Biomedical subjects

D L Patton

Publications and source records attributed to D L Patton.

At least 55 records · Page 3Linked to original sources

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↗

PubMed1989

Experimental Chlamydia trachomatis salpingitis in mice: initial studies on the characterization of the leukocyte response to chlamydial infection.

The murine biovar (mouse pneumonitis) of Chlamydia trachomatis was inoculated into the left oviduct of female Swiss Webster mice to establish acute salpingitis. Chlamydial inclusions were observed in secretory epithelial cells by both transmission electron microscopy and light microscopy using immunoperoxidase staining of deparaffinized sections. By days 5-8 after infection, a mixed polymorphonuclear and mononuclear cellular infiltrate was observed in the submucosa and mucosa. Epithelial cell deterioration occurred in the endosalpinx in areas of heavy mononuclear cellular infiltration. During the acute phase of the disease a cellular infiltrate consisting mainly of T cells was identified by staining frozen tissue sections with monoclonal antibodies to mouse lymphocyte antigens. Occasionally B lymphocytes were observed. Widespread deciliation of the mucosa was observed by scanning electron microscopy. No histopathologic or immunopathologic responses were observed in the control oviducts. These observations suggest an immunologic basis for the structural abnormalities seen in the infected oviducts.

Acute Disease↗

PubMed1989

A proposed mouse model for acute epididymitis provoked by genital serovar E, Chlamydia trachomatis.

This study was conducted to determine the efficacy of the male mouse as a model for epididymitis caused by human genital serovar E, Chlamydia trachomatis. C. trachomatis was reisolated from all tissues removed on Days 3, 5, and 7 post inoculation (pi). Although some infected epididymides removed on Days 10, 14, and 21 pi were positive, control tissues remained negative. Histopathology of tissues showed a heavy, mixed inflammatory infiltrate consisting of polymorphonuclear cells and lymphocytes. Serum antibody to C. trachomatis was detected in the infected mice only (titer greater than or equal to 1:32). Chlamydial inclusions and individual elementary bodies were confirmed by immunofluorescent and immunoperoxidase staining up to Day 7 pi. These data show that the male mouse is susceptible to C. trachomatis infection and is appropriate for studies dealing with the effect of C. trachomatis on male fertility.

Acute Disease↗

PubMed1989

Histopathology of Chlamydia trachomatis salpingitis after primary and repeated reinfections in the monkey subcutaneous pocket model.

Monkeys with subcutaneously autotransplanted salpingeal fimbrial tissues were subjected to primary and repeated infections with Chlamydia trachomatis. The inflammatory response after primary inoculation was characterized by infiltration with polymorphonuclear leucocytes in the acute phase and mononuclear cells in the chronic phase. However, the inflammatory response after repeated infections was dominated by a mononuclear cell infiltration with a conspicuous absence of the initial phase of polymorphonuclear leucocyte infiltration. The remarkable findings of repeated infections were plasma cell infiltration, lymphoid follicle formation, and increased fibroblast activity resulting in extensive fibrosis. These findings are similar to those described for monkeys inoculated directly into the oviducts with C. trachomatis and support our original hypothesis that, after chlamydial infection, the tissue damage is provoked by immune-mediated mechanisms.

Animals↗

PubMed1989

Time-lapse videomicroscopic study of in vitro wound closure in rabbit corneal cells.

This study used video time-lapse recording to characterize the dynamic features of corneal epithelial cells and keratocytes during in vitro wound closure. Confluent cultures of these two cell types from rabbits were established in Rose chambers. A wound 4 or 10 mm in diameter was produced in the center of each culture by mechanical removal of cells. Wound closure was recorded by videomicroscopy for 2-3 days and reviewed at a playback speed of 400 times normal. The epithelial cells at the wound margin initiated migration by extending lamellipodia with undulatory motions. Successive tiers of cells moved as a continuous sheet in a unified and coordinated manner while maintaining intercellular linkage. The migration was unidirectional, toward the wound center. The mean migration rate of the leading cells was 104 microns/hr. The trailing cells migrated at successively slower rates, inversely proportional to their distance from the wound margin. Mitosis was rare during migration but did occur simultaneously. The mitotic rate was 3.7 mitoses/100 cells. The relative mitotic frequency was 0.23 mitosis/hr. By contrast, in keratocyte cultures, the cells around the wound margin migrated individually and asynchronously without intercellular connection. Initially the cells moved generally toward the wound space, but later, different cells migrated in different directions. The mean migration rate was 15 microns/hr. Mitosis occurred frequently. The mitotic rate was 25.3 mitoses/100 cells, and the relative mitotic frequency was 1.33 mitoses/hr. The cell cycle duration was 9.9 hr. Thus corneal epithelial cells and keratocytes showed fundamentally different characteristics and mechanisms of wound closure in vitro.

