Large Numbers Of Leukocytes Research Articles

Two bicistronic DNA vaccines against Vibrio anguillarum and the immune effects on flounder Paralichthys olivaceus.

Chemokines are cytokines that can promote the activation and migration of immune cells, and increase the recognition of antigen by antigen-presenting cells (APC). Previous studies showed that a DNA vaccine can induce humoral and cellular immune responses of flounder after immunization. To explore the improvement of chemokines on the efficiency of OmpK vaccine, two bicistronic DNA candidate vaccines were constructed and the immune responses they induced in the flounder were investigated by reverse transcription polymerase chain reaction (RT-PCR), indirect immunofluorescent assay (IFA), H&E staining, flow cytometry (FCM), and quantificational real-time polymerase chain reaction (qRT-PCR). pBudCE4.1 plasmid as an expression vector, bicistronic DNA vaccines encoding OmpK gene and CC-motif ligand 4 gene (p-OmpK-CCL4), or Ompk gene and CC-motif ligand 19 gene (p-OmpK-CCL19) were successfully constructed. The results showed that two bicistronic DNA vaccines expressed Ompk protein of Vibrio anguillarum and CCL4/CCL19 proteins of flounder both in vitro and in vivo. After immunization, a large number of leucocytes in muscle were recruited at the injection site in treatment groups. The constructed vaccines induced significant increases in CD4-1+ and CD4-2+ T lymphocytes, and sIgM+ B lymphocytes in peripheral blood, spleen, and head kidney. The percentage of T lymphocytes peaked on the 14th post-vaccination day whereas that of B lymphocytes peaked in the 6th post-vaccination week. Moreover, the expression profiles of 10 immune-related genes increased in muscles around the injection site, spleen, and head kidney. After the challenge, p-OmpK-CCL4 and p-OmpK-CCL19 conferred a relative percentage survival (RPS) of 74.1% and 63.3%, respectively, higher than p-OmpK alone (40.8%). In conclusion, both CCL4 and CCL19 can improve the protection of p-OmpK via evoking local immune response and then humoral and cellular immunity. CCL4 and CCL19 will be potential molecular adjuvants for use in DNA vaccines.Electronic supplementary materialSupplementary material (Supplementary Material) is available in the online version of this article at 10.1007/s00343-021-1092-z.

PERFORATIVE PERITONITIS IN NEWBORNS: INSTRUMENTAL AND MORPHOLOGICAL EXAMINATION FINDINGS

The aim: To study and analyze the results of instrumental diagnostic measures and morphological verification of perforations of abdominal viscera in peritonitis in newborns, their importance in timely diagnosis, as well as to present authors’ experience of examining newborns with PP, taking into account the complexity of diagnosis and treatment of these patients. Materials and methods: The study is based on the results of examining 59 newborns with PP of various etiologies. Instrumental methods that were used in the diagnosis of PP and differential diagnosis of other emergencies involved the following: radiological examination (plain abdominal radiography), abdominal and retroperitoneal ultrasound, neurosonography, echocardiography, diagnostic abdominal paracentesis. Morphological verification of hollow viscus perforations was performed by methods of histologic examination in 54 newborns who underwent surgery. Results: It is emphasized that perforations in NEC and spontaneous gastrointestinal perforations have clinical and morphological differences. The analysis of morphological data showed absence of intestinal musculature or muscular wall defect in spontaneous gastrointestinal perforations. Perforations in NEC had massive gastric or bowel wall necrosis. Morphogenesis of gastrointestinal perforations in newborns is crucial for developing correct treatment strategy and choosing surgical approach. Conclusions: Diagnosis of PP in newborns should be comprehensive and include modern instrumental studies that enable to reliably establish the cause of peritonitis and indications for surgical treatment. Differential diagnosis of PP in newborns with other diseases aims to differentiate PP from a number of similar clinical symptoms of urgent conditions in order to conduct adequate preoperative preparation and appropriate surgery. Diagnostic markers of PP involve the following: pneumoperitoneum, free fluid in the abdomen, sentinel loop (intestinal distention), fixed bowel loop, cloudy brown or greenish intra-abdominal fluid with a large number of leukocytes and bacteria.

