Biomedical subjects

D L Montgomery

Publications and source records attributed to D L Montgomery.

At least 19 recordsLinked to original sources

PubMed2006

Comparison of two automated immunohistochemical procedures for the diagnosis of scrapie in domestic sheep and chronic wasting disease in North American white-tailed deer (Odocoileus virginianus) and mule deer (Odocoileus hemionus).

Two commercially available automated immunohistochemistry platforms, Ventana NexES and DakoCytomation Autostainer Universal Staining System, were compared for diagnosing sheep scrapie and cervid chronic wasting disease. Both automated platforms used the same antiprion protein monoclonal primary antibodies, but different platform-specific linker and amplification reagents and procedures. Duplicate sections of brainstem (at the level of the obex) and lymphoid tissue (retropharyngeal lymph node or tonsil) from the same tissue block were immunostained for the comparison. Examination of 1,020 tissues from 796 sheep revealed 100% concordance of results between the Ventana NexES and DakoCytomation platforms for diagnosing sheep scrapie from lymphoid tissue (103/103 positive; 405/405 negative) and brainstem (120/120 positive; 392/392 negative). Similarly, examination of 1,008 tissues from 504 white-tailed deer revealed 100% concordance between the Ventana NexES and DakoCytomation platforms for diagnosing chronic wasting disease from lymphoid tissue (104/104 positive; 400/400 negative) and brainstem (104/104 positive; 400/400 negative). Examination of 1,152 tissues from 482 mule deer revealed a concordance of 98.6% in lymphoid tissue and 99.9% in brainstem between the Ventana NexES and DakoCytomation platforms for diagnosing chronic wasting disease. The results indicate equivalence or near equivalence between the DakoCytomation and Ventana NexES autostainer platforms for identification of the disease-associated prion protein (PrPd)-positive and PrPd-negative brain and lymphoid tissues in sheep, white-tailed deer, and mule deer.

Animals↗

PubMed2006

Oral transmission of chronic wasting disease in captive Shira's moose.

Three captive Shira's moose (Alces alces shirasi) were orally inoculated with a single dose (5 g) of whole-brain homogenate prepared from chronic wasting disease (CWD)-affected mule deer (Odocoileus hemionus). All moose died of causes thought to be other than CWD. Histologic examination of one female moose dying 465 days postinoculation revealed spongiform change in the neuropil, typical of transmissible spongiform encephalopathy. Immunohistochemistry staining for the proteinase-resistant isoform of the prion protein was observed in multiple lymphoid and nervous tissues. Western blot and enzyme-linked immunosorbent assays provided additional confirmation of CWD. These results represent the first report of experimental CWD in moose.

Administration, Oral↗

PubMed2005

Status spongiosus of white matter in newborn Gelbvieh-cross calves.

Various forms of status spongiosus occur in neonatal cattle, the best characterized of which is due to mutations of the branched-chain alpha-keto acid dehydrogenase complex (BCKD), resulting in bovine maple syrup urine disease (MSUD, branched-chain ketoaciduria). A distinctive neurological syndrome was identified between 1998 and 2003 in 9 calves in a 250-cow stabilized Gelbvieh-Red Angus herd. Both sexes were affected (6 heifers, 3 bulls), with a low annual incidence (3 cases in 1998; no cases in 1999; 2 cases in 2000; 2 in 2001; 1 in 2002; 1 in 2003). Affected calves were born full-term, unable to stand, and had constant whole-body tremors when stimulated. Animals remained in lateral recumbency until death or euthanasia; the longest survival time was 10 days postpartum. The principal histological change in 2 affected calves was diffuse, moderately severe bilaterally symmetrical status spongiosus with Alzheimer type II cells throughout the white matter of the brain. Myelin deficits were not evident and vacuoles were due to cleaved myelin sheaths. Neither recognized mutation of MSUD was identified in the E1a subunit of BCKD in 2 affected calves, 8 dams that gave birth to affected calves, a grand-dam of 3 affected calves, or a sire of 1 calf. Amino acid analysis of serum from 1 affected calf revealed normal concentrations of branched-chain amino acids, indicating that this disease is distinct from MSUD. The genetic and biochemical basis for the disorder, provisionally named congenital status spongiosus of Gelbvieh-cross cattle, is undetermined. The pattern of inheritance was not established.

Animals↗

PubMed2005

Primitive neuroectodermal tumor in the spinal cord of a brahman crossbred calf.

