Relative Number Of Neutrophils Research Articles

Dampak paparan radiasi sinar x lingkungan terhadap leukosit dari petugas radiologi

White blood cells are part of the hematopoetic system that is radiosensitive and are markers for determining the effects and intensity of ionizing radiation. This quantitative study aims to see the effect of ionizing radiation (x-rays) from x-ray services on the white blood cells of radiology officers at advanced health facilities in Pare District, Kediri Regency between September 2022, and January 2023. Using the slovin formula, a total of 60 participants were determined, 30 radiology nurses and 30 comparison groups of administrative officers. When viewed as a whole, the relative number of white blood cells there was no significant difference between the radiology officer group and the administrative officer, but the administrative officer group tended to have a a little higher white blood cell count. Significant differences occurred in the relative number of neutrophils with a significance of 0.0001 (0.05) Neutrophils of radiology officer tended to be smaller than the dissident group. The number of lymphocytes of radiology officer is different from administrative officer and tends to have higher count. Based on the findings it can be indicated if x-ray exposure affects the white blood cell count.

Correlation between facial vascularized composite allotransplantation rejection and laboratory markers: Insights from a retrospective study of eight patients

The field of facial vascularized composite allotransplantation (fVCA) is still new and a limited number of patients have undergone the procedure. This has led to a lack of understanding about the impact of fVCA rejection on standard laboratory markers (e.g., CBC, BMP, CRP) for the acute management of these patients. It is not clear if rejection elicits a systemic inflammatory response that influences common inflammatory markers such as WBC and CRP. A comprehensive understanding of changes in these markers could help in the management of fVCA patients in the acute setting. The medical records of 8 fVCA patients with a total of 9 transplants were reviewed retrospectively, and data on standard laboratory values (CBC, BMP, LFTs, CRP) and vital signs were extracted. To examine the relationship between laboratory values and rejection status, linear mixed models were used to analyze the data, taking into account their longitudinal nature (repeated measures). A statistically significant relationship was found between the Banff grade of rejection and the relative number of basophils in the patient's blood during rejection (p=0.005). In addition, in patients with clinical signs of rejection (e.g., facial erythema, edema) and skin biopsy showing Banff ≥II, CRP was found to be significantly elevated (p=0.03). The WBC count remained stable during rejection, and the relative number of neutrophils was lower at the time of rejection, indicating possible consumption at the site of rejection. During fVCA rejection, most standard laboratory parameters and vital signs appear to be stable. However, the levels of CRP and basophils were elevated during rejection, while the neutrophil count was lower. Leukocytosis can likely be used as a marker of microbial infection in fVCA patients, as WBC does not seem to increase at the time of allograft rejection.

Antitumor immunity and therapeutic properties of marine seaweeds-derived extracts in the treatment of cancer

Marine seaweeds are important sources of drugs with several pharmacological characteristics. The present study aims to evaluate the antitumor and antitumor immunological potentials of the extracts from the brown alga Padinapavonica and the red alga Janiarubens, inhibiting the Egyptian marine coasts. Hep-G2 cell lines were used for assessment of the antitumor efficacy of Padinapavonica and Janiarubens extracts in vitro, while Ehrlich ascites carcinoma (EAC) cells were applied to gain more antitumor immunity and antitumor insights of P.pavonica and J.rubens extracts in vivo. In vitro antitumor potentials of P.pavonica and J.rubens extracts were analyzed against human liver cancer Hep-G2 cells by MTT and trypan blue exclusion assays. In vivo antitumor immunological potentials of P.pavonica and J.rubens extracts at low, high, and prophylactic doses were analyzed by blood counting and flow cytometry in mice challenged with Ehrlich ascites carcinoma (EAC) cells. In vitro results revealed that P.pavonica and J.rubens extracts caused significant decreases in the number and viability of Hep-G2 cells in a dose-dependent manner as compared to untreated Hep-G2 cells or Cisplatin®-treated Hep-G2 cells. In vivo findings showed that P.pavonica and J.rubens extracts at low, high, and prophylactic doses significantly reduced the number and viability of EAC tumor cells accompanied by increases in EAC apoptosis compared to naïve EAC mouse. Additionally, P.pavonica and J.rubens extracts at low and prophylactic doses remarkably increased both the total WBC count and the relative numbers of lymphocytes and decreased the relative numbers of neutrophils and monocytes. Flow cytometric analysis showed that P.pavonica and J.rubens extracts at the treatment and the prophylactic doses resulted in a significant increase in the phenotypic expressions of CD4+ T, CD8+ T, and CD335 cells compared to naïve EAC mouse. Overall, both extracts P.pavonica and J.rubens possess potential antitumor and antitumor immunological effects with less toxicity, opening new approaches for further studies of the chemical and biological mechanisms behind these effects.

