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Biomedical subjects

R L Boyd

Publications and source records attributed to R L Boyd.

At least 145 records · Page 8Linked to original sources

Stimulus-permeability coupling in rat pulmonary macrophages challenged by Pseudomonas aeruginosa. An X-ray microanalysis study.

Electron probe X-ray microanalysis (XRMA) of freeze-dried ultrathin sections provides the capability of measuring intracellular elemental content. This methodology was used to investigate the stimulus-permeability coupling responses associated with phagocytosis of Pseudomonas aeruginosa by cultured pulmonary alveolar macrophages (PAMs) of rats. PAMs were challenged with P. aeruginosa suspended in Gey's buffer at a bacteria to PAM ratio of 50:1 for 1 h at 37 degrees C. A 1-mm3 pellet of the unchallenged control PAMs, challenged PAMs and P. aeruginosa alone was quench-frozen in nitrogen-cooled, liquid propane, and 0.1-micron cryosections were cut at -100 degrees C. X-ray spectra were collected for nucleus and cytoplasm of 39 control PAMs, 36 challenged PAMs and 40 P. aeruginosa. Concentrations (mmole/kg dry weight) were obtained for Na, Cl, K, Ca, Mg, P, S for each cell. In the control PAMs, the content was similar to other mammalian cells. Moreover, there were no differences in elemental content between nucleus an cytoplasm. In the challenged PAMs, Na concentration was 4 times that of control PAMs (p less than 0.001) whereas Cl was double (p less than 0.001), K was 29% lower (p less than 0.001), and Ca was 4 times higher (p less than 0.05). The elemental concentration profile in the P. aeruginosa was distinctly different from that of the PAMs: higher Na, Ca, Mg, but lower Cl and K values. These results demonstrated elemental content changes in cultured PAMs challenged with P. aeruginosa that indicate a stimulus-permeability response by membranes associated with the phagocytic process.

Animals↗

Effect of self-administered daily irrigation with 0.02% SnF2 on periodontal disease activity.

To determine the effect on periodontal health of a daily self-administered irrigation with 0.02% stannous fluoride (SnF2) solution, 28 subjects who had moderate to advanced periodontitis were randomly divided into 3 groups: a control group (n = 9) which used no irrigation, a group (n = 8) which used a self-administered water irrigation device (Water Pik) daily with water (H2O group) and a group (n = 11) which used the Water Pik in a similar manner but with SnF2 solution (SnF2 group). All subjects were instructed in routine tooth brushing and flossing but received no other periodontal treatment. 4 study sites were selected from each patient which had pocket depths greater than 4 mm and bleeding upon probing. Plaque index, gingival index, bleeding tendency, pocket depth, loss of attachment, and microbiologic samples of subgingival plaque for morphologic determinations were collected from all study sites at baseline, 2, 6, and 10 weeks. A cross-over was then initiated for 2 additional monthly checks in which the H2O group changed to SnF2 and the SnF2 group was divided into 2 subgroups which either continued to use SnF2 or changed to H2O. The control group completed the study at the beginning of the crossover. The clinical data showed significantly more improvement in periodontal health during the first 10 weeks for the SnF2 group (p less than 0.01). After cross-over, the clinical data indicated the group that changed from H2O to SnF2 significantly improved their periodontal health, while the group that changed from SnF2 to H2O became worse. The microbiologic data showed trends which agreed with the clinical data during the first 10 weeks but were less significant. After cross-over, the %s of motile rods and spirochetes were too small (0-7%) to establish statistically significant changes considering the accuracy of the technique used.

Bacteria↗

Early/middle evoked potentials to tone bursts in quiet, white noise and notched noise.

Early/middle auditory-evoked potentials were recorded on 10 normal-hearing adults using 500-, 1 000, and 2 000-Hz tone bursts having 4-ms rise-fall and 1-ms plateau times. The stimuli were presented in quiet, white noise and notched noise at 50, 40, 30, 20 and 10 dB nHL. The noises were selected to improve frequency specificity by masking those regions of the cochlea that do not correspond to the test stimulus. The recording-amplifier system had a bandpass of 55-1 500 Hz; electric activity was measured during the 25-ms post-stimulus interval. The three waves observed during this time interval (P10, N15, and P20) were labeled to reflect polarity and latency. The results revealed no differences among stimulus conditions (quiet, white noise, notched noise) in the number of identifiable responses for any wave at any intensity. Moreover, no P10, N15 or P20 latency differences were observed among conditions at 40 dB nHL and below. When 500-Hz tone bursts were presented at 50 dB nHL, however, significantly shorter response latencies were observed in quiet than when the tone bursts were mixed with white noise or notched noise. It was concluded that frequency specificity can be improved when tone bursts are mixed with notched noise or white noise. White noise is preferred, however, because it can be implemented with less complex instrumentation and calibration than notched noise.

