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Objective: Increasing attention has been given to the assessment of patient satisfaction as a way to monitor quality of care in hospital settings. Postoperative patient satisfaction has been thought to be related to level of pain intensity, expectations of outcome, perceived concern by the staff, and helpfulness of treatments. The aim of this study is to develop a simple, reliable measure to assess pain and satisfaction in postsurgical patients and to examine factors related to patient satisfaction., Design: A satisfaction questionnaire was developed for this study and administered to 119 patients who had undergone a major orthopedic surgical procedure. The majority of the patients were diagnosed with osteoarthritis and reported moderate to severe preoperative pain. The 13-item measure was found to be reliable (test-retest r = .86; interexaminer r = .98), valid (exploratory factor analyses; intercorrelations), and easy to administer., Results: Results showed that the majority of the patients were satisfied with their care (91%), postoperative pain intensity (94%), and the way they were treated by the physicians and nurses (98%). Patients with low postoperative pain ratings who perceived that the physicians and nurses showed concern with how much pain they were feeling reported greatest satisfaction with their care (p < .001). In general, lower postoperative pain ratings were the best predictors of satisfaction and helpfulness of treatment. Preoperative pain status, expected level of postoperative pain, and time waiting for pain medication after a request was made were not significantly correlated with ratings of postoperative pain or satisfaction., Conclusions: These results highlight the important influence of adequate treatment of postoperative pain and perceived concern by the hospital staff on patient satisfaction.

During early cleavages of Sicyonia ingentis embryos, mitotic spindle orientations differ between blastomeres and change in a predictable manner with each successive mitosis. From 2nd through 7th cleavages, spindles orient at a 90 degrees angle with respect to the spindle of the parent blastomere. Thus, spindle orientation is parallel to the cleavage plane that formed the blastomere. To determine if specific spindle orientations were intrinsic properties of individual blastomeres, we altered blastomere associations and asked how mitotic spindle orientation was affected in successive cleavages using laser scanning confocal microscopy. Linear embryos were constructed by dissociating 4-cell embryos and recombining the blastomeres in a linear array. The ensuing cleavage (3rd embryonic cleavage) of these linear embryos was parallel to the long axis of the embryo, resulting in four parallel pairs of blastomeres which lay in a common plane that was parallel to the substratum. The 4th cleavage produced a linear embryo with the 16 blastomeres arranged in four parallel quartets. Then, in preparation for 5th cleavage, spindles oriented at a 45 degrees angle (not parallel as in normal development) with respect to the previous cleavage plane. When 8-cell linear embryos were separated into linear half-embryos, subsequent spindle orientations were not like those observed for intact 8-cell linear embryos, but rather regressed to the orientation seen in 4-cell linear embryos. We suggest that the reorientation of mitotic spindles during early cleavage of S. ingentis is neither an intrinsic property nor age dependent, but rather is cell contact related. Further, these results in conjunction with observations of non-manipulated embryos suggest that spindle poles (centrosomes) avoid cytoplasmic regions adjacent to where there is cell-cell contact during early development.

Using a murine respiratory infection model, we have demonstrated previously that infection with Bordetella pertussis or immunization with a whole-cell pertussis vaccine induced antigen-specific Th1 cells, which conferred a high level of protection against aerosol challenge. In contrast, immunization with an acellular vaccine, consisting of the B. pertussis components detoxified pertussis toxin, filamentous hemagglutinin, and pertactin adsorbed to alum, generated Th2 cells and was associated with delayed bacterial clearance following challenge. In this study, we demonstrated that addition of interleukin-12 (IL-12) either in vitro or in vivo enhanced type 1 T-cell cytokine responses induced with an acellular vaccine. Furthermore, the rate of bacterial clearance in mice coinjected with IL-12 and the acellular vaccine was similar to that observed following immunization with a potent whole-cell vaccine. Analysis of IL-12 secretion by murine macrophages suggested that this cytokine is produced in vivo following B. pertussis infection or immunization with the whole-cell vaccine. IL-12 was detected in the supernatants of lung, splenic, and peritoneal macrophages infected with live B. pertussis or stimulated with heat-killed whole B. pertussis or B. pertussis lipopolysaccharide. In contrast, IL-12 could not be detected following stimulation of macrophages with the bacterial antigens filamentous hemagglutinin, detoxified pertussis toxin, and pertactin, the components of acellular vaccines. Our findings suggest that induction of endogenous IL-12 may contribute to the high efficacy of pertussis whole-cell vaccines and also demonstrate that it is possible to attain these high levels of protection with a less reactogenic acellular vaccine incorporating IL-12 as an adjuvant.

