First Accurate Normalization of the ß-delayed α Decay of

The ^{12}C(α,γ)^{16}O reaction plays a central role in astrophysics, but its cross section at energies relevant for astrophysical applications is only poorly constrained by laboratory data. The reduced α width, γ_{11}, of the bound 1^{-} level in ^{16}O is particularly important to determine the cross section. The magnitude of γ_{11} is determined via sub-Coulomb α-transfer reactions or the ß-delayed α decay of ^{16}N, but the latter approach is presently hampered by the lack of sufficiently precise data on the ß-decay branching ratios. Here we report improved branching ratios for the bound 1^{-} level [b_{ß,11}=(5.02±0.10)×10^{-2}] and for ß-delayed α emission [b_{ßα}=(1.59±0.06)×10^{-5}]. Our value for b_{ßα} is 33% larger than previously held, leading to a substantial increase in γ_{11}. Our revised value for γ_{11} is in good agreement with the value obtained in α-transfer studies and the weighted average of the two gives a robust and precise determination of γ_{11}, which provides significantly improved constraints on the ^{12}C(α,γ) cross section in the energy range relevant to hydrostatic He burning.

Hypermethylation of the hMLH1 promoter in colon cancer with microsatellite instability.

Recent studies have demonstrated the presence of microsatellite instability (MSI) in tumors from patients with hereditary nonpolyposis colon cancer and in a subset of patients with sporadic colorectal cancer (CRC). In sporadic CRC, three tumor phenotypes have been defined: microsatellite stable (MSS), low-frequency MSI, and high-frequency MSI (MSI-H). Although defective mismatch repair, consisting primarily of alterations in hMSH2 and hMLH1, is believed to be responsible for the MSI phenotype in the majority of patients with hereditary nonpolyposis colon cancer, the genetic defect responsible for this phenotype in sporadic CRC has yet to be clearly delineated. Somatic or germ-line alterations in these two genes have been identified in only a minority of these cases. Analysis of the protein expression patterns of hMSH2 and hMLH1 in unselected CRC, however, suggests that alterations in hMLH1 may account for a majority of the MSI-H cases. In an effort to explore the underlying molecular basis for these findings, we have examined the methylation status of the presumptive hMLHI promoter region in 31 tumors that vary in regard to their MSI status (MSI-H or MSS), their hMLH1 protein expression (MLH- or MLH+), and their gene mutation (Mut+ or Mut-) status. Hypermethylation of the hMLH1 promoter occurred in all 13 MSI-H/ MLH- tumors that did not have a detectable mutation within the hMLH1 gene. Of those MSI-H tumors containing germ-line or somatic alterations in hMLH1 (n = 7, including 3 frameshift, 1 nonsense, 2 missense mutations, and 1 tumor containing multiple mutations: missense, splice-site alteration, and a frameshift), four had a normal methylation pattern, whereas three others demonstrated hypermethylation of the hMLH1 promoter region. Two of these cases had a missense alteration, the other a frameshift alteration. The single MSI-H/Mut+ tumor that had normal hMLH1 and hMSH2 expression, as well as 9 of the 10 MSS cases, lacked methylation of the hMLH1 promoter. Hypermethylation of the hMSH2 promoter was not observed for any of the cases. These results suggest that hypermethylation of the hMLH1 promoter may be the principal mechanism of gene inactivation in sporadic CRC characterized by widespread MSI.

HMSH6 alterations in patients with microsatellite instability-low colorectal cancer.

Two microsatellite instability (MSI) phenotypes have been described in colorectal cancer (CRC): MSI-H (instability at >30% of the loci examined) and MSI-L (MSI at 1-30% of the loci examined). The MSI-H phenotype, observed in both hereditary nonpolyposis colon cancer-associated CRC and approximately 15% of sporadic CRC, generally results from mutational or epigenetic inactivation of the DNA mismatch repair (MMR) genes hMSH2 or hMLH1. The genetic basis for the MSI-L phenotype, however, is unknown. Several other proteins, including hMSH3 and hMSH6, also participate in DNA MMR. Inactivating mutations of MSH6 in yeast and human tumor cell lines are associated with an impaired ability to repair single-base mispairs and small insertion-deletion loops but not large insertion-deletion loops. This suggests that hMSH6 mutations are more likely to be associated with a MSI-L phenotype than a MSI-H phenotype in CRC. To explore this possibility, we screened tumors from 41 patients with MSI-L CRC for hMSH6 mutations with conformation-sensitive gel electrophoresis (CSGE) and for hMSH6 protein expression by immunohistochemistry. Alterations found with CSGE were confirmed by DNA sequencing of normal and tumor tissue. One somatic (Asp389Asn) and 15 germ-line changes were found. Of the 15 germ-line changes, 9 were found in an intron (none involving splice junctions), and 6 were found in an exon (Gly39Glu, Leu395Val, and 4 silent alterations). Immunohistochemical staining for hMSH6 performed on 34 of the 41 tumors revealed strong nuclear hMSH6 expression in all of the cases. Overall, our results suggest that hMSH6 mutations do not play a major role in the development of sporadic CRC with a MSI-L phenotype.