THE STATE OF VAGINAL MICROBIOCENOSIS AFTER THE INFLUENZA IN THE EARLY PERIOD OF PREGNANCY

The aim of the study was to assess the state of the vaginal microbiocenosis in pregnant women after influenza in the early stages of pregnancy.Materials and research methods. We examined 120 women who had had influenza in the first trimester of pregnancy: the main group - 68 patients with placental dysfunction, the comparison group - 52 pregnant women without signs of placental insufficiency. Cultural, microscopic, auxiliary and clinical research methods were used.Research results. The microbial landscape of the vagina in pregnant women of the main group in terms of the frequency of detection of microorganisms with CFU 104 and above is characterized by an increased frequency of detection of pathogenic microflora: Staphylococcus epidermidis (22,1 %), Staphylococcus aureus (14,7 %), Staphylococcus haemolyticus (26,5 %), Streptococcus group В (26,5 %), Gardnerella vaginalis (27,9 %), Escherichia coli (33,8 %), Klebsiella (14,7 %), Chlamydia (32,4 %), Ureaplasma urealyticum (23,5 %) and Mycoplasma hominis (26.5%) ). The highest detection rate of fungi of the genus Candida (39.7%). The revealed violations occur against the background of suppression of normal microflora: Bifidobacterium were observed only in 17.6% of women, Lactobacterium in 25.0%. In most of the examined, vaginal contamination is associative in nature. In 64.7% of pregnant women, an inflammatory type of smear was detected, due to a variety of pathogenic microflora with a large number of leukocytes (41.2%) against the background of the absence of Dederlein sticks (Lactobacterium). 32.4% of patients report complaints of varying intensity, which may be caused by vaginal dysbiosis (excessive vaginal discharge, itching, edema and hyperemia of the mucous membrane), the pH of the vaginal contents is more than 4.5 - in 35.3% of patients, a positive amino test - in 23 1 %. Only in 14.7% of women the state of the vagina according to Kira’s classification is assessed as normocenosis, in 25.0% - moderate dysbiosis and in 27.9% - pronounced vaginal dysbiosis.Conclusions. Physiological immunosuppression during pregnancy in association with the immunosuppressive effect of influenza, changes in the acidity of the environment under the influence of gestational hormones disrupt the microbial balance of the biocenoses of the mucous membranes of the pregnant woman. Obstetric and perinatal complications in women after influenza may be partially mediated by abnormalities in the vaginal microbiome.

Expression of the G-protein-coupled estrogen receptor (GPER/GPR30) is characteristically altered in placenta pathophysiology

Objectives: During pregnancy the human decidua contains large numbers of leucocytes, predominantly very specialized natural killer (NK) cells and macrophages. These cells are considered to be implicated in the regulation of trophoblast invasion. In most previous studies, the number of decidual leucocytes was visually counted in mediumor high-power fields. These methods, however, cannot reveal the exact spatial relationship between leucocytes and the invasive trophoblast, and are prone to subjective errors. Thus, a more objective approach is required. Methods: Cryostat sections from tissues derived from first-trimester placentation sites were double-stained using either an anti-CD56 or an anti-CD163 antibody to detect NK cells or macrophages in combination with an anti-HLA-G antibody for labelling of invasive trophoblast cells. Digital images from the double-stained sectionswere evaluated applying a newmethod of morphometric image analysis. Distinct cell compartments as well as cell neighbourhood areas were defined. Finally, relative areal cell densities were calculated. The data were compared with those of tissue fragments derived from decidua parietalis without trophoblast cells. Results: At first-trimester placentation sites, a higher density of decidual NK cells as well as of decidual macrophages was found in close proximity to the invasive trophoblast, compared with the average areal cell density of decidual leucocytes in the tissue with exclusion of the trophoblast. The highest areal cell density was determined up to a distance of 20 m from the trophoblast cells, whereas in more distant regions it was even lower than average, indicating a migration of these cells towards the trophoblast invasion front. Conclusions: The new method of automated morphometric image analysis is well suited to rendering objective, observer-unbiased data on the distribution of mobile cells in tissues in relation to relatively immobile tissue structures. We anticipate that this method will also be useful for the investigation of recurrent miscarriage as well as for studies in the fields of tumour immunology and inflammation.