This study examines the expression of mRNA for the Th2 cytokine, interleukin-4 (IL-4). Peripheral blood mononuclear cells from deer infected with Mycobacterium bovis or vaccinated with live or killed M. bovis bacillus Calmette-Guérin (BCG) were cultured with mycobacterial antigens. IL-4 mRNA production was assayed using the polymerase chain reaction. Elevated levels of IL-4 mRNA were detected in response to at least one antigen preparation in all animals infected with M. bovis as compared with none of the non-infected control animals. After a primary immunization, elevated levels of IL-4 mRNA were detected in only a proportion of vaccinated animals and this did not correlate with whether the vaccine was live BCG or killed BCG in oil. After boosting, all the animals vaccinated with killed BCG in oil exhibited elevated IL-4 mRNA production whereas none of the animals vaccinated with live BCG showed elevated levels. The data suggest that IL-4 is turned off during the immune response to live BCG, that boosting of low-dose live BCG vaccine may be required to 'imprint' this signal and that this may be important in the development of protective immunity to tuberculosis. Killed BCG in adjuvant is not protective and as with experimental infection with virulent M. bovis it failed to switch off the IL-4 response. IL-4 may be useful as a diagnostic tool and as an in vitro marker of vaccine efficacy.

The efficacy of combining rate and pressure reduction during weaning by synchronous intermittent mandatory ventilation (SIMV) were compared with weaning by patient triggered ventilation (PTV) (pressure reduction alone) in two randomised trials. Regardless of ventilation mode, pressure was reduced to the same level according to the size of the infant. In the first trial, the SIMV rate was also reduced progressively to a minimum of 20 breaths/minute, and in the second to five breaths/minute. Forty premature infants aged 15 days of age or less were randomly allocated into each trial. No significant differences were found in the first trial between ventilation modes in either the duration of weaning or the number of infants in whom weaning failed. In the second trial, the duration of weaning was shorter by PTV than by SIMV (median 24 hours, range 7-432 v 50 hours, range 12-500; p < 0.05); weaning failed in two infants in the PTV group and in five in the SIMV group. It is concluded that weaning by a combination of pressure and rate reduction, such as can be achieved during SIMV, offers no significant advantage over pressure reduction alone.

We report the cloning and sequencing of the cervine interleukin-10 gene. Specific cDNA was amplified by PCR using primers based on the bovine sequence. This was cloned into pGEM 5Zf and several clones were sequenced. The 762 nucleotide product coded for a 179 amino acid protein which was 86% homologous with its bovine and 77% homologous with its human counterparts. There is a strongly hydrophobic signal sequence consisting of the first 20 amino acids and a potential glycosylation site at amino acids 134-136. There are three regions, comprising 34% of the protein, which show complete homology between the cervine, bovine and human sequences. The transcription of the gene was shown by Northern Blotting where a single, 1.8kb, mRNA transcript was detected 4-8 hours after activation of peripheral blood mononuclear cells with mitogen.

The author reviews the literature on factors which influence the health of African Americans. The concept of poverty as a health problem is discussed as well as the feminization of poverty. The author implores health care workers, to begin to implement the concepts of ethnicity and culture when giving care to clients of color.

An exploratory study utilizing ethnographic, participatory and Afrocentric research methodologies was conducted to identify causative factors related to the reluctance of a large percentage of African American women to avail themselves and their children to available health care in San Joaquin County, California. Although this qualitative study sought the responses of only ten African American women, the use of a windshield survey, a 20-item demographic questionnaire, five descriptive questions, a 128-item guide to dialogue, and transcription of dialogue from serial 60-90 minute individual and group interview sessions resulted in the identification of ten taxonomies and five prominent themes operating among this group of participants. The findings implied the need for nursing measures to enhance personal empowerment of this historically underserved population; as well as the need for nursing education to incorporate cultural-specific health care practices in curricula.

An outbreak of tuberculosis induced a mortality of 25 per cent in a captive herd of Arabian oryx (Oryx leucoryx). The diagnostic screening tests used on live animals included the comparative skin test, indirect and comparative ELISA tests and lymphocyte transformation tests. Difficulties in the interpretation of these tests stemmed principally from the facts that false negatives and false positives were encountered and that the threshold of positivity was difficult to establish with the ELISA test. The presence of other mycobacterial infections in the environment was almost certainly a complicating factor.

A patient with hypertensive crisis developed encephalopathy with a lumbar puncture revealing neutrophilic pleocytosis. No explanation for her case could be found, eg, no evidence of infection or vasculitis. It is concluded that the most probable cause of her neutrophilic cerebrospinal fluid pleocytosis is hypertensive encephalopathy.

