Reverse Translational Study of Fenofibrate's Observed Effects in Diabetes-Associated Retinopathy.

Clinical trials suggest that fenofibrate reduces the progression of retinopathies in patients with type 2 diabetes. Furthermore, patients with retinopathies have elevated levels of inflammatory chemokines and dysfunctional retinal angiogenesis. Therefore, we investigated the effects of fenofibrate on the production of inflammatory chemokines and genes associated with angiogenesis. Retinal pigment epithelial cells (RPECs) were cultured with IL-1ß and fenofibrate ranging from 1-50 µM. ENA-78, IL-8, and RANTES were measured in cell culture by ELISA. ENA-78, ABCA1, and ABCG1 gene expression were tested by RT-PCR. IL-1ß significantly induced the production of ENA-78, IL-8, and RANTES. Fenofibrate at concentrations of 25-50 uM blunted the IL-1ß induced production of ENA-78 (p < 0.05) with no significant effects on RANTES and IL-8. Fenofibrate also reduced the expression of the ENA-78 gene as well as ABCA1 and ABCG1, which are genes involved in angiogenesis. Fenofibrate decreases ENA-78 production and ABCA1/ABCG1 gene expression in RPECs.

Quantification of sodium pentobarbital residues from equine mortality compost piles.

Sodium pentobarbital, a euthanasia drug, can persist in animal carcasses following euthanasia and can cause secondary toxicosis to animals that consume the remains. This experiment was conducted to observe the effects of composting on euthanized horse carcass degradation and sodium pentobarbital residues in compost material up to 367 d. Six separate compost bins were constructed on pastureland. Three bins served as the control while 3 served as the treatment. The carbonaceous material, or bulking agent, consisted of hardwood chips mixed with yard waste wetted to approximately 50% moisture content. Bulking agent was added to each bin at a depth of 0.46 m, creating the pad. A licensed veterinarian provided 6 horse carcasses for use in the experiment. These horses had required euthanasia for health reasons. All horses were weighed and then sedated with an intravenous injection of 8 mL of xylazine. After sedation the 3 horses in the treatment group were euthanized by intravenous injection of 60 mL of sodium pentobarbital. The 3 control group horses were anesthetized by intravenous injection of 15 mL of ketamine hydrochloride and then humanely euthanized by precise gunshot to the temporal lobe. Following euthanasia, each carcass was placed on the center of the pad and surrounded with 0.6 m of additional bulking agent. Serum and liver samples were obtained immediately following death. Compost samples were obtained on d 7, 14, 28, 56, 84, 129, 233, and 367 while soil samples were obtained on d -1 and 367. Each sample was analyzed for sodium pentobarbital concentration. Compost pile and ambient temperatures were also recorded. Composting successfully degraded soft tissue with only large bones remaining. Data illustrate that sodium pentobarbital was detectable up to 367 d in compost piles with no clear trend of concentration reduction. Drug residues were detected in soil samples indicating that sodium pentobarbital leached from the carcass and through the pad. These findings confirm the persistence of sodium pentobarbital from equine mortality compost piles and emphasize the importance of proper carcass management of animals euthanized with a barbiturate to reduce environmental impact and secondary toxicosis.

A multiplex real-time polymerase chain reaction assay to diagnose Epiphyas postvittana (Lepidoptera: Tortricidae).

A molecular assay for diagnosis of light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae), in North America is reported. The assay multiplexes two TaqMan real-time polymerase chain reaction (RT-PCR) probe systems that are designed to target DNA segments of the internal transcribed spacer region 2 (ITS2) and 18S rRNA gene. The RT-PCR probe designed for the 18S target recognizes a DNA sequence conserved in all of the moths included in the study and functions as a control in the assay. The second probe recognizes a segment of the ITS2 specifically found in E. postvittana and not found in the other moths included in the study, i.e., this segment is not conserved. Inclusion of the two markers in a single multiplex reaction did not affect assay performance. The assay was tested against 637 moths representing > 90 taxa in 15 tribes in all three subfamilies in the Tortricidae. The assay generated no false negatives based on analysis of 355 E. postvittana collected from California, Hawaii, England, New Zealand, and Australia. Analysis of a data set including 282 moths representing 41 genera generated no false positives. Only three inconclusive results were generated from the 637 samples. Spike experiments demonstrated that DNA contamination in the assay can affect samples differently. Contaminated samples analyzed with the ITS2 RT-PCR assay and DNA barcode methodology by using the cytochrome oxidase I gene can generate contradictory diagnoses.

