Presence Of Polymorphonuclear Leukocytes Research Articles

The polymorphonuclear leukocyte: has it a role in fracture healing?

The aim of the present study was to assess whether the presence of polymorphonuclear leukocytes in the first stage of fracture repair was of any importance. In anesthetized male Wistar rats, a transverse osteotomy was performed at midshaft in one femur and immediately stabilized by an intramedullary Kirschner nail. The animals were allowed unprotected weight bearing immediately. Nine animals were made neutropenic before operation by injection of a specific rat polymorphonuclear leukocyte antiserum (antineutrophil serum, ANS) raised in sheep. The number of circulating polymorphonuclear leukocytes were kept below 20% of their normal value for 72 h postoperatively by intraperitoneal injections of ANS every 12 h. Control animals were injected with the same amount of normal sheep serum (NSS). All animals received cefuroxime (100 mg/kg day) concomitantly with the ANS injections. Six weeks after operation the animals were killed, the amount of callus formation was measured, and the bones were radiologically examined. The nails were then removed and both the healing fractures and the nonoperated femurs mechanically tested for bending. There were no differences in the amount of callus measured or in radiological healing. However, there was a significantly higher bending moment in femurs from animals treated with ANS than in those given NSS (P less than 0.02). No differences were observed in rigidity or total energy absorption.

Type 1 fimbriate strains of escherichia coli initiate renal parenchymal scarring

The renal scarring which characterizes chronic pyelonephritis is initiated by bacterial infection and is independent on the activation of an inflammatory response. Although the presence of polymorphonuclear leukocytes (PMN) is essential for initiating the scarring process, the bacterial structures responsible for their activation have not been investigated. In an animal model of chronic pyelonephritis the surface area of the renal scars produced by Type 1 fimbriate escherichia coli (E. coli) was significantly greater than that of those produced by P fimbriate and non-fimbriate strains (P less than 0.01). The activation of human PMN by the same Type 1 fimbriate organisms resulted in a significant release of lysosomal neutral protease activity (P less than 0.001) and activation of the respiratory burst (P less than 0.01). The neutral protease release in response to P fimbriate and non-fimbriate organisms was not significantly increased. The extent of renal scarring also correlated with the release of neutral protease activity (P less than 0.02) and with the degree of activation of the respiratory burst (P less than 0.05). These results demonstrate that the ability of E. coli strains to cause renal scars may be related to their capacity to express Type 1 fimbria, which may be a causative factor in the in vivo activation of the inflammatory response.

Role of polymorphonuclear leukocytes in collagenase production in chronic otitis media

The effect of polymorphonuclear leukocytes on collagenase production by fibroblast-like cells was studied in an in vitro cell culture system. An extract of rat intraperitoneal polymorphonuclear leukocytes induced significant collagenase production by rat skin fibroblast-like cells. Fibroblast-like cells cultured in the presence of polymorphonuclear leukocytes had many conspicuous microspikes on their surface and contained bundles of actin filaments in their cytoplasm. Polymorphonuclear leukocytes appears to be involved in bone resorption by inducing collagenase production by fibroblast-like cells, as well as by releasing their own enzymes involved in bone resorption.

A Polyclonal Human IgG Preparation Hyperimmune for Type III, Group B Streptococcus: In Vitro Opsonophagocytic Activity and Efficacy in Experimental Models

Neonates at risk for disease due to type III, group B Streptococcus (III-GBS) are those born with low serum levels of transplacentally derived, specific III-GBS antibody. Specific antibody is also required in vitro for opsonophagocytosis of III-GBS. Commercially available human immune serum globulins contain only moderate levels of III-GBS antibody. In the present study, IgG that was isolated from the serum of a human volunteer after vaccination with III-GBS polysaccharide contained very high levels of III-GBS antibody. Small amounts of this hyperimmune preparation added in vitro significantly increased the opsonophagocytosis of III-GBS in neonatal sera in the presence of polymorphonuclear leukocytes. Furthermore, small volumes of the preparation were protective in mouse and neonatal rat models of III-GBS disease. The administration of such hyperimmune human IgG preparations should be considered for preventing or treating III-GBS disease in infants.