Method for sequential staining of GTL-banded metaphases with fluorescent-labeled chromosome-specific paint probes.

We describe a method for use of fluorescent-labeled whole chromosome-specific paint probes on GTL-banded metaphases to utilize the combined potential of these techniques for defining chromosome abnormalities. The efficacy of this method was tested on 6 cases involving different chromosome abnormalities and various tissues, including blood, amniotic fluid, skin fibroblasts, and bone marrow.

Fluorescence in situ hybridization with X and Y chromosome probes for cytogenetic studies on bone marrow cells after opposite sex transplantation.

We investigated the efficacy of fluorescence-labelled chromosome probes (CEP-X/Y) for the X and Y chromosomes to study patients who have had opposite sex BMT. These probes hybridize to the centromere region of the X chromosome and nearly the entire long arm of the Y chromosome. These probes are direct-labelled and produce X and Y signals that can be simultaneously viewed and readily distinguished from each other by color and size after only five brief washes. We investigated BM specimens from 20 normal donors and 16 patients who had undergone an opposite sex BMT. We found no significant interinvestigator differences with respect to scoring XX or XY interphase cells. The 'normal range' for XX cells in males was up to 0.628% and for XY cells in females it was up to 0.299%. Each of the specimens from the patients who underwent BMT had a significant number of donor cells compared with normal range. We suggest that an economical, rapid and accurate cytogenetic test can be achieved by using these probes as an adjunct to conventional cytogenetics.

The application of fluorescent in situ hybridization to detect Mbcr/abl fusion in variant Ph chromosomes in CML and ALL.

We investigated the usefulness of fluorescent in situ hybridization with different-colored major breakpoint cluster region (Mbcr) and Abelson oncogene (abl) probes in clinical practice. In standard Ph chromosomes with a Mbcr breakpoint, these probes produced a fusion of Mbcr and abl signals that was visible in interphase and metaphase cells. The normal range for apparent Mbcr/abl fusion signals in interphase nuclei was established in bone marrow from 25 normal controls. We tested the probes on 35 bone marrow specimens from five normal subjects and 29 patients with various kinds of Ph chromosomes and chronic myelogenous leukemia or acute lymphocytic leukemia. This method produced Mbcr/abl fusion signals in patients with a standard Ph chromosome, simple or complex variants of Ph chromosomes, and "Ph-negative chronic myelogenous leukemia." In metaphase cells of patients with acute lymphocytic leukemia, this method established Ph chromosomes with minor bcr (mbcr) breakpoints. Fluorescent in situ hybridization is a relatively inexpensive and rapid method. When this method is used in conjunction with conventional chromosome analysis, the cytogeneticist can combine the power of complete karyotype studies and the resolution of molecular techniques for patients suspected of having a Ph chromosome.

A novel three-color, clone-specific fluorescence in situ hybridization procedure for monoclonal gammopathies.

We have developed a three-color cytoplasmic immunoglobulin (cIg) and fluorescence in situ hybridization (FISH) technique to detect plasma cell chromosomal aneuploidy in patients with multiple myeloma (MM), monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), and amyloidosis (AL). Immunofluorescent-labeled antibodies to detect light chain expression and six directly labeled alpha-satellite chromosome specific enumeration probes (CEP) were used simultaneously to detect aneuploidy of the plasma cells. The six probes were specific for chromosomes 7, 9, 11, 15, 18, and X. The technique was tested in 12 consecutive patient samples (5 MM, 2 MGUS, 3 SMM, and 2 AL). Based on the alpha-satellite signals, we found trisomic clones for CEP 7 (4 of 12), CEP 11 (4 of 12), CEP X (1 of 12), CEP 9 (8 of 12), CEP 15 (7 of 12), and CEP 18 (5 of 12). Trisomic clones of at least one of the six chromosomes were present in 9 of 12 patients. We believe that this technique efficiently identifies monotypic plasma cells and permits simultaneous analysis of numeric chromosome anomalies by FISH in emerging neoplastic cells. We are in the process of applying this technique to a series of about 100 newly diagnosed monoclonal gammopathy patients.

Detection of RB1 deletions by fluorescence in situ hybridization in malignant hematologic disorders.