Development of a ribosomal DNA ITS2 marker for the identification of the thrips, Scirtothrips dorsalis.

The thrips Scirtothrips dorsalis Hood (Thysanoptera: Thripidae) is an invasive pest that poses a significant economical threat to U.S. agriculture and trade. In this study, DNA sequence data and polymerase chain reaction (PCR) were utilized to develop a molecular diagnostic marker for S. dorsalis. The DNA sequence variation from the internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA (rDNA) was analyzed from various thrips species, including S. dorsalis. A primer set and polymerase chain reaction cycling parameters were designed for the amplification of a single marker fragment of S. dorsalis ITS2 rDNA. Specificity tests performed on ten thrips species, efficacy tests performed on fifteen S. dorsalis populations, and tests on primer sensitivity and robustness all demonstrated the diagnostic utility of this marker. This diagnostic PCR assay provides a quick, simple, and reliable molecular technique to be used in the identification of S. dorsalis.

The Ribosomal Database Project: improved alignments and new tools for rRNA analysis.

The Ribosomal Database Project (RDP) provides researchers with quality-controlled bacterial and archaeal small subunit rRNA alignments and analysis tools. An improved alignment strategy uses the Infernal secondary structure aware aligner to provide a more consistent higher quality alignment and faster processing of user sequences. Substantial new analysis features include a new Pyrosequencing Pipeline that provides tools to support analysis of ultra high-throughput rRNA sequencing data. This pipeline offers a collection of tools that automate the data processing and simplify the computationally intensive analysis of large sequencing libraries. In addition, a new Taxomatic visualization tool allows rapid visualization of taxonomic inconsistencies and suggests corrections, and a new class Assignment Generator provides instructors with a lesson plan and individualized teaching materials. Details about RDP data and analytical functions can be found at http://rdp.cme.msu.edu/.

The ribosomal database project (RDP-II): introducing myRDP space and quality controlled public data.

Substantial new features have been implemented at the Ribosomal Database Project in response to the increased importance of high-throughput rRNA sequence analysis in microbial ecology and related disciplines. The most important changes include quality analysis, including chimera detection, for all available rRNA sequences and the introduction of myRDP Space, a new web component designed to help researchers place their own data in context with the RDP's data. In addition, new video tutorials describe how to use RDP features. Details about RDP data and analytical functions can be found at the RDP-II website (http://rdp.cme.msu.edu/).

Deep-vein thrombosis in high-energy skeletal trauma despite thromboprophylaxis.

We report the incidence and location of deep-vein thrombosis in 312 patients who had sustained high-energy, skeletal trauma. They were investigated using magnetic resonance venography and Duplex ultrasound. Despite thromboprophylaxis, 36 (11.5%) developed venous thromboembolic disease with an incidence of 10% in those with non-pelvic trauma and 12.2% in the group with pelvic trauma. Of patients who developed deep-vein thrombosis, 13 of 27 in the pelvic group (48%) and only one of nine in the non-pelvic group (11%) had a definite pelvic deep-vein thrombosis. When compared with magnetic resonance venography, ultrasound had a false-negative rate of 77% in diagnosing pelvic deep-vein thrombosis. Its value in the pelvis was limited, although it was more accurate than magnetic resonance venography in diagnosing clots in the lower limbs. Additional screening may be needed to detect pelvic deep-vein thrombosis in patients with pelvic or acetabular fractures.

The Ribosomal Database Project (RDP-II): sequences and tools for high-throughput rRNA analysis.

The Ribosomal Database Project (RDP-II) provides the research community with aligned and annotated rRNA gene sequences, along with analysis services and a phylogenetically consistent taxonomic framework for these data. Updated monthly, these services are made available through the RDP-II website (http://rdp.cme.msu.edu/). RDP-II release 9.21 (August 2004) contains 101,632 bacterial small subunit rRNA gene sequences in aligned and annotated format. High-throughput tools for initial taxonomic placement, identification of related sequences, probe and primer testing, data navigation and subalignment download are provided. The RDP-II email address for questions or comments is rdpstaff@msu.edu.

