Absence Of White Blood Cells Research Articles

The Basic Science of Platelet-rich Plasma (PRP)

Platelet-rich plasma (PRP) has been advocated for the biological augmentation of tissue healing and regeneration through the local introduction of increased levels (above baseline) of platelets and their associated bioactive molecules. In theory, the increased levels of autologous growth factors and secretory proteins provided by the concentrated platelets may enhance the wound healing process, especially in degenerative tissues or biologically compromised individuals. Although PRP has been increasingly utilized in the treatment of a variety of sports-related injuries, improvements in healing and clinical outcomes have not been universally reported. One reason for this may be the fact that all PRP preparations are not the same. Variations in the volume of whole blood taken, the platelet recovery efficacy, the final volume of plasma in which the platelets are suspended, and the presence or absence of white blood cells, and the addition of exogenous thrombin to activate the platelets or calcium chloride to induce fibrin formation, can all affect the character and potential efficacy of the final PRP product. This article will review the basic principles involved in creating PRP and examine the potential basic scientific significance of the individual blood components contained in the various forms of PRP currently used in sports medicine.

Chronic pelvic pain syndrome and voiding dysfunction.

Chronic prostatitis/chronic pelvic pain syndrome is a disease that is mainly characterized by three parameters: pain in the suprapubic and pelvic area, presence or absence of white blood cells in expressed prostatic secretions, and voiding disorders of various degrees. The causative factors underlying this very common condition are poorly understood. Therapeutic options (ie, antimicrobial treatment) often are based on the presence of an inflammatory reaction in the expressed prostatic secretions, but the benefit of recurring or prolonged courses of antimicrobial agents is highly variable. Observations have been made regarding functional and structural changes in the lower urinary tract that are suggestive to have an impact on the pathogenesis of chronic pelvic pain syndrome.

Male accessory gland infection: standardization of inflammatory parameters including cytokines

Assessment of infection of the male accessory glands is usually based on the search for white blood cells in different specimens to document an inflammatory reaction. This widely used practice allows to establish the diagnosis of inflammation in many cases. However, clinical symptoms do not always correlate with the presence or absence of white blood cells. This is particularly true for chronic prostatitis/chronic pelvic pain syndrome. In the last few years different research efforts have been made to look for markers of inflammation other than elements of the white blood cell line. Several studies suggest that humoral rather than cellular parameters are involved in male accessory gland infections. Substances such as reactive oxygen species, nerve growth factor and cytokines seem to be important not only in the pathogenesis of the inflammatory reaction but may also serve as diagnostic markers to indicate the presence of inflammation.

Cardiology patient page. Bacterial endocarditis: the disease, treatment, and prevention.

Endocarditis is a disease characterized by inflammation or infection of the inner surface of the heart (the endocardium). This article will focus on endocarditis that is the result of infection (infective endocarditis). Endocarditis commonly affects heart valves, but may also involve nonvalvular areas or mechanical devices that are implanted in the heart, such as artificial heart valves, pacemakers, or implantable defibrillators. Infective endocarditis is an uncommon, but not rare, disease affecting about 10 000 to 20 000 persons in the United States each year.1 Although uncommon, endocarditis is important because, despite antimicrobial therapy, it can result in serious complications such as stroke, the need for open heart surgery, or even death. When endocarditis occurs, small masses called vegetations form at the site of infection (Figure 1 and Figure 2). When vegetations are viewed under a microscope, generally one sees the microorganism that causes the infection embedded in a meshwork of fibrin and other cellular material similar to that used by the body to form blood clots. White blood cells that the body uses to fight infection are uncommon, a finding which explains the need to give antibiotics over many weeks to kill the infecting organism and cure endocarditis. The absence of white blood cells in vegetations is not fully explained but likely relates in part to the dense nature of the vegetation tissue, which in turn restricts the migration of these cells. Also, the bacteria causing endocarditis are buried in a nongrowing state deep in the vegetation. In this state they do not generate the intense chemical signals that usually promote the migration of white cells to a site of infection. Figure 1. Mitral valve vegetation shown by echocardiogram. The vegetation is the mass seen in the dark space between the left atrium (LA) and left ventricle (LV). RA …

Symptomevaluation beim chronischen Beckenschmerzsyndrom des Mannes

According to the National Institutes of Health classification system, chronic non-bacterial prostatitis/chronic pelvic pain syndrome (CPPS) is subdivided into an inflammatory (category IIIa) and a non-inflammatory (category IIIb) form. The difference is based on the presence or absence of white blood cells in expressed prostatic secretions, urine after prostatic massage, or semen. This is the only criterion which allows a differentiation between the IIIa and IIIb forms. The symptoms, i.e. pain and urinary complaints of various degrees, are thought to be similar in both forms. These symptoms can be assessed with the Chronic Prostatitis Symptom Index (CPSI) and the International Prostate Symptom Score (IPSS), which are both available in a validated German translation. One hundred and six patients with CPPS were evaluated with CPSI and IPSS. Urinary symptoms troubled all patients. Men with category IIIa had significantly more urinary symptoms when compared to men with category IIIb. There was no difference between the two groups regarding pain and impact on the quality of life. Although pain is thought to be the hallmark of CPPS, the contribution of urinary troubles to the symptoms must not be underestimated.