We evaluated the usefulness of fluorescence in situ hybridization (FISH) using different-colored commercial RB1 and 13qter DNA probes to identify RB1 deletions in interphase nuclei of bone marrow from 24 patients with agnogenic myeloid metaplasia (AMM), 20 patients with multiple myeloma (MM), 21 patients with other hematologic malignancies, and 25 normal bone marrow transplant (BMT) donors. Based on the 25 normal BMT donors, the upper boundary for the normal percentage of nuclei with one RB1 signal was 6.5%. Based on eight specimens known to have a deletion of 13q14 by cytogenetic studies, the lower limit of abnormal for the percentage of nuclei with one RB1 signal was 12.5%. More than 12.5% of nuclei had a single RB1 signal in 7/24 (29%) patients with AMM and 3/20 (15%) patients with MM. None of the 21 patients with hematologic malignancies other than AMM or MM had more than 12.5% nuclei with loss of RB1. The results of this study suggest that FISH with RB1 probes is useful to detect loss of RB1 in interphase nuclei from patients with hematologic disorders who have chromosome abnormalities involving 13q14. Thus, FISH with probes for RB1 is efficacious to investigate the pathogenesis of RB1 in malignant neoplasms and is a useful adjunct to conventional cytogenetic studies in clinical practice when abnormalities of 13q14 are involved.

Water quality before and after deep tunnel operation in Milwaukee, Wisconsin.

The mitigative effects of the deep tunnel for temporary storage of storm water and sewage, on the water quality of the Milwaukee, Menomonee, and Kinnickinnic Rivers are investigated. The analysis is based on data from the Milwaukee Metropolitan Sewerage District's overflow and surface-water quality monitoring program. Statistical analysis of water quality parameters (BOD, phosphorus, suspended solid, fecal coliform counts, zinc, and chloride) in the three rivers indicates that Menomonee River benefits the most from the deep tunnel. Fecal coliform counts inside the CSO area, and to a certain extent BOD and zinc levels, exhibit the most significant decline after 1994 when the tunnel came on line. These conclusions are based on t-test comparisons of regional averages incorporating spatial and temporal correlations from 1991 to 1993 and 1994 to 1997. The results from t-tests are complemented and confirmed with results from Mann-Kendall tests for trend. Suspended solids and chloride do not decrease after 1994.

Primary epiphyseal osteomyelitis in children. Report of three cases and review of the literature.

In the literature 21 children have been reported with haematogenous osteomyelitis involving only the epiphyses of long bones; in 20 the epiphyses of the knee were involved and the radiographs showed a lytic lesion. Most patients gave a history of pain, limp or refusal to walk for weeks or months without general illness. The erythrocyte sedimentation rate and white cell count were of little diagnostic value, but a bone scan was usually positive. All the patients were cured by antibiotics alone or in combination with curettage.

Determination of PAH sources in dated sediments from Green Bay, Wisconsin, by a chemical mass balance model.

Six sediment cores were collected from Green Bay, Wisconsin, in order to identify possible sources of polycyclic aromatic hydrocarbons (PAHs) by a chemical mass balance (CMB) model. The cores which were obtained in 1995 had total PAH concentrations between 8.04 and 0.460 ppm. 210Pb and 137Cs dating was used to determine historical trends of PAH inputs, and elemental carbon particle analysis was done to characterize particles from combustion of coal, wood and petroleum. The results show that coke burning, highway dust, and wood burning are likely sources of PAHs to Green Bay. The contribution of coke oven emissions (CB) for the Green Bay cores is in the range of 5 to 90%. The overall highway dust (HWY) contribution is between 5 and 70%. There is a maximum (approximately 67%) contribution of HWY around 1988 which is in agreement with the historical US petroleum consumption. The wood burning (WB) contribution is between 1 to 30%, except in core GB-A where a maximum (approximately 50%) is found around 1994. The average relative errors of measurement for x2 equal to the number of degrees of freedom, are 52.5, 56.2, 36.2, 52.3, and 42.8 (df = 3) for the Green Bay cores A, B, C, E, and F, respectively. The sums of the contribution factors are less than one, indicating gain of inert biological or other bulk material between source and receptor. The results of carbon particles for Green Bay core D show that coal, oil, and wood burning are consistent with the CMB modeling results.

Joint action of chemicals in algal toxicity tests: influence of response level and dose-response regression model.

