Search PubMedSearch

Biomedical subjects

D M Stark

Publications and source records attributed to D M Stark.

At least 19 recordsLinked to original sources

Hemodynamic effects of nasal and face mask continuous positive airway pressure.

Studies of the hemodynamic effects of nasal continuous positive airway pressure (n-CPAP) in normal subjects have had conflicting results. The largest study (n = 19) found no effect of up to 15 cm H2O on heart rate (HR), cardiac stroke volume (SV), or cardiac index. We hypothesized that n-CPAP, by increasing intrathoracic pressure, should decrease SV and cardiac output (CO) in a dose-dependent fashion in normal subjects. We also hypothesized that mouth position, i.e., open or closed, could affect intrathoracic pressure and thus SV and CO. Six normal subjects were tested with four levels of CPAP (5, 10, 15, and 20 cm H2O) under three mask conditions-face mask and nasal mask with the mouth open (mo) or with the mouth closed (mc). Noninvasive pulsed Doppler measurements of SV and HR were made under each condition. N-CPAP (mc) and face mask CPAP (f-CPAP) resulted in significant dose-dependent decreases of SV-24 +/- 5 ml (21%) and 33 +/- 5 ml (28%), respectively--from baseline to 20 cm H2O (p < 0.05). HR were unchanged and CO significantly decreased with n-CPAP(mc) and with f-CPAP, 1.6 +/- 0.38 L/min (23%) and 2.29 +/- 0.54 L/min (31%), respectively, from baseline to 20 cm H2O (p < 0.05). Esophageal pressure measurements verified increasing intrathoracic pressure with increasing levels of f-CPAP and n-CPAP (mc) but not with n-CPAP (mo). In conclusion, n-CPAP (mc) and f-CPAP resulted in significant and similar dose-dependent decreases in SV and CO.

Adult

Hyperoxic training increases work capacity after maximal training at moderate altitude.

High-intensity training may be difficult to sustain due to limitations in systemic oxygen transport, particularly at high altitudes. The purpose of this study was to examine the effects of a high-intensity training protocol using hyperoxic gas breathing in athletes "maximally trained" at an altitude of 1,600 m. Five subjects underwent progressive cycle training until they reached a plateau of aerobic capacity, maximal workload, and endurance time at 85 percent maximal workload. Significant decreases (2 to 6 percent) in arterial oxygen saturation were found after the 85 percent maximal workload tests. Training intensity was then increased to 95 percent maximal workload while the subjects breathed a gas mixture containing at least 70 percent oxygen. After 6 weeks of hyperoxic training, exercise parameters were compared with the plateau values obtained during the baseline training period. Total time during maximal cycle testing increased from 19.1 to 19.6 min (p = 0.015), heart rate at 85 percent maximal workload decreased from 168 to 163 bpm (p = 0.047), and endurance time at 85 percent maximal workload increased from 6.2 to 8.2 min (p = 0.012). There was a trend toward improvement of maximal workload. We conclude that hyperoxic training increases work capacity after attainment of "maximal training" at moderate altitude.

Adult

cDNA cloning and nucleotide sequence of the wheat streak mosaic virus capsid protein gene.

The 3'-terminal region of wheat streak mosaic virus (WSMV) genomic RNA was cloned and a cDNA sequence of 1809 nucleotides upstream of the poly(A) tract was determined. The sequence contains a single open reading frame of 1662 nucleotides and a 3' untranslated region of 147 nucleotides. Translation products from WSMV RNA and WSMV cDNA transcripts were immunoprecipitated by WSMV capsid protein antiserum, indicating that the 3'-terminal region of WSMV RNA encodes the capsid protein. Five potential N-terminal capsid protein protease cleavage sites were identified, which would yield proteins ranging from 31.7K to 46.8K. Alignment of the deduced amino acid sequence of the WSMV capsid protein with those of other potyviruses showed significant, but limited, identity as compared to the alignment of two or more aphid-transmitted potyviruses. Although WSMV has characteristics distinct from potyviruses, because of its particle morphology, translation strategy apparently based on polyprotein processing, the ability to form cytoplasmic cylindrical inclusions and the degree of capsid protein homology with aphid-transmitted potyviruses, it should be considered a member of the potyvirus group.

Amino Acid Sequence

Rigid internal fixation for shoulder arthrodesis.