Apparent Effect of Catalase on Airway Edema in Guinea Pigs

The airway edema that develops in guinea pigs after exposure to toluene diisocyanate (TDI) requires the presence of polymorphonuclear leukocytes (PMN). To determine whether this airway edema is mediated by the release of hydrogen peroxide from PMN, we treated animals intravenously with catalase bound to polyethylene glycol and examined the extravasation of Evans blue dye into the tracheal wall after exposure to air or 3 ppm TDI for 1 h. Catalase (25,000, 100,000, and 300,000 IU/kg) caused a dose-dependent inhibition of the TDI-induced increase in dye extravasation. However, treatment with catalase, inactivated at the peroxide binding site with 3-aminotriazole, inhibited dye extravasation after exposure to TDI as effectively as the equimolar 100,000 IU/kg dose of active catalase. Injection of polyethylene glycol alone was without effect. Dose-dependent decreases in extravascular migration of PMN and in circulating PMN also were noted after catalase treatment. These results suggest that the catalase preparations used in these studies inhibited the PMN-dependent airway edema by an effect other than hydrogen peroxide scavenging. Examination of this and other commercially available catalase preparations revealed trace concentrations of endotoxin at levels that could be responsible for the observed effects on PMN function. Treatment of animals with doses of Escherichia coli endotoxin similar to those inadvertantly administered to the catalase-treated groups (0.1 ng/kg to 100 ng/kg, intravenously) inhibited TDI-induced extravasation of Evans blue dye in a dose-dependent manner. These results suggest that contaminating endotoxin may contribute to some of the protective effects of preparations of catalase observed in previous studies of vascular permeability.

Intraepidermal accumulation of polymorphonuclear leukocytes in chronic stable plaque psoriasis.

The intraepidermal penetration of polymorphonuclear leukocytes is an early event in the pathogenesis of psoriasis. In the present study we quantified these cells in biopsies from lesional epidermis of chronic stable plaque psoriasis. In this phase intraepidermal penetration of polymorphonuclear leukocytes was shown in 41% of the lesions. Micropustules of Kogoj were seen in 10% of the lesions. Pronounced parakeratosis was associated with the presence of polymorphonuclear leukocytes. Microabscesses of Munro were located exclusively within areas of parakeratosis. The absence of polymorphonuclear leukocytes in the majority of skin biopsies from chronic stable plaque psoriasis is compatible with the cyclic pattern of the exocytosis of these cells.

Evaluation of cultural techniques for isolating Campylobacter pyloridis from endoscopic biopsies of gastric mucosa.

One hundred and three gastroscopic biopsies from 80 patients were cultured for Campylobacter pyloridis and studied histologically. Active chronic gastritis, as shown by the presence of polymorphonuclear leucocytes, was diagnosed in 51 biopsies and C pyloridis was found in 47. Sixteen gastric biopsies showed normal histology (no inflammation); C pyloridis was detected in only one of these, and a second biopsy taken from this patient at the same time showed active gastritis. Biopsies could be kept at 4 degrees C for five hours without loss of viability of C pyloridis. An inoculum made by grinding the biopsy in a ground glass grinder consistently gave a much heavier growth of C pyloridis than one made by mincing the specimen. The campylobacter supplement ferrous sulphate, sodium metabisulphite, sodium pyruvate (FBP) (Oxoid) was inhibitory for some isolates; the inhibitory component was found to be sodium metabisulphite. Contaminants, but not C pyloridis, were inhibited by the incorporation of vancomycin 6 mg/l, nalidixic acid 20 mg/l, and amphotericin 2 mg/l, but higher concentrations inhibited C pyloridis. Undried plates kept in a plastic container at room temperature for up to two weeks were as satisfactory as freshly poured plates for the isolation of C pyloridis.

Campylobacter cinaedi (sp. nov.) and Campylobacter fennelliae (sp. nov.): two new Campylobacter species associated with enteric disease in homosexual men.

In a recent study in Seattle, we isolated Campylobacter-like organisms (CLOs) significantly more often from homosexual men with intestinal symptoms than from asymptomatic heterosexual and homosexual control subjects. Biochemical tests, morphology, and guanine-plus-cytosine content of whole-cell DNA showed that these organisms were similar to, but distinguishable from, other species in the genus Campylobacter. DNA homology tests showed that the Campylobacter-like organisms belong to four unique genetic groups, two of which were phenotypically identical. We developed a rapid screening test to genetically group these organisms and to assess the correlation of each genotype with enteric disease in homosexual men. Three of the four CLO groups were isolated only from men with proctitis, proctocolitis, and/or enteritis. The remaining group, isolated from both symptomatic and asymptomatic homosexual men, was significantly correlated with the presence of polymorphonuclear leukocytes in the rectal secretions of asymptomatic men, a finding that suggests subclinical disease. We have tentatively classified two of the CLO groups into species, Campylobacter cinaedi sp. nov. and Campylobacter fennelliae sp. nov.

Influence of migratory blood cells on the attachment of tumor cells to vascular endothelium.