The successful treatment of osteomyelitis with commercially prepared gentamicin-polymethylmethacrylate (PMMA) (Septopal) beads and surgical debridement has led to the use of this technique in the United States. However, commercially prepared gentamicin-PMMA beads are not currently available to orthopedic surgeons in the United States. Therefore, these surgeons commonly manufacture their own antibiotic-containing cement beads in the operating room at the time of surgery. There is little data that compare the antibiotic elution characteristics of such preparations to commercially prepared gentamicin-PMMA beads. This study compares the measured amount of antibiotic elution of either gentamicin or tobramycin from laboratory manufactured Zimmer, Simplex, or Palacos beads to commercially prepared gentamicin-PMMA (Septopal) beads. During a 30-day study period, commercially prepared gentamicin-PMMA beads eluted more total antibiotic and maintain higher concentrations than did antibiotic acrylic composites manufactured in the authors' laboratory.

We investigated the relative role of C3bi-CR3 interaction in the binding and phagocytosis of EAC43 by mouse peritoneal macrophages. Anti-Mac-1 F(ab')2 markedly inhibited the binding and lymphokine-induced phagocytosis of both EAC43b and EAC43bi. Fifty per cent inhibition of attachment and phagocytosis occurred at 1 microgram/ml of anti-Mac-1 F(ab')2 in the incubation media. On the other hand, EIgG binding and phagocytosis were not inhibited at all even at a concentration of 10 micrograms/ml. Depletion of divalent cations from the incubation media abolished EAC43b and EAC43bi rosettes but not EIgG rosettes or phagocytosis. These data suggested that both EAC43b and EAC43bi binding to macrophages were mediated via CR3. Because a drastic decrease of EAC43bi rosettes was observed in the case of EAC43bi cells prepared with smaller amounts of C3, a small contamination of C3bi molecules on EAC43b, itself, cannot explain the efficient attachment of EAC43b. We propose that EAC43b on the macrophage surface can be quickly converted to EAC43bi, forming EAC43bi rosettes, and that those erythrocytes are vigorously ingested by lymphokine-activated macrophages. In accordance with this hypothesis, we demonstrated that EAC43b was converted to EAC43bi in the medium in which macrophages had been incubated.

Macrophage C3 receptors are normally immobilized in the plane of the cells' plasma membrane and are unable to promote phagocytosis even though they promote avid particle binding. We have previously identified a lymphokine that activates macrophage C3 receptors for phagocytosis both in vitro and in vivo, and others have found that certain types of nonimmunologically mediated inflammation are also able to activate mononuclear phagocyte C3 receptors. These findings raised the possibility that macrophage C3 receptor activation is a universal consequence of inflammation. We sought in the present experiments to determine whether or not inflammation induced by microbial infection in a nonimmune host resulted in activation of macrophage C3 receptors. We injected mice i.p. with either viable microorganisms, microbe-containing immune complexes, or bacterial LPS. Macrophages were harvested by peritoneal lavage 4 days later; nearly all lavage fluids grew the microorganism with which the mouse had been injected, indicating that an infection had been established. Monolayers of macrophages were established and their interaction with sheep E coated with C3 (EIgMC) was determined. All macrophages bound EIgMC, but only macrophages from mice injected with either very high concentrations of LPS or microbe-containing immune complexes ingested them. C3 receptors of macrophages that ingested EIgMC were mobile; others were not. Thus, inflammation induced by microbial infection does not commonly, if at all, activate macrophage C3 receptors; microbe-containing immune complexes and high concentrations of LPS do. The mechanism of receptor activation in each case is C3 receptor mobilization, which is probably mediated by a lymphokine.

The ongoing epidemic of acquired immune deficiency syndrome (AIDS) has affected homosexual men, intravenous (IV) drug abusers, Haitians, hemophiliacs, and others. Defects in cell-mediated immunity place these patients at risk for opportunistic infections. We recently saw three men from Alabama with disseminated infection due to Histoplasma capsulatum. Two of these men were homosexual and the other was an IV drug abuser. These three patients had evidence of depressed cellular immunity consistent with a diagnosis of AIDS. Infection caused by organisms indigenous to certain geographic areas of the United States may become more common in patients with AIDS as the epidemic continues.