The Ribosomal Database Project (RDP-II): previewing a new autoaligner that allows regular updates and the new prokaryotic taxonomy.

The Ribosomal Database Project-II (RDP-II) pro-vides data, tools and services related to ribosomal RNA sequences to the research community. Through its website (http://rdp.cme.msu.edu), RDP-II offers aligned and annotated rRNA sequence data, analysis services, and phylogenetic inferences (trees) derived from these data. RDP-II release 8.1 contains 16 277 prokaryotic, 5201 eukaryotic, and 1503 mitochondrial small subunit rRNA sequences in aligned and annotated format. The current public beta release of 9.0 debuts a new regularly updated alignment of over 50 000 annotated (eu)bacterial sequences. New analysis services include a sequence search and selection tool (Hierarchy Browser) and a phylogenetic tree building and visualization tool (Phylip Interface). A new interactive tutorial guides users through the basics of rRNA sequence analysis. Other services include probe checking, phylogenetic placement of user sequences, screening of users' sequences for chimeric rRNA sequences, automated alignment, production of similarity matrices, and services to plan and analyze terminal restriction fragment polymorphism (T-RFLP) experiments. The RDP-II email address for questions or comments is rdpstaff@msu.edu.

The RDP-II (Ribosomal Database Project).

The Ribosomal Database Project (RDP-II), previously described by Maidak et al. [Nucleic Acids Res. (2000), 28, 173-174], continued during the past year to add new rRNA sequences to the aligned data and to improve the analysis commands. Release 8.0 (June 1, 2000) consisted of 16 277 aligned prokaryotic small subunit (SSU) rRNA sequences while the number of eukaryotic and mitochondrial SSU rRNA sequences in aligned form remained at 2055 and 1503, respectively. The number of prokaryotic SSU rRNA sequences more than doubled from the previous release 14 months earlier, and approximately 75% are longer than 899 bp. An RDP-II mirror site in Japan is now available (http://wdcm.nig.ac.jp/RDP/html/index.h tml). RDP-II provides aligned and annotated rRNA sequences, derived phylogenetic trees and taxonomic hierarchies, and analysis services through its WWW server (http://rdp.cme.msu.edu/). Analysis services include rRNA probe checking, approximate phylogenetic placement of user sequences, screening user sequences for possible chimeric rRNA sequences, automated alignment, production of similarity matrices and services to plan and analyze terminal restriction fragment polymorphism experiments. The RDP-II email address for questions and comments has been changed from curator@cme.msu.edu to rdpstaff@msu.edu.

Lymphoscintigraphic technique to image canine esophageal lymph nodes.

Current noninvasive methods of imaging esophageal lymph nodes have an accuracy, specificity, and sensitivity of 70%. Using a flexible esophagoscope, technetium-99m antimony sulfide colloid was injected in the esophageal submucosa of six dogs who then underwent nuclear scans to identify lymph-node location. The euthanized animals underwent dissection of cervical, thoracic, and abdominal lymph nodes. Student's t-test showed no statistical difference in the number of lymph nodes visualized in the neck (3.5 +/- 0.6), parietal thorax (1.2 +/- 0.4), visceral thorax (2.2 +/- 0.7), and abdomen (1.0 +/- 0.0) on premorbid nuclear scans and in the number of radiolabeled lymph nodes found in the neck (3.2 +/- 0.9), parietal thorax (1.2 +/- 0.2), visceral thorax (1.8 +/- 1.0), and abdomen (1.2 +/- 0.2) on dissection of the carcass. The positions of the lymph nodes based on the premorbid nuclear scans matched the locations of the radiolabeled lymph nodes at dissection. Dissected tissue was pathologically confirmed as lymph node. The position and number of lymph nodes in the cervical, intrathoracic, and abdominal regions on nuclear scan correlated with the position and number of lymph nodes found on anatomic dissection. This technique may have a higher sensitivity and specificity than current noninvasive techniques in the staging of esophageal lymphatic metastasis.