A variety of embryonal tumors of the central nervous system, typically malignant and occurring in young individuals, are recognized in humans and animals. This report describes an invasive subdural but predominantly extramedullary primitive neuroectodermal tumor developing at the lumbosacral junction in a 6-month-old Brahman crossbred calf. The tumor was composed of spindloid embryonal cells organized in interlacing fascicles. The cells had oval to elongate or round hyperchromic nuclei, single to double nucleoli, and scant discernible cytoplasm. Immunohistochemical staining for neuron-specific enolase, synaptophysin, and S-100 protein and formation of pseudorosettes suggested neuronal and possibly ependymal differentiation.

Animals↗

PubMed2003

Effect of hyperbaric oxygen on oxygen uptake and measurements in the blood and tissues in a normobaric environment.

OBJECTIVE: To examine venous partial pressure of oxygen (PvO(2)), transcutaneous oxygen tension (tcPO(2)), and VO(2)MAX in a normobaric environment after a single hyperbaric oxygen (HBO(2)) treatment. METHODS: This was a prospective study of conditions after the intervention compared with baseline. The participants were 10 moderately trained (VO(2)MAX = 57.6 ml/kg/min) men. Two HBO(2) treatments consisting of breathing 95% oxygen at 2.5 atmospheres absolute (ATA) for 90 minutes were administered on non-consecutive days. Baseline testing included measures of VO(2)MAX, tcPO(2), and anthropometry. At 6.0 (1.0) minutes after the first HBO(2) treatment, a VO(2)MAX test was performed. After the second HBO(2) treatment, leg and chest tcPO(2) and PvO(2) were monitored for 60 minutes. RESULTS: VO(2)MAX, running time, and peak blood lactate were not altered after the HBO(2) treatment. Leg tcPO(2) was lower (p = 0.003) and chest tcPO(2) was unchanged after the HBO(2) treatment compared with baseline values. PvO(2) was significantly (p<0.001) lower in the first three minutes after treatment than subsequent values, but no other differences were found. CONCLUSIONS: A single HBO(2) treatment at 2.5 ATA for 90 minutes does not raise PvO(2), tcPO(2), or VO(2)MAX in a normobaric, normoxic environment.

Adult↗

PubMed2003

A comparison of skating economy on-ice and on the skating treadmill.

The purpose of this study was to compare skating economy and oxygen uptake (VO2) on-ice and on the skating treadmill (TM). Male varsity hockey players (n = 15, age = 21.0 yr) performed skating tests on a TM and on-ice. The subjects skated for 4 min at each of 3 submaximal velocities (18, 20, and 22 km . h(-1)), separated by 5 min of passive recovery. A VO2max test followed the submaximal tests and commenced at 24 km . h(-1) with the velocity increasing by 1 km . h(-1) every minute until volitional fatigue. VO2 was 39.7, 42.9, 46.0, and 53.4 ml . kg(-1) . min(-1) at 18, 20, 22, and maximum speed (km . h(-1)) on the TM. VO2 was significantly lower (p < .05) 31.5, 36.9, and 42.7 ml . kg(-1) . min(-1) at 18, 20, and 22 km . h(-1) on-ice. The on-ice VO2max (54.7 ml . kg(-1) . min(-1)) was similar to TM. Stride rate, stride length and heart rate (HR) were significantly different on-ice compared to TM. These results show that at submaximal velocities, VO2, HR, and stride rate are higher on TM compared to on-ice. VO2max was similar while HRmax was higher on the skating treadmill compared to on-ice

Adult↗

PubMed2002

Malignant catarrhal fever in a bison (Bison bison) feedlot, 1993-2000.