T helper cell subsets and related target cells in acute COVID-19

Current review presents a brief overview of the immune system dysregulation during acute COVID-19 and illustrates the main alterations in peripheral blood CD4+ T-cell (Th) subsets as well as related target cells. Effects of dendritic cell dysfunction induced by SARS-CoV-2 exhibited decreased expression of cell-surface HLA-DR, CCR7 as well as co-stimulatory molecules CD80 and CD86, suggesting reduced antigen presentation, migratory and activation capacities of peripheral blood dendritic cells. SARS-CoV-2-specific Th cells could be detected as early as days 24 post-symptom onset, whereas the prolonged lack of SARS-CoV-2-specific Th cells was associated with severe and/or poor COVID-19 outcome. Firstly, in acute COVID-19 the frequency of Th1 cell was comparable with control levels, but several studies have reported about upregulated inhibitory immune checkpoint receptors and exhaustion-associated molecules (TIM3, PD-1, BTLA, TIGIT etc.) on circulating CD8+ T-cells and NK-cells, whereas the macrophage count was increased in bronchoalveolar lavage (BAL) samples. Next, type 2 immune responses are mediated mainly by Th2 cells, and several studies have revealed a skewing towards dominance of Th2 cell subset in peripheral blood samples from patients with acute COVID-19. Furthermore, the decrease of circulating main Th2 target cells basophiles and eosinophils were associated with severe COVID-19, whereas the lung tissue was enriched with mast cells and relevant mediators released during degranulation. Moreover, the frequency of peripheral blood Th17 cells was closely linked to COVID-19 severity, so that low level of Th17 cells was observed in patients with severe COVID-19, but in BAL the relative number of Th17 cells as well as the concentrations of relevant effector cytokines were dramatically increased. It was shown that severe COVID-19 patients vs. healthy control had higher relative numbers of neutrophils if compared, and the majority of patients with COVID-19 had increased frequency and absolute number of immature neutrophils with altered ROS production. Finally, the frequency of Tfh cells was decreased during acute COVID-19 infection. Elevated count of activated Tfh were found as well as the alterations in Tfh cell subsets characterized by decreased regulatory Tfh1 cell and increased pro-inflammatory Tfh2 as well as Tfh17 cell subsets were revealed. Descriptions of peripheral blood B cells during an acute SARS-CoV-2 infection werev reported as relative B cell lymphopenia with decreased frequency of nave and memory B cell subsets, as well as increased level of CD27hiCD38hiCD24 plasma cell precursors and atypical CD21low B cells. Thus, the emerging evidence suggests that functional alterations occur in all Th cell subsets being linked with loss-of-functions of main Th cell subsets target cells. Furthermore, recovered individuals could suffer from long-term immune dysregulation and other persistent symptoms lasting for many months even after SARS-CoV-2 elimination, a condition referred to as post-acute COVID-19 syndrome.

Variations in immune parameters with age in a wild rodent population and links with survival.

Recent findings suggest that immune functions do not unidirectionally deteriorate with age but that a potentially adaptive remodeling, where functions of the immune system get downregulated while others get upregulated with age could also occur. Scarce in wild populations, longitudinal studies are yet necessary to properly understand the patterns and consequences of age variations of the immune system in the wild. Meanwhile, it is challenging to understand if the observed variations in immune parameters with age are due to changes at the within‐individual level or to selective (dis)appearance of individuals with peculiar immune phenotypes. Thanks to a long‐term and longitudinal monitoring of a wild Alpine marmot population, we aimed to understand within‐ and between‐individual variation in the immune phenotype with age, in order to improve our knowledge about the occurrence and the evolutionary consequences of such age variations in the wild. To do so, we recorded the age‐specific leukocyte concentration and leukocyte profile in repeatedly sampled dominant individuals. We then tested whether the potential changes with age were attributable to within‐individual variations and/or selective (dis)appearance. Finally, we investigated if the leukocyte concentration and profiles were correlated to the probability of death at a given age. The leukocyte concentration was stable with age, but the relative number of lymphocytes decreased, while the relative number of neutrophils increased, over the course of an individual's life. Moreover, between individuals of the same age, individuals with fewer lymphocytes but more neutrophils were more likely to die. Therefore, selective disappearance seems to play a role in the age variations of the immune parameters in this population. Further investigations linking age variations in immune phenotype to individual fitness are needed to understand whether remodeling of the immune system with age could or could not be adaptive.