Effects of platelets on the adhesion between leukocytes and liver sinusoidal endothelium cells as well as on the transendothelial migration of leukocytes undergoing hypoxia-reoxygenation

To investigate the effects of platelet on intercellular adhesion between leukocyte and liver sinusoidal endothelial cell(LSEC) and the transendothelial migration under the hypoxia-reoxygenation condition, as well as the role of relevant adhesion molecules. LSEC was cultured for 24 hours under hypoxia condition and then reoxygenated for 2 hours (hypoxia-reoxygenation, HR). This hypoxia-reoxygenation model was used to simulate the clinical liver ischemia-reperfusion injury process (IRI). Platelets and leukocytes were labeled with fluorescence dye, and then the adhesion was detected by fluorescence microscope, fluorescence plate reader and laser scanning confocal microscope. Antibody blockage experiment was used to analyze the relevant adhesion molecules. The adhesion between platelets and LSEC was increased significantly after HR. The fluorescence intensity of adherent platelets increased from 142.10 ± 7.53 to 289.17 ± 20.00(P < 0.01). After H-R treatment and the addition of platelets, the number of adherent leukocytes increased markedly, and a significant statistical difference (360.71 ± 23.47 and 186.39 ± 17.96, P < 0.01) was found in comparing with the platelet deficient group. These adhesion processes could be blocked respectively by anti-GPIb, anti-GPIIb, anti-GPIIIa, anti-P-selectin, anti-CD31, anti-ICAM-1, anti-VCAM-1 and anti-ELAM-1. Confocal microscopy showed that platelets located between leukocytes and LSEC, and mediated their adhesion process. However, the adhesion of platelets to LSEC decreased the transendothelial migration of leukocytes (227.79 ± 16.51 and 167.27 ± 10.92, P < 0.05). During ischemia-reperfusion condition platelets adhere to the surface of LSEC, and then further mediate more adhesion processes between leukocytes and endothelial cells, as well as inhibit the transendothelial migration of leukocytes. The consequence is that large numbers of leukocytes were sequestrated within hepatic sinus, and deteriorate liver ischemia-reperfusion injury.

1574 MOLECULAR SIGNATURES ASSOCIATED WITH PROSTATIC INFLAMMATION

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research1 Apr 20121574 MOLECULAR SIGNATURES ASSOCIATED WITH PROSTATIC INFLAMMATION Yasuhito Funahashi, Zhou Wang, Katherine J. Willet, Ryosuke Takahashi, Momokazu Gotoh, and Naoki Yoshimura Yasuhito FunahashiYasuhito Funahashi Pittsburgh, PA More articles by this author , Zhou WangZhou Wang Pittsburgh, PA More articles by this author , Katherine J. WilletKatherine J. Willet Pittsburgh, PA More articles by this author , Ryosuke TakahashiRyosuke Takahashi Pittsburgh, PA More articles by this author , Momokazu GotohMomokazu Gotoh Nagoya, Japan More articles by this author , and Naoki YoshimuraNaoki Yoshimura Pittsburgh, PA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1346AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Prostatic inflammation is reportedly associated with symptomatic BPH with LUTS and/or prostate cancer development. We therefore investigated the effects of prostatic inflammation on expression levels of androgen receptor-related genes and growth factors in the rat prostate. METHODS Prostatic inflammation was induced by either formalin (5%; 50 μl per lobe) injection into bilateral ventral lobes of the prostate or E. coli (strain 1677) injection (0.2 mL of 1x 108 CFU/mL) into the prostatic urethra of male SD rats. After 5 days, ventral lobes of the prostate were harvested, and histopathology and real-time RT-PCR for pro-inflammatory cytokines were performed. Using laser-capture microdissection (LCM), glandular epithelial cells and stromal regions were separately isolated. Real-time RT-PCR was performed to examine mRNA levels of androgen-responsive (AR) genes in the epithelium and TGF-β cascade genes in the stroma. RESULTS (1) Histopathology: In the formalin-injected prostate, inflammatory changes were observed focally. In the severe inflammation area, a large number of leukocytes were present in a slightly edematous stromal region and the epithelium shrank while other regions had almost normal appearance. In the E.coli-treated prostate, inflammation distributed diffusely throughout the prostate. Polymorphonuclear cells were filled in the ductal spaces and infiltrated into the highly edematous stroma, without morphological changes in the epithelium. (2) Cytokine levels: In the formalin-treated prostate, IL-1β, IL-6, and COX-2 mRNA were increased 3, 9, and 4-fold compared to control, respectively. In the E.coli-treated prostate, TNF-α, IL-1β, IL-6, and COX-2 mRNA increased 42, 149, 18 and 12-fold, respectively. (3) LCM: Epithelial cells from the area with no/little inflammation of formalin-treated prostate showed 2-3 fold increases in androgen receptors and AR genes such as Eaf2, ELL2, FKBP5, calreticulin and orinthine decarboxylase compared to control. In contrast, epithelial cells from E. coli-treated prostate or the severe inflammatory area of formalin-treated prostate showed the reduction in these genes. In the stromal area from formalin or E. coli-treated prostate, TGF-β cascade genes such as TGF-β1, fibronectin and collagen-1 were up-regulated 2-5 fold. CONCLUSIONS Prostatic inflammation changes expression of AR genes in the epithelium and TGF-β cascade genes in the stroma. Activation of TGF-β cascade genes in the inflammatory stroma as well as altered AR gene expression in the epithelium might be involved in the development of symptomatic BPH and/or prostate cancer. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e637-e638 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Yasuhito Funahashi Pittsburgh, PA More articles by this author Zhou Wang Pittsburgh, PA More articles by this author Katherine J. Willet Pittsburgh, PA More articles by this author Ryosuke Takahashi Pittsburgh, PA More articles by this author Momokazu Gotoh Nagoya, Japan More articles by this author Naoki Yoshimura Pittsburgh, PA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Actinobaculum schaalii, a cause of urinary tract infections in children?