These experiments were designed to evaluate the role of macrophage plasma membrane receptors for the third component of complement (C) and for the Fc portion of IgG in the ingestion phase of phagocytosis. Sheep erythrocyte (E) were coated with anti-E IgG [E(IgG)]; these E(IgG) were then attached to cultivated monolayers of mouse peritoneal macrophages under conditions which reversibly inhibit ingestion of E(IgG). The E(IgG)-macrophage complexes were further incubated under similar conditions with an antimacrophage IgG fraction which blocks Fc receptor-mediated ingestion but has no effect upon ingestion mediated by other phagocytic receptors. When these cultures were subsequently incubated under conditions optimal for particle ingestion, phagocytosis of the IgG-coated erythrocytes did not occur; the erythrocytes remained bound to the Fc receptors of the macrophage plasma membrane. To determine whether ligands must cover the entire surface of an attached particle to permit ingestion of that particle, C-coated E [E(IgM)C] were bound to the C receptors of thioglycollate-induced (activated) macrophages at 4 degrees C. E(IgM)C-macrophage complexes were then trypsinized at 4 degrees C, a procedure which resulted in cleavage of erythrocyte-bound C3b molecules to a form of C3 not recognized by the macrophage receptors for C3b. Under the conditions used, trypsin did not affect the attachment of E(IgM)C to the macrophage surface or the macrophage receptors for C3b. When these trypsin treated E(IgM)C-macrophage complexes were incubated at 37 degrees C, the bound E(IgM)C were not ingested; the erythrocytes remained attached to the macrophage plasma membrane via the macrophage's C receptors. These results indicate that attachment of a particle to specific receptors on the macrophage plasma membrane is not sufficient to trigger ingestion of that particle. Rather, ingestion requires the sequential, circumferential interaction of particle-bound ligands with specific plasma membrane receptors not involved in the initial attachment process.

Moderate concentrations of sodium azide (0.1-0.2%) significantly inhibited guinea-pig and human complement-mediated lysis of both IgM- and IgG-sensitized sheep erythrocytes. The reduction in cytolysis was not attributable to non-specific ionic effects, to inactivation of native complement components by azide, or to irreversible interactions of azide with sensitized erythrocytes. Mouse complement-dependent opsonization of sensitized erythrocytes, as judged by macrophage complement receptor-mediated attachment and phagocytosis of the erythrocytes, was comparably inhibited by sodium azide, suggesting that azide acted within the sequence of the first four components of the classical complement pathway.

We have identified a human T lymphocyte cell line, the Mo line, that constitutively elaborates a lymphokine that activates macrophage receptors for the third complement component (C3) for phagocytosis. The molecule is physically, functionally, and immunologically very similar, if not identical, to a previously characterized murine lymphokine. Both molecules were inactivated by the same physical and chemical treatments, activated macrophage C3 receptors for phagocytosis, and freed macrophage C3 receptors to move within the cell's plasma membrane. In addition, both the human and the murine products were recognized by the same monoclonal antibodies. Treatment with monoclonal antibodies abolished both the ability of lymphokine-containing supernatants to mobilize macrophage C3 receptors and the supernatants' ability to activate C3 receptor-mediated phagocytosis. The dose-response curves for both effects of the antibodies were identical, strongly suggesting that a single molecule was responsible for both effects and that the lymphokine activated marophage C3 receptors for phagocytosis by freeing the anchored receptors and allowing them to diffuse within the cell's plasma membrane. These findings strengthen the hypothesis that, for a receptor to promote phagocytosis, it must be able to diffuse within the macrophage plasma membrane.

The uncontrolled exposure of Fischer's medium to cool white fluorescent (CWF) light or other sources emitting near-ultraviolet or visible light absorbance by riboflavin is a crucial random variable in experiments which utilize L5178Y cells and this medium. The radiation effects of CWF light result in the rapid development of toxic photoproducts in the medium which are cytostatic at lower doses of radiation and cytotoxic at higher doses. After a 24-hr suspension in medium irradiated for 3 or 48 hr, the cloning efficiencies of cells subsequently plated in light-protected medium were 87 and 3%, respectively. The corresponding near-ultraviolet doses for these periods of exposure to CWF light were 0.22 x 10(4) for a 3-hr exposure and 3.47 x 10(4) J/sq m for a 48-hr exposure. Cells incubated in lightly irradiated medium resumed growth at nearly normal rates following a 24- to 48-hr period in which no increase in cell numbers occurred. Exposure of medium containing riboflavin, but not tryptophan or tyrosine, to CWF light also produces toxic medium. Tryptophan enhances riboflavin-induced phototoxicity, whereas tyrosine diminishes this effect. As photosusceptibility of this system is very high, Fischer's medium must be fully protected from all sources of light absorbable by riboflavin.

Sperm of the shrimp, Sicyonia ingentis, undergo a biphasic acrosome reaction consisting of acrosomal exocytosis and acrosomal filament formation. These events are temporally separated by 10-20 min, in vivo. Using egg water preparations the complete reaction can be induced, in vitro, albeit the temporal separation of the two phases is lengthened. External Ca ++ is required for the exocytotic phase, while a cytoplasmic acidification and K + efflux are associated with polymerization of the acrosomal filament.