Adult↗

Interactions and quantitative analysis of immunoregulatory cells in the chicken thymus.

Step-wise dilution of chicken thymus cell suspensions has been used to sequentially reveal suppressor, effector, and helper cells in these suspensions. The cells were tested either alone or in autologous mixture combinations with peripheral blood lymphocytes (PBL) as a source of effector cells. The assays studied were graft-vs-host reaction (GvHR) and mixed lymphocyte (MLR) reaction, spontaneous cellular cytotoxicity and antibody-dependent cell-mediated cytotoxicity, and mitogen responsiveness to Con A, PHA, and PWM. When tested alone, high numbers of thymus cells (1 X 10(7) gave weak or low responses, with the exception of GvHR, which was high. When this number of thymocytes was mixed with a strongly responding PBL effector population, there was marked suppression of the latter. Nonspecific crowding was excluded as a cause for the decreased responsiveness, and the data therefore demonstrated the presence of suppressor cells in the thymus. With gradual reduction of the thymus cell number in the mixtures, the suppressor activity was lost, but concomitant with this was the appearance of, or a gradual increase in, thymus effector cells giving good responses. Further dilutions of the thymus (to, e.g., 1 X 10(5) cells) depleted the suspension of effector cells, but helper cells capable of markedly amplifying the effector potential of PBL were revealed. The suppressor/helper function of the thymus was not only dependent on the absolute numbers of thymus cells present, but also on the degree of inherent responsiveness of the effector PBL. If the response of PBL alone was strong, a thymus suspension containing both helper and suppressor cells (e.g., 1 X 10(6) cells) caused suppression of the PBL; if the PBL alone were weak, this same thymus cell suspension caused enhancement. The outcome of an immune response is therefore dependent not only on the presence or absence of particular cell types, but also on the ratios between these cells. An imbalance in these ratios in vivo may underlie diseases of immunologic origin, e.g., autoimmunity.

Animals↗

Ontogeny of surface markers on functionally distinct T cell subsets in the chicken.

Three subsets of chicken peripheral T cells (T1, T2 and T3) have been identified in peripheral blood of adult chickens on the basis of fluorescence intensity after staining with certain xenogeneic anti-thymus cell sera (from turkeys and rabbits). They differentiate between 3-10 weeks of age in parallel with development of responsiveness to the mitogens concanavalin A (Con A), phytohemagglutinin (PHA) and pokeweed mitogen (PWM). Functional tests on the T subsets, sorted with a fluorescence-activated cell sorter, have shown that T2, 3 cells respond to Con A, PHA and PWM and are capable of eliciting a graft-vs.-host reaction (GvHR). In contrast, although T1 cells respond to Con A, they respond poorly to PHA and not at all to PWM or in GvHR. There was some indication of cooperation between T1 and T2,3 cells for the PHA response. Parallels between these chicken subsets and helper and suppressor/cytotoxic subsets in mammalian systems are discussed.

Aging↗

MHC- and non-MHC-encoded surface antigens of chicken lymphoid cells and erythrocytes recognized by polyclonal xeno-, allo- and monoclonal antibodies.