The joint toxicity of nonylamine and decylamine and of atrazine and decylamine was evaluated in assays with the green alga Selenastrum capricornutum based on an isobologram method. In this method, curves of constant response, isoboles, are plotted versus concentrations of two toxicants. The response parameter was growth rate based on biomass, and several response levels were used. Dose-response curves were developed for dilution series using fixed ratios between concentrations in toxic units of the compounds. Probit and Weibull dose-response curves were then determined by nonlinear regression. A model for isoboles for partially similar action was used when applicable. The no-effect concentration (NEC or EC0) was estimated based on a newly proposed model containing median effective concentration (EC50) and EC0 as explicit variables. Results show that nonylamine and decylamine are nearly concentration additive at EC50 and EC10 (similarity parameter lambda = 0.70-0.76) and to a lesser extent at EC0. By contrast, the mixtures of atrazine and decylamine show antagonism in that atrazine acts as an antidote to decylamine. The shapes of these isoboles are independent of response level. The EC50 values (mg/L) for chemicals acting singly were 0.090 (nonylamine), 0.039 to 0.044 (decylamine), and 0.225 (atrazine). In order to determine NEC effectively, the level of inhibition must be fairly low, with observed growth rates between 0.6 and 1.0 times the average growth rate of the controls.

PCB sources and degradation in sediments of Ashtabula River, Ohio, USA, determined from receptor models.

The PCB pollution in Ashtabula River sediments was evaluated using a factor analysis (FA) model with non-negative constraints, and a chemical mass balance (CMB) model. The FA model identified Aroclor 1248 as the major PCB source, and also a congener pattern significantly different from that of any Aroclor. The CMB model that uses linear combinations of Aroclors, failed to reproduce the sample congener profiles with good statistical fit. The findings from both models indicate that the PCBs in Ashtabula River sediments have undergone significant alterations changing their profiles from those of the original sources. These alterations may be explained by the anaerobic dechlorination of highly chlorinated congeners, according to dechlorination activities H/H'.

Behavior therapy versus "will power" in the management of obesity.

In a study of the management of obesity 43 patients were randomly assigned to behavior therapy, will power, and no-treatment control groups. The behavioral treatment involved contingency contracting, stimulus control, self-monitoring, energy expenditure, and group reinforcement procedures over an 18-week period. The will power patients were told to do the same thing as the behavior therapy patients; however, instead of having formal contingencies and meeting regularly, they were told to apply "will power" on their own, as this was the most important aspect of losing weight. The third group was a standard no-treatment control group. Analyses of covariance indicated that (a) the behavior therapy group lost significantly more weight than the will power (p less than .05) and no-treatment control (p less than .01) groups, and (b) the will power and no-treatment control groups did not differ significantly from each other. An 18-week maintenance follow-up of the behavior therapy group indicated that there was no significant weight gain from the end of treatment to follow-up. Also, an analysis of time spent per patient suggested that the behavioral treatment employed in this study was more efficient than previous treatments.

A general noninteractive multiple toxicity model including probit, logit, and Weibull transformations.

A multiple toxicity model for the quantal response of organisms is constructed based on an existing bivariate theory. The main assumption is that the tolerances follow a multivariate normal distribution function. However, any monotone tolerance distribution can be applied by mapping the integration region in the n-dimensional space of transforms on the n-dimensional space of normal equivalent deviates. General requirements to noninteractive bivariate tolerance distributions are discussed, and it is shown that bivariate logit and Weibull distributions, constructed according to the mapping procedure, meet these criteria. The univariate Weibull dose-response model is given a novel interpretation in terms of reactions between toxicant molecules and a hypothetical key receptor of the organism. The application of the multiple toxicity model is demonstrated using literature data for the action of gamma-benzene hexachloride and pyrethrins on flour beetles (Tribolium castaneum). Nonnormal tolerance distributions are needed when the mortality data include extreme response probabilities.

PAHs in dated sediments of Ashtabula River, Ohio, USA.

Polycyclic aromatic hydrocarbons (PAHs) in dated sediments from Ashtabula River, Ohio, were determined, and a chemical mass balance (CMB) model was used to apportion sources. Three cores (AR-1,AR-2,AR-3) were dated by correlating uranium-supported 210Pb peaks with 1964, 1972, 1977, and 1979 maxima in the discharge record for Ashtabula River. These cores had sedimentation rates between 7.1 and 4.4 cm/year, while a fourth (AR-4) exhibited a much higher rate of 27.8 +/- 18 cm/year. The highest PAH concentration was 11,500 ng/g found in layer 6 of AR-1 (1986), and the lowest was 621 ng/g found in layer 8 of AR-2 (1982). The source contributions to the total PAH concentrations estimated by the CMB model are 0.1-2.2%, 16.8-22.8%, and 78.1-83.8% for wood burning (WB), coke oven (CO), and highway dust (HWY), respectively. Petroleum generated PAHs have maximal contribution during 1977-79, and wood burning PAHs show minimal emissions during 1975-77 in accordance with U.S. consumption records and other studies. Among six PAH markers, only phenanthrene may be subjected to aerobic biodegradation or photolysis with an apparent half-life of 0.005-0.025 year. No anaerobic degradation was observed based on the CMB model. The model works well for the nonmarker compounds, fluoranthene, and benzo[b]fluoranthene/benzo[k]fluoranthene.