Serum samples were collected from 3 mature female African elephants once each week for 15-18 months. Circulating concentrations of progesterone, oestradiol and LH were determined by radioimmunoassay (RIA). The LH RIA was validated by demonstrating parallel cross-reaction with partly purified elephant LH pituitary fractions. Changing serum progesterone concentrations indicated an oestrous cycle length of 13.3 +/- 1.3 weeks (n = 11). The presumed luteal phase, characterized by elevated serum progesterone values, was 9.1 +/- 1.1 weeks (n = 11). Two abbreviated phases of progesterone in serum lasting 2-3 weeks were observed in 2 elephants, indicating short luteal phases. Oestradiol concentrations in serum were variable, with no clear pattern of secretion. More frequent blood samples were collected during periovulatory periods and 9 distinct LH peaks were detected; all were followed by rises in serum progesterone concentrations. Periovulatory changes in progesterone and LH in sera correlated with external signs of oestrus and mating behaviour.

We have previously reported that treatment with a unique lymphokine enables resident mouse peritoneal macrophages to phagocytize via their complement receptors and we have presented evidence that the lymphokine act by enabling complement receptor engagement by C3b ligands to generate a phagocytic signal, thereby linking the cell surface binding event with the intracellular phagocytic machinery. In the present experiments, we used immobilized immune complexes to study the topography of C3b receptors of resident mouse peritoneal macrophages treated with the lymphokine. Our results indicate that lymphokine treatment enables the macrophages' C3b receptors to migrate within the plane of the cells' plasma membrane and that manipulations of macrophages that abrogate one response to the lymphokine, complement receptor mobility, also abrogate the other response, complement receptor-mediated phagocytosis. These findings strongly suggest that lateral mobility of a ligand-bound receptor within the macrophage plasma membrane is an essential component of the phagocytic signal. Moreover, our results indicate that the difference in complement receptor function among various populations of macrophages is not due to the expression of different types of complement receptors by the different macrophage populations but rather to a difference in the relationship of the C3b receptor with other plasma membrane or intracellular components.

A method for the isolation of 8-10 mg of human C3 from 20 ml of plasma is described. The procedure is simple and rapid with excellent yields in hemolytic activity (74%) and antigenic activity (68.7%). It consists of polyethylene glycol precipitation, DEAE-Sephacel chromatography, and immunoadsorption. The final product is free of contaminating proteins as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The simplicity and speed of this procedure allow for the continual availability of hemolytically active C3.

A method of attaching mouse RBCs to mouse macrophages is described. Both cell types were coated with rabbit anti-mouse macrophage F(ab')(2), and cross-linkage of cells was effected with sheep F(ab')(2) directed against rabbit F(ab')(2). 98% of macrophages attached an average of 11 RBCs each. Attachment occurred at 37 degrees C and was stable for at least 4 h. Less than 0.1% of macrophages ingested RBCs under these conditions. Latex particles and opsonized pneumococci were ingested as avidly by RBC-coated macrophages as by native macrophages. Ingestion of these particles did not prompt ingestion of attached RBCs. When anti-RBC IgG was added, however, over 90% of macrophages ingested an average of six RBCs each. Thus, ingestion of one particle does not trigger generalized phagocytosis of all particles attached to the cell's plasma membrane, and the phagocytic stimulus is confined to the segment of the cell's plasma membrane immediately adjacent to the particle being ingested.

The ability of several immunologic ligands to mediate extracellular cytolysis of sheep erythrocytes (E) by mouse macrophages was studied. E coated with rabbit IgG (EIgG), but not nonsensitized E or E coated with rabbit IgM (EIgM), were lysed and phagocytized by resident peritoneal macrophages as well as by macrophages activated in vivo with either BCG or Corynebacterium parvum or in vitro with lymphokine. EIgM incubated with mouse serum to deposit the third component of complement (C3) onto E (EIgMC) were ingested by thioglycolate-elicited and lymphokine-treated macrophages, but not by the other macrophage populations examined. However, none of the macrophages performed lysis of EIgMC, suggesting that engagement of macrophage C3 receptors by target cell-bound C3 was not a sufficient trigger for cytolysis. Lysis of E coated with both IgG and C3 (EIgGC) was moderately elevated over that of EIgG; this enhancement was not abolished after proteolytic destruction of the macrophage C3-rosetting capacity, indicating that C3 receptors were not responsible. EIgGC and EIgMC were more susceptible to hypotonic lysis than were either E, EIgG, or EIgM, suggesting that enhanced lysis of serum-treated E may be partially explained by increased E fragility.