Surface antigens on chicken thymus and bursa cells were analyzed by immunoprecipitation using polyclonal and monoclonal antisera raised against (and specific for) thymus (ATS) or bursa (ABS) cells, respectively. The antigens identified were compared with those governed by the B-F, B-L and B-G regions of the chicken major histocompatibility complex (B complex). Four proteins were precipitated from thymus cells by 2 polyclonal ATS: both antisera recognized molecules of apparent molecular mass of 172-182, 132-135, 75-76 kDa, and one antiserum in addition recognized a protein of 102 kDa. The 172-182 and 102-kDa peaks were still demonstrable under reducing conditions indicating that they are composed of a single polypeptide chain, the other 2 were lost under reducing conditions, therefore, must be composed of smaller subunits. Of the 2 monoclonal ATS tested, one identified a single protein of 186 kDa and the other a 135-kDa protein (in addition to 2 smaller molecules); whether these are the same as those precipitated by the polyclonal antisera remains to be determined as they behaved differently under reducing conditions. Proteins of 162 and 78-84 kDa were revealed by 2 polyclonal ABS under nonreducing conditions but the former may in one case be a polymer (it disappeared under reducing conditions) and in the other a single molecule. In addition molecules of 182 kDa were identified by one antiserum and of 84 and 60 kDa by the other under nonreducing conditions. Of the 4 monoclonal ABS only one identified a 200-kDa protein: molecules of 115-125, 90-100, 48-52 and 40-43 kDa were also precipitated, all of which were reduced to smaller molecules. With 2 specific anti-B-F alloantisera we were able to precipitate the "conventional" B-F antigen from red blood cell lysates of CB-strain chickens resolving into a 40-kDa peak and a light chain of about 12 kDa corresponding to beta 2 microglobulin. Precipitates from peripheral blood lymphocytes, bursa and thymus cells revealed an additional protein of 22 kDa. With 2 specific B-L alloantisera two peaks of 33 kDa and 31 kDa were obtained from peripheral blood lymphocytes. Using anti-B-G alloantisera a double band corresponding to 47 and 42 kDa was seen under reducing conditions. There is no evidence from these data to indicate that the polyclonal and monoclonal antibodies are directed towards major histocompatibility complex antigens.

Animals↗

Lymphoid antigenic determinants of the chicken: ontogeny of bursa-dependent lymphoid tissue.

The ontogenic development of the bursa-dependent lymphoid tissue in the chicken has been studied using rabbit antisera specific for B lymphocyte sub-populations and two elements of the bursal microenvironment. The antigens investigated were: the chicken B lymphocyte antigen ( CBLA ); mature B lymphocyte antigen ( CMBLA ); a foetal-associated antigen ( CFAA ) present on embryonic haemopoietic cells and adult bone marrow and bursa cells; immunoglobulin (Ig) and IgG; a bursa-specific cortical reticulin fibre antigen ( CBRFA ); a gut-associated mucin antigen ( CGAMA ) present on bursal medullary reticular epithelial (REp) cells. The development of suspected precursor cells was examined using a rabbit antiserum specific for foetal spleen cells. The major finding was the interrelationship between developing B cells and the bursal microenvironment. The first CBLA - and CFAA -positive cells were detected in the bursa at day 8 of incubation but their precise localization was difficult to assess. In 12-15 day embryos, both these cells were found predominantly in the tunica-propria in close proximity to cells bearing the reticulin fibre antigen CBRFA . This close association between CBLA -, CFAA - and CBRFA determinants represents the earliest stages of B cell differentiation and mimics that found in the adult bursa cortex. By day 18, the majority of bursa cells expressed CBLA and Ig and were localized in the developing medullary follicles, the REp cells of which were CGAMA -positive, demonstrating a very early interaction between intestinal tract contents, bursal REp cells and B cell maturation. Around hatching some bursa cells showed a marked increase in the membrane expression of CBLA and Ig, and the simultaneous expression of CMBLA and IgG. These cells were present in medullary follicles; CBRFA was present on cortical reticulin fibres which provided a supporting framework for the more immature CFAA -positive cells.

Animals↗

The B-L (Ia-like) antigens of the chicken. Lymphocyte plasma membrane distribution and tissue localization.

Specific antisera reacting with B-L (Ia-like) antigens were prepared by reciprocal immunization of animals from the congeneic lines CB and CC. The resulting antisera were tested either in direct or indirect immunofluorescence tests and stained 10-16% of peripheral blood cells (PBL). Of the B-L+ cells, 90% were B cells and 8% were T cells. After in vitro stimulation of PBL with ConA, 58% were B-L+ and 91% of these were T cells. B and T cells were defined by means of rabbit antisera raised against bursa and thymus cells made specific by absorption with the relevant tissues. Antigens determined by anti-B-L antisera, rabbit anti-bursa (ABS) and rabbit anti-thymus (ATS) sera showed an independent distribution on the membrane of PBL. The tissue distribution of B-L+ cells, defined by means of allo-antisera and monoclonal antibodies, was studied by direct and indirect immunofluorescence on sections of skin, liver, kidney and brain. In all organs, in addition to B cells and a small number of, presumably activated, T cells, macrophages and dendritic cells were positive. Notably, glia cells in the brain were also shown to express B-L antigen.