A fatal enteric syndrome was identified in American bison (Bison bison) at a large feedlot in the American Midwest in early 1998. An estimated 150 bison died of the syndrome between January 1998 and December 1999. The syndrome was identified as malignant catarrhal fever (MCF), primarily the alimentary form. Clinical onset was acute, and most affected bison died within 1-3 days; none recovered. Consistent lesions were hemorrhagic cystitis, ulcerative enterotyphlocolitis, and arteritis-phlebitis. Vasculitis was milder and more localized than that in cattle with MCF, and in contrast to the situation in cattle, lymphadenomegaly was minimal. Virtually all affected bison examined were positive for ovine herpesvirus 2 (OvHV-2) by polymerase chain reaction (PCR) assay. A retrospective study of archived tissues established that MCF occurred in the yard as early as 1993. A prospective study was undertaken to establish the importance of MCF relative to other fatal diseases at the feedlot. The fate of a group of 300 healthy male bison in a consignment of 1,101 animals was followed for up to 7 months to slaughter. At entry, 23% (71/300) of bison were seropositive for MCF viruses, and 11% (8/71) of these seropositive bison were PCR positive for OvHV-2. Forty seronegative bison were selected at random from the group, and all were PCR negative for OvHV-2. There was no change in seroprevalence in the group during the investigation. The minimum infection rate for MCF virus was 36.3% (93/256). Twenty-two (7.3%) of the 300 bison in the feedlot died. Of these, 15 had MCF, 4 had acute or chronic pneumonia, and 3 were unexamined. Losses in the entire consignment were higher (98/1,101; 8.8% death loss); 76% of deaths were attributable to MCF. The study failed to reveal a relationship between subclinical infection and development of clinical disease.

Animals↗

PubMed2001

Antigen levels and antibody titers after DNA vaccination.

DNA vaccination generates strong cellular and humoral immunity in animal models. The mechanisms by which plasmid DNA uptake and expression after intramuscular injection lead to immune responses are not well understood. In particular, the importance of antigen expression levels on subsequent antibody immune responses has not been established. We found that a chemiluminescent assay for alkaline phosphatase allows measurement of antigen levels of secreted alkaline phosphatase (SEAP) in vivo after intramuscular injection of a wide range of plasmid doses. The mice produced antibodies to the alkaline phosphatase reporter gene and both antigen levels and antibody titers were measured over time. We found that the correlation between initial antigen level and antibody response was high (r = 0.74, p < 0.001) and remained high even after accounting for the dose of plasmid injected (r = 0.61, p < 0.001). The correlation between DNA dose and antibody titer was statistically significant (r = 0.53, p < 0.001) but was reduced to almost zero after we accounted for initial antigen levels.

Alkaline Phosphatase↗

PubMed2000

Structural insights into substrate binding by the molecular chaperone DnaK.

How substrate affinity is modulated by nucleotide binding remains a fundamental, unanswered question in the study of 70 kDa heat shock protein (Hsp70) molecular chaperones. We find here that the Escherichia coli Hsp70, DnaK, lacking the entire alpha-helical domain, DnaK(1-507), retains the ability to support lambda phage replication in vivo and to pass information from the nucleotide binding domain to the substrate binding domain, and vice versa, in vitro. We determined the NMR solution structure of the corresponding substrate binding domain, DnaK(393-507), without substrate, and assessed the impact of substrate binding. Without bound substrate, loop L3,4 and strand beta3 are in significantly different conformations than observed in previous structures of the bound DnaK substrate binding domain, leading to occlusion of the substrate binding site. Upon substrate binding, the beta-domain shifts towards the structure seen in earlier X-ray and NMR structures. Taken together, our results suggest that conformational changes in the beta-domain itself contribute to the mechanism by which nucleotide binding modulates substrate binding affinity.

Adenosine Triphosphatases↗

PubMed2000

Acute effects of intense interval training on running mechanics.

The aims of this study were to determine if there are significant kinematic changes in running pattern after intense interval workouts, whether duration of recovery affects running kinematics, and whether changes in running economy are related to changes in running kinematics. Seven highly trained male endurance runners (VO2max = 72.3+/-3.3 ml x kg(-1) x min(-1); mean +/- s) performed three interval running workouts of 10 x 400 m at a speed of 5.94+/-0.19 m x s(-1) (356+/-11.2 m x min(-1)) with a minimum of 4 days recovery between runs. Recovery of 60, 120 or 180 s between each 400 m repetition was assigned at random. Before and after each workout, running economy and several kinematic variables were measured at speeds of 3.33 and 4.47 m x s(-1) (200 and 268 m x min(-1)). Speed was found to have a significant effect on shank angle, knee velocity and stride length (P < 0.05). Correlations between changes pre- and post-test for VO2 (ml x kg(-1) x min(-1)) and several kinematic variables were not significant (P > 0.05) at both speeds. In general, duration of recovery was not found to adversely affect running economy or the kinematic variables assessed, possibly because of intra-individual adaptations to fatigue.

Adaptation, Physiological↗

PubMed2000

Tuberculosis vaccine design: influence of the completed genome sequence.

