Biomedical subjects

D L Patton

Publications and source records attributed to D L Patton.

At least 19 recordsLinked to original sources

PubMed1996

Failure to detect Chlamydia pneumoniae in coronary atheromas of patients undergoing atherectomy.

To further investigate a proposed relationship between Chlamydia pneumoniae and coronary heart disease, coronary atheromas were collected from patients undergoing percutaneous atherectomy. Fifty-eight atheroma specimens were examined by culture and polymerase chain reaction (PCR) and 22 by electron microscopy. All were negative for C. pneumoniae, except a single specimen that was PCR-positive. These results differ from studies in other populations, in which this organism was identified by nonculture methods within coronary atheromas obtained at autopsy. Anti-C. pneumoniae antibody titers from 65 of the patients were compared with those of 28 asymptomatic controls. IgG titers were higher in controls than in patients. There is no evidence that C. pneumoniae exists within atheromas in this study population, nor does seroprevalence correlate with the presence of coronary disease in these patients.

Adult↗

PubMed1996

Experimental rabbit models of Chlamydia pneumoniae infection.

Chlamydia pneumoniae (TWAR), a common cause of acute respiratory disease in humans, has recently been associated with coronary and aortic atherosclerosis. In this study, we evaluated rabbit models of chlamydial infection to investigate the pathogenesis of C. pneumoniae infection. New Zealand White rabbits were inoculated intranasally and intratracheally with C. pneumoniae, strain AR-39, and primary and repeated infection were assessed. After a single inoculation, lung pathology was characterized by a moderate self-resolving interstitial pneumonia with bronchiolitis of 21 days in duration. Chlamydial DNA was detected by polymerase chain reaction (PCR) intermittently in the upper respiratory tract and lung tissue through day 21 postinoculation, spleen tissue at day 14, and peripheral blood mononuclear cells at days 3 and 21. After repeated inoculations, chlamydial DNA was detected by PCR in the upper respiratory tract and lung tissue through day 42. Lung lesions consisted of multifocal interstitial mononuclear cell aggregates that persisted up to day 42. Watanabe heritable hyperlipidemic rabbits were less susceptible to C. pneumoniae infection. After multiple inoculations of Watanabe rabbits, C. pneumoniae was detected by PCR and/or immunocytochemistry until day 21. In conclusion, C. pneumoniae induced a moderate respiratory infection in these rabbit models.

Animals↗

PubMed1995

Detection of Chlamydia pneumoniae TWAR in human coronary atherectomy tissues.

Chlamydia pneumoniae TWAR has been associated with coronary heart disease by seroepidemiologic studies and direct detection of the organism in atheromatous lesions of coronary arteries and aorta. In this study, 38 fresh tissue specimens from patients with coronary artery lesions that were treated by directional coronary atherectomy were tested for C. pneumoniae. Twenty-three specimens were from patients with primary lesions and 15 were from patients with restenoses. C. pneumoniae was detected by polymerase chain reaction (PCR), immunocytochemical stain (ICC), or both in 20 of 38 specimens. Using cell identity markers, the organism was localized to macrophages. Ultrastructural evidence of the organism was found in the 2 specimens examined by transmission electron microscopy, which were also positive by both ICC and PCR. C. pneumoniae was found more frequently in tissues from restenoses than in primary lesions (P = .17). There was no relation between the frequency of detection of the organism and C. pneumoniae-specific antibody titers.

Adult↗

PubMed1994

Detection of Chlamydia trachomatis in fallopian tube tissue in women with postinfectious tubal infertility.

OBJECTIVE: Biopsy tissues from women with postinfectious tubal infertility were studied for the presence of Chlamydia trachomatis. STUDY DESIGN: Tubal biopsy specimens from 25 women with postinfectious tubal infertility undergoing laparoscopy for repair of fallopian tubes were evaluated by culture, in situ hybridization. Immunocytochemistry, and transmission electron microscopy for the presence of Chlamydia trachomatis. Serum was also tested for Chlamydia trachomatis antibodies. RESULTS: Chlamydia trachomatis was detected in postinfectious tubal biopsy specimens in three of 25 patients by culture, 12 of 24 by in situ hybridization, 15 of 22 by immunoperoxidase stain, and two of 10 by transmission electron microscopy. Serum antibody against Chlamydia trachomatis was detected in 15 of 21 patients. CONCLUSION: Chlamydia trachomatis deoxyribonucleic acid or antigens were detected at a high percentage (19/24 women) in the biopsy tissues of the fimbrial and peritubal adhesions by in situ hybridization or immunoperoxidase stain, suggesting a persistent infection in these women even after antibiotic treatment.