Animals↗

PubMed1989

Experimentally induced ocular chlamydial infection in infant pig-tailed macaques.

Four Macaca nemestrina monkeys were inoculated in the conjunctiva with Chlamydia trachomatis (strain E) at 6 weeks of age. A fifth monkey was inoculated with HeLa cell materials only. Ten weeks later, all monkeys were reinoculated with either strain E or strain C. All inoculated monkeys were susceptible to infection with C. trachomatis as documented by fluorescent antibody staining of smears and reisolation of the organism from conjunctival and nasopharyngeal swab specimens. Rectal and vaginal swab specimens remained negative throughout the study. Three of four inoculated animals responded with IgM titers reaching a peak of 1:16 (M#3) and 1:32 (M#1, M#4) 2 weeks after the primary inoculation. IgG appeared in all inoculated animals and titers rose to peak levels of 1:64 (M#2), 1:128 (M#1, M#3), and 1:256 (M#4). Histopathology documented a dramatic difference in immunological response following secondary inoculation. Primary inoculation elicited a typical inflammatory response characterized by moderate stromal infiltration of polymorphonuclear and mononuclear leukocytes. Plasma cells appeared by week 3 postinoculation (pi). Following a secondary inoculation, classic follicle formation was evident by 1 week pi. Mononuclear markers identified a germinal center composed of B cells and a T cell cap. Epithelial thinning near the cap of the follicle was accompanied by a complete loss of goblet cells. This model may be useful for studying the immunopathology of infant chlamydial infections.

Animals↗

PubMed1989

Chlamydia trachomatis oculogenital infection in the subcutaneous autotransplant model of conjunctiva, salpinx and endometrium.

The subcutaneous pocket model of salpingeal, endometrial, and conjunctival autografts for studying Chlamydia trachomatis infection in monkeys is described. Portions of the salpinx that were transplanted included fimbria, ampulla, and isthmus. The model is an extension of the original model which consists of either salpingeal fimbria or conjunctive autografts. Transplantation of the ampulla portion of the Fallopian tube enabled us to increase the number of pockets or test sites. Salpingeal and conjunctival autografts could be established during a single surgery. In addition, it is possible to autotransplant endometrium and provoke endometritis. The autografts were shown to be susceptible to C. trachomatis infection. Preliminary rechallenge experiments showed infection of the subcutaneous transplants may induce immunity, indicating the model may be used for immunity and vaccine studies. Simultaneous transplantation of different parts of the oviduct, endometrium, and conjunctive should expand the usefulness of the subcutaneous model in other studies on mixed infections or immune responses to infection.

Animals↗

PubMed1989

A comparison of the fallopian tube's response to overt and silent salpingitis.

We examined two groups of infertile women who underwent microsurgical repair of their fallopian tube(s) for distal tubal obstruction, one with a history of salpingitis (overt pelvic inflammatory disease) (N = 34) and one without (silent pelvic inflammatory disease) (N = 25). Nine women with normal tubes undergoing hysterectomy and salpingectomy served as controls. Tubal biopsy specimens were studied by light, scanning, and transmission electron microscopy to assess tubal damage. Morphologic damage was scored 0-9, with 0 representing normal tubal architecture and 9 assigned to severe tubal damage. The mean score (+/- SEM) in women with overt pelvic inflammatory disease was 4.2 +/- 0.4; in women with silent pelvic inflammatory disease, 4.3 +/- 0.4; and in the control group, 0.76 +/- 0.2 (P less than .001). Fallopian tube abnormalities seen in women with overt and silent pelvic inflammatory disease included flattened mucosal folds, extensive deciliation, and degeneration of secretory epithelial cells, morphologic changes that are similar to the cellular changes observed in our experimental Chlamydia trachomatis infections in monkeys. Laser light-scattering spectroscopy was used to measure the ciliary activity of the epithelial cells. Ciliary beat frequency was significantly reduced in women with overt pelvic inflammatory disease (N = 13; f = 6.4 +/- 1.2 Hz) and in women with silent pelvic inflammatory disease (N = 11; f = 7.2 +/- 1.2 Hz) as compared with the controls (N = 5; f = 23.4 +/- 1.5 Hz) (P less than .001).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1988

