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PubMed2003

Sampling natural viral communities from soil for culture-independent analyses.

An essential first step in investigations of viruses in soil is the evaluation of viral recovery methods suitable for subsequent culture-independent analyses. In this study, four elution buffers (10% beef extract, 250 mM glycine buffer, 10 mM sodium pyrophosphate, and 1% potassium citrate) and three enumeration techniques (plaque assay, epifluorescence microscopy [EFM], and transmission electron microscopy [TEM]) were compared to determine the best method of extracting autochthonous bacteriophages from two Delaware agricultural soils. Beef extract and glycine buffer were the most effective in eluting viable phages inoculated into soils (up to 29% recovery); however, extraction efficiency varied significantly with phage strain. Potassium citrate eluted the highest numbers of virus-like particles from both soils based on enumerations by EFM (mean, 5.3 x 10(8) g of dry soil(-1)), but specific soil-eluant combinations posed significant problems to enumeration by EFM. Observations of virus-like particles under TEM gave confidence that the particles were, in fact, phages, but TEM enumerations yielded measurements of phage abundance (mean, 1.5 x 10(8) g of dry soil(-1)) that were about five times lower. Clearly, the measurement of phage abundance in soils varies with both the extraction and enumeration methodology; thus, it is important to assess multiple extraction and enumeration approaches prior to undertaking ecological studies of phages in a particular soil.

Aeromonas↗
PubMed2004

Transport of the harmful bloom alga Aureococcus anophagefferens by oceangoing ships and coastal boats.

It is well established that cyst-forming phytoplankton species are transported in ships' ballast tanks. However, there is increasing evidence that other phytoplankton species which do not encyst are also capable of surviving ballast transit. These species have alternative modes of nutrition (hetero- or mixotrophy) and/or are able to survive long-term darkness. In our studies of no-ballast-on-board vessels arriving in the Great Lakes, we tested for the presence of the harmful algal bloom species Aureococcus anophagefferens (brown tide) in residual (i.e., unpumpable) ballast water using methods based on the PCR. During 2001, the brown tide organism was detected in 7 of 18 ballast water tanks in commercial ships following transit from foreign ports. Furthermore, it was detected after 10 days of ballast tank confinement during a vessel transit in the Great Lakes, a significant result given the large disparity between the salinity tolerance for active growth of Aureococcus (>22 ppt) and the low salinity of the residual ballast water (approximately 2 ppt). We also investigated the potential for smaller, recreational vessels to transport and distribute Aureococcus. During the summer of 2002, 11 trailered boats from the inland bays of Delaware and coastal bays of Maryland were sampled. Brown tide was detected in the bilge water in the bottoms of eight boats, as well as in one live-well sample. Commercial ships and small recreational boats are therefore implicated as potential vectors for long-distance transport and local-scale dispersal of Aureococcus.

DNA, Ribosomal↗
PubMed1996

Enzyme-linked immunosorbent assay-based detection of antibodies to antigenic subtypes of infectious bursal disease viruses of chickens.

An enzyme-linked immunosorbent assay (ELISA) using a fragment of the infectious bursal disease virus (IBDV) VP2 gene expressed in baculovirus was developed. A 944-bp portion of the VP2 gene from the Del-A strain of IBDV was ligated into the pAc360 transfer vector and transfected into baculovirus, Recombinant baculoviruses were identified by dot blot hybridization. The recombinant baculovirus 9A5 expressed a 57-kDa VP2 fusion protein, which was immunoprecipitated. This baculovirus-expressed VP2 was used as an antigen in an ELISA (Ohio State University [OSU]-ELISA). Titers of sera from specific-pathogen-free chickens infected with different strains of IBDV in our laboratory were determined by the OSU-ELISA, commercial ELISAs, and a virus neutralization assay. The results indicate that all sera from the specific-pathogen-free chickens infected with IBDV strains contained high titers of neutralizing antibodies. Each of these antisera also tested positive with the commercial ELISA kits. The OSU-ELISA did not detect antibodies to all strains of IBDV tested. This ELISA detected antibodies to the antigenically similar Delaware variant strains Del-A, Del-E, and GLS but did not detect or detected very poorly antibodies to antigenically heterologous classic IBDV strains STC, D78, and BVM. Although the antiserum to the IN strain of IBDV contained a virus-neutralizing antibody titer, the OSU-ELISA did not detect antibodies in this serum, suggesting IN is antigenically heterologous to Del-A. Titers to the IN strain were detected with the commercial ELISA kits. The OSU-ELISA detected antibodies to the SAL strain, suggesting this virus is antigenically similar to Del-A, which is supported by previously reported vaccination and challenge studies. In conclusion, the OSU-ELISA could be used to detect antibodies to a subgroup of IBDV strains, while commercially available ELISA kits detected antibodies to all the antigenic subtypes of IBDV tested.