Adult↗

PubMed1994

Detection of Chlamydia trachomatis deoxyribonucleic acid in monkey models (Macaca nemestrina) of salpingitis by in situ hybridization: implications for pathogenesis.

OBJECTIVE: Our purpose was to determine whether Chlamydia trachomatis persists in tubal tissues from monkey pocket models of chlamydial salpingitis and tubal infertility and to relate its presence to disease progression and histopathologic mechanisms. STUDY DESIGN: In situ hybridization was used to detect Chlamydia deoxyribonucleic acid in a monkey pocket model of salpingitis and in the direct tubal inoculation monkey model of tubal infertility. Results were correlated with culture and immunocytochemistry results and histopathologic characteristics. RESULTS: Chlamydia deoxyribonucleic acid was detected in the mucosa, submucosa, and deep tissues in the pocket model. In addition, it was present in peritubal adhesions from the direct inoculation model. Deoxyribonucleic acid was found at sites of inflammation and when culture and immunocytochemistry studies were negative. CONCLUSION: The presence of Chlamydia trachomatis deoxyribonucleic acid at sites of inflammation and tissue damage in monkey models of chlamydial salpingitis and tubal infertility suggests that Chlamydia persists and may be directly involved in the stimulation of the immune-mediated tissue destruction associated with Chlamydia trachomatis infections.

Animals↗

PubMed1994

Demonstration of delayed hypersensitivity in Chlamydia trachomatis salpingitis in monkeys: a pathogenic mechanism of tubal damage.

The role of delayed hypersensitivity in the pathogenesis of Chlamydia t trachomatis salpingitis was studied in the monkey "pocket" model. Pigtailed monkeys (Macaca nemestrina) were sensitized by inoculation of live C. trachomatis organisms (E/UW-5/Cx) into subcutaneous pockets containing salpingeal autotransplants. At 21 days, affinity-purified recombinant C. trachomatis heat-shock protein (rhsp60) was injected into pockets either previously sensitized with C. trachomatis or not sensitized in the same monkey. Delayed-type hypersensitivity reaction was observed, characterized by mononuclear cell infiltration with peak reaction at 48 h. Injection of rhsp60 into the pockets of a naive animal did not induce inflammation. This study showed that C. trachomatis infection in monkeys induced delayed hypersensitivity, which is mediated by hsp60. Histologic findings of the salpinx were consistent with delayed hypersensitivity reaction observed in ocular C. trachomatis infection, further suggesting a similar pathogenesis for both salpingitis and trachoma.

Animals↗

PubMed1994

Ultrastructural lung pathology of experimental Chlamydia pneumoniae pneumonitis in mice.

The ultrastructural lung pathology of Swiss Webster mice on days 2, 4, 7, 11, 15, and 21 after intranasal inoculation of Chlamydia pneumoniae AR-39 is described. The inflammatory infiltrate was predominantly polymorphonuclear leukocytes on day 2. By day 7, mononuclear cells were most prominent in the infiltrate. On day 2, chlamydial inclusions were found frequently in the bronchial ciliated epithelial cells and less frequently in the interstitial cells that appeared to be macrophages. Free particles of all developmental forms of the chlamydial microorganism were found in the bronchial lumen and alveolar space. These particles were likely organisms released from infected cells. Inclusions as well as free particles were difficult to find after day 4. These ultrastructural observations suggest an immunopathologic basis for the acute phase of the disease process.

Acute Disease↗

PubMed1993

Demonstration of Chlamydia pneumoniae in atherosclerotic lesions of coronary arteries.

Chlamydia pneumoniae is a human respiratory pathogen that causes acute respiratory disease and approximately 10% of community-acquired pneumonia. The infections are geographically widespread. Antibody prevalence studies have shown that virtually everyone is infected with the C. pneumoniae organisms at some time and that reinfection is common. In addition to respiratory disease, seroepidemiologic studies have shown an association of this organism with coronary artery disease. C. pneumoniae was detected in coronary artery atheromas by immunocytochemistry (15/36) and by polymerase chain reaction (PCR) (13/30) in 20 of 36 autopsy cases from Johannesburg, South Africa. Sequence analysis of the C. pneumoniae rRNA genes amplified by PCR confirmed that the amplified gene products were C. pneumoniae. Electron microscopy revealed typical pear-shaped C. pneumoniae elementary bodies in 6 of 21 atheromatous plaques. These findings support the seroepidemiologic studies and offer further evidence that C. pneumoniae may be involved in the atherosclerotic process.