Fifteen patients underwent a shoulder arthrodesis utilizing standard dynamic compression plate fixation, but with limited postoperative immobilization with only an abduction pillow. In each case, the position of the extremity relative to the scapula and trunk was recorded immediately postoperatively, at regular intervals until fusion, and at follow-up evaluations. Thirteen of 15 shoulders fused without change of intraoperative position after an average postoperative period of 4 months. One patient lost position in the early postoperative period secondary to inadequate fixation, but subsequently fused. Another who demonstrated a persistent non-union at 2 1/2 years was subsequently explored and underwent a bone graft. Four patients complained of residual symptomatic hardware, with two requiring surgical removal of the plate and screws. All but one patient were satisfied with the clinical result at follow up. Only two patients were within 5 degrees of the preoperatively determined position of 30 degrees abduction, 30 degrees forward flexion, and 30 degrees internal rotation. However, almost all were able to function satisfactorily. The authors concluded that shoulder arthrodesis utilizing rigid internal fixation without postoperative cast or brace immobilization maximizes patient comfort without compromising the success of arthrodesis. However, control of arm position remains inexact and additional modifications are needed to ensure fusion position and to minimize disability.

Adolescent

Screening rabbit colonies for antibodies to Pasteurella multocida by an ELISA.

Rabbit serum samples from eleven different research facilities were evaluated for the presence of immunoglobulin G against Pasteurella multocida by using an enzyme-linked immunosorbent assay (ELISA). Each facility which submitted serum samples also provided a brief history of each rabbit colony tested. Rabbits from colonies reported to have endemic P. multocida or of undetermined status had 83 (58.9%) of 141 rabbits that were positive. Colonies reported to be free from P. multocida had 110 (92.4%) of 119 rabbits that were negative by ELISA. The ELISA test described here showed a high degree of agreement (92-94%) with two other P. multocida ELISAs at different diagnostic facilities. This study confirms that an ELISA testing for serum antibodies against the P. multocida is a reliable diagnostic tool to screen colonies for P. multocida.

Animals

Prevalence of feline immunodeficiency virus and feline leukemia virus infections in random-source cats.

Retroviral serologic profiles were generated for 506 random-source cats (Felis catus) that were received by our facility during a twenty-month period. Feline leukemia virus antigens were detected in plasma samples from 26 (5.1%) of the cats. Antibodies to feline immunodeficiency virus were present in 24 (4.7%) of the samples tested. A single cat (0.2%) was positive for both viruses. Neither gender nor vendor correlation with retroviral seropositivity could be demonstrated.

Animals

Protection against tobacco mosaic virus in transgenic plants that express tobacco mosaic virus antisense RNA.

Transgenic tobacco plants that express RNA sequences complementary to the tobacco mosaic virus (TMV) coat protein (CP) coding sequence with or without the tRNA-like structure at the 3' end of the TMV RNA were produced. Progeny of self-pollinated plants were challenged with TMV to determine their resistance to infection. Plants that expressed RNA sequences complementary to the CP coding region and the 3' untranslated region, including the tRNA-like sequences, were protected from infection by TMV at low levels of inoculum. However, plants that expressed RNA complementary to the CP coding sequence alone were not protected from infection. These results indicate that sequences complementary to the terminal 117 nucleotides of TMV, which include a putative replicase binding site, are responsible for the protection. However, the level of protection in these plants was considerably less than in transgenic plants that expressed the TMV CP gene and accumulated CP. Since the mechanisms of protection in the two systems are different, it may be possible to increase protection by introducing both sequences into transgenic plants.

Molecular Weight

The nucleotide sequence of a soybean mosaic virus coat protein-coding region and its expression in Escherichia coli, Agrobacterium tumefaciens and tobacco callus.

A DNA complementary to the 3'-terminal 1168 nucleotides of the genome of the N strain of soybean mosaic virus (SMV) has been cloned and sequenced. cDNA sequence and coat protein analyses indicate that the SMV coat protein-coding region is at the 3' end of the genome, and that the coat protein is processed from a larger protein. The coat protein-coding sequence is predicted to be 795 nucleotides in length, encoding a protein of 265 amino acids with a calculated Mr of 29,857. The 3' untranslated region is 259 nucleotides in length and is followed by a polyadenylate tract. The SMV coat protein-coding region, along with a small amount of upstream sequence, has been expressed in Escherichia coli as a beta-galactosidase fusion protein. The size of the protein was less than predicted for the fusion protein, suggesting processing in E. coli. The coat protein-coding region has also been expressed in Agrobacterium tumefaciens and transgenic tobacco callus as an unfused protein under the control of the cauliflower mosaic virus 35S promoter. The coat protein produced in transgenic tobacco callus had an electrophoretic mobility identical to that of SMV coat protein and constituted approximately 0.05% (w/w) of the total extracted protein.

Amino Acid Sequence

Identification of nonspecific reactions in laboratory rodent specimens tested by Rotazyme rotavirus ELISA.