Animals↗
PubMed2003

Evidence of Borrelia lonestari DNA in Amblyomma americanum (Acari: Ixodidae) removed from humans.

We used a nested PCR with Borrelia flagellin gene (flaB) primers and DNA sequencing to determine if Borrelia lonestari was present in Amblyomma americanum ticks removed from military personnel and sent to the Tick-Borne Disease Laboratory of the U.S. Army Center for Health Promotion and Preventive Medicine. In our preliminary investigation, we detected Borrelia sequences in 19 of 510 A. americanum adults and nymphs from Ft. A. P. Hill, Va. During the 2001 tick season, the flaB primers were used to test all A. americanum samples as they were received, and 29 of 2,358 A. americanum samples tested individually or in small pools were positive. PCRs with 2,146 A. americanum samples in 2002 yielded 26 more Borrelia-positive samples. The positive ticks in 2001 and 2002 were from Arkansas, Delaware, Kansas, Kentucky, Maryland, New Jersey, North Carolina, Tennessee, and Virginia. The last positive sample of the 2001 season was a pool of larvae. To further investigate larval infection, we collected and tested questing A. americanum larvae from Aberdeen Proving Ground, Md.; 4 of 33 pools (40 larvae per pool) were positive. Infection of unfed larvae provides evidence of the maintenance of B. lonestari by means of transovarial transmission. Sequence analysis revealed that the amplicons were identical to sequences of the B. lonestari flaB gene in GenBank. Despite the low prevalence of infection, the risk of B. lonestari transmission may be magnified because A. americanum is often abundant and aggressive, and many tick bite victims receive multiple bites.

Animals↗
PubMed1992

Two types of mouse (Mus musculus domesticus) Y chromosomes in Quebec.

A Y chromosomal repetitive sequence identified two types of Y chromosomes in mice (Mus musculus domesticus) caught near Ste. Anne de Bellevue, Quebec. One type is apparently identical to the Y chromosome found in Maryland, Delaware, and California, whereas the other type is similar, but not identical, to the Y chromosome present in M.m. poschiavinus, an Alpine race of M.m. domesticus. These findings suggest that the domesticus Y chromosome is highly polymorphic and thus useful for elucidating the relationships among American and European house mouse populations.

Animals↗
PubMed1991

Corporate-sponsored breast cancer screening at the work site: results of a statewide program.

Mobile screening mammography was offered to 3,627 employees of a large corporation in Pennsylvania and Delaware. The examination was available to women employees or employee spouses aged 35 years and older. Women were charged $30 for a standard two-view examination. They also received health education materials on mammography and breast self-examination. The remaining costs of the program were underwritten by the corporation. During this program, 3,627 mammographic studies were performed; 63 biopsies were recommended. Fifty-seven biopsies were performed, and nine cancers were diagnosed. Costs of this program are presented in detail. The authors conclude that mobile screening programs at the work site provide an inexpensive, convenient alternative to more traditional screening programs. The inherent advantages of this program are the low cost, the relative ease with which the examination can be performed, and the positive role that corporate medical personnel assume in encouraging individual and group participation.

Adult↗
PubMed2006

Chicken genomics resource: sequencing and annotation of 35,407 ESTs from single and multiple tissue cDNA libraries and CAP3 assembly of a chicken gene index.

Its accessibility, unique evolutionary position, and recently assembled genome sequence have advanced the chicken to the forefront of comparative genomics and developmental biology research as a model organism. Several chicken expressed sequence tag (EST) projects have placed the chicken in 10th place for accrued ESTs among all organisms in GenBank. We have completed the single-pass 5'-end sequencing of 37,557 chicken cDNA clones from several single and multiple tissue cDNA libraries and have entered 35,407 EST sequences into GenBank. Our chicken EST sequences and those found in public databases (on July 1, 2004) provided a total of 517,727 public chicken ESTs and mRNAs. These sequences were used in the CAP3 assembly of a chicken gene index composed of 40,850 contigs and 79,192 unassembled singlets. The CAP3 contigs show a 96.7% match to the chicken genome sequence. The University of Delaware (UD) EST collection (43,928 clones) was assembled into 19,237 nonredundant sequences (13,495 contigs and 5,742 unassembled singlets). The UD collection contains 6,223 unique sequences that are not found in other public EST collections but show a 76% match to the chicken genome sequence. Our chicken contig and singlet sequences were annotated according to the highest BlastX and/or BlastN hits. The UD CAP3 contig assemblies and singlets are searchable by nucleotide sequence or key word (http://cogburn.dbi.udel.edu), and the cDNA clones are readily available for distribution from the chick EST website and clone repository (http://www.chickest.udel.edu). The present paper describes the construction and normalization of single and multiple tissue chicken cDNA libraries, high-throughput EST sequencing from these libraries, the CAP3 assembly of a chicken gene index from all public ESTs, and the identification of several nonredundant chicken gene sets for production of custom DNA microarrays.