Adult↗

PubMed1993

Effects of quinolone analog CI-960 in a monkey model of Chlamydia trachomatis salpingitis.

Several quinolones have been shown to have antichlamydial activity in vitro and in vivo. We evaluated the effects of the quinolone CI-960 (Parke-Davis) on primary or repeated chlamydial infection in the monkey salpinx pocket model. The antichlamydial effect was evaluated in the tissues, and we tested for the presence of the organism by culture, immunocytochemical stains, in situ hybridization, and histopathology. An intravenous dosage of 5 mg/kg of body weight produced therapeutic concentrations in plasma (blood sera and pocket fluids) of at least 0.25 microgram/ml at 2 h posttreatment. In monkeys with primary infections, treatment was started 2 days after inoculation and was continued for 7 days. After CI-960 treatment, all animals became culture negative. One of two control animals was culture positive through day 10 postinoculation. In monkeys with repeated infections five inoculations were given within 2 weeks. A 7-day intravenous treatment was started on day 2 postinoculation following the last inoculation. Isolation of Chlamydia trachomatis prior to treatment was positive intermittently for all monkeys. After treatment, isolation of C. trachomatis was negative for all monkeys. In monkeys with both primary and repeated infections, no significant differences were noted in the inflammatory responses in the tissues of treated and untreated animals. All tissues tested were positive by immunoperoxidase staining and/or in situ hybridization. After CI-960 therapy, C. trachomatis organisms were no longer recoverable by cell culture. The persistent finding of chlamydial DNA throughout the observation periods following drug therapy may indicate the presence of dead organisms or viable organisms in an unculturable state.

Animals↗

PubMed1993

Detection of Chlamydia trachomatis deoxyribonucleic acid in women with tubal infertility.

OBJECTIVE: To determine the presence of Chlamydia trachomatis by in situ hybridization in fallopian tube tissues of women with distal tubal occlusion. PATIENTS: Subjects were selected from a Seattle-based study of infertility in women with distally occluded fallopian tubes undergoing reconstructive surgery. For comparison, six specimens were obtained from women undergoing surgery for noninfertility-related conditions who had normal appearing tubes. METHODS: Tissue specimens from 16 of these patients were selected for analysis by in situ hybridization and immunoperoxidase (IP) staining. RESULTS: C. trachomatis was detected in 9 of 16 women by either in situ hybridization or IP. Six of the nine were seropositive with titers ranging from 1:16 to 1:64. Tissue from 6 women with normal appearing fallopian tubes were negative by in situ hybridization. CONCLUSIONS: The demonstration of C. trachomatis deoxyribonucleic acid and/or antigens in fallopian tube tissue from infertile women with distal tubal disease suggests that C. trachomatis can persist in these tissues in an uncultivable state.

Biopsy↗

PubMed1992

In vitro activity of nonoxynol 9 on HeLa 229 cells and primary monkey cervical epithelial cells infected with Chlamydia trachomatis.

Nonoxynol 9 (non-9) is the active ingredient in a wide variety of vaginal contraceptive preparations. The manufacturer recommendation for optimal contraceptive practice is repeated application every 6 h. We studied the in vitro activity of non-9 against Chlamydia trachomatis (E/UW-5/Cx) and its toxicity against HeLa 229 cells and monkey cervical epithelial cells. With a contact time of 6 h, non-9 was toxic to HeLa cells at concentrations of 50 micrograms/ml or greater and to monkey cervical cells at 100 micrograms/ml or greater. Inhibitory effects of non-9 on extracellular chlamydiae were observed at concentrations of 50 micrograms/ml or greater. Inhibition of intracellular growth of chlamydiae in monkey cervical cells was observed at a nontoxic concentration of 50 micrograms/ml. Our studies show that non-9 has antichlamydial activity. However, owing to its toxicity to cervical cells in vitro, the effects of prolonged use of non-9 in vivo should be further studied.

Animals↗

PubMed1992

Effects of ascorbic acid on Chlamydia trachomatis infection and on erythromycin treatment in primary cultures of human amniotic cells.