We investigated the likelihood that human peripheral blood mononuclear cells which incorporate tritiated thymidine in the presence of 6-thioguanine (6-TG) are mutants by comparing findings for cells from the same blood specimen cultured in the presence or absence of phytohemagglutinin (PHA). After 42 h in culture, autoradiography revealed that about 0.1% of cells from healthy control donors and 1% of cells from patients receiving chemotherapy for breast cancer labeled in the absence of PHA. About 5% of this number labeled when 6-TG was added to the medium. This high frequency of cells labeling in the presence of 6-TG suggested that many cells were resistant to 6-TG for reasons other than mutation at the HGPRT locus. Despite a labeling index of about 30% in the presence of PHA alone, the addition of PHA to cells cultured with 6-TG did not increase the labeling index over that observed with 6-TG alone. Also, in the presence of 6-TG there was a significant correlation between labeling indices for cells from the same blood specimen cultured with or without PHA. These findings suggest that the two conditions measured the same population of non-mutant cells. These studies reinforce observations made by the developers of this technique and other laboratories which indicate that refinements will be required before the assay can be used to study somatic cell mutation in human populations. Such refinements are underway in several laboratories; although not definitive, the approach and procedures used in this report which investigated the original assay procedure provide methods and criteria to judge whether modifications of assay techniques improve the quantification of in vivo human somatic cell mutation.

Invasive pulmonary aspergillosis occurs predominantly in individuals who are neutropenic or who have severe defects in cell-mediated immunity. The isolation of Aspergillus from respiratory secretions of normal hosts usually signifies tracheobronchial colonization, not disease. Recent experience with three nonimmunocompromised patients who had invasive pulmonary aspergillosis, each of whom had Aspergillus isolated from respiratory secretions early in his illness, led to a reassessment of the significance of the isolation of Aspergillus from tracheobronchial secretions. Two of 10 nonimmunocompromised, nonleukopenic individuals who had pulmonary infiltrates and whose sputum yielded Aspergillus had invasive pulmonary aspergillosis, whereas two of five individuals who had pulmonary infiltrates and whose bronchial washings grew Aspergillus had invasive disease. These findings indicate that invasive pulmonary aspergillosis should be considered when Aspergillus is isolated from the respiratory secretions of anyone who has pneumonia, regardless of host defense status.

We have examined the roles of Fc receptors and complement receptors in mediating the interaction of sensitized sheep erythrocytes (E) with activated and with nonactivated mouse peritoneal macrophages. Both activated and nonactivated macrophages ingest IgG-coated erythrocytes [E(IgG)]; activated cells intest 1.5-2 times as man E(IgG) as do nonactivated macrophages. Thus, there is a quantitative difference in Fc receptor-mediated ingestion between activated and nonactivated macrophages. There is, however, a qualitative difference in function of complement receptors of activated and nonactivated macrophages. Nonactivated macrophages avidly bind complement-coated E [E(IgM)Ia1, but do not ingest them to a significant degree. Activated macrophages, on the other hand, bind and ingest E(IgM)C. The possibility of Fc receptor participation in mediating ingestion of E(IgM)C by activated macrophages was eliminated by blocking Fc receptors with an antimacrophage IgG fraction. Activated macrophages treated with antimacrophage IgG did not ingest E(igG) but did ingest both E(IgM)C AND E(IgM)C. Nonactivated macrophages treated with antimacrophage IgG did not interact at all with E(IgG). These cells bound, but did not ingest, E(IgM)C and E(IgM)C. Complement receptor-mediated ingestion is a marker for macrophage activation and may be physiologically important in the elimination of complement-coated particles.

The function of complement receptors of mouse peritoneal macrophages was converted in vitro from mediating only attachment of macrophage complement receptor function was achieved by treating freshly explanted macrophages with supernates from cultures containing T lymphocytes and appropriately triggered macrophages. Fc receptor-mediated phagocyctosis by macrophages was required for the production of active supernates, for neither ingestion via the cells' complement receptors nor ingestion via nonimmunologic means was a sufficient stimulus for the macrophages' participation in the generation of supernatant activity. Fc receptor-triggered macrophages interacted by a contact dependent, but histocompatibility independent, mechanism with T lymphocytes, thereby signalling the lymphocytes to elaborate the active product. The possible significance of enhanced macrophage complement receptor function in inflammation, host defense against microbial pathogens, immune complex disease, and neoplasia is discussed.

Macrophage complement receptors, while innately incapable of promoting phagocytosis, can be activated to do so by a number of inflammatory stimuli and by several immunologic mechanisms. Studies with a complement receptor-activating lymphokine reveal that activation occurs as a result of mobilization of innately immobile receptors and suggest that receptor mobility is a prerequisite for phagocytosis. Since Fc receptors are susceptible to blockade by immune complexes at inflammatory sites, phagocytosis mediated by macrophage complement receptors may be of prime importance in vivo.