Prognostic role of systemic inflammation in oral cavity squamous cell carcinoma

Background . The inflammatory is an integral part of the formation and development of malignant neoplasms, including oral cavity squamous cell carcinoma (OCSCC). The aim. Identification of the prognostic role of the factors of systemic inflammation in the peripheral blood of patients with OCSCC, as well as the relative indicators: neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and lymphocyte-to-monocyte ratio (LMR), with an assessment of their effect on overall survival (OS) and progression-free survival (PFS). Materials and methods . A prospective study was included 165 patients with OCSCC. At the initial assessment, all patients underwent counting of peripheral blood parameters: leukocytes, lymphocytes, monocytes, eosinophils, platelets, as well as calculation of NLR, PLR and LMR. Results. Due to univariate analysis a negative effect on OS is exerted by: the relative number of neutrophils > 61.18 % (HR = 0.66; p = 0.0280), the absolute number of lymphocytes ≤ 2.12 × 10 9 /L (HR = 0.65; p = 0.0025), relative number of monocytes > 9.1 % (HR = 0.67; p = 0.0313), NLR > 2.30 (HR = 0.63; p = 0.0130) and LMR ≤ 3.47 (HR = 0.64; p = 0.0157). The PFS in univariate analysis was significantly negatively influenced by: the level of peripheral blood leukocytes ≤ 5.23 × 10 9 /L (HR = 0.58; p = 0.0492), the absolute number of neutrophils > 2.86 × 10 9 /L (HR = 0.62; p = 0.0386), the relative number of neutrophils > 61.18 % (HR = 0.53; p = 0.0028), the absolute number of lymphocytes ≤ 2.88 × 10 9 /L (HR = 0.55; p = 0.0097), relative lymphocyte number 29.71 % (HR = 0.52; p = 0.0006), absolute number of eosinophils ≤ 0.09 × 10 9 /L (HR = 0.55; p = 0.0244), relative number of monocytes > 6.96 % (HR = 0.58; p = 0.0087), NLR > 1.74 (HR = 0.52; p = 0.0006), PLR > 111.39 (HR = 0.63; p = 0.0196) and LMR ≤ 3.71 (HR = 0.55; p = 0.0019). According to the results of multivariate analysis, a significant negative effect on PFS was exerted by the relative number of monocytes > 6.96 %, and the absolute number of eosinophils in the peripheral blood ≤ 0.09 × 10 9 /L. Conclusion. The analysis allows us to consider the factors of systemic inflammation recorded in the peripheral blood, as well as the NLR, PLR and LMR can serve as reliable additional factors in predicting the course of OCSCC.

The influence of recombinant interferon on morphological and biochemical parameters of calves’ blood in early postnatal period