Animals↗
PubMed2004

Deregulated cyclin D1 expression is associated with decreased efficacy of the selective epidermal growth factor receptor tyrosine kinase inhibitor gefitinib in head and neck squamous cell carcinoma cell lines.

PURPOSE: Despite promising initial results, recent Phase III trials of the selective epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor gefitinib ("Iressa"; AstraZeneca, Wilmington, Delaware) in advanced head and neck squamous cell carcinoma (HNSCC) have been equivocal. Cyclin D1, an EGFR target gene, is frequently overexpressed in HNSCC, has been implicated in its pathogenesis, and is strongly associated with poor prognosis in this disease. Therefore, we examined the relationship between deregulated cyclin D1 expression and sensitivity to gefitinib to determine whether this frequently occurring oncogenic change affected the cellular response to gefitinib. EXPERIMENTAL DESIGN: A panel of six EGFR-overexpressing HNSCC cell lines was used to correlate CCND1 gene copy number, cyclin D1 expression, and response to gefitinib. The effect of constitutive overexpression of cyclin D1 was assessed by establishing stably transfected clonal SCC-9 cell lines. RESULTS: Three of six cell lines displayed cyclin D1 amplification and/or overexpression, and these cell lines were resistant to gefitinib. SCC 9 clones overexpressing cyclin D1 continued to proliferate and maintained their S-phase fraction when treated with gefitinib, whereas empty vector control clones and the parental SCC 9 cells were profoundly inhibited and displayed marked reductions in S-phase. The resistance of cyclin D1-overexpressing clones and cyclin D1-amplified cell lines was associated with maintenance of cyclin D1 expression after gefitinib treatment. CONCLUSIONS: These data suggest that deregulated cyclin D1 overexpression may be associated with resistance of HNSCC to EGFR inhibitors. Therefore, the role of cyclin D1 as a marker of therapeutic response and its utility as a prognostic marker in HNSCC warrant additional analysis.

Antineoplastic Agents↗
PubMed1996

An assessment tool for evaluating hospital breastfeeding policies and practices.

In 1991, UNICEF and WHO launched the Baby Friendly Hospital Initiative, a global initiative promoting breastfeeding within hospitals. This article describes the development and use of a detailed questionnaire to acquire baseline data on hospital breastfeeding policies in the southeastern Pennsylvania Delaware Valley and to classify hospitals by their level of implementation of the Baby Friendly Hospital Initiative. The purpose of the questionnaire is twofold: a) to describe and analyze hospitals' breastfeeding policies and practices, and b) to classify hospitals with respect to their level of implementation of each of the Ten Steps and the implementation of the Ten Steps overall.

Breast Feeding↗
PubMed1994

Cardiac patients and spouses: family functioning and emotions.

This study examined the relationships among satisfaction with family function and emotional states of a nonprobability sample of sixty-six myocardial infarction (MI) patients and their spouses who were recruited from hospitals in the Delaware Valley. Patients and spouses completed the Family APGAR and the Affects Balance Scale. It was found that both patients and spouses experienced substantial emotional distress when compared with the norms for nonclinical subjects. They were highly correlated on both measures, particularly for positive emotion and satisfaction with family function (p < .0001). Satisfaction with family function was significantly correlated with positive affect for spouses (p < .05). These results suggest the importance of assessment of satisfaction with family function and emotional status of both MI patient and spouse, with spouses with lower satisfaction with family function being at greater risk for emotional distress.

Adaptation, Psychological↗
PubMed1998

Centralization of histocompatibility laboratories: impact on organ allocation efficiency and outcomes of cadaveric renal transplantation.

