Development, testing, and deployment of an air sampling manifold for spiking elemental and oxidized mercury during the Reno Atmospheric Mercury Intercomparison Experiment (RAMIX).

The Reno Atmospheric Mercury Intercomparison Experiment (RAMIX) was in Reno, NV from August 22, 2011 to September 16, 2011. The goals of the experiment were to compare existing and new methods for measurements of ambient elemental and oxidized Hg, and to test these with quantitative spikes of Hg(0), HgBr2, O3 and water vapor. In this paper we describe the design, testing, and deployment of a high flow manifold system designed to deliver ambient air and spiked compounds to multiple instruments simultaneously. The manifold was constructed of 1" OD PFA tubing and heated to 115 °C for the entire active zone. Manifold flow was controlled at ∼200 LPM using a blower and a velocity sensor in a feedback control system. Permeation tubes in controlled ovens were used to deliver Hg(0) and HgBr2. Ozone was generated from a small UV lamp in a flow of high purity O2. Water vapor was generated by pumping a flow of purified N2 through heated, high purity water. The spiking delivery for Hg(0), HgBr2, O3, and water vapor after dilution in the manifold ranged up to 20 ng m(-3), 0.64 ng m(-3), 100 ppbv, and 20 g kg(-1), respectively. During laboratory tests the average transmission efficiencies for Hg(0), HgBr2, and O3 were found to be 92%, 76%, and 93%, respectively.

Identification of genes regulated during osteoblastic differentiation by genome-wide expression analysis of mouse calvaria primary osteoblasts in vitro.

Although several independent studies of gene expression patterns during osteoblast differentiation in cultures from calvaria and other in vitro models have been reported, only a small portion of the mRNAs expressed in osteoblasts have been characterized. We have previously analyzed the behavior of several known markers in osteoblasts, using Affymetrix GeneChip murine probe arrays (27,000 genes). In the present study we report larger groups of transcripts displaying significant expression modulation during the culture of osteoblasts isolated from mice calvaria. The expression profiles of 601 such regulated genes, classified in distinct functional families, are presented and analyzed here. Although some of these genes have previously been shown to play important roles in bone biology, the large majority of them have never been demonstrated to be regulated during osteoblast differentiation. Despite the fact that the precise involvement of these genes in osteoblast differentiation and function needs to be evaluated, the data presented herein will aid in the identification of genes that play a significant role in osteoblasts. This will provide a better understanding of the regulation of osteoblast differentiation and maturation.

Behavior of osteoblast, adipocyte, and myoblast markers in genome-wide expression analysis of mouse calvaria primary osteoblasts in vitro.

Several genes, such as alkaline phosphatase, osteocalcin, and Cbfa1/Osf2, are known to be regulated during osteoblastic differentiation and are commonly used as "osteoblast markers" for in vitro or in vivo studies. The number of these genes is very limited, however, and it is of major interest to identify new genes that are activated or repressed during the process of osteoblast differentiation and bone formation as well as to extend the available information on gene families relevant to this particular differentiation pathway. To identify such genes, we have implemented a genome-wide analysis by determining changes in expression levels of 27,000 genes during in vitro differentiation of primary osteoblasts isolated from mouse calvaria. This study focuses on the description of the analytical and filtering process applied; on the transcriptional analysis of well-established "bone," "adipocyte," and "muscle" pathway markers; and on a description of the regulation profiles for genes recently described in the Skeletal Gene Database. We also demonstrate that new array technologies constitute reliable and powerful tools to monitor the transcription of genes involved in osteoblastic differentiation, allowing a more integrated vision of the biological pathways regulated during osteoblast commitment, differentiation, and function.

Are rural residents less likely to obtain recommended preventive healthcare services?

BACKGROUND: This study examined rural-urban differences in utilization of preventive healthcare services and assessed the impact of rural residence, demographic factors, health insurance status, and health system characteristics on the likelihood of obtaining each service. METHODS: National data from the 1997 Behavioral Risk Factor Surveillance System (BRFSS) and the 1999 Area Resource File were used to evaluate the adequacy of preventive services obtained by rural and urban women and men, using three sets of nationally accepted preventive services guidelines from the American Cancer Society, U.S. Preventive Services Task Force, and Healthy People 2010. Logistic regression models were developed to control for the effect of demographic factors, health insurance status, and health system characteristics. RESULTS: Rural residents are less likely than urban residents to obtain certain preventive health services and are further behind urban residents in meeting Healthy People 2010 objectives. CONCLUSIONS: Efforts to increase rural preventive services utilization need to build on federal, state, and community-based initiatives and to recognize the special challenges that rural areas present.