In vitro growth of Chlamydia trachomatis in conjunctival and corneal epithelium.

This study used primary cultures of conjunctival and corneal epithelial cells from rabbit, monkey and humans to investigate the infection process and host interactions of chlamydia. The epithelial cells were isolated from bulbar and palpebral conjunctivae and the cornea following incubation with EDTA or dispase and microdissection. The cells were trypsinized and grown in microtest wells or Rose Chambers (gelatin-coated glass substratum). The cell origin of the cultured epithelial cells was verified by immunofluorescence staining with anticytokeratins. The cells were nonreactive when stained with antivimentin, a fibroblast marker. The growth kinetics of the cultured epithelial cells were determined by morphological criteria. Cells reached confluency by 5-7 days, and remained as such for 18 days. The cells were maintained as long as 32 days in primary culture. After the cells had attained confluency (5-7 days), they were infected with ocular strains B/TW-5/OT or C/TW-3/OT C. trachomatis. Chlamydial inclusions were identified by light and electron microscopy. Direct FITC C. trachomatis staining (species-specific monoclonal antibody) of the inclusions was used to determine the infectivity of the various types of epithelial cells. The dynamics of the infection process was documented by time lapse photomicroscopy. Small inclusions were identified by 18 hr postinoculation (pi). By 48 hr pi, the inclusions had dramatically increased in size, occupying much of the host perikarya. The reticulate bodies were very active at this time. By 70 to 100 hr pi, the inclusions contained the highly infectious elementary bodies. At this time, rupture of the inclusion and lysis of the infected cells usually occurred.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

A comparison between argon laser and microsuture anastomosis of the rat uterine horn.

For assessment of the use of the argon laser for tubal anastomosis, the uterine horns of 12 Sprague-Dawley rats were surgically divided and then anastomosed, 6 by argon laser photocoagulation and 6 by the conventional technique of microsurgery. After a 4- to 6-week postsurgical period subjects were reexamined. All microsutured anastomoses were fully patent and continuous, with no apparent fibrosis. Four of six laser subjects had complete occlusion; the other two exhibited patencies between 10% and 20% of normal luminal area. Although initially producing satisfactory union, argon laser photocoagulation proved highly tissue traumatic, resulting in poor regeneration of the anastomotic site.

Animals↗

PubMed1987

Chlamydial infection of subcutaneous fimbrial transplants in cynomolgus and rhesus monkeys.

Acute infection with Chlamydia trachomatis serotype E was established in monkey fallopian tube fimbriae by subcutaneous implantation. Depending upon monkey species, from eight to 20 implants could be established in each animal. Animals were given estrogen before percutaneous inoculation of the autografts with Chlamydia. Acute inflammatory changes were found in homografts examined in the first week after infection, with chronic inflammatory changes noted thereafter. Chlamydial inclusions were detected within fimbrial epithelial cells up to seven days postinoculation by fluorescent-antibody staining and immunoperoxidase staining with C. trachomatis-specific monoclonal antibody. Organisms were recovered from autografts up to five days after infection. Analysis of serum antibody by microimmunofluorescence revealed that serotype E-specific antibody of both IgM and IgG classes was produced after infection. We conclude that subcutaneously implanted fallopian tube autografts may provide a useful primate model for kinetic studies of chlamydial infection and immunity.

Animals↗

PubMed1987

Distal tubal obstruction induced by repeated Chlamydia trachomatis salpingeal infections in pig-tailed macaques.