Ascorbic acid (vitamin C) is an essential nutrient for humans. It may also be needed by Chlamydia trachomatis, an intracellular bacterium. We investigated the effects of vitamin C on the growth of C. trachomatis E/UW-5/Cx in a primary culture of human amniotic epithelial cells. The results showed that vitamin C enhances C. trachomatis infection at concentrations of 0.2, 0.6, and 1.2 mg/dl (P less than 0.001). These three concentrations represent the in vivo concentrations of deficiency, normal, and overload levels in serum, respectively. The enhancement was dose dependent. However, the growth of C. trachomatis was inhibited at vitamin C concentrations of 120 and 1,200 mg/dl. The inhibitory effect of erythromycin against C. trachomatis was shown to be reduced in the presence of vitamin C at the three concentrations tested (P less than 0.025-0.001), and MICs were four times greater (1.6 versus 0.4 micrograms/ml). Human amniotic cells were tolerant to vitamin C concentrations of up to 1,200 mg/dl. The results show that vitamin C may be an important nutrient for C. trachomatis and that incorporation of vitamin C in the culture medium may enhance the isolation and propagation of C. trachomatis in cell cultures.

Amnion↗

PubMed1992

Detection of Chlamydia pneumoniae in coronary arterial fatty streaks and atheromatous plaques.

There is serological evidence that a recently described new strain of Chlamydia, namely Chlamydia pneumoniae TWAR, is associated with coronary artery disease. This study describes the detection of TWAR organisms in the coronary arterial fatty streaks and atheromatous plaques in 7 autopsy cases. TWAR-like organisms were observed ultrastructurally in the lipid-rich core area of fibrolipid plaques and in intimal smooth-muscle cells. In 5 cases, immunoperoxidase staining showed positive reactions to Chlamydia genus- and C. pneumoniae species-specific monoclonal antibodies.

Adult↗

PubMed1991

Salt-split human skin substrate for the immunofluorescent screening of serum from patients with cicatricial pemphigoid and a new method of immunoprecipitation with IgA antibodies.

In patients with cicatricial pemphigoid, immunoglobulins (usually IgG) and complement are deposited within the dermoepidermal junction and are detected by direct immunofluorescent staining of perilesional mucous membrane and/or skin with fluorescein-labeled antibodies to human immunoglobulins. Although rare, some patients also have circulating low-titer, anti-basement membrane zone autoantibodies. In this study, we report 11 patients with the clinical, histologic, and immunologic criteria for cicatricial pemphigoid who had circulating anti-BMZ autoantibodies as demonstrated by positive indirect staining of a normal human skin that had been fractured through the dermoepidermal junction by prolonged incubation in a cold, 1 mol/L sodium chloride solution. On this salt-split human skin substrate, 9 of the 11 patients (82%) had autoantibodies that bound to the epidermal roof, one serum stained only the dermal floor, and one serum stained both sides of the separation. The predominant class of immunoglobulin in the patients' sera that bound to the substrate was IgA; IgA was the single immunoglobulin in 55% and was associated with IgG in 18%. IgG was the only immunoglobulin detected in 27% of the cases. No specific protein was detected by either Western immunoblot or a new IgA immunoprecipitation procedure.

Adult↗

PubMed1991

Salpingoscopy: light microscopic and electron microscopic correlations.

In order to examine the ability of salpingoscopy to diagnose intratubal pathology, 32 fallopian tubes were evaluated salpingoscopically and histologically. Both flexible and rigid salpingoscopes were used, and observations were documented by intratubal photography. Salpingoscopic criteria were established and each criterion assigned a numerical value. Each tube was evaluated for patency, mucosal fold architecture, erythema, adhesions, and dilatation. Based on these criteria, tubes were graded as normal or as abnormal with mild, moderate, or severe changes. Histologically, each tube was evaluated for patency, epithelial changes, vascularity, dilatation, adhesions, and active inflammation. Six tubes with significant histologic findings and two histologically normal fallopian tubes were also examined by transmission electron microscopy. In five discordant cases, histology revealed epithelial and stromal changes not detected by salpingoscopy. Fallopian tubes with severe disease were diagnosed by both methods. Transmission electron microscopy of histologically abnormal tubes showed flattening of the epithelium with markedly reduced ciliary distribution, degenerating secretory epithelial cells with large intracellular vacuoles, and swollen nuclei containing sparse chromatin. Our results indicate that salpingoscopic observations are consistent with histologic findings when endotubal disease is severe. However, moderate pathologic changes as documented by light microscopy and transmission electron microscopy were frequently not diagnosed salpingoscopically, even with magnification.