The main mortality of young cattle due to infectious diseases mostly occurs in the first two months of the life, which is resulted from the calves' immune system immaturity. In order to correct the immunological status, various immunomodulators are used, interferons in particular. The aim of the study was to assess the effect of recombinant bovine interferon-α2d on the biochemical and morphological parameters of the calves' blood in the early postnatal period. Two groups of young animals were formed. Calves from the 1st group (G1,n=10) was twice injected interferon intramuscularly with the interval between injections - 7 days (500 th. MU). The 2nd group was a control (G2, n=10), the injection of the immunostimulant was replaced with saline. Before the beginning (on the 7th day of life) and at the end of the experiment (on the 21st day of life), the blood was taken for the biochemical and morphological analysis. In G1 and G2, the concentration of unconjugated bilirubin at the beginning of the experiment was 2.2 ± 0.2 and 2.1 ± 0.3 mmol/L respectively, in the G1 interferon concentration decreased to 1.1 ± 0.1 mmol/L, which was 2 times less than in the G2 (P ≤ 0.01). The activity of alkaline phosphatase at the beginning of the study was 613.8 ± 54.2 in the G2 and 514.6 ± 22.4 units/L in the G1. By the 21st day of life, the level of it decreased by 16.2% (431, 1 ± 38.0 units/L) in the G1, and it was less by 31.3% (P ≤ 0.05) than the G2 (627.3 ± 77.6 units/ L). The ALT activity at the beginning of the experiment was 4.0 ± 0.8 unit/ L in the G2 and 6.1 ± 1.2 units/L in the G1. After two weeks in the G2 level of ALT increased to 6.0 ± 2.0 U / L, while in calves from the G1, on the contrary, it was decreased in 2.4 times (P≤0.05) - 2.5 ± 0.1 U / L. The AST activity in the intact group by the 21st day was 16.0 ± 8.0 units/ L, which was 8.4 times higher (P≤0.05) than in the G1 (1.9 ± 0.4). Under the influence of interferon, there was the increase in the average concentration of leukocytes (by 13.2%), as well as the decrease in the relative number of neutrophils by 36.8% (P≤0.05). The absolute number of agranulocytes increased by 37.7% (P≤0.05) compared to the control one (5.3 ± 0.5 * 109/ L) and it was 7.3 ± 0.7 * 109/ L. Thus, parenteral injection of recombinant bovine interferon-α2d to calves in the early postnatal period causes the decrease in transaminase activity and bilirubin concentration, as well as the increase in the number of cells responsible for the humoral immune response.

Blood variables associated with survival in canine congestive heart failure patients

Our study aimed to investigate the blood variables: coenzyme Q10, N-terminal pro-B-type natriuretic peptide (NT-proBNP), electrolytes, cholesterol, triglyceride, glutathione peroxidase, white blood cell, neutrophil, lymphocyte and monocyte counts and relative numbers of neutrophils, lymphocytes and monocytes that may be associated with survival of dogs in congestive heart failure (CHF). Twenty-one client-owned dogs in CHF were included in the study. Cox regression analysis showed significant association only between NT-proBNP blood concentration and survival.

IMPACT OF CHEMOTHERAPY ON THE STANDARD SOLVENTS AND AUTOLOGOUS LIQUID TISSUES ON CYTOKINE STATUS AND NEUTROPHILS OF PERIPHERAL BLOOD OF OVARIAN CANCER PATIENTS

Chemotherapeutic treatment using standard solvents is stopped in every fourth ovarian cancer patient because of significant side effects. The autohemochemotherapy method alters the pharmacokinetics of the drug, and significantly reduces the frequency and severity of side effects. In order to compare the effects of neoadjuvant chemotherapy using autologous liquid tissues or standard solvents on cytokine status and neutrophils in the peripheral blood we evaluated the number of neutrophils and their ability to generate reactive oxygen species, and levels of interleukin-1p, IL-1Ra, IL-6, IL-10, IL-18 and TNF-a in 244 primary stage III-IV ovarian cancer patients during chemotherapy according to the AP scheme (cisplatin 75 mg/m2, doxorubicin 50 mg/m2) with infusion by standard intravenous method or as autohemochemotherapy. We established that the effectiveness of chemotherapy was associated with the redox status of neutrophils and cytokine status of peripheral blood, registered prior to neoadjuvant chemotherapy. For patients with tumor progression, increased absolute and relative number of neutrophils with reduced ability to produce reactive oxygen species is significant. For patients with a lack of tumor response, significant are the high levels of serum IL-10 and IL-18. For patients with complete or partial tumor response, significant is the decrease of IL-6 level, compared to the norm. No significant differences in neutrophil and cytokine status in both modes of infusion of chemodrugs have been established. Thus, serum IL-10, IL-18 and IL-6 may be used to predict tumor response to neoadjuvant chemotherapy according to the AP scheme.

Methods for In Vivo/Ex Vivo Analysis of Antimicrobial Peptides in Bacterial Keratitis: siRNA Knockdown, Colony Counts, Myeloperoxidase, Immunostaining, and RT-PCR Assays.