To inform that Actinobaculum schaalii can colonize the urine and cause urinary tract infection in children. Urine samples were examined by wet smear microscopy, incubated in 5% CO(2) for 1-2 days, and species-specific real-time polymerase chain reaction (PCR) for A. schaalii was performed. In 5 of the 29 screened urines, A. schaalii was found only by real-time PCR in quantities equivalent to ≥ 10(4) -10(5) CFU/mL. In addition, A. schaalii was found in quantities equivalent to ≥ 10(6) CFU/mL by both culture and PCR in two children with a urinary tract infection and large numbers of leucocytes in the urine. Actinobaculum schaalii is CO(2)-dependent. Therefore, if there are clinical symptoms and/or a negative culture despite the presence of leucocytes in the urine, Gram staining and incubation in 5% CO(2) or species-specific real-time PCR should be performed to identify A. schaalii.

Direct Leukocyte Migration across Pulmonary Arterioles and Venules into the Perivascular Interstitium of Murine Lungs during Bleomycin Injury and Repair

During acute lung injury and repair, leukocytes are thought to enter the lung primarily across alveolar capillaries and postcapillary venules. We hypothesized that leukocytes also migrate across pulmonary arterioles and venules, which serve as alternative sites for leukocyte influx into the lung during acute lung injury and repair. Lung sections from C57BL/6J mice up to 14 days after intratracheal bleomycin (3.33 U/kg) or saline instillation were assessed by light, fluorescence, confocal, and transmission electron microscopy for evidence of inflammatory cell sequestration and transmigration at these sites. After bleomycin treatment, large numbers of leukocytes (including neutrophils, eosinophils, and monocytes) were present in the vascular lumina and in perivascular interstitia of pulmonary arterioles and venules, as well as within the vascular walls. Leukocytes were observed within well-defined pathways in arteriolar walls and much less structured pathways in venular walls, apparently in the process of transmigration. Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were expressed at sites of leukocyte interaction with the luminal surface, especially in arterioles. Leukocytes appeared to exit from the vessels near collagen fibers into the perivascular interstitium. Results indicate that leukocytes can directly migrate across arteriolar and venular walls into the perivascular interstitium, which may represent an important but under-recognized pathway for leukocyte influx into the lung during injury and repair.

Dr William Coley and tumour regression: a place in history or in the future

Spontaneous tumour regression has followed bacterial, fungal, viral, and protozoal infections. This phenomenon inspired the development of numerous rudimentary cancer immunotherapies, with a history spanning thousands of years. Coley took advantage of this natural phenomenon, developing a killed bacterial vaccine for cancer in the late 1800s. He observed that inducing a fever was crucial for tumour regression. Unfortunately, at the present time little credence is given to the febrile response in fighting infections-no less cancer. Rapidly growing tumours contain large numbers of leucocytes. These cells play a part in both defence and repair; however, reparative functions can also support tumour growth. Intratumoural infections may reactivate defensive functions, causing tumour regression. Can it be a coincidence that this method of immunotherapy has been "rediscovered" repeatedly throughout the centuries? Clearly, Coley's approach to cancer treatment has a place in the past, present, and future. It offers a rare opportunity for the development of a broadly applicable, relatively inexpensive, yet effective treatment for cancer. Even in cases beyond the reach of conventional therapy, there is hope.