Do employees use report cards to assess health care provider systems?

OBJECTIVE: To investigate consumers' use of report cards that provide information on service quality and satisfaction at the provider group level. DATA SOURCES: In 1998 we conducted a telephone survey of randomly selected employees in firms aligned with the Buyers Health Care Action Group (BHCAG) in the Minneapolis-St. Paul market. STUDY DESIGN: Univariate probit models were used to determine report card utilization, perceived helpfulness of the report card, and ease of selecting a provider group. The characteristics used in the models included health status, age, gender, education, residency, job tenure, marital status, presence of dependent children, household income, and whether consumers changed provider groups. DATA COLLECTION: Our sample consists of survey responses from 996 single individuals (a response rate of 91 percent) and 913 families (a response rate of 96 percent). The survey was supplemented with data obtained directly from employers aligned with BHCAG. PRINCIPLE FINDINGS: Consumers who changed to a new provider group are more likely to use report card information and find it helpful, consumers employed in large firms are less likely to use the report card, and families who use information from their own health care experiences are less likely to find the report card helpful. In addition, individuals who changed to a new provider group are more likely to find the selection decision difficult. CONCLUSION: The findings show that health care consumers are using satisfaction and service-quality information provided by their employers.

Disenrollment from Medicare HMOs.

BACKGROUND: Since the program's inception, there has been great interest in determining whether beneficiaries who enter and subsequently leave Medicare health maintenance organizations (HMOs) are more or less costly than those remaining in fee-for-service (FFS) Medicare. OBJECTIVES: To examine whether relatively high-cost beneficiaries disenroll from Medicare HMOs (disenrollment bias) and whether disenrollment bias varies by Medicare HMO market characteristics. In addition, we compare rates of surgical procedures and hospitalizations for ambulatory care-sensitive conditions for disenrollees and continuing FFS beneficiaries. DESIGN: Cross-sectional analysis of 1994 Medicare data. PARTICIPANTS AND METHODS: Medicare beneficiaries were first sampled from the 124 counties with at least 1000 Medicare HMO enrollees. From this pool, HMO disenrollees and a sample of continuing FFS beneficiaries were drawn. The FFS beneficiaries were assigned dates of "pseudodisenrollment." Expenditures and inpatient service use were compared for 6 months after disenrollment or pseudodisenrollment. RESULTS: The HMO disenrollees were no more likely than the continuing FFS beneficiaries to have positive total expenditures (Part A plus Part B) or Part B expenditures in the first 6 months after disenrollment. However, disenrollees were more likely to have Part A expenditures. Among beneficiaries with spending, disenrollees had higher total and Part B expenditures than continuing FFS beneficiaries. Moreover, the disparity in total and Part B spending between disenrollees and continuing FFS beneficiaries increased with HMO market penetration. Although Part A spending was higher for disenrollees with spending, it was not sensitive to changes in market share. The HMO disenrollees received more surgical procedures and were hospitalized for more of the ambulatory care-sensitive conditions than the FFS beneficiaries. CONCLUSIONS: On several measures, Medicare HMOs experienced favorable disenrollment relative to continuing FFS beneficiaries as recently as 1994, which increased as HMO market share increased.

Comparing mortality and time until death for medicare HMO and FFS beneficiaries.