Antimicrobial peptides (AMPs) are essential components of the innate immune response. They have direct killing ability as well as immunomodulatory functions. Here, we describe techniques to identify specific AMPs involved in the protection against microbial keratitis, a vision threatening infection of the cornea of the eye which is the most serious complication of contact lens wear. Specifically we detail the use of siRNA technology to temporarily knockdown AMP expression at the murine ocular surface in vivo and then describe ex vivo assays to determine the level of bacteria, relative number of neutrophils, and levels of cytokines, chemokines, and AMPs in infected corneas.

The evaluation of comparativeness of results of hematologic analyses obtained form vacuum test-tubes with K2EDTA of various manufacturers.

The article deals with estimate of comparativeness of 20 parameters of hematologic analyses performed from vacuum test-tubes, containing anti-coagulant in the form of di-potassium salt of ethylenediaminetetraacetic acid by manufacturers Vacuette (Austria) and Lind-Vac (Estonia) according methods of protocols of the Institute of clinical laboratory standards (CLSI) GP-34A and EP9-A2-IR. The studies were implemented in the St. Petersburg Saint Luke clinical hospital using the analyzer Hemalit 5500 Corway (China) in 40 planned patients with measurement of samples in duplicates from every type of test-tube. The sampling of blood was administered by medical nurses according protocol CLSI H3-A6. The comparing was applied to significant differences between the results from test-tubes of different manufacturer and concerned to coefficient of variation (CV%), bias (B%) and total error (TE%) with regard to criteria of quality, based on biological variation. The results of implemented study revealed no significant differences between measurements from test-tubes of analyzed brands. The CV% was higher in test-tubes Lind-Vac in six indices: erythrocytes, hematoсrit, leukocytes, absolute and relative number of neutrophils, PLT (p<0.05). In the test-tubes Vacuette CV% was higher for absolute and relative number of monocytes (p<0.05). The given differences were not higher than value of allowable total error (ATE) because of minimal bias between results from different test-tubes on all measured parameters except hematoсrit. The actual study revealed no significant differences in the results of hematologic analyses performed from Lind-Vac test-tubes as compared with Vacuette. The variability of indices of convergence for every type of test-tubes is to be considered in case of clinical interpretation of results under diagnostic of anemic conditions.

Anti-inflammatory Dietary Interventions and Supplements to Improve Performance during Athletic Training

Despite the numerous positive effects of physical exercise, some negative physiological changes occur in long-lasting heavy training with transient dysfunction of the immune system, increased inflammation, and oxidative stress. This is the case of elite athletes, who train intensively to compete at the highest levels. However, these athletes can counteract the negative effects of heavy training, reducing acute and chronic inflammations and supporting the immune system, with nutritional and supplementation countermeasures. For this purpose, macronutrient manipulation with an appropriate use of certain supplements can be considered as an intervention to reduce exercise-induced immune changes and inflammatory risk. For example, branched-chain amino acid (BCAA) supplementation may promote such immune responses in skeletal muscle. Furthermore, micronutrients play an important role in immune function; in particular, the antioxidant capacity of several dietary micronutrients (e.g., tocopherols, docosahexaenoate, and flavonoids) is very interesting to support the endogenous antioxidant defense systems of the athletes, counterbalancing the negative effects of oxidative damage due to free radicals. Some of these nutrients have potential anti-inflammatory properties as assessed by the attenuated levels of interleukin-6 (IL-6) and C-reactive protein (CRP).Key Teaching Points• Long-lasting heavy training plan and competition can lead to chronic immune suppression in athletes, increasing infection risk.• Chronic exercise increases mobilization of neutrophils, decreases mobilization of lymphocytes, and decreases the absolute and relative numbers of neutrophils at rest.• Nutritional deficiencies alter the immuno-system and increase infection risk.• Nutrition can influence exercise-induced immune suppression.• Elite athletes competing at the highest levels can benefit from nutritional and supplementation support to improve immunity and reduce acute and chronic inflammations.

