Blood Leukocyte Concentrations Research Articles

Non-invasive detection of the content of white blood cells in the blood of humans based on dynamic spectrum

Objective. Changes in white blood cell content have been shown to be useful in determining whether the body is in a healthy state. We propose an improved data processing and modeling approach, which helps to accommodate blood component content detection and improve prediction accuracy. A pproach. In this experiment, the finger-end transmission method was used for spectral measurement, and we collected a total of 440 sample data. In this paper, we first use the method of CEEMDAN combined with wavelet threshold to denoise the PPG signal, and then use the integral method to extract the spectral features, which makes up for the defects of the single-edge method using incomplete data and the deviation of the slope of the rising segment from the actual signal. We further improve the screening of samples and wavelengths, and used PLS regression modeling combine the double nonlinear correction method to build the most stable and universal model. Main results. The model has been applied to 332 subjects’ finger transmission spectral data to predict the concentration of leukocytes. The correlation coefficient of the final training set result was 0.927, and the root mean square error (RMSE) is 0.569×109 l−1, the correlation coefficient of the prediction set result is 0.817, and the RMSE is 0.826×109 l−1, which proves the practicability of the proposed method. Significance. We propose a non-invasive method for detecting leukocyte concentration in blood that can also be generalized to detect other blood components.

Intertwined leukocyte balances in tumours and peripheral blood as robust predictors of right and left colorectal cancer survival.

BACKGROUNDColorectal cancer (CRC) accounts for 9.4% of overall cancer deaths, ranking second after lung cancer. Despite the large number of factors tested to predict their outcome, most patients with similar variables show big differences in survival. Moreover, right-sided CRC (RCRC) and left-sided CRC (LCRC) patients exhibit large differences in outcome after surgical intervention as assessed by preoperative blood leukocyte status. We hypothesised that stronger indexes than circulating (blood) leukocyte ratios to predict RCRC and LCRC patient outcomes will result from combining both circulating and infiltrated (tumour/peritumour fixed tissues) concentrations of leukocytes.AIMTo seek variables involving leukocyte balances in peripheral blood and tumour tissues and to predict the outcome of CRC patients.METHODSSixty-five patients diagnosed with colon adenocarcinoma by the Digestive Surgery Service of the La Paz University Hospital (Madrid, Spain) were enrolled in this study: 43 with RCRC and 22 with LCRC. Patients were followed-up from January 2017 to March 2021 to record overall survival (OS) and recurrence-free survival (RFS) after surgical interventions. Leukocyte concentrations in peripheral blood were determined by routine laboratory protocols. Paraffin-fixed samples of tumour and peritumoural tissues were assessed for leukocyte concentrations by immunohistochemical detection of CD4, CD8, and CD14 marker expression. Ratios of leukocyte concentration in blood and tissues were calculated and evaluated for their predictor values for OS and RFS with Spearman correlations and Cox univariate and multivariate proportional hazards regression, followed by the calculation of the receiver-operating characteristic and area under the curve (AUC) and the determination of Youden’s optimal cutoff values for those variables that significantly correlated with either RCRC or LCRC patient outcomes. RCRC patients from the cohort were randomly assigned to modelling and validation sets, and clinician-friendly nomograms were developed to predict OS and RFS from the respective significant indexes. The accuracy of the model was evaluated using calibration and validation plots.RESULTSThe relationship of leukocyte ratios in blood and peritumour resulted in six robust predictors of worse OS in RCRC: CD8+ lymphocyte content in peritumour (CD8pt, AUC = 0.585, cutoff < 8.250, P = 0.0077); total lymphocyte content in peritumour (CD4CD8pt, AUC = 0.550, cutoff < 10.160, P = 0.0188); lymphocyte-to-monocyte ratio in peritumour (LMRpt, AUC = 0.807, cutoff < 3.185, P = 0.0028); CD8+ LMR in peritumour (CD8MRpt, AUC = 0.757, cutoff < 1.650, P = 0.0007); the ratio of blood LMR to LMR in peritumour (LMRb/LMRpt, AUC = 0.672, cutoff > 0.985, P = 0.0244); and the ratio of blood LMR to CD8+ LMR in peritumour (LMRb/CD8MRpt, AUC = 0.601, cutoff > 1.485, P = 0.0101). In addition, three robust predictors of worse RFS in RCRC were found: LMRpt (AUC = 0.737, cutoff < 3.185, P = 0.0046); LMRb/LMRpt (AUC = 0.678, cutoff > 0.985, P = 0.0155) and LMRb/CD8MRpt (AUC = 0.615, cutoff > 1.485, P = 0.0141). Furthermore, the ratio of blood LMR to CD4+ LMR in peritumour (LMRb/CD4MRpt, AUC = 0.786, cutoff > 10.570, P = 0.0416) was found to robustly predict poorer OS in LCRC patients. The nomograms showed moderate accuracy in predicting OS and RFS in RCRC patients, with concordance index of 0.600 and 0.605, respectively.CONCLUSIONEasily obtainable variables at preoperative consultation, defining the status of leukocyte balances between peripheral blood and peritumoural tissues, are robust predictors for OS and RFS of both RCRC and LCRC patients.