Cervical Lactobacillus and leukocyte infiltration in preterm premature rupture of membranes

Objectives: To evaluate the possible association between microorganisms present in the cervical secretions and amniotic fluid of pregnant women with preterm premature rupture of membranes (PPROM), and histologic chorioamnionitis. Methods: Thirty-seven pregnant women with PPROM and 21 healthy pregnant women were studied. Secretions from the cervical canal and amniotic fluid were collected to isolate microorganisms present in the genital tract. Cervical smears were Gram stained and evaluated microscopically. At delivery, chorioamniotic membranes were collected for histopathologic analysis. Results: Microscopic examination of the cervical secretion smears obtained from the PPROM group showed a low rate of Lactobacillus species, large numbers of leukocytes, and a wide diversity of microorganisms compared with the control group. The PPROM group presented an 80% rate of chorioamnionitis. Staphylococcus aureus isolation in cervical secretion was associated with intense inflammatory infiltrate in the membranes and might play a role in the pathogenesis of PPROM. Conclusions: The low colonization of cervical flora by Lactobacillus species associated with an intense leukocyte infiltrate detected in Gram-stained cervical smears can be considered a rapid method of detecting chorioamnionitis in pregnant women with PPROM.

Donor leukocyte infusion for Japanese patients with relapsed leukemia after allogeneic bone marrow transplantation: indications and dose escalation.

To clarify the role of dose escalation of donor leukocyte infusion (DLI) in the treatment of relapsed leukemia after allogeneic bone marrow transplant (BMT), data from 100 patients were collected from 46 facilities in Japan and analyzed with respect to indications and infused cell dose. Complete remission (CR) was achieved in 11 of 12 (91%) patients with relapsed chronic myelogenous leukemia (CML) in the chronic phase, 3 of 11 (27%) with CML in the acute phase, 8 of 21 (38%) with acute myelogenous leukemia (AML), 6 of 23 (25%) with acute lymphoblastic leukemia (ALL), and 5 of 11 (45%) with myelodysplastic syndrome (MDS). The probability of remaining in CR at 3 years was 82% in CML patients in the chronic phase, but 0% in those with CML in the acute phase, 7% in those with AML, 0% with ALL, and 33% with MDS. Acute graft-versus-host disease (GVHD) (> or = 2) developed in 31 of 89 (34%) patients with human leukocyte antigen identical related donors and was fatal for 7 (7%). A leukocyte dose of 1 x 10(7)/kg of recipient body weight with CML in the chronic phase, 3 x 10(7)/kg of recipient body weight with MDS, and 1 x 10(8)/kg of recipient body weight with acute leukemia appeared to be optimal as an initial dose of DLI. However, the minimal dose of leukocyte developing fatal GVHD was 7 x 10(7)/kg of recipient body weight. These suggest that a relatively small dose of DLI ranging from 1 x 10(7)/kg to 5 x 10(7)/kg of recipient body weight should be administered initially then the infused escalating dose 2 or 3 months later in patients with CML in the chronic phase and MDS. However, a large number of leukocytes around 1 x 10(8)/kg are needed to induce graft versus leukemia effects in patients with acute leukemia despite a 7% fatality in GVHD.

Responses of male brushtail possums to sterile females: implications for biological control

Summary 1. Increasingly immunocontraception is being considered as a potential method for the biological control of mammalian pests. Although several studies have investigated or modelled its demographic consequences, there have been few studies of the possible effects of the presence of sterile females on local males. 2. In New Zealand, the brushtail possum Trichosurus vulpecula is an important mammalian pest, for which immunocontraception has been proposed as a control method. 3. We surgically sterilized female brushtail possums in two populations to study the potential effects of immunocontraception on body condition and local sex ratios. 4. Large numbers of leucocytes (indicating oestrus) were present in the reproductive tracts of sterile female possums several weeks after most control (non‐sterile) females were pregnant and had ceased mating activity. This indicates that the sterilization of females prolonged the mating season of local possums. 5. The body condition of males was significantly poorer in the presence of sterilized females in the winter post‐mating period following the treatment. In contrast, the body condition of females showed no change following the sterilization treatment, either during the autumn mating period or in the winter post‐mating period. 6. More adult male possums were recorded at both study sites after the sterilization treatment, resulting in a change to the originally female‐biased sex ratio. This may have been caused by the prolonged presence of oestrous females attracting males from surrounding areas. 7. The implications of our results for the control of possums by sexually transmitted immunocontraception are twofold. First, the reduced body condition of males in the presence of sterilized females might result in increased male mortality. Secondly, the attraction of males from surrounding areas to those containing sterilized females might facilitate the local spread of an immunocontraceptive agent. 8. We advocate further assessments of the effects of sterilization and/or immunocontraception on the population demography, individual condition and survival of pest animals.

