glucose oxidase (GO), an enzyme that generates H2O2, accelerated the onset of the AR. Direct addition of H2O2 also stimulated the AR; catalase inhibited both the stimulation by GO and by H2O2. The onset of the AR was always preceded by the appearance of hyperactivated motility. The stimulation of the AR by H2O2 was manifest 1-2 h after the addition of H2O2. Catalase added at 3 h of incubation was less effective in inhibiting the AR than catalase added at the beginning. Incubation of sperm with catalase prevented the induction of the AR by the membrane-perturbing lipid, lysophosphatidyl choline. Taken together, these results suggest that H2O2 produced by hamster sperm plays a significant role during capacitation, possibly in membrane reorganization to facilitate the fusion that takes place during exocytosis of the acrosomal contents.

This paper presents the final results of the application of a probabilistic approach for evaluating the water quality impact of highway stormwater runoff to receiving waters. A brief review of the development of the probabilistic characterization of runoff water quality data is presented along with a description of the probabilistic stream impact method. In this paper we present the interactive Decision Support program that takes the procedure user through the Federal Highway Administration's highway runoff analysis procedure. The procedure requires the input of fixed site data, rainfall and streamflow statistics, highway runoff water quality, and target concentrations. The methodology user is provided guidance in entering the required data. Output of the procedure is a return frequency of the exceedances of the target concentration. This methodology is intended for planning level analyses.

18547 Background: Little is known about how information needs of cancer survivors/patients (S/P), caregivers (CG), and non-caregiver relatives (NCG) evolve during the cancer experience. Methods: We conducted structured interviews of 21 leukemia, lymphoma, and multiple myeloma S/P, 13 CG, and 19 NCG, and asked them to prioritize information needs during the 4 cancer experience phases from a list of 13 topics. This research was supported by the Division of Cancer Prevention and Control of the Centers for Disease Control and Prevention. Results: Information preferences varied (Friedman’s test, P < 0.0001) by phase of the cancer experience. Topics ranked highest (median rank in parentheses) before treatment, during treatment, after treatment, and at relapse, respectively, were diagnostic tests (2), cancer/ cancer types (2.5), treatment options (3.4), hospitals/cancer centers (3.5), and insurance/financial issues (4); coping with treatment side effects (3), insurance/financial issues (4.5), treatment options (5), hospitals/cancer centers (5), and long term side effects (5); long term side effects (2), tests to detect recurrence (2), risk factors (3), diagnostic tests (4), and support groups (5); and tests to detect recurrence (3), insurance/financial issues (3), cancer/cancer types (4), diagnostic tests (5), and treatment options (5). Across the cancer experience, the highest priority topics for S/P, CG, and NCG, were cancer/ cancer types (4), treatment options (4), long term side effects (5), and risk factors (5); diagnostic tests (4.5), coping with side effects of treatment (5), and treatment options (5); and treatment options (3), diagnostic tests (4), and 5 other topics tied for 3rd place (6). In-depth, semi-structured interviews conducted with the surveys provided additional detail regarding responses collected in the prioritization exercise. Conclusions: Clinicians can help S/P, CG, and NCG by offering role- and cancer continuum-targeted information. No significant financial relationships to disclose.

5055 Objectives: COX-2 has been implicated as tumorogenic in various tumor models. Our previous immunohistochemistry work has shown that normal ovarian epithelium, normal inclusion cysts, serous cy...

9673 Background: Cancer-testis antigens (CTA), are selectively expressed in various types of human neoplasms but not in normal tissues other than testis. This characteristic feature of CTA makes th...

5034 Background: Approximately 50% of ovarian tumors have copy number increases in chromsome20q. The 20q ovarian amplicon has yet to be precisely mapped, but is known to include part of 20q12 and 2...

2583 Background: Cancer/testis (CTA) antigens expression is highly tissue-restricted, immunogenic in cancer patients, and are novel targets for cancer vaccine.These CTA are expressed in different t...