Animals↗

Distribution and functional analysis of B-L/Ia-positive cells in the chicken: expression of B-L/Ia antigens on thyroid epithelial cells in spontaneous autoimmune thyroiditis.

The B-L region of the chicken major histocompatibility complex (MHC), the so-called B-locus, corresponds to the murine H-2 I-region. Using alloantibodies and monoclonal antibodies to B-L we analyzed: (a) the tissue distribution of B-L+ cells, (b) the function of B-L+ cells, and (c) the possible role of B-L+ cells in the development of spontaneous autoimmune thyroiditis (SAT) in Obese strain (OS) chickens. The tissue distribution of B-L+ cells in peripheral blood and various lymphoid and nonlymphoid organs corresponds to what is known for mammals. In the bursa of Fabricius most lymphoid cells and the dendritic cells carry the B-L antigen; B-L+ thymic nurse cells (TNC) first appear on day 17 of embryonic life; chickens possess dendritic B-L+ cells in the skin resembling mammalian Langerhans cells; in addition we found that the microglia is unequivocally B-L+. B-L+ peripheral blood lymphocytes (PBL) were separated with a fluorescence-activated cell sorter. Ten percent of unstimulated PBL and 60% of phytohemagglutinin (PHA) stimulated T-cell blasts are B-L+. In graft-vs-host (GvH) assays B-L- cells were identified as the effector cells. These cells respond to PHA and concanavalin A (Con A), but not to pokeweed mitogen (PWM). B-L+ cells cannot be stimulated by Con A and PHA, but respond to PWM. They possess only a very low activity in GvH assays which can be inhibited by anti-T-cell sera. In OS chickens B-L+/non-B/, non-T and B-L+ T (blasts?) cells are found in the "first line" of mononuclear cell infiltration in the thyroid glands. Most interesting, thyroid epithelial cells--which are normally B-L- -become B-L+ in the neighbourhood of B-L+ infiltrating mononuclear cells. This observation may be of significance for autoantigen presentation and perpetuation in autoimmune thyroiditis. Finally, OS thymuses contain significantly less TNC than normal controls.

Animals↗

Clinical assessment of injuries in orthodontic movement of impacted teeth. II. Surgical recommendations.

Adverse effects on periodontal tissues have been associated with surgical techniques used to expose impacted teeth that are to undergo orthodontic tooth movement. The purpose of this study was to compare the effects on the periodontium of two of the more commonly used surgical techniques for labially impacted canines--(1) exposure of the entire labial aspect of the anatomic crown with total excision of all keratinized tissue (the window approach) and (2) a technique which exposes only 4 to 5 mm of the most superficial portion of the labial aspect of the cusp tip while maintaining 2 to 3 mm of keratinized tissue. Twelve patients were selected who had undergone comprehensive orthodontic therapy for a unilateral labial impaction of an upper canine, and who had the impacted canine surgically exposed with a window approach. This group's periodontal status after orthodontic therapy was compared to that of another group of twelve patients who had also undergone comprehensive orthodontic therapy for a unilaterally impacted labial upper canine but in whom the surgical exposure of the impacted tooth had been performed in a manner which preserved 2 to 3 mm of keratinized tissue on the labial aspect. The results indicated that the twelve subjects in whom the window approach had been used had statistically more gingival recession, gingival inflammation, and loss of attachment on the labial and labioproximal surfaces of the impacted teeth than the other group that had 2 to 3 mm keratinized tissue maintained after surgical exposure (p less than 0.05).

Adolescent↗

Functional analysis of B-L (Ia-like) antigen-bearing chicken peripheral blood cells.