THE DYNAMICS OF THE IMMUNE STATUS IN CHILDREN WITH RECURRENT RESPIRATORY TRACT INFECTIONS DURING IMMUNE REHABILITATION IN REMISSION

Objective: to assess the immune status of children who frequently and continuously suffer from recurrent respiratory tract infections with diverse clinical manifestations in dynamics in the course of immune rehabilitation. Materials and methods. Neutrophil and lymphocyte subpopulations of peripheral blood were studied by flow cytometry method based on monoclonal antibodies (CD3, CD8, CD11a, CD18, CD19, CD28, CD154, LPS) as well as G, M, A, E immunoglobulins of peripheral blood were studied by turbodymetric method in 78 children aged 2-6. The group of the children included those who frequently and chronically suffered from respiratory infections. They were distributed into 3 groups according to the clinical variants of acute respiratory infections. All the studies were performed three times: before the immune rehabilitation, 10 days later and 2 months later after the course of the immune rehabilitation. Results. The study detected an increase in the relative number of neutrophils expressing CD18 + receptor in all the clinical subgroups and CD11a + receptor in the subgroups with recurrent upper respiratory tract infections without complications and with ENT complications. The groups with ENT (LPS + CD19 + ) and bronchopulmonary complications (LPS + CD19 + , LPS + CD3 + ) observed reduction of the lipopolysaccharide-binding activity of lymphocytes. The group with recurrent upper respiratory tract infections without complications observed reduction of CD154 + -subpopulation of lymphocytes. The immune rehabilitation performed in remission brought positive effect on the immunological characteristics of the patients from the group of frequently and chronically infected: increased LPS + and CD154 + lymphocyte level and decreased excessive activation of the immune system in the form of lower overexpression of adhesion CD11a + and CD18 + receptors on neutrophils. Conclusion. The detected changes can serve as a criterion of forming risk groups of relapse cases of acute upper respiratory tract infections with ENT and bronchopulmonary complications as well as justify the need for further immune rehabilitation and laboratory monitoring in this group of the patients.

Selective phosphodiesterase 3 inhibitor olprinone attenuates meconium-induced oxidative lung injury

Since inflammation and oxidation play a key role in the pathophysiology of neonatal meconium aspiration syndrome, various anti-inflammatory drugs have been tested in the treatment. This study evaluated whether the phosphodiesterase (PDE) 3 inhibitor olprinone can alleviate meconium-induced inflammation and oxidative lung injury. Oxygen-ventilated rabbits intratracheally received 4 ml/kg of meconium (25 mg/ml) or saline. Thirty minutes after meconium/saline instillation, meconium-instilled animals were treated by intravenous olprinone (0.2 mg/kg) or were left without treatment. All animals were oxygen-ventilated for an additional 5 h. A bronchoalveolar lavage (BAL) of the left lungs was performed and differential leukocyte count in the sediment was estimated. The right lungs were used to determine lung edema by wet/dry weight ratio, as well as to detect oxidative damage to the lungs. In the lung tissue homogenate, total antioxidant status (TAS) was determined. In isolated lung mitochondria, the thiol group content, conjugated dienes, thiobarbituric acid-reactive substances (TBARS), dityrosine, lysine-lipid peroxidation products, and activity of cytochrome c oxidase (COX) were estimated. To evaluate the effects of meconium instillation and olprinone treatment on the systemic level, TBARS and TAS were determined in the blood plasma, as well. Meconium instillation increased the relative numbers of neutrophils and eosinophils in the BAL fluid, increased edema formation and concentrations of oxidation markers, and decreased TAS. Treatment with olprinone reduced the numbers of polymorphonuclears in the BAL fluid, decreased the formation of most oxidation markers in the lungs, reduced lung edema and prevented a decrease in TAS in the lung homogenate compared to non-treated animals. In the blood plasma, olprinone decreased TBARS and increased TAS compared to the non-treated group. Conclusion, the selective PDE3 inhibitor olprinone has shown potent antioxidative and anti-inflammatory effects in the meconium-induced oxidative lung injury.