Patterns in child and adolescent consumption of fruit and vegetables: effects of gender and ethnicity across four sites.

OBJECTIVES: Few studies have examined the association of gender and ethnicity with fruit and vegetable consumption. We examined these associations using baseline data from four school-based sites funded under the National Cancer Institute's 5 A Day for Better Health Program. METHODS: Diet was measured using 24-hour recalls at three sites and seven-day food records at one site. Demographics were obtained via self-report or school records. Regression analyses for clustered data were employed with fruit and vegetables combined and fruit and vegetables separately. RESULTS: Girls ate more fruit, more vegetables and more fruit and vegetables combined than boys at the Georgia site. Ethnicity was significant in two sites: In Georgia, African-Americans ate more fruit and more fruit and vegetables combined than European-Americans; in Minnesota, Asian-American/Pacific Islanders and African-Americans ate more fruit than European-Americans, and European-Americans and African-Americans ate more vegetables than Asian-Americans. No significant effects were found at the Alabama or Louisiana sites. CONCLUSIONS: Ethnicity was related to fruit and vegetable consumption in Georgia and Minnesota. Consistent with prior studies, gender was related to fruit and vegetable consumption, with girls consuming more servings than boys; however, this was observed at one site only, Georgia. Consumption levels were similar to national estimates for children and varied by region. Further studies are needed using a single methodology to facilitate regional comparisons.

Nutrient intake and food group consumption of 10-year-olds by sugar intake level: the Bogalusa Heart Study.

OBJECTIVE: The effect of total sugar intake on nutrient intake and food group consumption was examined in children. METHODS: Twenty-four hour dietary recalls were collected on a sample of 568 ten-year-olds from two cross-sectional surveys. The population was stratified according to total sugar intake quartiles: < or = 25th (46 g/1000 kcal), 25-50th (67 g/1000 kcal), 50-75th (81 g/1000 kcal) and > or = 75th (106 g/1000 kcal). RESULTS: No significant difference was shown in energy intakes across the quartiles and no significant race or gender differences were observed in mean total sugar intakes. However, with increasing total sugar intake, there was a significant linear decrease in mean intakes of protein, fat, saturated fat, starch, cholesterol, sodium, vitamins B6 and E, thiamin, niacin, iron, and zinc; and a significant linear increase in mean intakes of carbohydrate, fructose, lactose, sucrose, vitamin D, and calcium. Eating patterns reflected the differing nutrient intakes, with high sugar consumers having significantly higher intakes of total g of candy, beverages and milk and lower intakes of total g of meats, and cheese than lower sugar consumers. The nutritional quality of children's diets high in total sugar appear to be adequate regarding vitamin and mineral intakes and are closer to meeting current dietary fat recommendations.

Outcomes of a high school program to increase fruit and vegetable consumption: Gimme 5--a fresh nutrition concept for students.

This paper describes an intervention to increase high school students' fruit and vegetable consumption. Twelve schools were randomized to intervention or control conditions. The cohort (2,213 students; 56% females, 84% Caucasian) were followed from 9th to 12th grades. Interventions comprised a media campaign, classroom workshops, school meal modification, and parental support. Usual daily servings of fruit/vegetables increased 14% in the intervention compared to the control group (p > 0.001) the first three years. At follow-up, consumption within the control group also increased, resulting in no significant difference between groups. Intervention group knowledge scores and awareness indicators were significantly higher than those of the control group (p < 0.0001). Gimme 5 provided a first model to show that dietary habits of high school students can be influenced by positive media messages relative to that age group, increased exposure to a variety of tasty products, and minimal classroom activity.

Information source preferences for selecting dental services among elderly consumers.

This manuscript presents the results of a primary research study conducted among the elderly to determine the sources of information they prefer for the selection of dental services. The results of the study are presented in terms of source preferences by dental need (generalist or specialist), source preferences by gender (male or female), source preferences by level of education (low or high), and source preferences by income level (low or high). Conclusions are drawn from the results of the research, and suggestions are made regarding the use of information source preferences for the marketing of dental services to the elderly.