This project was undertaken to determine whether centralization of histocompatibility laboratory services for renal transplants performed within eastern Pennsylvania could improve the efficiency of allograft allocation and short-term allograft function. A nonconcurrent cohort study was performed comparing renal allografts transplanted between September 15, 1993, and September 14, 1994, to those transplanted between September 15, 1994, and September 14, 1995. All allografts were procured and allocated by the Delaware Valley Transplant Program, the organ procurement agency in eastern Pennsylvania. Cold preservation time and delayed allograft function were used to measure efficiency of allograft allocation and short term allograft function, respectively. The mean cold preservation time was reduced from 25.08 hours to 20.68 hours (P < 0.001). The percentage of delayed allograft function was 19.9 and 17.4 for the pre- and postcentralization groups, respectively (P = 0.5). Therefore, centralization of histocompatibility tissue typing was a regionally effective process intervention for reducing cold preservation time without adversely impacting short-term graft function. The magnitude of this reduction varied between individual centers. Further investigation is required to determine the effect on long-term allograft function and system wide costs.

Adult↗
PubMed2004

Building a community network for a cancer education/screening program: the process and lessons learned (or relearned).

Similar to many communities around the United States in the late 1990s, Muncie, Indiana, and the surrounding county created a year 2000 committee, called, "You and Year 2000, Inc.: The Muncie and Delaware County Millennium Project" to sponsor, plan, and conduct activities to celebrate the new millennium and ponder the community's past, present, and path to the future. Part of the Committee's planning was to create a legacy of the year-long celebration that would attack a community problem. Partnering with the local hospital, the Committee decided to attack the problem of cancer through a fund-raising campaign to build a cancer center and cancer education/screening program. The purpose of this article is to describe the application of a community organizing/building model used to create a community-wide cancer education/screening program and share the lessons learned (or relearned). The planning process used to develop the education/screening program is one that could be duplicated elsewhere.

Adolescent↗
PubMed2004

ATECAR: An Asian American community-based participatory research model on tobacco and cancer control.

In the past few decades, community-based participatory research, which underscores the indispensable role of the community in all phases of the research process, has been recognized as a viable approach to working constructively with communities to achieve mutually beneficial goals. This article presents a history of the Asian Tobacco Education, Cancer Awareness and Research's pioneering efforts in conducting community-based participatory research among Asian Americans in the Delaware Valley region of Pennsylvania and New Jersey. Information about project background, target populations, and the rationale for the conduct of community-based participatory research in American communities is provided. It also delineates the manner in which the principles of community-based participatory research were applied as guides for the development of partnership infrastructures, research programs, and the challenges and barriers that were encountered. Facilitating factors in partnership building, and implications of employing this model in this ethnically and racially diverse population, are further discussed.

Asian↗
PubMed2005

Development of a cDNA array for chicken gene expression analysis.

BACKGROUND: The application of microarray technology to functional genomic analysis in the chicken has been limited by the lack of arrays containing large numbers of genes. RESULTS: We have produced cDNA arrays using chicken EST collections generated by BBSRC, University of Delaware and the Fred Hutchinson Cancer Research Center. From a total of 363,838 chicken ESTs representing 24 different adult or embryonic tissues, a set of 11,447 non-redundant ESTs were selected and added to an existing collection of clones (4,162) from immune tissues and a chicken bursal cell line (DT40). Quality control analysis indicates there are 13,007 useable features on the array, including 160 control spots. The array provides broad coverage of mRNAs expressed in many tissues; in addition, clones with expression unique to various tissues can be detected. CONCLUSIONS: A chicken multi-tissue cDNA microarray with 13,007 features is now available to academic researchers from http://genomics@fhcrc.org. Sequence information for all features on the array is in GenBank, and clones can be readily obtained. Targeted users include researchers in comparative and developmental biology, immunology, vaccine and agricultural technology. These arrays will be an important resource for the entire research community using the chicken as a model.

Animals↗
PubMed1984

Influence of place of treatment on diagnosis, treatment, and survival in three pediatric solid tumors.

This study examines differences between cancer centers (CC) and noncancer centers (NCC) in terms of management procedures and outcomes for three pediatric solid tumors: Wilms' tumor (N = 147), rhabdomyosarcoma (N = 87), and medulloblastoma (N = 76). Data were derived for the period 1970-1979 from the population-based Greater Delaware Valley Pediatric Tumor Registry maintained at the Children's Cancer Research Center, which routinely collects data on all childhood neoplasms that occur in a 31-county region. Management measures reviewed included the degree to which important pretreatment evaluations were performed, types of therapy used, and extent of follow-up examinations conducted. Outcome variables were three-year disease-free survival and frequency of deaths related to complications of therapy. Differences in three-year disease-free survival between CC and NCC were noted for medulloblastoma (52% v 24%) and rhabdomyosarcoma (48% v 10%), but not for Wilms' tumor (79% v 68%). Among medulloblastoma patients, differences were detected in the frequency of pretreatment evaluations and in the therapy used. The principal management contrast found in rhabdomyosarcoma was that multiagent chemotherapy was used less often in NCC. Wilms' tumor patients were evaluated and treated similarly in the community versus the CC, except for some contrasts in the surgical approach and the frequency of follow-up for the detection of late complications.