OBJECTIVE: To compare adjusted mortality rates of TEFRA-risk HMO enrollees and disenrollees with rates of beneficiaries enrolled in the Medicare fee-for-service sector (FFS), and to compare the time until death for decedents in these three groups. DATA SOURCE: Data are from the 124 counties with the largest TEFRA-risk HMO enrollment using 1993-1994 Medicare Denominator files for beneficiaries enrolled in the FFS and TEFRA-risk HMO sectors. STUDY DESIGN: A retrospective study that tracks the mortality rates and time until death of a random sample of 1,240,120 Medicare beneficiaries in the FFS sector and 1,526,502 enrollees in HMOs between April 1, 1993 and April 1, 1994. A total of 58,201 beneficiaries switched from an HMO to the FFS sector and were analyzed separately. PRINCIPAL FINDINGS: HMO enrollees have lower relative odds of mortality than a comparable group of FFS beneficiaries. Conversely, HMO disenrollees have higher relative odds of mortality than comparable FFS beneficiaries. Among decedents in the three groups, HMO enrollees lived longer than FFS beneficiaries, who in turn lived longer than HMO disenrollees. CONCLUSIONS: Medicare TEFRA-risk HMO enrollees appear to be, on average, healthier than beneficiaries enrolled in the FFS sector, who appear to be in turn healthier than HMO disenrollees. These health status differences persist, even after controlling for beneficiary demographics and county-level variables that might confound the relationship between mortality and the insurance sector.

Uncovering the missing Medicaid cases and assessing their bias for estimates of the uninsured.

General population surveys of health insurance coverage are thought to undercount Medicaid enrollment, which may bias estimates of the uninsured. This article describes the results of an experiment undertaken in conjunction with a general population survey in Minnesota. Responses to health insurance questions by a known sample of public program enrollees are analyzed to determine possible reasons for the undercount and the amount of bias introduced in estimates of uninsured people. While public program enrollees often misreport the type of coverage they have, the impact on estimates of those without insurance is negligible. Restrictions to generalizing the finding beyond this study are discussed.

Sequence analysis and functional characterization of the violacein biosynthetic pathway from Chromobacterium violaceum.

Violacein is a purple-colored, broad-spectrum antibacterial pigment that has a dimeric structure composed of 5-hydroxyindole, oxindole and 2-pyyrolidone subunits formed by the condensation of two modified tryptophan molecules. The violacein biosynthetic gene cluster from Chromobacterium violaceum was characterized by DNA sequencing, transposon mutagenesis, and chemical analysis of the pathway intermediates produced heterologously in Escherichia. coli. The violacein biosynthetic gene cluster spans eight kilobases and is comprised of the four genes, vioABCD, that are necessary for violacein production. Sequence analysis suggests that the products of vioA, vioC and vioD are nucleotide-dependent monooxygenases. Disruption of vioA or vioB completely abrogates the biosynthesis of violacein intermediates, while disruption of the vioC or vioD genes results in the production of violacein precursors.

Rural beneficiaries with chronic conditions: does prevalence pose a risk to Medicare managed care?

One of several possible barriers to the growth of Medicare managed care in rural areas is the fear of adverse selection (i.e., the perception that rural beneficiaries are less healthy and have pent-up demand for services). Using 1993 Medicare Current Beneficiary Survey data, we conclude that specific chronic conditions common among the elderly are not more prevalent among rural than urban beneficiaries. Medicare reimbursements for beneficiaries with chronic conditions are generally lower in rural counties. However, the difference between actual Medicare reimbursements and projected capitated payments to managed care organizations is similar in magnitude for rural and urban beneficiaries with these conditions.

Selection experiences in Medicare HMOs: pre-enrollment expenditures.

Using 1993 and 1994 data, the authors examine whether beneficiaries who enroll in a Medicare health maintenance organization (HMO), including those enrolling for only a short period of time, have lower expenditures than continuous fee-for-service (FFS) beneficiaries the year prior to enrollment. We also test whether biased selection varies by the level of HMO market penetration and the rate of market-share growth. We find favorable selection associated with enrollment into Medicare HMOs, which declines as market share increases but does not disappear. Among short-term enrollees, we find unfavorable selection, however, selection bias was not sensitive to market characteristics.

Mapping an endometrial cancer tumor suppressor gene at 10q25 and development of a bacterial clone contig for the consensus deletion interval.

Frequent loss of chromosome 10q sequences in endometrial cancers suggests the involvement of a tumor suppressor gene. Previous loss-of-heterozygosity (LOH)studies have pointed to the 10q25-q26 region as the likely site of a tumor suppressor involved in endometrial tumorigenesis (S. L. Peiffer et al., 1995, Cancer Res. 55: 1922-1926; S. Nagase et al., 1996, Br. J. Cancer 74: 1979-1983; S. Nagase et al.,1997, Cancer Res. 57: 1630-1633). In an attempt to define further the localization of a tumor suppressor gene at 10q25, we screened a panel of 123 endometrioid adenocarcinomas for loss of heterozygosity of 10q25.3 sequences. Forty-three (35%) revealed LOH at one or more loci. The observed patterns of allelic loss define a minimum consensus region of deletion between D10S221 and D10S610. A sequence-ready bacterial clone contig and a long-range restriction map for a 1-Mb interval spanning the deletion region were developed as the first step in experiments directed toward the discovery the 10q25 tumor suppressor.