Integrin activation by chemokines: relevance to inflammatory adhesion cascade during T cell migration.

The adhesive function of integrins is regulated through cytoplasmic signaling induced by several stimuli, whose process is designated "inside-out signaling". A large number of leukocytes are rapidly recruited to the sites of inflammation where they form an essential component of the response to infection, injury, autoimmune disorders, allergy, tumor invasion, atherosclerosis and so on. The recruitment of leukocytes into tissue is regulated by a sequence of interactions between the circulating leukocytes and the endothelial cells. Leukocyte integrins play a pivotal role in leukocyte adhesion to endothelial cells. During the process, the activation of integrins by various chemoattractants, especially chemokines, is essential for integrin-mediated adhesion in which a signal transduced to the leukocyte converts the functionally inactive integrin to an active adhesive configuration. We have proposed that H-Ras-sensitive activation of phosphoinositide 3 (PI 3)-kinase and subsequent profilin-mediated actin polymerization, can be involved in chemokine-induced integrin-dependent adhesion of T cells. The present review documents the relevance of cytoplasmic signaling and cytoskeletal assembly to integrin-mediated adhesion induced by chemoattractants including chemokines during inflammatory processes. In contrast, various adhesion molecules are known to transduce extracellular information into cytoplasm, which leads to T cell activation and cytokine production from the cells, designated "outside-in signaling". Such a bi-directional "cross-talking" among adhesion molecules and cytokines is most relevant to inflammatory processes by augmenting immune cell migration from circulation into inflamed tissue such as rheumatoid arthritis, tumor invasion, Behçet's disease and atherosclerosis.

Chronic Prostatitis Is Not Commonly Seem in Men with Spinal Cord Injury (SCI)

Objectives: Recent evidence indicates that impaired sperm motility and viability in men with SCI may be due to factors in their seminal plasma. Men with SCI have been shown to have elevated serum and decreased seminal plasma PSA concentrations, indicating that prostatic secretory dysfunction is present (J Urol, 1999; 162:89–91). The objective of this study was to determine if a pathological process could be found in the prostate gland of men with SCI to explain the secretory dysfunction, which in turn may be contributing to their poor sperm quality. Design: A prospective analysis was performed on prostate biopsies in men with SCI and age-matched controls. Materials and Methods: Seven men with SCI (mean age 26.5 years) and four controls, including three healthy men and one brain-dead organ donor (mean age 35.0 years), underwent a standard transrectal ultrasound guided prostate biopsy procedure. The semen analysis of all SCI subjects was typical for this population, with normal sperm concentration, low sperm motility, low sperm viability, and large numbers of leukocytes. The semen analysis of three control subjects was within normal ranges. (Semen was not obtained from the brain dead control subject). In all except the brain dead subject, at least 3 cores were obtained and underwent routine H&E staining. The brain dead organ donor’s prostate was harvested and random sections, as well as a needle biopsy core were taken. Each patient’s biopsy was reviewed with a pathologist. Results: Only two SCI subjects had histologic evidence of prostatic inflammation. Both of these subjects had a small focus of periductal and interstitial chronic prostatitis found in only one core. No evidence of prostatic inflammation was seen in the control subjects. Two different SCI subjects and one control had histological evidence of focal benign nodular hyperplasia. No malignancies were encountered in any subject. Conclusions: The prostate biopsies did not reveal any significant inflammatory process, chronic or acute. Our findings indicate that the prostate tissue of men with SCI is not the source of the increased number of seminal fluid inflammatory cells. Future research should be targeted at identifying the origin of these cells. This project was supported by the Miami Project to Cure Paralysis and the State of Florida Specific Appropriations.