The effects of repeated infections with Chlamydia trachomatis, serovars F, D, and J, were examined in pig-tailed macaques. The fallopian tubes of three experimental monkeys were inoculated at the middle of the menstrual cycle on three consecutive months. Monkey 1 received homologous F; monkey 2 received heterologous F, D, and J; and monkey 3 received homologous inoculations in the right and heterologous inoculations in the left fallopian tubes. One control monkey (4) received repeated inoculations of HeLa cell materials only and remained normal throughout the experiment. Infection was confirmed by isolating the microorganism from both the endosalpinx and endocervix. Antibody to the infecting strains was demonstrated in sera, tears, and cervical secretions by using microimmunofluorescence. Mild chronic salpingitis developed in monkeys 1 and 3, and chronic follicular salpingitis developed in monkey 2. Peritubal and periadnexal scarring and endosalpingeal adhesion formation were produced after reinfection. The right fallopian tube of monkey 3 was distally obstructed (confirmed by hysterosalpingography). Systemic complications, including perihepatitis and conjunctivitis, were also documented in these monkeys. We conclude that repeated infections produced extensive tubal scarring, chronic salpingitis, and distal tubal obstruction, findings not apparent in primary infection.

Animals↗

PubMed1987

Experimental trachoma in subcutaneous conjunctival autografts in macaques.

To study chlamydial conjunctivitis, conjunctival autografts subcutaneously implanted in pockets on the abdomens of rhesus (Macaca mulatta) and pig-tailed (M. nemestrina) monkeys were inoculated percutaneously (4-10 X 10(4) inclusion-forming units per pocket) with trachoma strains of Chlamydia trachomatis (serovars B & C). These conjunctival pockets were removed on days 2, 3, 4, 5, 6, 7, 8, 10, 14 and 16 post-inoculation (pi). Tissues (at least two samples at each time interval) were prepared either for reisolation of the organisms by cell culture or histologic examination by light and electron microscopy. Control tissue, inoculated with either HeLa cell material or UV-inactivated organisms, were prepared in parallel. Bloods were drawn and tear strips taken at weekly intervals for 8 weeks. A total of 221 bulbar and 99 palpebral conjunctival pockets were established over time with the success rate of 75% and 88%, respectively, in six rhesus and ten pig-tailed monkeys. Histological examination revealed widespread infiltration of mixed polymorphonuclear and mononuclear cells on days 2 and 3 pi. By day 5 pi, lymphocytes had migrated into the thinned epithelial layer. Patches of inflammatory infiltrate similar to trachoma follicles were observed, although distinct germinal centers were absent. Surface morphologies of normal and infected pocket conjunctiva were examined by scanning electron microscopy. Normal tissue was characterized by a regular mosaic pattern of closely packed epithelial cells containing numerous microvilli. Infected tissue was edematous, and the continuity of the mucosal surface had been altered. Chlamydiae were reisolated from the pockets on days 2, 3, 6, 8 and 10 pi.(ABSTRACT TRUNCATED AT 250 WORDS)

Abdomen↗

PubMed1986

The histopathology of experimental trachoma: ultrastructural changes in the conjunctival epithelium.

Experimental acute conjunctivitis was produced in cynomolgus monkeys by ocular inoculation with serovar B of Chlamydia trachomatis. The cellular responses to chlamydial conjunctivitis infection were examined by light, transmission, and scanning electron microscopy. A self-limited, acute conjunctivitis resulted from a single primary inoculation. A moderate lymphocytic infiltrate was present in the conjunctiva. After repeated inoculation, a chronic conjunctivitis (trachoma) developed. Prominent lymphoid follicles with distinct germinal centers were present in these tissues. Scanning electron microscopy revealed patchy areas of cellular alteration and loss of microvilli. Intracellular injury, documented by transmission electron microscopy, included disruption of the plasmalemmal membranes and rupture of the cytoplasmic organelles. The inflammatory infiltrate consisted of plasma cells, polymorphonuclear cells, lymphocytes, eosinophils, and degranulating mast cells. The immune response following single and repeated chlamydial infections was characterized by immunoperoxidase staining with monoclonal antibodies to pan-leukocytes, macrophages, and B cells. The center of the follicle was comprised of B cells, with T cells in the cap region. Large macrophages were also found in the germinal center. Further study is required to determine which cellular mechanisms are involved in the histopathologic and immunologic alterations induced in the conjunctiva after chlamydial infection.

Animals↗