The immature platelet fraction affects the efficacy of platelet rich plasma therapy for knee osteoarthritis.

IntroductionPlatelet-rich plasma (PRP) therapy is used to treat pathological conditions such as degenerative inflammatory diseases including osteoarthritis (OA) by enhancing tissue repair and promoting anti-inflammatory effects. Although PRP therapy for patients with knee OA improved pain and functional scores, the association of clinical outcomes and quality of PRP including cell composition and concentration is unclear.MethodsTherefore, this study analyzed blood cell counts, including the immature platelet fraction (IPF), in peripheral blood and PRP of 144 patients with knee OA who underwent PRP therapy. The mean leukocyte and platelet concentrations in whole blood and PRP were analyzed using an XN-1000 automated hematology analyzer. Visual analogue scale (VAS) scores and knee injury and osteoarthritis outcome scores (KOOS) before and 1 month after a single PRP injection were also determined.ResultsHigher platelet and lower leukocyte concentration rates were observed in PRP compared with whole blood. The platelet concentration in whole blood was negatively correlated with VAS improvement. The percentage of IPF (IPF%) in whole blood was positively correlated with VAS improvement and KOOS (pain) improvement, whereas the IPF% in PRP tended to correlate with VAS improvement. Furthermore, multivariate logistic regression demonstrated the high IPF% in whole blood was significantly associated with VAS improvement. The low percentage of neutrophil (neutrophil%) in PRP was significantly associated with the VAS improvement and KOOS (ADL) improvement.ConclusionsTherefore, PRP efficacy for OA might depend on the patient's biological status.

Assessment with clinical data of a coupled bio-hemodynamics numerical model to predict leukocyte adhesion in coronary arteries

Numerical simulations of coupled hemodynamics and leukocyte transport and adhesion inside coronary arteries have been performed. Realistic artery geometries have been obtained for a set of four patients from intravascular ultrasound and angiography images. The numerical model computes unsteady three-dimensional blood hemodynamics and leukocyte concentration in the blood. Wall-shear stress dependent leukocyte adhesion is also computed through agent-based modeling rules, fully coupled to the hemodynamics and leukocyte transport. Numerical results have a good correlation with clinical data. Regions where high adhesion is predicted by the simulations coincide to a good approximation with artery segments presenting plaque increase, as documented by clinical data from baseline and six-month follow-up exam of the same artery. In addition, it is observed that the artery geometry and, in particular, the tortuosity of the centerline are a primary factor in determining the spatial distribution of wall-shear stress, and of the resulting leukocyte adhesion patterns. Although further work is required to overcome the limitations of the present model and ultimately quantify plaque growth in the simulations, these results are encouraging towards establishing a predictive methodology for atherosclerosis progress.