Dynamic Expression of IL-6 Trans-Signalling Molecules in the Lungs of Preterm Baboons Undergoing Mechanical Ventilation

Background: Interleukin (IL)-6, when complexed with soluble IL-6 receptor (sIL-6R), has emerged as an important modulator of chemokine expression and leukocyte recruitment during inflammation and in this state can be specifically antagonised by soluble gp130 (sgp130). The expression of these modifiers of IL-6 activity during ventilator-induced inflammation remains poorly understood. Objectives: To ascertain the expression pattern of IL-6, sIL-6R and sgp130 in response to mechanical ventilation in the preterm neonatal lung and define its relationship to associated markers of inflammation. Methods: Inflammatory cell recruitment and expression of IL-6, sIL-6R, sgp130, IL-8 and monocyte chemotactic protein-1 (MCP-1) were quantified in tracheal aspirate fluid collected over a 14-day period from preterm (125 days) baboons undergoing mechanical ventilation. Results: Over the period of ventilation, the ratio of agonistic IL-6/sIL-6R increased 4.3-fold between days 3 and 10–11 (p < 0.01) while the ratio of antagonistic sgp130/IL-6 decreased 2.6-fold over the same period (p < 0.05). Over the same period, the relative numbers of neutrophils compared to mononuclear cells shifted from an excess of 1.8 on day 1 to 0.6 on day 14 (p < 0.01). Both IL-8 and MCP-1 were elevated between days 1 and 10–11 of ventilation (p < 0.01). Conclusions: In the ventilated preterm baboon lung, expression of sIL-6R and dynamic modulation of sgp130 expression appear to modulate the activity and inflammatory potential of IL-6.

Respiratory Disease Markers in Porcine Bronchoalveolar Lavage Fluid

SummaryIn bronchoalveolar lavage fluid (BALF) of pigs originating from different herds bacteria, cells and the antibacterial peptide PR‐39 were examined to gain information about the lung health status. In a high health nucleus herd 56% and in low health herds 20–100% of the examined pigs were found positive for potentially pathogenic bacteria. Based on these findings, a novel definition for bacterial respiratory tract disease was established using an 8% cut‐off for the relative number of neutrophils in bronchoscopic and a 40% cut‐off in transtracheal BALF in combination with the occurrence of potentially pathogenic microorganisms. The antibacterial peptide PR‐39 was highly correlated to this definition of respiratory disease. An assessment of the bacteriological respiratory health status appears to be possibly based on the determination of PR‐39 concentrations in BALF using different cut‐off values according to the lavage method (2.5 nM for bronchoscopic and 5 nM for transtracheal BALF).

Ingestion of low doses of deoxynivalenol does not affect hematological, biochemical, or immune responses of piglets1

Deoxynivalenol (DON), a mycotoxin produced by Fusarium spp., is a frequent contaminant of cereals. Because of their rich cereal diet, pigs could be exposed to this mycotoxin. Pigs are among the animal species showing the greatest sensitivity to DON. Effects of intermediate to high levels of DON on pigs are well known and include feed refusal, decreased feed intake, and alteration of the immune response. Effects of low levels of DON, which are commonly detected in contaminated feed, remain unknown. The aim of this study was to investigate the effect of a diet naturally contaminated with a low concentration of DON (0, 280, 560, or 840 microg/kg of feed) on performance of weanling piglets and on 34 hematological, biochemical, and immune variables. Low doses of DON did not alter the animal performances (feed intake and BW gain). Such low levels of DON did not modify the 9 hematological variables measured (including white blood cell, red blood cell, and platelet counts, relative numbers of neutrophils and lymphocytes, and hematocrit and hemoglobin concentrations) or the 18 biochemical variables tested (including cations, glucose, urea, creatinine, bilirubin, cholesterol and triglyceride concentrations, and plasma enzyme activity). Similarly, no effect of low doses of DON was observed on the immune responses of the animals (immunoglobulin subset concentration, lymphocyte proliferation, and cytokine production).