Endoscopy↗

PubMed1990

Cinematographic observations of growth cycles of Chlamydia trachomatis in primary cultures of human amniotic cells.

Time-lapse cinematography was used to study the growth cycle of Chlamydia trachomatis in primary cell cultures of human amnion. Twelve preterm and twelve term placentas were obtained within 8 h of delivery, and epithelial cells were dissociated from the amniotic membranes by trypsinization and grown in Rose chambers. The epithelial nature of the cultured cells was documented by morphology and by immunofluorescence staining for cytoskeletal proteins, which matched the staining of intact amnion. With regular feedings, uninfected cultures remained healthy for up to 30 days. Confluent cultures (7 to 10 days) were infected with a genital strain (E/UW-5/CX) of C. trachomatis at 10(5) infectious units per chamber. Infections were done in culture medium without cycloheximide, which is often used to induce susceptibility of the cells. Between 66 and 90% of the cells were infected. Intracytoplasmic inclusions were visible by 18 h post infection (p.i.) and grew larger as the organisms inside multiplied. By 72 h p.i., the inclusions occupied the entire cytoplasm of the host cells. Further growth of the inclusions overdistended and ruptured the host cells on days 3 to 7. Cells not infected by the original inoculum became infected on day 5 or 6 p.i. by the chlamydial particles released from the ruptured cells. No amniotic cell was ever observed to survive the infection. The data presented support the hypothesis that amniotic epithelium is susceptible to infection and damage by C. trachomatis. This culture system provided detailed and dynamic observations of chlamydial infection under conditions more nearly physiologic than previously reported.

Amnion↗

PubMed1990

The effects of Chlamydia trachomatis on the female reproductive tract of the Macaca nemestrina after a single tubal challenge following repeated cervical inoculations.

The effects of repeated cervical infections followed by a single direct tubal inoculation with Chlamydia trachomatis, serovars D and F, were examined in 11 pig-tailed macaques to test the hypothesis that tubal inoculation after cervical priming causes a more severe disease than primary tubal inoculation alone. Animals were cervically inoculated between two and five times. Fallopian tubes were inoculated with serovar D or F 1 week after the last cervical challenge. Three control monkeys received only one direct tubal inoculation without previous cervical inoculation. Infection was confirmed by isolating the microorganism from the endocervix in 13 of 14 monkeys and from the endosalpinx in four only after the tubal inoculation. Antibody was detected in post-infection sera of all 14. Tubal edema occurred in seven of 11 animals after the first cervical inoculation, and uterine erythema occurred in 11 of 11 after the second cervical inoculation. Peritubal adhesions were induced before the tubal inoculation in zero of seven given three or fewer cervical inoculations and four of four given five cervical inoculations (P less than .01). After direct tubal inoculation, peritubal adhesions became more prominent, and the 11 hysterectomy specimens showed plasma cell endometritis in nine and salpingitis in nine. Two control monkeys developed minor adhesions, the other none. One tube in two of three controls showed mild plasma cell infiltrates, whereas no evidence of endometritis was observed in controls. Histopathology in these monkeys was characteristic of chlamydial endometritis and salpingitis. However, the pathogenesis of these changes is uncertain because C trachomatis was not isolated from the endosalpinx after cervical inoculations alone.

Animals↗

PubMed1989

Studies of polyglycolic acid hollow self-retaining vasal stent in vasovasostomy.

A simple method of vasovasostomy using a unique hollow, biflanged, hydrolyzable, self-retaining intraluminal stent made of polyglycolic acid (PGA) was evaluated in rat and dog animal models. The stented vasovasostomy was compared with a modified two-layer technique in a randomized prospective trial in rats. Twenty of 22 vasovasostomies using the 0.8890 mm and 0.9398 mm stents were patent, whereas only 4 of 22 vasovasostomies using a modified two-layer anastomosis without a stent were patent (P less than 0.01). Eight dogs underwent implantation of the absorbable vasovasostomy stent on one side and two-layer vasovasostomy on the other. Scanning electron microscopy showed superior re-epithelialization and closer return to normal morphology in the stented vasovasostomies than with the conventional two-layer technique.

Animals↗