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D Paine

Publications and source records attributed to D Paine.

13 recordsLinked to original sources

Molecular characterization of fluoroquinolone-resistant Campylobacter spp. isolated from poultry.

Campylobacteriosis, an infectious disease caused by Campylobacter jejuni and Campylobacter coli, is treated by fluoroquinolone antibiotics in clinical practices. However, use of these drugs in animal husbandry may select for fluoroquinolone-resistant campylobacters and, thereby, compromise the clinical treatment of infection. In this study, 21 fluoroquinolone-resistant campylobacters were isolated from poultry samples. Morphological and biochemical characteristics indicated that 19 isolates were C. jejuni and two were C. coli. All isolates were resistant to multiple antibiotics but sensitive to chloramphenicol and gentamicin. These isolates were characterized at the molecular level by amplifying the flagellin gene (flaA) by PCR. The PCR protocol amplified a 1.7-kb flaA gene from all isolates. RFLP analysis of the 1.7-kb amplicons after digestion with DdeI yielded four distinct patterns. The 21 fluoroquinolone-resistant campylobacter isolates were further characterized by pulsed-field gel electrophoresis (PFGE) and compared with the PFGE patterns of nine fluoroquinolone-sensitive campylobacter strains. Four of the 21 fluoroquinolone-resistant isolates were untypable by the PFGE protocol. The PFGE analysis with SalI or SmaI indicated that seven or five, respectively, of the 17 resistant isolates had identical macrorestriction profiles (mrps). However, PFGE analysis with a combination of SalI and SmaI indicated that four of the 17 isolates had similar macrorestriction profiles. The PFGE patterns of the 17 fluoroquinolone-resistant isolates were different from the nine sensitive campylobacter strains.

Animals↗

Ankle biomechanics during four landing techniques.

PURPOSE: An understanding of landing techniques is important for the prevention of injuries in a number of athletic events. There is a risk of injury to the ankle during landings, and the kinematics and forces involved in different landing strategies may be related to the occurrence of trauma. METHODS: In the current study, four drop conditions from a 30.48-cm (12-inch) height were tested. The conditions were a) BN: Bent knee (self-selected), Natural (self-selected) plantar flexor contraction; b) SN: Stiff-knee, Natural plantar flexors; c) SP: Stiff-knee, Plantar flexors absorbing the impact; and d) SH: Stiff-knee, absorbing most of the impact in the Heels. Peak vertical forces and accelerations were measured, and Achilles tendon forces and stiffnesses were calculated. RESULTS: Peak vertical forces and peak tibial accelerations were highest for the SH condition (2418 N and 20.7 G), whereas peak Achilles tendon force was highest for SP drops. The overall average AT stiffness was 166,345 N x m(-1). CONCLUSIONS: The results from the study were used in an extensive cadaver study to investigate ankle injuries. The data from the current study indicate that athletes may not use their full energy absorbing potential in landings during sporting activities.

Achilles Tendon↗

Validation of a polymerase chain reaction method for the detection of rendered bovine-derived materials in feedstuffs.

This study validated a polymerase chain reaction-based method for the detection of a specific bovine mitochondrial gene derived from rendered bovine tissues and admixed with complete animal feed. Four laboratories participated in this effort: one state laboratory and three Food and Drug Administration (FDA) laboratories, including one FDA field laboratory. The protocol used a statistical approach of 90% probability, with a 95% confidence interval for determining acceptable rates of false-positive and false-negative samples. Each participating laboratory analyzed 30 samples of feed each containing 0, 0.125, and 2.0% bovine meat and bone meal (BMBM), for a total of 90 feed samples. The samples were randomized such that the analysts were unaware of the true identity of the test samples. The results demonstrated that all laboratories met the acceptance criteria established for this protocol. The overall rates of false-negative results were 0.83% (1/120) at the level of 0.125% BMBM and 1.67% (2/120) at the level of 2% BMBM. The overall rate of false-negative results for all levels of BMBM was 1.25% (3/240). The rate for false-positive results was 0.83%.

Animal Feed↗

A rapid method for PCR detection of bovine materials in animal feedstuffs.

Rapid identification of bovine materials in animal feedstuffs is essential for effective control of a potential source of bovine spongiform encephalopathy. We have developed a rapid method for the detection of the presence of bovine materials in animal feeds. Animal feed samples were prepared by a Chelex-100 treatment method, then subjected to polymerase chain reaction (PCR) detection. The assay can be completed in 2 h including 30 min for sample preparation, 35-65 min for PCR cycling and 30 min for gel electrophoresis. This method is not only rapid, simple and consistent, but also avoids a hazardous waste disposal issue associated with a previously described guanidine thiocyanate (GuSCN) extraction-PCR method.