Development of a school-based nutrition intervention for high school students: Gimme 5.

PURPOSE: To describe a 4-year intervention targeting fruit/vegetable consumption by high school students. DESIGN: This is a cohort study involving six pairs of schools (n = 12) matched on gender, race, enrollment, and location with schools randomly assigned within pairs to intervention or control conditions. SETTING: Twelve Archdiocese of New Orleans high schools. SUBJECTS: Cohort was defined as students (n = 2339) who were ninth-graders in the 1993-94 school year who provided baseline data. INTERVENTION: Four components of the intervention are: (1) school-wide media-marketing campaign, (2) school-wide meal and snack modification, (3) classroom workshops and supplementary subject matter activities, and (4) parental involvement. MEASURES: Focus groups were conducted for target population input and program development. Process evaluation included student feedback on media-marketing intervention materials and activities reported here. Process measures also included school meal participation, student characteristics, and verification of intervention activities. RESULTS: Focus groups identified barriers to increased consumption of fruit and vegetables as lack of availability, variety, and inconsistency in taste. Student attitudes were favorable regarding a school program to improve diet and parental involvement. Low consumption of fruits/vegetables was reported. After a 2-month school-wide program introduction utilizing various media-marketing materials and activities, 93% of students were aware of the program and 96% could identify the healthy eating message. CONCLUSIONS: Program development can be guided and enriched by student input via focus groups. Media-marketing activities effectively delivered health messages and attracted students' attention. Materials and activities used were acceptable channels for increasing awareness, positive attitudes, and knowledge about fruits/vegetables.

Exon 2 containing myelin basic protein (MBP) transcripts are expressed in lesions of experimental allergic encephalomyelitis (EAE).

Exon 2 containing myelin basic protein (MBP) transcripts are expressed during developmental myelination in mice and humans, and during remyelination subsequent to virally induced demyelination in adult mice. Since remyelination characterizes CNS lesions during experimental allergic encephalomyelitis (EAE) and multiple sclerosis (MS), we investigated whether exon 2 containing isoforms of MBP are expressed in EAE lesions during relapsing disease. Exon 2 containing MBP transcripts were detected by in situ hybridization in 17 of 52 EAE mice and in 16 of 30 mice at the peak of the first or second episode of paralysis. Thus exon 2 containing MBP transcripts are expressed in lesions of the CNS during active phases of chronic relapsing autoimmune disease. Implications of these findings with respect to future therapies aimed toward enhancing remyelination in EAE and, possibly MS, are discussed.

Epitope spreading occurs in active but not passive EAE induced by myelin basic protein.

Using experimental allergic encephalomyelitis, EAE, as a model for the study of autoimmune demyelinating disease in the CNS, previous studies have indicated that spread may occur with respect to the specificity of T cell responses during disease. This phenomenon, known as epitope spreading, is central to therapeutic strategies in multiple sclerosis (MS). However, in EAE, the clinical course, neuropathology and immunopathogenesis vary depending upon host factors and the method of disease induction. Since passive EAE in SJL/J mice resembles MS clinically and neuropathologically, this model was chosen to study the immune phenomenon of epitope spreading. T cells specific for whole 18.5 kDa MBP were used to initiate disease since MBP or one of its naturally occurring cleavage fragments may initiate a more physiological immune response than one generated to an artificially designed synthetic peptide. While a progressive increase in T cell responsiveness specific for the immunodominant MBP 87-106 region was observed during disease, there was no evidence of either intermolecular epitope spreading to the immunodominant region of proteolipid protein (PLP) 139-151 or of intramolecular epitope spreading to the exon 2 encoded region of MBP, which is spliced out of 18.5 kDa MBP. In addition there was no shift in immunodominance toward the subdominant MBP 16-35 region during disease. In contrast during active EAE induced by MBP, epitope spreading to the immunodominant epitope of PLP, 139-151, was observed. These data demonstrate that immune responses generated during passive versus active EAE may differ, and suggest that significant epitope spreading does not occur in chronic relapsing demyelinating disease initiated with T cells specific for whole MBP in the absence of exogenous antigen, complete Freund's adjuvant and pertussis. Implications of these findings with regard to epitope spreading in MS are discussed.