Adolescent↗
PubMed1984

Reanalysis of central cervical cord injury management.

Early investigators indicated that conservative management was superior to operative intervention in the treatment of central cord injuries. Their clinical data suggested that operative treatment, in fact, worsened the condition. Recent experience with this clinical entity, however, indicates that in selected patients operative intervention may be of value in improving the rate and degree of motor recovery. A retrospective study of all individuals admitted to our hospital (Delaware Valley Spinal Cord Injury Center) with central cervical spinal cord injury was done (28 patients). One-half had been treated with medical therapy alone (Group I); the others were treated both medically and surgically (Group II). Medical therapy consisted of intravenous mannitol, dexamethasone, and sodium bicarbonate given during the acute phase of the injury. Both groups were immobilized using either a halo or a Philadelphia collar. Criteria for entry into the surgical group were one or both of the following: (a) failure to improve progressively after an initial period of improvement, with persistent compression of neural tissue visualized on myelography and (b) unacceptable instability of the spinal bony elements. The patients were given neurological scores based on the motor power of the major muscle groups. The stability of the spine was scored using the Panjabi-White scale. The two groups were compared using Student's t-test and the two-factor analysis of variance. There was no significant difference in initial neurological scores between the groups. The surgical groups had a higher incidence of instability of the bony elements of the cervical spine, as judged by the Panjabi-White scale.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed2001

Environmental public health surveillance: possible estuary-associated syndrome.

Public health surveillance involves the collection, analysis, and dissemination of data for use in public health practice. A surveillance system includes the capacity to collect and analyze data as well as the ability to disseminate the data to public health agencies that can undertake effective prevention and control activities. An emerging issue in environmental public health surveillance involves human exposure to the toxins produced by microorganisms present in oceans and estuaries. One of these organisms is Pfiesteria piscicida Steidinger & Burkholder, a dinoflagellate found in estuaries along the Atlantic and gulf coasts of the United States. There have been reports of both human illness associated with occupational exposures to concentrated laboratory cultures of P. piscicida and massive fill kills associated with the presence of the organism in rivers and estuaries. These reports, and anecdotal reports from people who worked on rivers where the organism has been found, generated concern that environmental exposures to P. piscicida, similar organisms, or perhaps a toxin or toxins produced by the organism(s), could cause adverse human health effects. To begin to evaluate the public health burden associated with P. piscicida, investigators from the National Center for Environmental Health at Centers for Disease Control and Prevention and health agencies from states along the Atlantic coast collaborated to develop a passive surveillance system for collecting, classifying, and tracking public inquiries about the organism. Specifically, the group developed exposure and symptom criteria and developed data collection and reporting capabilities to capture the human health parameters collectively referred to as possible estuary-associated syndrome (PEAS). The surveillance system was implemented in six states (Delaware, Florida, Maryland, North Carolina, South Carolina, Virginia) beginning in June 1998. From 1 June 1998 through 30 June 2001, the six state health agencies participating in the PEAS surveillance system received 3,859 calls: 3,768 callers requested information and 91 callers reported symptoms. Five individuals have been identified as meeting PEAS criteria.

Animals↗
PubMed2003

Estrogens from sewage in coastal marine environments.

Estrogens are ancient molecules that act as hormones in vertebrates and are biologically active in diverse animal phyla. Sewage contains natural and synthetic estrogens that are detectable in streams, rivers, and lakes. There are no studies reporting the distribution of steroidal estrogens in marine environments. We measured estrogens in sewage, injection-well water, and coastal tropical and offshore tropical water in the Pacific Ocean, western Atlantic Ocean, and Caribbean Sea. Concentrations of unconjugated estrone ranged from undetectable (< 40 pg/L) in the open ocean to nearly 2,000 pg/L in Key West, Florida, and Rehoboth Bay, Delaware (USA); estrone concentrations were highest near sources of sewage. Enzymatic hydrolysis of steroid conjugates in seawater samples indicated that polar conjugates comprise one-half to two-thirds of "total estrone" (unconjugated plus conjugated) in Hawaiian coastal samples. Adsorption to basalt gravel and carbonate sand was less than 20% per week and indicates that estrogens can easily leach into the marine environment from septic fields and high-estrogen groundwater. Of 20 sites (n = 129 samples), the mean values from 12 sites were above the threshold concentration for uptake into coral, indicating that there is a net uptake of anthropogenic steroidal estrogen into these environments, with unknown impacts.

Atlantic Ocean↗