Tuberculosis continues to be a major health problem, with more adults dying from Mycobacterium tuberculosis than any other pathogen world-wide. With the onset of the HIV epidemic and an increase in drug-resistant M. tuberculosis strains, the need for an improved vaccine has become an international priority. The recent completion of the genome sequences for two M. tuberculosis strains provides a wealth of information that can be used to design new strategies for vaccine development. The challenge comes in making rational choices from among the 4,000 genes of the most probable candidate immunogens or virulence genes.Thus, a well-designed screen is needed to reduce the number of candidates that must be tested. Presently, the most valuable role that bioinformatics can play is to provide such a screen.

Animals↗

PubMed2000

Leukoencephalitis and vasculitis with perivascular demyelination in a Weimaraner dog.

Aseptic and noninfectious diseases of the central nervous system are being recognized with increasing frequency. After multiple episodes of neurologic illness, this 7-year-old Weimaraner dog was euthanatized and submitted for postmortem examination. Lesions in the central nervous system were found mainly in the white matter of the cerebral cortex and cervical spinal cord and represented acute and more chronic injury. Necrotizing vasculitis with fibrinoid change and a marked neutrophilic infiltrate dominated the acute lesions. More chronic changes consisted of perivascular demyelination and accumulation of foamy macrophages with positive staining for myelin. An immune-complex (Arthus-type) vasculitis is suspected.

Animals↗

PubMed1999

Mutations in the substrate binding domain of the Escherichia coli 70 kDa molecular chaperone, DnaK, which alter substrate affinity or interdomain coupling.

In Escherichia coli, DnaK is essential for the replication of bacteriophage lambda DNA; this in vivo activity provides the basis of a screen for mutations affecting DnaK function. Mn PCR was used to introduce mutations into residues 405-468 of the C-terminal polypeptide-binding domain of DnaK. These mutant proteins were screened for the ability to propagate bacteriophage lambda in the background of a dnaK deficient cell line, BB1553. This initial screen identified several proteins which were mutant at multiple positions. The multiple mutants were further dissected into single mutants which remained negative for lambda propagation. Four of these single-site mutants were purified and assayed for biochemical functionality. Two single-site mutations, F426S and S427P, are localized in the peptide binding site and display weakened peptide binding affinity. This indicates that the crystallographically determined peptide binding site is also critical for in vivo lambda replication. Two other mutations, K414I and N451K, are located at the edge of the beta-sandwich domain near alpha-helix A. The K414I mutant binds peptide moderately well, yet displays defects in allosteric functions, including peptide-stimulated ATPase activity, ATP-induced changes in tryptophan fluorescence, ATP-induced peptide release, and elevated ATPase activity. The K414 position is close in tertiary structure to the linker region to the ATPase domain and reflects a specific area of the peptide-binding domain which is necessary for interdomain coupling. The mutant N451K displays defects in both peptide binding and allosteric interaction.

Adenosine Triphosphatases↗

PubMed1999

Effects of hyperbaric oxygen on aerobic performance in a normobaric environment.

The purpose was to examine the acute effects of a hyperbaric oxygen (HBO2) treatment on a) recovery following prolonged exercise and b) aerobic performance in a trained population. Subjects were six male and six female, trained runners with mean Vo2max values of 64.6 +/- 5.6 and 51.9 +/- 6.6 ml x kg(-1) x min(-1), respectively. Subjects performed four exercise-HBO2 conditions in random order: a) control, b) exercise-no HBO2, c) no exercise-HBO2; and d) exercise-HBO2. Exercise was a 90-min run at 75-80% of Vo2max x HBO2 treatments consisted of breathing 95% O2 at 2.5 atm abs for 90 min. At the end of each condition, aerobic performance was assessed with a VO2max test and by the oxygen cost of running on a treadmill at three submaximal velocities. Recovery was not enhanced following a single HBO2 treatment at 2.5 atm abs for 90 min, nor did it alter submaximal or maximal running performance.

Adult↗

PubMed1998

DNA vaccines against tuberculosis.

DNA plasmids encoding Mycobacterium tuberculosis antigen 85 (Ag85) were tested as vaccines in animal models. Ag85 DNA induced relevant immune responses (i.e. T helper (Th) cells, Th1 cytokines and cytotoxic T lymphocytes) and was protective in mouse and guinea pig models of mycobacterial disease. Therefore, DNA vaccination holds promise as an effective means of preventing tuberculosis in humans. Furthermore, this technique is amenable to identifying the protective antigens of M. tuberculosis.

Acyltransferases↗

PubMed1998

Effect of intense interval workouts on running economy using three recovery durations.