Endotracheal Aspiration in the Diagnosis of Ventilator-Associated Pneumonia

T he time-honored method of identifying bacterial pathogens that are potentially responsible for nosocomial pneumonia in patients receiving mechanical ventilation is the microscopic examination of specimens obtained by endotracheal aspiration (EA). This technique is the simplest noninvasive means of obtaining respiratory secretions from patients receiving mechanical ventilation; it is readily performed at the bedside and requires minimal training by health-care providers. This section focuses on clinical studies evaluating diagnostic procedures using endotracheal specimens (ie, cytologic examination, antibody coating, elastin fibers, Gram’s stain, and culture) in immunocompetent adults with suspected VAP. The cytologic examination of specimens containing a large number of leukocytes and a paucity of epithelial cells is likely to produce the most valid representation of infectious organisms. A test for the detection of the presence of antibody coating on bacteria has been developed in an attempt to distinguish organisms that are colonizing the lower respiratory tract from those that actually are infecting it. The test is based on the premise that an infection will elicit an antibody response in the host and that this response will be detectable on the microorganism. In addition, using 40% potassium hydroxide to detect elastin fibers has been promoted as a rapid and inexpensive way to demonstrate the destruction of lung parenchymal tissue that is caused by pneumonia. However, an analysis of endotracheal specimens obtained by aspiration has been diagnostically inadequate. Several qualitative articles have reviewed the use of endotracheal specimens to diagnose VAP. Among the challenges for investigators and clinicians are the following: distinguishing upper from lower respiratory tract infection; distinguishing infection from colonization and contamination; standardizing aspiration collection methods and microbiological techniques; and interpreting test properties in light of the host’s immune status, the pathogenic load, and the effect of prior antimicrobial therapy. Newer bronchoscopic methods for diagnosing VAP have become the focus of recent investigations, conferences, and professional documents. Invasive approaches have not necessarily been adopted by clinicians,2 at least in part because of procedural access, cost, and the absence of compelling evidence that treatment based on the derived data changes clinical or economic outcomes. Thus, many physicians continue to use endotracheal specimens and other clinical features in diagnosing VAP.

Leucocyte populations and cytokine regulation in human uteroplacental tissues.

Human endometrial tissue and the decidualized endometrium in pregnancy contain relatively large numbers of leucocytes, the proportions of which vary during both the menstrual cycle and early pregnancy. CD3+ T-cells, CD14+ macrophages and a population of phenotypically unusual CD3-CD16-CD56++ large granular lymphocytes (LGL) are present, whereas B-cells are virtually absent. Relative T-cell numbers decrease in first-trimester decidua; T-cells are therefore unlikely to have an important role in the immunological maintenance of pregnancy but could be more important in implantation, when their relative numbers are greater. Extensive numbers of class II MHC-positive tissue macrophages in both the endometrium and placenta will provide an immediate antigen non-specific host defence to infection at this important site. Nevertheless, most attention has focused on a role for the predominant LGL endometrial cell population in the implantation and maintenance of pregnancy because, at the time of implantation, these LGLs comprise 70-80% of all leucocytes in the endometrium. It is now well recognized that there is substantial and complex cytokine activity within human uteroplacental tissues; both leucocytic and non-leucocytic cells have been shown to be capable of producing a significant array of cytokines. However, to avoid excessive pathological sequelae, such cytokine activity must be locally regulated. This has been highlighted by recent reports indicating that abnormal Th1-type cytokine responses could be a reason for immunological reproductive failure in women. Key cytokines controlling differentiation into a Thl (interleukin 12) or Th2 (interleukin 4) type pattern both exist in unusual molecular forms at the human maternal-fetal tissue interface and hence might be fundamental regulatory elements controlling cytokine action locally by an antagonistic action.

Late-Onset Chronic Inflammatory Encephalopathy in Immune-Competent and Severe Combined Immune-Deficient (SCID) Mice with Astrocyte-Targeted Expression of Tumor Necrosis Factor

To examine the role of tumor necrosis factor (TNF)-alpha in the pathogenesis of degenerative disorders of the central nervous system (CNS), transgenic mice were developed in which expression of murine TNF-alpha was targeted to astrocytes using a glial fibrillary acidic protein (GFAP)-TNF-alpha fusion gene. In two independent GFAP-TNFalpha transgenic lines (termed GT-8 or GT-2) adult (>4 months of age) animals developed a progressive ataxia (GT-8) or total paralysis affecting the lower body (GT-2). Symptomatic mice had prominent meningoencephalitis (GT-8) or encephalomyelitis (GT-2) in which large numbers of B cells and CD4+ and CD8+ T cells accumulated at predominantly perivascular sites. The majority of these lymphocytes displayed a memory cell phenotype (CD44high, CD62Llow, CD25-) and expressed an early activation marker (CD69). Parenchymal lesions contained mostly CD45+ high, MHC class II+, and Mac-1+ cells of the macrophage microglial lineage with lower numbers of neutrophils and few CD4+ and CD8+ T cells. Cerebral expression of the cellular adhesion molecules ICAM-1, VCAM-1, and MAdCAM as well as a number of alpha- and beta-chemokines was induced or upregulated and preceded the development of inflammation, suggesting an important signaling role for these molecules in the CNS leukocyte migration. Degenerative changes in the CNS of the GFAP-TNFalpha mice paralleled the development of the inflammatory lesions and included primary and secondary demyelination and neurodegeneration. Disease exacerbation with more extensive inflammatory lesions that contained activated cells of the macrophage/microglial lineage occurred in GFAP-TNFalpha mice with severe combined immune deficiency. Thus, persistent astrocyte expression of murine TNF-alpha in the CNS induces a late-onset chronic inflammatory encephalopathy in which macrophage/microglial cells but not lymphocytes play a central role in mediating injury.