Animal Feed↗

Potential role for hyaluronan and the hyaluronan receptor RHAMM in mobilization and trafficking of hematopoietic progenitor cells.

Although the mechanism(s) underlying mobilization of hematopoietic progenitor cells (HPCs) is unknown, detachment from the bone marrow (BM) microenvironment and motility are likely to play a role. This work analyzes the motile behavior of HPCs and the receptors involved. CD34(+)45(lo/med)Scatterlo/med HPCs from granulocyte colony-stimulating factor (G-CSF)-mobilized blood and mobilized BM were compared with steady-state BM for their ability to bind hyaluronan (HA), their expression of the HA receptors RHAMM and CD44, and their motogenic behavior. Although RHAMM and CD44 are expressed by mobilized blood HPCs, function blocking monoclonal antibodies (MoAbs) identified RHAMM as a major HA binding receptor, with a less consistent participation by CD44. Permeabilization of mobilized blood HPCs showed a pool of intracellular (ic) RHAMM and a smaller pool of icCD44. In contrast, steady-state BM HPCs have significantly larger pools of icRHAMM and icCD44. Also, in contrast to mobilized blood HPCs, for steady-state BM HPCs, MoAbs to RHAMM and CD44 act as agonists to upregulate HA binding. The comparison between mobilized and steady-state BM HPCs suggests that G-CSF mobilization is associated with depletion of intracellular stores of HA receptors and modulates HA receptor usage. To confirm that mobilization alters the HA receptor distribution and usage by HPCs, samples of BM were collected at the peak of G-CSF mobilization in parallel with mobilized blood samples. HA receptor distribution of mobilized BM HPCs was closely matched with mobilized blood HPCs and different from steady-state BM HPCs. Mobilized BM HPCs had lower pools of icHA receptors, similar to those of mobilized blood HPCs. Treatment of mobilized BM HPCs with anti-RHAMM MoAb decreased HA binding, in contrast to steady-state BM HPCs. Thus, G-CSF mobilization may stimulate an autocrine stimulatory loop for HPCs in which HA interacts with basal levels of RHAMM and/or CD44 to stimulate receptor recycling. Consistent with this, treatment of HPCs with azide, nystatin, or cytochalasin B increased HA binding, implicating an energy-dependent process involving lipid rafts and the cytoskeleton. Of the sorted HPCs, 66% were adherent and 27% were motile on fibronectin plus HA. HPC adherence was inhibited by MoAbs to beta1 integrin and CD44, but not to RHAMM, whereas HPC motility was inhibited by MoAb to RHAMM and beta1 integrin, but not to CD44. This finding suggests that RHAMM and CD44 play reciprocal roles in adhesion and motility by HPCs. The G-CSF-associated alterations in RHAMM distribution and the RHAMM-dependent motility of HPCs suggest a potential role for HA and RHAMM in trafficking of HPCs and the possible use of HA as a mobilizing agent in vivo.

Blood Component Removal↗

Secondary prevention of coronary heart disease in primary care: a healthy heart initiative.

We assessed the effectiveness of secondary prevention of coronary heart disease (CHD) in primary care, in a cross-sectional study of 1015 patients aged < 75 years with documented CHD. Patients records were examined for documentation of CHD risk factors; 722 patients then attended education sessions where blood pressure and cholesterol were measured, a supervised questionnaire detailing modifiable risk factors was completed, and advice on lifestyle modification was given. Management of risk factors was generally poor, and was worse in women. Approximately 20% of subjects remained hypertensive, with half of these receiving anti-hypertensive medication. Examining the primary care records, serum cholesterol was documented in 17.5% of men and 26.5% of women. Of the 722 subjects who had cholesterol measured, 30% of men and 25% of women had cholesterol < 5.2 mmol/l. Mean cholesterol was significantly higher in the women (6.1 mmol/l vs. 5.6 mmol/l, p = 0.001). Lifestyle risk management was also poor, with significant numbers smoking and drinking more than recommended. Women were more overweight than men (mean BMI 27.9 kg/m2 vs. 26.9 kg/m2, p = 0.006). Aspirin was being taken by 56% of patients.

Antihypertensive Agents↗

Multidrug transporter P-glycoprotein 170 as a differentiation antigen on normal human lymphocytes and thymocytes: modulation with differentiation stage and during aging.