Sperm from a marine shrimp, Sicyonia ingentis, were frozen to -196 degrees C using a variety of cooling rates and cryoprotectants. A cooling rate of 1 degree C/min resulted in minimal cell breakage. Sperm samples were frozen in solutions of known membrane stabilizers--trehalose, sucrose, proline, and glycerol. These compounds were somewhat effective but a dramatic increase in sperm viability was seen when DMSO was present in the freezing medium. Sperm viability was assessed using the in vitro acrosome reaction technique of Griffin et al. (1987). The highest sperm survival (56%) was obtained with samples frozen at 1 degrees C/min in a 5% (v/v) DMSO solution. No decrease in viability was seen in sperm samples stored in liquid nitrogen (-196 degrees C) for 1 month.

The complement receptor of the macrophage membrane recognizes particle-bound C3b but does not recognize particle-bound C3d. C3-b-coated sheep erythrocytes were bound to macrophages via their C3b receptors, and the preparations were then incubated with either latex particles or opsonized pneumococci (test particles). Macrophages ingested the test particles, but erythrocytes were not ingested; they remained bound to C3b receptors of the macrophage plasma membrane. Thus, a signal initiating ingestion via one type of receptor is not transmitted to all receptors which have the potential to mediate phagocytosis.

In this study the lymphocyte function of therapy matched rheumatoid and non-rheumatoid patients in the mitogen mediated in vitro whole blood microculture system has been analyzed. Normal responses to phytohemagglutinin (PHA) and poke weed mitogen (PWM) and depressed responses to concanavalin A (Con A) were found. The PHA and Con A responses were influenced by drug therapy. Differences in the relative activity of PHA to Con A among the groups studied further supports the contention that rheumatoid arthritis (RA) patients have functional abnormalities in their T cell subpopulations. These differences were not due to a decrease in circulating lymphocytes or in the ratio of T lymphocyte numbers but likely reflects a disproportionate representation of Con A reactive T cells in the circulating T cell pool of RA patients.

The effects of ingestion of soluble immune complexes upon macrophage phagocytic function was studied. Ingestion of immune complexes severely impaired the macrophage's ability to ingest IgG-coated particles but did not alter its ability to interact with particles by means other than its Fc receptors. Treatment of macrophages that had ingested immune complexes with supernates containing the previously described lymphokine that augments macrophage complement receptor function failed to enhance the cells' interaction with either IgG-coated erythrocytes or zymosan particles but markedly enhanced their ability to phagocytize via their complement receptors. The possible significance of these findings in immunologically mediated inflammation is discussed.

I have studied the roles of macrophage Fc and C3b receptors in the cell's interaction with encapsulated Cryptococcus neoformans and have defined the effects of a lymphokine that enhances macrophage complement receptor function, the effects of ingestion of soluble immune complexes, and the effects of corticosteroid treatment upon the ability of macrophages to phagocytize cryptococci via these receptors. Neither uncoated nor C3-coated cryptococci were phagocytized, whereas IgG-coated cryptococci were avidly phagocytized by mouse peritoneal macrophages. Treatment of macrophages with the lymphokine enabled them to ingest C3-coated cryptococci. Prior ingestion of soluble immune complexes severely compromised macrophages' ability to phagocytize cryptococci via their Fc receptors but did not affect their ability to ingest cryptococci via their complement receptors. Corticosteroid treatment severely impaired the ability of macrophages to respond to the lymphokine. Based upon these experimental observations, I have constructed a model for normal host defense mechanisms against disease due to C. neoformans.

Macrophage receptors for the third component of complement (C3) are normally immobilized and unable to diffuse within the cell's plasma membrane and, even though they promote avid particle binding, are unable to promote phagocytosis of C3-coated particles. We have previously identified a lymphokine that activates macrophage C3 receptors for phagocytosis and have found that it acts by freeing the receptors so that they can diffuse within the macrophage plasma membrane. It seemed likely to us that the initial lymphokine-macrophage interaction would occur at the macrophage surface, perhaps via a specific lymphokine receptor. Since the binding of many ligands to cells is mediated by cell surface glycoproteins, we examined the protein and sugar requirements for murine peritoneal macrophages to respond to the lymphokine. Macrophages treated with the neutral protease Dispase lost the ability to respond to the lymphokine, and inclusion of L-fucose in the incubation medium containing lymphokine and macrophages inhibited markedly the macrophages' response to the lymphokine, suggesting that the lymphokine exerts its effects by first binding to fucose residues on a glycoprotein receptor on the macrophage surface. Further evidence for the essential role of macrophage surface fucose was obtained by demonstrating that pretreatment of macrophages with either fucosidase or gorse lectin, a fucose-binding lectin, strikingly disabled the cells from responding to the lymphokine. All treatments that prevented lymphokine activation of macrophage C3 receptors for phagocytosis also prevented lymphokine-induced C3 receptor mobility. These results strongly suggest that the lymphokine binds to a fucose-bearing macrophage surface glycoprotein, perhaps a specific lymphokine receptor. They also strengthen our hypothesis that, for a receptor to be able to promote phagocytosis, it must be able to diffuse within the macrophage plasma membrane.