Allelic imbalance, including deletion of PTEN/MMACI, at the Cowden disease locus on 10q22-23, in hamartomas from patients with Cowden syndrome and germline PTEN mutation.

Cowden disease (CD) is a rare, autosomal dominant inherited cancer syndrome characterized by multiple benign and malignant lesions in a wide spectrum of tissues. While individuals with CD have an increased risk of breast and thyroid neoplasms, the primary features of CD are hamartomas. The gene for CD has been mapped by linkage analysis to a 6 cM region on the long arm of chromosome 10 at 10q22-23. Loss of heterozygosity (LOH) studies of sporadic follicular thyroid adenomas and carcinomas, both component tumors of CD, have suggested that the putative susceptibility gene for CD is a tumor suppressor gene. Somatic missense and nonsense mutations have recently been identified in breast, prostate, and brain tumor cell lines in a gene encoding a dual specificity phosphatase, PTEN/MMACI, mapped at 10q23.3. Furthermore, germline PTEN/MMACI mutations are associated with CD. In the present study, 20 hamartomas from 11 individuals belonging to ten unrelated families with CD have been examined for LOH of markers flanking and within PTEN/MMACI. Eight of these ten families have germline PTEN/MMACI mutations. LOH involving microsatellite markers within the CD interval, and including PTEN/MMACI, was identified in two fibroadenomas of the breast, a thyroid adenoma, and a pulmonary hamartoma belonging to 3 to 11 (27%) of these patients. The wild-type allele was lost in these hamartomas. Semi-quantitative PCR performed on RNA from hamartomas from three different tissues from a CD patient suggested substantial reduction of PTEN/MMACI RNA levels in all of these tissues. The LOH identified in samples from individuals with CD and the suggestion of allelic loss and reduced transcription in hamartomas from a CD patient provide evidence that PTEN/MMACI functions as a tumor suppressor in CD.

Who is still uninsured in Minnesota? Lessons from state reform efforts.

OBJECTIVE: To describe Minnesota's health care system reform efforts and their implications for other state and national reform initiatives, document the rate of uninsurance in 1990 and 1995 with special attention to childrens' access to health insurance, and examine the effectiveness of MinnesotaCare, a voluntary state-subsidized health care plan, in serving its target population. DESIGN: Three cross-sectional telephone surveys: 2-stage random samples of Minnesotans of all ages in 1990 and 1995 and a stratified random sample of MinnesotaCare enrollees in 1994. PARTICIPANTS: For the 2 statewide surveys, 10310 respondents participated in 1990 and 11519 in 1995; more detailed information was collected on approximately 1600 respondents in each survey. Eight hundred MinnesotaCare enrollees participated in the third survey conducted in 1994. MAIN OUTCOME MEASURE: Changes in rates of uninsurance. RESULTS: While the rate of uninsurance increased at the national level, the point-in-time Minnesota rate remained stable and low at 6% between 1990 and 1995. The proportion of children uninsured for 12 months or more decreased from 5.2% in 1990 to 3.1% in 1995, while the proportion of uninsured single adults remained stable at approximately 11%. There was no evidence that MinnesotaCare enrollees are gaming the program, or that the program has resulted in significant erosion from the private market. CONCLUSIONS: MinnesotaCare has enabled the state to maintain a low rate of uninsurance and has reduced this rate among its primary target: children. The program has been less effective in enrolling single adults, although it may be too early to witness the effects of recent expansions targeting this group. Minnesota's experience suggests that other state and national reform efforts aimed at reducing uninsurance, particularly among children, are likely to be successful.

Germline mutations of the PTEN gene in Cowden disease, an inherited breast and thyroid cancer syndrome.