Fecal specimens from several laboratory animal species were tested for rotavirus antigen by Rotazyme II, a commercially available enzyme-linked immunosorbent assay (ELISA) that is widely used in human diagnostic studies. Fecal samples from rabbits, hamsters, guinea pigs, dogs, and cats tested negative; whereas those from rats and mice yielded a high proportion of positive results. Rats had the highest rate with 82% of the samples being positive. However, the presence of rotavirus in positive rodent samples could not be confirmed by virus isolation, electron microscopy or blocking ELISA using anti-EDIM mouse rotavirus serum. Several lines of evidence indicated that these positive reactions were false positives, apparently due to a non-specifically reacting substance in the diet of rats and mice. All the positive fecal samples were from rats and mice that had been fed nonautoclaved diet. Samples from rodents fed autoclaved diet were consistently negative in the Rotazyme test. When rats fed autoclaved diet were subsequently fed nonautoclaved diet, their stool converted from negative to positive within 6 hours. Conversely, rats with positive stool samples converted to negative within 15 hours when fed autoclaved diet. Similar results were found with mice. Positive fecal specimens and nonautoclaved rodent diet both contained a substance that apparently attached nonspecifically to the antibody coated beads used in the ELISA and reacted directly with the substrate in the absence of the conjugate. This substance was heat labile and trypsin sensitive, suggesting that it was a protein.

Animal Feed

Serum albumin treatment to eliminate false-positive results with laboratory rodent specimens tested by rotazyme enzyme-linked immunosorbent assay.

A high proportion of positive results was found in fecal specimens from mice and rats tested by Rotazyme II enzyme-linked immunosorbent assay, but the presence of virus could not be confirmed. Positive specimens and nonautoclaved rodent diet contained a substance that apparently attached nonspecifically to the antibody-coated beads used in the test and reacted directly with the substrate. Pretreatment of the beads with 0.1% bovine serum albumin eliminated this nonspecific activity.

Animal Feed

Prevalence of feline leukemia virus infection in random source laboratory cats.

Three years of quarantine data involving the prevalence of Feline Leukemia Virus (FeLV) in laboratory cats (Felis catus) was evaluated. Testing was performed using a commercially available ELISA system. Annual prevalence of infection ranged from 5.4 to 10.7% of the 937 cats tested. Results obtained with the enzyme linked immunosorbent assay (ELISA) system compared well with tests performed using an immunofluorescent antibody assay system (IFA). Seasonal periodicity was noted with higher infection rates among animals received in April. Holding period at the supplier and gender did not influence disease prevalence. The availability of a simple test system, the unacceptability of FeLV infected cats for research, the danger of transmission and a higher prevalence of infection than earlier reports indicated that supplier and user evaluation of research cats for FeLV would be prudent.

Animals

A battery of potential alternatives to the Draize test: uridine uptake inhibition, morphological cytotoxicity, macrophage chemotaxis and exfoliative cytology.

Four assays that may serve as components of a battery of alternatives to the conventional Draize test were described. The exfoliative cytology assay is a refinement of the Draize test that may provide a more sensitive and more objective end-point. The macrophage migration assay addresses the inflammatory aspects of the physiological response to irritation. The uridine uptake inhibition assay uses a quantitative, reversible end-point to detect the short-term action of agents on cell membranes and cell phosphorylative potential. Finally, the cytological assay serves as a rapid, easily performed general indicator of cytotoxic action. The two latter assays have been demonstrated to correlate very well with Draize test results and with each other for a wide range of test agents.

Animal Testing Alternatives

Susceptibility of laboratory rats, hamsters, and mice to wound infection with Staphylococcus aureus.

Response of rodents to experimentally induced subcutaneous infection was examined to determine whether laboratory rats used in invasive procedures have a superior ability to withstand wound infection than do hamsters and mice. Rats, hamsters, and mice were injected subcutaneously with 10(9), 10(7), and 10(5) colony-forming units of Staphylococcus aureus. Quantitative counts of viable S aureus from the injection site, bacteriologic cultures of heart blood, and histologic examinations of the subcutaneous tissues were performed. Multiple linear regression of the quantitative data and equality of regression lines among groups were determined. Results indicated that the ability to eliminate bacteria varied between species and depended on the dose injected within each species. Compared with hamsters and mice at all doses, rats eliminated bacteria faster and had the mildest and most rapidly organized inflammatory response after inoculation. Experimental bacteremia developed in 3.7% of all animals evaluated, with no species-specific pattern. The rat was more resistant to localized wound infection with S aureus than were hamsters and mice.

Abscess

Comparison of three serologic techniques for detection of antibody to Mycoplasma pulmonis.

A comparison of three micro-assay detection systems for naturally occurring and experimentally induced rat antibody to Mycoplasma pulmonis was performed. The micro-enzyme linked immunoabsorbent assay was found superior to counter immunoelectrophoresis or complement fixation techniques when comparing sensitivity, duration of antibody response, or technical requirements. The primary immune response was routinely detected at 3 weeks after antigen exposure when using the enzyme linked or complement fixation assays.

Animals