We have examined the effect of the distribution of anti-immunoglobulin IgG molecules on the surface of bone marrow-derived lymphocytes upon the interaction of these cells with macrophages. Lymphocytes which were diffusely coated with antibodies to surface immunoglogulin were ingested by macrophages. Lymphocytes which had the same number of anti-immunoglobulin IgG molecules redistributed to one pole of the surface bound to the macrophages' Fc receptors but were not ingested. These results confirm our previous hypothesis that ingestion of an immunologically coated particle requires the sequential, circumferential binding of specific receptors on the plasma membrane of a phagocytic cell to immunologic ligands distributed over the entire particle surface. Macrophages which had bound capped lymphocytes by the macrophages' Fc receptors removed the immune complex caps from the lymphocyte surface without destroying the lymphocytes. These lymphocytes remained attached to the macrophage surface. The finding that macrophages can phagocytize immune complexes from the surface of a cell without destroying the cell to which these complexes are attached may be important in understanding the effects of antigens and antibodies on cells participating in a humoral immune response, in identifying the mechanisms by which chronic viral infections are established, and in defining the roles of blocking antibodies in tumor immunity.

The phagocytic functions of polymorphonuclear leukocytes and mononuclear phagocytes are crucial elements in host defence against a variety of invading microorganisms. Phagocytosis is a highly selective process, requiring specific interactions between the surface of the particle to be ingested and the plasma membrane of the phagocytic cell. The phagocyte can therefore discriminate between 'ingestible' and 'non-ingestible' particles even when the different particles are located in close proximity on the plasma membrane of the phagocytic cell. Previous work has demonstrated that these specific interactions between particle and phagocyte are required not only for the initiation of phagocytosis--that is, attachment of the particle to the phagocytic cells and generation of a signal to trigger phagocyte pseudopod extension--but also for the subsequent progression of pseudopods over the entire surface of the particle. We present evidence here that the continued interactions between phagocyte plasma membrane receptors and particle-bound ligands do not function merely to direct otherwise random phagocyte pseudopod movement, but instead are required for the repeated generation of intracellular phagocytic signals during the entire ingestion process.

We studied the phagocytosis by human polymorphonuclear leukocytes (PMN) of sheep erythrocytes (E) passively sensitized with pneumococcal C-polysaccharide (E-PnC), E-PnC coated with C-reactive protein (E-PnC-CRP), and E coated with rabbit antisheep E IgG (E-IgG). PMN isolated from the blood of normal individuals failed to ingest either E-PnC or E-PnC-CRP; however, after incubation with supernatants from stimulated peripheral blood mononuclear leukocytes, glass-adherent PMN ingested E-PnC-CRP with a mean phagocytic index of 47.8 +/- 14.9 (means +/- SD, n = 9) and E-PnC to a lesser extent with a phagocytic index of 9.6 +/- 2.9 (mean +/- SD, n = 4). We also observed a statistically significant increase in the ingestion of E-IgG by lymphokine-stimulated PMN with phagocytic indices of 85.2 +/- 21.2 (mean +/- SD, n = 12) for unstimulated PMN and 158 +/- 37.1 (mean +/- SD, n = 9) for stimulated PMN. The best conditions for stimulating release of this phagocytosis-promoting mediator included exposure to phytohemagglutinin (PHA) in the presence of monocytes that had ingested IgG-coated sheep erythrocytes. The induction of phagocytosis of E-PnC-CRP was rapid, reaching a maximal level after stimulation of the adherent PMN with the conditioned media for 30 min. The factor(s) responsible for the induction of the ingestion of E-PnC-CRP was less than 10,000 daltons and was heat stable (56 degrees C for 45 min). These data are similar to earlier results obtained with PMN activated by 12-0-tetradecanoyl-phorbol-13-acetate (PMA), an important contrast being that in the current studies PMN were activated by a soluble factor released from stimulated mononuclear cells under conditions simulating those in vivo.