Cowden disease (CD) is an autosomal dominant cancer predisposition syndrome associated with an elevated risk for tumours of the breast, thyroid and skin. Lhermitte-Duclos disease (LDD) cosegregates with a subset of CD families and is associated with macrocephaly, ataxia and dysplastic cerebellar gangliocytomatosis. The common feature of these diseases is a predisposition to hamartomas, benign tumours containing differentiated but disorganized cells indigenous to the tissue of origin. Linkage analysis has determined that a single locus within chromosome 10q23 is likely to be responsible for both of these diseases. A candidate tumour suppressor gene (PTEN) within this region is mutated in sporadic brain, breast and prostate cancer. Another group has independently isolated the same gene, termed MMAC1, and also found somatic mutations throughout the gene in advanced sporadic cancers. Mutational analysis of PTEN in CD kindreds has identified germline mutations in four of five families. We found nonsense and missense mutations that are predicted to disrupt the protein tyrosine/dual-specificity phosphatase domain of this gene. Thus, PTEN appears to behave as a tumour suppressor gene in the germline. Our data also imply that PTEN may play a role in organizing the relationship of different cell types within an organ during development.

Molecular studies of an ependymoma-associated constitutional t(1;22)(p22;q11.2).

We previously described a patient with a de novo constitutional translocation, t(1;22)(p22;q11.2), who developed a malignant ependymoma at age 5, and we proposed that the translocation predisposed the child to the development of the tumor. As a step toward isolation of a putative cancer gene, we have characterized the breakpoints of the (1;22) translocation at the molecular level. The chromosome 22 breakpoint has been narrowed to a region between ARVCF and D22S264. The chromosome 1 breakpoint has been mapped onto a doubly-linked Whitehead YAC contig by PCR analysis of the STS contents of the patient's derivative chromosomes isolated in somatic cell hybrids. Loss-of-heterozygosity (LOH) studies of the patient's ependymoma and of sporadic ependymomas showed no evidence of consistent loss in the breakpoint regions, suggesting that activation of an oncogene, rather than inactivation of a tumor suppressor gene, is the more likely molecular mechanism involved in this case. The gene for Edg-1, a neurally expressed, seven-segment transmembrane receptor, maps to the region of the chromosome 1 breakpoint but does not appear to be interrupted by the translocation. Molecular characterization of the breakpoint regions reported here represents an important step in the identification of the gene(s) affected by this translocation.

A common region of 10p deleted in DiGeorge and velocardiofacial syndromes.

DiGeorge (DGS, MIM 188400) and velocardiofacial (VCFS, MIM 192430) syndromes may present many clinical problems including cardiac defects, hypoparathyroidism, T-cell immunodeficiency and facial dysmorphism. They are frequently associated with deletions within 22q11.2, but a number of cases have no detectable molecular defect of this region. A number of single case reports with deletions of 10p suggest genetic heterogeneity of DGS. Here we compare the regions of hemizygosity in four patients with terminal deletions of 10p (one patient diagnosed as having hypoparathyroidism and three as DGS) and one patient with a large interstitial deletion (diagnosed as VCFS). Fluorescence in situ hybridization (FISH) analysis demonstrates that these patients have overlapping deletions at the 10p13/10p14 boundary. A YAC contig spanning the shortest region of deletion overlap (SRO) has been assembled, and allows the size of SRO to be approximated to 2 Mb. As with deletions of 22q11, phenotypes vary considerably between affected patients. These results strongly support the hypothesis that haploinsufficiency of a gene or genes within 10p (the DGSII locus) can cause the DGS/VCFS spectrum of malformation.

The effects of work intensity on adolescent mental health, achievement, and behavioral adjustment: new evidence from a prospective study.

This article examines the effects of work intensity on adolescent mental health, academic achievement, and behavioral adjustment. Questionnaire data were collected yearly from an initial panel of 1,000 randomly selected ninth graders (14-15 years old). Consistent with other studies, students who worked at higher intensity engaged in more alcohol use. The methodological strengths of this research (a representative panel studied prospectively over a 4-year period with minimal attrition and an analysis incorporating key control and lagged variables) provide strong evidence that adolescent work fosters alcohol use. The contention that work of high intensity has deleterious effects on mental health, academic achievement, and 2 other indicators of behavioral adjustment did not withstand our stringent tests. However, high school seniors who worked at moderate intensity (1-20 hours per week) had higher grades than both nonworkers and students who worked more hours per week.