The functional of B-L (Ia-equivalent)-positive (B-L+) adn -negative (B-L-) chicken peripheral blood lymphocytes (PBL) was studied in vitro and in vivo. The PBL were first stained in direct immunofluorescence tests with a fluorescein isothiocyanate-labelled anti-B-L alloantiserum and then separated by means of a fluorescence-activated cell sorter. In agreement with our previous findings, B-L- cells showed functional properties of T lymphocytes, responding to concanavalin A and phytohaemagglutinin-P in vitro and inducing a graft-versus-host (GVH) reaction when injected into allogeneic embryos. Sorted B-L+ gave no responses in any of these assays. Neither B-L+ nor B-L- cells, when tested alone, responded significantly to pokeweed mitogen, but mixtures of the two restored the responsiveness to that of the original unsorted suspension. Of the B-L+ PBL, 10% were T cells, which may account for the low GVH reactivity given by this population.

Animals↗

Pulmonary function of the reserpine and isoproterenol models of cystic fibrosis.

Two experimental animal models exhibiting functional and morphologic changes of exocrine glands similar to those seen in patients with cystic fibrosis (CF) have been reported in the rat: chronic stimulation with reserpine (Martinez et al. 1973 Pediatr. Res. 9:463, 470) and chronic stimulation with isoprenaline (Sturgess and Reid 1973 Br. J. Exp. Pathol. 54:388). We have studied the pulmonary function of these models induced by injecting rats subcutaneously with reserpine (RES, 0.5 mg/kg/day), isoproterenol (ISO, 25 mg/kg/day), or saline (Con, 1.0 ml/kg/day) for 6 days. Plethysmographic measurements were made for functional residual capacity (FRC), airways resistance (Raw), specific airways conductance (sGaw), phase difference between air flow rate and mean alveolar pressure (PD), frequency of breathing (f), and tidal volume (VT) of the anesthetized rats. In the RES and ISO rats, the FRC, Raw and f were not different from Con values. The PD was greater and the VT was less than Con values (p less than 0.05). The results of both studies indicate uneven ventilation (increased PD) and penduluft (decreased VT) consistent with maldistribution of resistance and/or compliances of the peripheral airways and/or alveolar compartments. These physiologic effects can be related to the morphologic changes reported for the airways of rats under chronic adrenergic stimulation (ISO) and expected for rats under chronic catecholamines depletion (RES). Since peripheral airways involvement is usually the earliest pulmonary lesion found in CF, these studies indicate that the RES and ISO models may be representative of the early pulmonary involvement of CF.

Airway Resistance↗

Obese strain (OS) chickens with spontaneous autoimmune thyroiditis have a deficiency in thymic nurse cells.

Chicken thymic nurse cells (TNC) were isolated and their plasma membrane antigens were characterized by immunofluorescence with a panel of allo-, hetero-, and monoclonal antibodies. Analogous to the murine system, chicken TNC are localized in the subcapsular region of the cortex and were strongly B-L and B-F positive (MHC Ia and H-2 D, K equivalents). The TNC were non-phagocytic and lacked fc receptors, but expressed an antigen specific for thymic epithelial cells, and their cytoplasm was stained by an anti-keratin antiserum. The internalized lymphocytes were all intact T cells of a distinct differentiation stage, their antigenic profile consistent with that of immature cortical thymocytes. To determine whether TNC may play a role in the activation of autoimmune disease, Obese strain chickens with spontaneous autoimmune thyroiditis were investigated. These chickens were found to have a twofold defect in their TNC content: the thymuses contained approximately one-half of the number of TNC found in normal chickens beginning as early as 17 days of embryogenesis, and the lymphocyte content of the Obese strain TNC was one-half that of the normals. The deficiency is specific for TNC and may be a major reason for the abnormal T cell differentiation in the Obese strain.

Animals↗

In vitro effect of captopril and three captopril metabolites on neutrophilic progenitor cells from normal mice and mice given busulfan.