The purpose of this study was to determine whether the presence of migratory blood cells in association with tumor emboli had the capacity to alter the attachment of tumor cells to vascular endothelium. Highly metastatic RT7-4bs rat hepatocarcinoma cells were labelled with [125I]UdR before being allowed to form mixed cellular spheroids incorporating resident peritoneal macrophages, activated peritoneal macrophages, polymorphonuclear leukocytes, splenic T lymphocytes, or splenic B lymphocytes derived from both normal and tumor-bearing animals. The presence of polymorphonuclear leukocytes or activated macrophages led to a considerable increase in the number of tumor cells attaching to endothelial cell monolayers in vitro. The presence of T or B lymphocytes from either normal or tumor-bearing rats was without effect on tumor attachment to endothelium. Increased tumor cell retention in the lungs was evident for mixed spheroids containing tumor cells and polymorphonuclear leukocytes compared to homotypic tumor spheroids composed of tumor cells alone. Furthermore, preinjection of polymorphonuclear leukocytes intravascularly or inoculation of tumor cells as heterotypic spheroids containing polymorphonuclear leukocytes increased lung colony formation over that obtained after inoculation of tumor cells alone. Several simple sugars were tested for their ability to block tumor cell, polymorphonuclear leukocyte or activated macrophage binding to endothelium in vitro. The results indicate that the glycosylated cell surface components mediating tumor cell attachment to endothelium are not identical with those mediating attachment of either polymorphonuclear leukocytes or activated macrophages. Medium conditioned during mixed spheroid formation was without effect on tumor cell attachment to endothelium. We conclude that the presence of some, but not all classes of leukocytes can modulate tumor cell attachment to vascular endothelium, an effect most likely mediated by a mechanism involving direct contact between the leukocytes and the endothelial cell monolayer.

Pathogenesis of bacterial meningitis: contributions by experimental models in rabbits.

Rabbits models of bacterial meningitis have contributed substantially to our understanding of the disease, although the technical characteristics of these models only allow the study of specific aspects of the disease. Bacterial multiplication in the subarachnoidal space is not substantially influenced by host defense mechanisms, mainly because of the lack of sufficient amounts of specific antibodies and functional complement in infected CSF. The multiplying bacteria induce profound changes in the blood-brain barrier, an influx of serum proteins into the CSF and the invasion of polymorphonuclear leukocytes at the site of the infection. The presence of polymorphonuclear leukocytes in CSF not only appears to be of limited value in combating the infection, but also seems to produce deleterious effects on the central nervous system. Components of the leukocytes, such as unsaturated fatty acids, arachidonic metabolites and free oxygen radicals, may contribute to the profound hydrodynamic, structural and metabolic changes that are currently under study in experimental models of the disease. A better understanding of the pathophysiology of bacterial meningitis may allow us to design more effective therapeutic strategies and improve the outcome of this disease.

Prospective study of gram-stained stool smears in diagnosis of Clostridium difficile colitis.

Gram stains of stools from patients with diarrhea and control patients with no diarrhea were examined for a predominance of gram-positive rods and the presence of polymorphonuclear leukocytes. Results were compared with those from lower gastrointestinal endoscopy for pseudomembranes. Clostridium difficile culturing, and C. difficile toxin assay. The Gram stain was moderately difficult to interpret and was not useful in diagnosing diarrheal disease associated with C. difficile.

Pathogenesis of foreign body infection: description and characteristics of an animal model.

An animal model involving the subcutaneous implantation of tissue cages into guinea pigs and subsequent infection with Staphylococcus aureus was used for study factors pertinent to foreign body infection. Whereas 10(8) colony-forming units (cfu) of S. aureus strain Wood 46 did not produce any abscesses in the absence of foreign material, 10(2) cfu was sufficient to infect 95% of the tissue cages despite the presence of polymorphonuclear leukocytes (PMNLs) in sterile tissue cage fluid. Opsonization of S. aureus by tissue cage fluid was adequate during the first hour of infection, but opsonic coating of the organisms decreased at 20 hr after the induction of infection. PMNLs from sterile tissue cage fluid showed decreased phagocytic and bactericidal activities when compared with PMNLs from either blood or peritoneal exudate obtained after short- or long-term stimulation (P less than 0.001).

Bacteriology of Tracheal Aspirates in Intubated Newborn

The bacteriology of tracheal aspirates from 28 intubated newborn babies was studied. There was no correlation between such colonization and the respiratory distress syndrome. Endotracheal suction of intubated infants did provide a reliable specimen source for determining the etiology of perinatal pneumonia. Presence of polymorphonuclear leukocytes in the aspirate correlated well with infection. Anaerobic bacteria were found to play a role in three of the five cases of pneumonia.