235 MORPHOMETRIC DETERMINATION OF ENDOMETRIAL LEUKOCYTE MIGRATION DURING DIFFERENT STAGES OF THE EQUINE ESTROUS CYCLE

Uterine defenses against bacterial challenge are more efficient during estrus than during diestrus. The aim of the present study was to test the hypothesis that the magnitude of endometrial leukocyte migration after instillation of semen into the uterine lumen is greater during estrus than during diestrus. Eight Nooitgedacht mares of proven fertility were used in the study. Each mare received a different treatment (Rx) during each of 4 estrous cycles, with a rest cycle after each Rx. Two Rxs were performed during diestrus and 2 during estrus. One Rx for each stage of the cycle was a control Rx (no challenge to the endometrium). At time zero (T0) of challenged cycles, a single aliquot of 13 mL raw semen, frozen-thawed without addition of any cryoprotectant or extender, was instilled into the uterus. An endometrial biopsy was taken 6 and 48 h after T0 and endometrial cytology and culture were performed at 48 and 120 h after T0. An image analyzer was used to record the total number of cells, round cells, neutrophils, and eosinophils per unit surface area of epithelium, stratum compactum (SC), and stratum spongiosum (SS). The relative number of round cells, neutrophils, and eosinophils were expressed as proportions of the number of each cell type per total number of cells. The use of an image analyzer made the collection of quantitative data from histologic sections possible. Without respect to challenge or time, the total numbers of epithelial cells per mm epithelium and the total number of cells (fibroblasts, round cells, neutrophils, and eosinophils) and of round cells per mm2 of SS were significantly greater during diestrus than during estrus; however, no such difference could be demonstrated for the SC. The stage of the estrous cycle had no meaningful influence on any other (measured or calculated) variable. During challenged cycles, absolute and relative numbers of neutrophils were significantly greater in the epithelium, SC, and SS than during control cycles. There was an interaction (not always reaching significance) between Rx and time with regard to the absolute and relative numbers of neutrophils in epithelium and SS and round cells in the epithelium. Numbers of neutrophils and round cells were significantly higher 6 h after Rx than 48 h after Rx in challenged cycles, but did not differ during control cycles. During challenged cycles, the stage of the estrous cycle when Rx occurred had no effect on the duration of the induced endometritis, the occurrence of positive cytology or culture results, or the type of bacteria that were cultured. Regardless of the stage of their cycles when they were challenged, all mares rid themselves of the opportunistic pathogens placed into the uterine lumen within one estrous cycle. It is concluded that the stage of the estrous cycle did not influence the magnitude of the endometrial leukocyte response to a standardized challenge with semen in these reproductively sound mares. A similar study will be required to test whether this conclusion also holds true for mares that are susceptible to endometritis.

Limitations on the use of dihydrorhodamine 123 for flow cytometric analysis of the neutrophil respiratory burst

Intracellular oxidation of dihydrorhodamine 123 (DHR) to the fluorescent compound rhodamine 123 (Rho123) was used to detect the production of oxygen metabolites in activated neutrophils. Total leukocyte preparations can be used in this assay, which is a great advantage when priming of the respiratory burst is studied. We have defined the conditions that should be taken into account when priming is studied with this assay. We found that neither the extent nor the kinetics of DHR oxidation match those of NADPH oxidase activity. In addition, DHR oxidation is influenced by the absolute and relative number of neutrophils in the leukocyte suspension, by the DHR concentration and by myeloperoxidase availability. The results presented in this study emphasize the need for carefully designed experiments when DHR is used to study the respiratory burst in neutrophils.

Neutrophil influx into guinea-pig airway lumen during cholinergic and non-cholinergic bronchoconstriction.

Sensory nerve activation will produce adherence of neutrophils to tracheobronchial microvessels. The aim of the present study was to investigate whether this adherence would lead to an influx of neutrophils into the airway lumen. To do this, we studied the effects of 20 minutes of vagal stimulation (1 Hz, 5 ms, 5 V) in anaesthetized and tracheostomized guinea-pigs on both lung resistance, and the cell picture in bronchoalveolar lavage. Any changes were compared to those of intravenous methacholine infusion, producing similar changes in lung resistance. Since high pressure ventilation could produce lung damage, we also studied the effects of ventilation through an extracorporeal resistor, producing a similar change in transpulmonary pressure (45 +/- 2 cmH2O) as vagal stimulation (42 +/- 4 cmH2O). The total number of cells recovered in the lavage was not increased by vagal stimulation, methacholine infusion or the extracorporeal resistor. However, both vagal stimulation and methacholine infusion significantly increased the relative number of neutrophils in the lavage compared to sham stimulated animals (21 +/- 11%, 13 +/- 4% and 4 +/- 1% respectively), but the extracorporeal resistor had no effect (4 +/- 2%). Our data suggests that prolonged bronchoconstriction per se may induce an influx of neutrophils into the airway lumen of the guinea-pig.