The relation between reactive oxygen species and cytokines in andrological patients with or without male accessory gland infection.

The presence of various cytokines, namely hepatocyte growth factor (HGF), interleukin-1 receptor antagonist (IL-1 RA), and interleukins (IL-1 alpha, IL-6, and IL-8), as well as the production of reactive oxygen species (ROS) was investigated in seminal plasma of fertile and infertile patients in order to evaluate the possible value of measuring these substances for the diagnosis of male accessory gland infection, and to assess the possible relationship between oxidative stress and cytokines during leucocytospermia and male accessory gland infection (MAGI). Our findings indicate that all of the measured cytokines seem to be produced locally as well as by white blood cells (WBC) and that, due to the presence of higher numbers of WBC, accessory gland infection may exert a deleterious effect on sperm quality through the production of ROS and/or of particular cytokines such as IL-1 alpha, IL-1 RA, and IL-8. The most specific marker for a sensitivity of 95% in discriminating between cases with or without MAGI is the measurement of IL-6 in seminal plasma. In the absence of WBC several cytokines are constitutively produced and correlate with sperm concentration (HGF, IL-8), alpha-glucosidase (IL-6), and gamma-glutamyltransferase activity (HGF). The measurement of these cytokines in semen may provide clinically useful information for the diagnosis of male accessory gland infection, as well as in the absence of WBC where it can provide information about certain mechanisms of male reproductive function and dysfunction.

Interferon gamma inhibits luteinized human granulosa cell steroid production in vitro

OBJECTIVE: The purpose of this study was to determine whether interferon gamma affects luteinized human granulosa cell progesterone, estrone, and estradiol production in the presence and absence of associated white blood cells by either cytotoxic or antiproliferative mechanisms. STUDY DESIGN: Luteinized granulosa cells were isolated by Percoll centrifugation from women during in vitro fertilization cycles. Some cell suspensions were further treated with anti-CD45 magnetic immunobeads to remove associated white blood cells. Granulosa cells with and without white blood cells were cultured in the presence of interferon gamma (0.5 to 50 ng/ml) for 48 hours. Medium was changed at 24-hour intervals, and spent medium was assayed for progesterone, estrone, and estradiol. In separate experiments granulosa cell viability was assessed with the tetrazolium salt reduction assay. RESULTS: Interferon gamma significantly inhibited granulosa cell progesterone production in both basal and human chorionic gonadotropin stimulated cells cocultured with white blood cells in a concentration-dependent manner, whereas cells cultured free of white blood cells demonstrated less inhibition. In the absence of interferon gamma a more profound increase in granulosa cell progesterone synthesis was found in human chorionic gonadotropin-stimulated cultures without associated white blood cells. Interferon gamma inhibited granulosacell estrone and estradiol production in basal cultures containing white blood cells in both a time- and concentration-dependent manner. Estrone production was not affected by interferon gamma in human chorionic gonadotropin-stimulated granulosa cell cultures containing white blood cells, whereas estradiol secretion was decreased at 48 hours with 50 ng/ml interferon gamma. Both estrone and estradiol synthesis were inhibited by 50 ng/ml interferon gamma in granulosa cel cultures free of white blood cells. In cultures free of interferon gamma, granulosa cell estrone and estradiol secretion was not affected by human chorionic gonadotropin stimulation compared with basal controls regardless of the presence or absence of white blood cells. All concentrations of interferon gamma used had no effect on granulosa cell viability at any time point tested. CONCLUSION: Our data suggest that interferon gamma affects granulosa cell steroid production both independently and in synergy with associated white blood cells and further supports the hypothesis that interferon gamma may be an important intraovarian regulator of ovarian steroid production during the luteal phase.

A method of human semen centrifugation to minimize the iatrogenic sperm injuries caused by reactive oxygen species.

Current techniques of sperm preparation for in vitro fertilization or intrauterine insemination require centrifugation of human semen to separate spermatozoa from the seminal plasma. Centrifugation increases reactive oxygen species (ROS) formation in semen. Moreover, high levels of ROS are associated with sperm membrane injury through spontaneous lipid peroxidation, which may alter sperm function. We investigated the relationship between centrifugation variables (time and g-force) and ROS production to establish an optimal centrifugation protocol for sperm preparation techniques. Semen from 38 men (24 patients and 14 normal volunteers) was evaluated for the formation of ROS before centrifugation and after centrifugation at 200 g for 2 or 10 min and after 500 g for 2 or 10 min. The absence of white blood cells in semen which can also produce ROS was determined with the myeloperoxidase technique (Endtz test). All specimens were negative (< 1 x 10(6)/ml) by the Endtz test. The formation of ROS was measured by chemiluminescence. ROS formation was regarded as high (positive) when the chemiluminescence response was at least 10 x 10(4) counted photons/min (cpm). The sperm concentration in each sample was adjusted to 15-20 x 10(6) cells/ml before analysis. Eight specimens (7 patients and 1 donor) exhibited high levels of ROS before centrifugation. All 8 showed further, significant increases in ROS formation regardless of g-force or time. The increase in ROS was significantly less when semen was centrifuged for 2 as compared to 10 min (p < 0.001). Six specimens previously ROS-negative became ROS-positive after centrifugation for 10 min at 200 and 500g. We conclude that the time of centrifugation is more important than g-force for inducing ROS formation in semen. Based on these results, we recommend a shorter centrifugation period in the preparation of sperm for assisted reproductive techniques.