Pneumonia associated withTorulopsis glabratain an aborted bovine fetus

The second case of bovine abortion caused by Torulopsis glabrata observed in examining more than 5000 fetuses is reported. The yeast was cultured from fetal abomasal content and fetal lung; attempts to isolate viruses and bacteria were unsuccessful. The yeast was seen in association with diffuse fetal pneumonia characterized by presence of polymorphonuclear leukocytes and macrophages within bronchioles, alveolar lumina and alveolar septa. Macrophages containing many yeast cells were observed within bronchioles.

Microbial ecology of the female lower genital tract during pregnancy

Summary The posterior fornices of the vagina of 280 unselected pregnant women were sampled, using a sponge swab, squeezed out into Stuart’s Transport Medium. Other specimens were also taken. Eleven primary media were inoculated, wet swabs were examined, and Gramstained smears were also made. Nineteen groups or species of microbes were identified. Nearly 80% of women harbour pathogenic, or potentially pathogenic microbes; 52% harbour streptococci of various sorts. The incidence of the various groups, and species of microbes is tabulated. There is a statistically significant correlation between the presence of polymorphonuclear leucocytes in excess of one per high power field, and the presence of lesions of the cervix.

Ultrastructure of Canine Renal Autografts Following 24-Hour Hypothermic Preservation

29 canine kidneys were preserved for 24 h by hypothermic, pulsatile or nonpulsatile perfusion with albumin or cryoprecipitated plasma and were autotransplanted. Electron microscopy of biopsies taken at 60 min posttransplantation revealed no significant ultra-structural differences between surviving kidneys preserved with pulsatile or nonpulsatile flow, or between organs preserved with plasma or the albumin solution. Comparison of surviving and nonsurviving grafts, however, revealed conspicuous glomerular ultrastructural changes and the presence of polymorphonuclear leukocytes in the capillaries of the nonsurvivors. These changes appear to be useful in determining graft viability following preservation.

Benign mucous membrane pemphigoid. An unusual case with electron microscopic findings.

An 85-year-old woman developed complete esophageal obstruction from benign mucous membrane pemphigoid. Electron microscopic examination of nonscarring cutaneous lesions revealed two patterns. The first pattern showed early degenerative changes of epidermal cells with loss of tonofilaments and hemidesmosomes; the second showed frank dermoepidermal cleavage within the intermembranous space in the presence of polymorphonuclear leukocytes and without any epidermal degeneration.

Polymorphonuclear leukocytes in immunologic reactions. The destruction of vascular basement membrane in vivo and in vitro.

Vascular basement membrane was disrupted in the presence of polymorphonuclear leukocytes (PMN's) during two immunologic reactions: The Arthus phenomenon and the reaction to locally injected antibody to vascular basement membrane. This disruption was evidenced by (a) the inability of the basement membrane to retain circulating carbon, by (b) loss of antigenic constituents, and by (c) electron microscopic observation showing actual gaps in the structure of the vascular basement membrane. The factors within PMN's responsible for damage to isolated glomerular basement membrane in vitro were found by isolation procedures to be cathepsins D and E. Cationic proteins of PMN's were separable from the cathepsins. While inducing vascular permeability upon injection, these basic proteins failed to inflict the severe damage to the basement membrane observed in Arthus and antibasement membrane reactions. It is concluded that the full expression of these immunologic lesions requires destruction of the basement membrane possibly brought about by cathepsins D and E. Some of the physicochemical properties of these pathologically active leukocytic factors are given.

The influence of bacteria and irritation in the initiation of periodontal disease in germfree and conventional rats.

Within the conditions of the experimental design, this study has provided laboratory evidence that microorganisms were necessary for the occurrence of inflammatory periodontal disease. Furthermore, this investigation has shown that the presence of both local irritation and microorganisms was necessary to produce periodontal inflammation. However, neither factor alone was capable of producing the inflammation. The results of this study have shown that a type of dental calculus may be formed in the absence of microorganisms. However, the same results have demonstrated that the presence of microorganisms greatly facilitates the formation of calculus. The presence of polymorphonuclear leukocytes in the gingival crevices and tissues could not be explained on the basis of bacteria and/or irritation. It was postulated that these cells are a normal component of the gingival tissues of these animals and that their presence does not constitute a pathologic process. The findings of the present study were considered with those of other studies. From this comparison it was suggested that differences in animal species and strains should be considered when evaluations and extrapolations of experimental results are made.

Passive Uveal Anaphylaxis in Guinea Pigs

Summary Passive anaphylactic reactions were obtained in albino guinea pigs by injection of antibody into the vitreous body or into the anterior chamber of the eye. When the antibody is injected into both the vitreous body and the anterior chamber, only the anterior part of the uvea participates in the reaction. Edema of the tissues, changes in the permeability of the capillaries (especially small veins), presence of polymorphonuclear leukocytes in large numbers in the anterior chamber and around the ciliary body are observed. The posterior uvea does not seem to participate in these anaphylactic reactions.