The purposes of this study were to determine whether running economy (RE) is adversely affected following intense interval bouts of 10 x 400-m running, and whether there is an interaction effect between RE and recovery duration during the workouts. Twelve highly trained male endurance athletes [maximal oxygen consumption; VO2max = 72.5 (4.3) ml x kg(-1) x min(-1) mean (SD)] performed three interval running workouts of 10 x 400 m with a minimum of 4 days between runs. Recovery duration between the repetitions was randomly assigned at 60, 120 or 180 s. The velocity for each 400-m run was determined from a treadmill VO2max test. The average running velocity was 357.9 (9.0) m x min(-1). Following the workout, the rating of perceived exertion (RPE) increased significantly (P < 0.01) as recovery duration between the 400-m repetitions decreased (14.4, 16.1, and 17.7 at 180s, 120s, and 60 s recovery, respectively). Prior to and following each workout, RE was measured at speeds of 200 and 268 m x min(-1). Changes in RE from pre- to post-workout, as well as heart rate (HR) and respiratory exchange ratio (R) were similar for the three recovery conditions. When averaged across conditions, oxygen consumption (VO2) increased significantly (P < 0.01) from pre- to post-test [from 38.5 to 40.5 ml x kg(-1) x min(-1) at 200 m x min(-1), and from 53.1 to 54.5 ml x kg(-1) x min(-1) at 268 m x min(-1), respectively]. HR increased (from 124 to 138, and from 151 to 157 beats x min(-1) respectively) and R decreased (from 0.90 to 0.78, and from 0.93 to 0.89, respectively) at 200 and 268 m x min(-1), respectively (P < 0.01). This study showed that RE can be perturbed after a high-intensity interval workout and that the changes in VO2, HR and R were independent of the recovery duration between the repetitions.

Adolescent↗

PubMed1998

Analysis of plasmid DNA from a pharmaceutical perspective.

The advent of gene therapy and polynucleotide-based vaccines has resulted in the use of plasmid DNA as a drug substance. Although biologically (cell or animal) based assays must currently be employed to establish the identity and potency of such drugs, we argue that in the future, a combination of microchip-based mutation detection devices combined with an array of chromatographic, electrophoretic, hydrodynamic, and spectroscopic methods can be employed to rigorously establish these properties. We review a variety of such methods in this context and also consider the issue of the chemical stability of plasmids. Extensive comparison is made to protein-based pharmaceuticals with the unique importance of polynucleotide sequence emphasized in comparison to protein tertiary structure.

Chemistry Techniques, Analytical↗

PubMed1998

A randomized controlled trial of the effect of naproxen on delayed onset muscle soreness and muscle strength.

OBJECTIVE: To determine the effect of naproxen in attenuating the symptoms (muscle soreness level) and signs (plasma creatine kinase [CK] activity and muscular strength decrement) of delayed onset muscle soreness (DOMS) induced by repeated bouts of eccentric exercise. DESIGN: The design was a randomized, double-blind, placebo-controlled, crossover trial with two testing phases of 8 days' duration that were separated by a "washout" period of 7 days. SETTING: University-based sports science center. PARTICIPANTS: Twenty healthy male volunteers who responded to a notice in the university's athletic complex. INTERVENTIONS: Eccentric single-leg exercises were performed on days 1, 3, and 4 to induce DOMS in the quadriceps muscles. Naproxen or placebo tablets (500 mg) were taken orally twice per day beginning on day 2 and continuing until the end of the testing phase. MAIN OUTCOME MEASURES: Perception of muscle soreness and knee extensor torque were evaluated daily throughout each phase. Plasma CK levels were evaluated on days 1, 3, 6, and 8 of each phase. RESULTS: After the eccentric exercise, plasma CK levels were similarly elevated in both naproxen and placebo conditions (F=1.42; p=0.25). After DOMS developed, naproxen reduced the perception of soreness on day 3, when muscle soreness was highest (F=2.20; p=0.04). After treatments with naproxen, peak quadriceps torque during leg extension at 60 degrees/s was higher than that after treatment with the placebo (F=4.77; p=0.04). There were no significant differences between the naproxen and placebo conditions for leg extension at 180 degrees/s (F= 1.66; p=0.21) and 300 degrees/s (F=0.71; p=0.41). CONCLUSION: The data indicate that therapeutic doses of naproxen do not prevent CK release into the plasma but decrease the perception of muscle soreness and positively influence quadriceps peak torque.

Adult↗