P-glycoprotein 170 (P-gp), the multidrug transport pump, excludes drugs from the interior of cells and is inhibited by agents such as cyclosporin A (CsA), verapamil, and FK-506, which are also substrates for the P-gp pump. This work documents the age- and differentiation-related changes in P-gp on T and B lymphocytes from human blood or spleen, and its absence on most thymus and bone marrow cells. Analysis of rhodamine 123 (Rh123) dye efflux, and its inhibition by cyclosporin A, was used as a quantitative measure of functional P-gp, and reactivity with MRK-16 was used as a measure of P-gp surface expression. The dye efflux and phenotypic expression of P-gp+ PBMC appeared equivalent to that of a moderately drug-resistant cell line, although efflux is prolonged. The sensitivity to inhibition by CsA, cyclosporin G (CsG), and PSC833 of P-gp on PBMC, thymocytes, or T-cell lines varied with apparent cell-type specificity. Normal blood and splenic T- or B-cells included 50-80% of cells with surface P-gp (MRK-16+), which mediated CsA-sensitive dye export. The proportion of P-gp+ T- and B-cells was lowest among children under age 10 years, increased in adulthood, and decreased after age 60. Thymus included 30% of P-gp+ cells mediating CsA-sensitive dye export, including CD3-4-8- progenitors and mature CD3hi CD4+8- or CD4-8+ thymocytes. Mature T-cells in cord or adult blood, spleen, and bone marrow included a large proportion (50-60%) with efficient CsA-sensitive dye export, preferentially among the CD45RA+ subset. Monocytes from all tissue sources, and most bone marrow cells, expressed surface P-gp but retained Rh123, suggesting the absence of a functional dye export mechanism. In vitro mitogen-stimulated PBMC T and B lymphocytes lost P-gp function within 4-24 hr, consistent with the observation that P-gp was reduced on antigen-experienced CD45R0+ T-cells in vivo. Drug export by P-gp may protect lymphocytes from toxic effects of CsA, and may contribute to the immunosuppressive effects of such drugs. The developmentally regulated expression of P-gp function on lymphocytes, and its modulation on activated T- or B-cells, suggest an important role in normal immune development.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Impact on empiric treatment of group A streptococcal pharyngitis using an optical immunoassay.

The potential impact of using a rapid diagnostic test (Strep A OIA) on detection and treatment of group A beta-hemolytic streptococcal (GABHS) pharyngitis in a large-volume pediatric and adolescent clinic was examined. Of 519 swabs processed for both culture and the OIA test, 114 were culture-positive for GABHS compared with 133 positive by the OIA test, for an agreement of 94%. OIA test sensitivity compared with culture was 96%, and specificity was 94%. Forty-seven percent of all study patients were empirically placed on antibiotics. In-clinic OIA testing could have reduced inappropriate therapy and been a cost-effective alternative to culture.

Adolescent↗

Training in urology.

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Education, Medical, Graduate↗

Effects of Naturally Occurring Aquatic Organic Fractions on Am Uptake by Scenedesmus obliquus (Chlorophyceae) and Aeromonas hydrophila (Pseudomonadaceae).

Naturally occurring organics were extracted from water collected from Skinface Pond near Aiken, S.C. Organics were separated into four nominal diameter size fractions (I, >0.0183; II, 0.0183 to 0.0032; III, 0.0032 to 0.0009; IV, <0.0009 mum) by membrane ultrafiltration and introduced into Scenedesmus obliquus and Aeromonas hydrophila cultures to determine their effects on Am availability for uptake. Effects on Am uptake were determined in actively growing S. obliquus cultures after 96 h of growth and in dense cultures of nongrowing cells after 4 h. Uptake by A. hydrophila was determined after 4 and 24 h in actively growing cultures. All organic fractions stimulated S. obliquus growth, with the most pronounced effects due to larger organic fractions, whereas no apparent growth stimulation of A. hydrophila was observed for any organic fraction. For both long-term and short-term studies, cellular Am concentration (picocuries/cell) increased with increasing Am concentration for S. obliquus and A. hydrophila. Fraction IV increased Am uptake by both S. obliquus and A. hydrophila during 4-h incubations. During 96-h incubations fraction I was flocculated and cosedimented, with S. obliquus and A. hydrophila cells causing an apparent increase in Am uptake. Fractions II and III reduced apparent Am uptake by S. obliquus as a result of biological dilution caused by increased algal growth due to the organics. Fraction IV caused a reduction in Am uptake by S. obliquus not attributable to biological dilution. Organics increased Am uptake by A. hydrophila during 4- and 24-h incubations. A. hydrophila also caused flocculation of fraction I during 96-h incubations.

Journal Article↗