The effect of endotoxin on neonatal erythrocyte intracellular calcium concentration

Erythrocyte membrane deformability is dependent on the maintenance of “normal” intracellular calcium (Ca) levels. Red cell flexibility is known to be altered in the septic neonate. In turn, this adversely affects viscosity and compromises flow in the microcirculation. It has been suggested that this may play a role in the mesenteric hypoperfusion associated with necrotizing enterocolitis. This study was designed to determine the effect of endotoxin on erythrocyte Ca homeostasis in the neonate. Paired specimens were obtained from the umbilical cord of 10 healthy neonates. The samples were incubated with either buffered saline (control) or 2 μg/mL of Escherichia coli endotoxin (LPS). Erythrocytes were then isolated, washed, and loaded with the fluorescent Ca chelator, FURA-2. The free cytosolic Ca concentration was determined by spectroscopic analysis of the ratio of fluorescent intensities at 340 nm and 380 nm. Results were obtained every 1.8 seconds for 1 minute, and the mean value was used for analysis. In 10 additional neonates, the white blood cells were removed before incubation in saline and LPS, and the erythrocytes were evaluated as described above. Differences were analyzed statistically by the paired t test. In the presence of white blood cells, endotoxin resulted in a significant increase in free cytosolic Ca concentration (LPS, 42.602 ± 5.166 nmol; control, 31.661 ± 4.002 nmol; P < .02). However, no significant difference were detected when cells were incubated in the absence of white blood cells (LPS, 32.374 ± 2.479 nmol; control, 34.021 ± 2.549 nmol). Endotoxin induces a significant increase in neonatal free cytosolic Ca concentration. This is dependent on white blood cells and/or their mediators. These endotoxin-induced alterations have an adverse affect on erythrocyte flexibility and may play a role in the perfusional abnormalities noted in neonatal sepsis.

Leucocyte-dependent platelet activation: an alternative pathway for initiation of blood clotting in inflammation.

Fibrin deposition is a prominent feature of several inflammatory diseases but the extract mechanism(s) leading to fibrinogen-fibrin conversion has not been completely clarified. We describe here a new cellular pathway for initiation of blood clotting resulting from a leucocyte-platelet interaction. Human washed platelet suspensions, free of leucocytes, isolated from whole blood or leucocyte-enriched platelet-rich plasma (PRP) after four hours' incubation with bacterial endotoxin, had strong procoagulant activity (40-100-fold that of control platelets). When platelets were challenged with endotoxin in the absence of white blood cells (i.e. in PRP) the subsequently washed platelets were devoid of procoagulant activity indicating that leucocytes are essential mediators in the development of platelet coagulant activity induced by endotoxin. This property is mostly confined to the mononuclear fraction. 'Stimulated' platelets have the peculiarity that they trigger blood coagulation by activating factor X independently of both the intrinsic and extrinsic pathways. These findings add a new function to circulating mononuclear cells and provide experimental evidence for an unrecognized cellular pathway of fibrin formation in inflammatory diseases.

Pulmonary Injury Elicited by Blood: An Experimental Study

To examine the role of cellular and humoral (serotonin, histamine) changes of blood after contact with foreign surfaces in the production of pulmonary injury, homologous whole blood (WB), leukocyte-poor blood (LPB) and leukocyte- and platelet-poor blood (LPPB) were perfused through the isolated rabbit lung or recirculated through the perfusion circuit without lung. Perfusion with WB was accompanied by an increase in pulmonary vascular resistance and deterioration of pulmonary gas exchange function within 60 min. Microscopic examination of WB-perfused lung revealed perivascular and alveolar hemorrhage, edema and microaggregates. Morphometrically, pulmonary arteries and arterioles exhibited narrowed vascular lumina and thickened muscular walls, indicating precapillary vasoconstriction as a primary event underlying the observed pathological changes. With LPPB or LPB no significant functional or morphological changes were seen. Granulocytes, which were sequestered by the isolated lung, represent an essential factor in eliciting significant pulmonary damage. No direct damaging effects of serotonin or histamine could be demonstrated in the absence of white blood cells.

STUDIES UPON EXPERIMENTAL PNEUMONIA IN RABBITS

STUDIES UPON EXPERIMENTAL PNEUMONIA IN RABBITS