CD18-dependent leukocyte adherence and vascular injury in pig cerebral circulation after ischemia.

Recent accumulating evidence indicates that leukocytes contribute importantly to ischemic brain injury. Although large numbers of leukocytes are present in the ischemic territory of reperfused brain 24-48 h after the ischemic insult, little is known of the acute inflammatory response to cerebral ischemia, particularly regarding the time course and magnitude of leukocyte adherence to cerebrovascular endothelium and the functional consequences of such adherence. To study these issues, we developed an epifluorescence videomicroscopy system for observing and quantifying the dynamic behavior of rhodamine-labeled leukocytes in the cerebrovascular microcirculation. Anesthetized piglets equipped with closed cranial windows were used in these investigations. During the initial 2 h of reperfusion after 9 min of asphyxia (n = 6), a marked, progressive increase in adherent leukocytes was noted in cerebral postcapillary venules that was significantly greater in magnitude than that seen in nonasphyxic, time-matched controls (n = 8). A similar response was observed after complete global ischemia of 10 min duration. A significant increase in sodium fluorescein permeability was also measured at 2 h of reperfusion in asphyxic animals. Pretreating a separate asphyxic animal group (n = 7) with a monoclonal antibody to the leukocyte adhesion glycoprotein complex CD11/CD18 severely attenuated both leukocyte adherence and the increase in vascular permeability. These results provide evidence that adherent leukocytes contribute to disruption of endothelial integrity during early reperfusion after global ischemic insults, the inhibition of which may reduce the vasogenic edema that occurs early during reperfusion after birth asphyxia, stroke, and cardiac arrest.

Do the Leukocytes in the Surface Secretion on the Adenoid have an Immunological Function?

To elucidate whether leukocytes in the surface secretion on the adenoid have a potential immunological function, imprint samples of surface secretion were obtained from the adenoid of 11 children and the corresponding mucosal area of 10 adults. May-Grunewald-Giemsa staining was used for evaluation of cell morphology and spatial relations, and immuno-histochemical staining for identification of granulocytes, B cells, T cells and macrophages. The children's imprints showed large numbers of leukocytes, with mononuclear cells in the majority. The adults' imprints were characterized by moderate numbers of epithelial cells and few leukocytes. In six out of seven adenoid secretions successfully analysed with all four CD antigens, there was the simultaneous presence of granulocytes, B cells, T cells and macrophages. This was the case in one of nine analysable adult secretions. The CD-positive cells were often seen in juxtaposition, in clusters of two to 10 cells, as well as in contact with leukocytes of unknown CD antigenicity and epithelial cells. The simultaneous presence in the secretion of morphologically intact and CD antigen-presenting leukocytes in juxtaposition could indicate a potential immunological function of these cells.

Demodex Nycticeii: a new species of hair follicle mite (Acari: Demodecidae) from the evening bat,Nycticeius humeralis(Chiroptera: Vespertilionidae)

Abstract Demodex nycticeii sp. nov. is described, in all life stages, from papule-forming colonies in the skin of the Evening bat, Nycticeius humeralis. The papules, measuring to one mm in diameter and containing many mites, develop following initial invasion by the mites of the hair follicles of the general body fur. Papules are surrounded by multinucleate giant cells and large numbers of leucocytes, the latter especially if mites penetrate through the epithelial lining of the papule into the surrounding dermis. Demodex nycticeii is most similar to D. novazelandica of the New Zealand bat, Mystacina tuberculata, but differs in size, shape of opisthosoma and structure of podosomal shield.