Neutrophilic progenitor cells (CFU-NM) in mice with busulfan-induced latent bone marrow hypoplasia have been shown to be selectively suppressed by captopril in vivo. To study this further, marrow from mice given busulfan was used to test for myelotoxicity in vitro. It was found that myelotoxic agents, such as doxorubicin, cytarabine, 1,3-bis(2-chloroethyl)-1-nitrosourea, and chloramphenicol have the same suppressive effect in vitro on CFU-NM obtained from normal marrow as obtained from marrow of mice given busulfan. Our results suggest that in vitro, the CFU-NMs from mice given busulfan are not excessively sensitive to the action of at least these myelotoxic agents. It was also found that neither captopril nor three known captopril metabolites suppress CFU-NM from mice given busulfan. This result suggests either that the myelotoxicity induced in vivo by captopril is not caused by the parent compound or any of the three metabolites tested, or more likely, that the suppressive action of captopril on CFU-NM in vivo is caused by the suppression of an earlier myeloid progenitor cell.

Animals↗

Longitudinal evaluation of a system for self-monitoring plaque control effectiveness in orthodontic patients.

This study was undertaken to determine the effectiveness of the Plaklite as an adjunct to plaque control instruction in orthodontic patients. 24 subjects, aged 9-14, who were to receive full banded Edgewise orthodontic appliances, were divided randomly into 3 groups. The first group was a control and did not receive structured plaque control instructions. The second group received plaque control instructions using the modified Bass brushing technique with no disclosant. The third group received a similar type of plaque control instructions supplemented with instructions in the use of a Plaklite to self-assess the effectiveness of plaque control. Both plaque control groups received monthly reinforcement of their initial instructions for the first 5 months of orthodontic treatment. The Plaque Index and Gingivitis Index were utilized to assess plaque and gingival inflammation before and after banding, and at 6-week intervals for the first 9 months of treatment. These indices were recorded single blind. The results indicated that plaque and gingivitis scores progressively increased during the test period for the control group. Also those subjects who received a plaque control program which included the Plaklite had a more sustained improvement in plaque and gingivitis scores for the test period than the group receiving plaque control instructions only.

Adolescent↗

Chronic colonization of rat airways with Pseudomonas aeruginosa.

Colonization of the airways of rats by Pseudomonas aeruginosa was established by treating the animals with hexamethylphosphoramide (HMPA) and inoculating with P. aeruginosa. Male Sprague-Dawley rats were given tap water (controls) or HMPA in the drinking water at 2 or 4 mg/ml. The ciliated cells of the airway epithelium were denuded, and microulcerative lesions in the epithelium were induced in the HMPA-treated rats. After 2 weeks of treatment, the rats were inoculated by transoral intratracheal instillation with 5 X 10(7) CFU of P. aeruginosa obtained from a cystic fibrosis patient. Two weeks after inoculation, P. aeruginosa was cultured from the airways, and scanning and transmission electron microscopy showed bacilli adhering to or invading the injured airway epithelium. P. aeruginosa was present in tracheal and intrapulmonary tissue homogenates of 9% of the P. aeruginosa-inoculated control rats (n = 22) as compared with 61% of the 2-mg/ml (n = 18) and 65% of the 4-mg/ml (n = 20) HMPA-treated rats (P less than 0.05). No dose-response relationship was found between 2 and 4 mg of HMPA per ml and colonization. Contamination of 47% of all of the rats with Mycoplasma pulmonis, as indicated by a positive enzyme-linked immunosorbent assay for immunoglobulin G, had no discernible significant effect on colonization by P. aeruginosa. These results indicate that colonization of the rat airway by P. aeruginosa can be achieved experimentally by treating the animals with HMPA. This research supports the hypothesis that colonization by P. aeruginosa may occur in airways where the ciliated epithelium has been injured and epithelial lesions exist.

Animals↗

Genetically-controlled severity of autoimmune thyroiditis in Obese strains (OS) chickens is expressed at both the humoral and cellular effector mechanism levels.

Two lines of Obese strain (OS) chickens of identical MHC (B) genotype, B5B5, bred over 10 years with different selection parameters, differ in their severity of spontaneous autoimmune thyroiditis. To determine whether alterations in immune responsiveness underly this discrepancy, the two lines were compared for their thyroiditis effector mechanisms. The OS B5B5 chickens, selected for high levels of serum thyroglobulin autoantibody, had correspondingly higher levels of thyroid-specific cytotoxic cells and also antibody dependent cellular cytotoxicity (ADCC) than the equivalent B5B5 line selected solely for the phenotypic trait of hypothyroidism. These results thus emphasize the importance of the non-MHC locus controlling immune responsiveness, in the 3 locus-model for this autoimmune disorder.

Animals↗