Volutrauma Increases Exhaled Pentanal in Rats: A Potential Breath Biomarker for Ventilator-Induced Lung Injury.

Mechanical ventilation injures lungs, but there are currently no reliable methods for detecting early injury. We therefore evaluated whether exhaled pentanal, a lipid peroxidation product, might be a useful breath biomarker for stretch-induced lung injury in rats. A total of 150 male Sprague-Dawley rats were investigated in 2 substudies. The first randomly assigned 75 rats to 7 hours of mechanical ventilation at tidal volumes of 6, 8, 12, 16, and 20 mL·kg-1. The second included 75 rats. A reference group was ventilated at a tidal volume of 6 mL·kg-1 for 10 hours 4 interventional groups were ventilated at a tidal volume of 6 mL·kg-1 for 1 hour, and then for 0.5, 1, 2, or 3 hours at a tidal volume of 16 mL.kg-1 before returning to a tidal volume of 6 mL·kg-1 for additional 6 hours. Exhaled pentanal was monitored by multicapillary column-ion mobility spectrometry. The first substudy included cytokine and leukocyte measurements in blood and bronchoalveolar fluid, histological assessment of the proportion of alveolar space, and measurements of myeloperoxidase activity in lung tissue. The second substudy included measurements of pentanal in arterial blood plasma, cytokine and leukocyte concentrations in bronchoalveolar fluid, and cleaved caspase 3 in lung tissue. Exhaled pentanal concentrations increased by only 0.5 ppb·h-1 (95% confidence interval [CI], 0.3-0.6) when rats were ventilated at 6 mL·kg-1. In contrast, exhaled pentanal concentrations increased substantially and roughly linearly at higher tidal volumes, up to 3.1 ppb·h-1 (95% CI, 2.3-3.8) at tidal volumes of 20 mL·kg-1. Exhaled pentanal increased at average rates between 1.0 ppb·h-1 (95% CI, 0.3-1.7) and 2.5 ppb·h-1 (95% CI, 1.4-3.6) after the onset of 16 mL·kg-1 tidal volumes and decreased rapidly by a median of 2 ppb (interquartile range [IQR], 0.9-3.2), corresponding to a 38% (IQR, 31-43) reduction when tidal volume returned to 6 mL·kg-1. Tidal volume, inspiratory pressure, and mechanical power were positively associated with pentanal exhalation. Exhaled and plasma pentanal were uncorrelated. Alveolar space decreased and inflammatory markers in bronchoalveolar lavage fluid increased in animals ventilated at high tidal volumes. Short, intermittent ventilation at high tidal volumes for up to 3 hours increased neither inflammatory markers in bronchoalveolar fluid nor the proportion of cleaved caspase 3 in lung tissue. Exhaled pentanal is a potential biomarker for early detection of ventilator-induced lung injury in rats.

MicroRNA expression levels in lung recipients: correlations with clinical and laboratory data

Objective: to evaluate the expression levels of miRNA (miR-27, miR-101, miR-142, miR-339 and miR-424) and its relationship with clinical and laboratory parameters in lung transplant recipients. Materials and methods. The study included 57 lung recipients aged 10 to 74 years (35 ± 15), including six children (9%) – four boys 10, 12, 13 and 17 years and girls 13 and 14 years old – and 51 adult recipients, including 30 men (62.5%). The control group was made up of 14 healthy individuals that were not significantly different by gender and age. Expression levels of the microRNAs studied in blood plasma were determined via quantitative polymerase chain reaction (PCR). Correlations of miRNA expression levels with complete blood count and biochemical blood test indicators were analyzed. Results. Patients with end-stage chronic respiratory failure (potential lung recipients) were found to have significantly higher expression levels of miR-27, miR-101 and miR-339 in plasma than the healthy individuals (p = 0.02, p = 0.03 and p = 0.01, respectively). The expression level of miR-339 correlated with the age of potential lung recipients (p = 0.04). It was a negative correlation (r = –0.46). The expression levels of the other four miRNAs were age independent. The average expression level of miR-424 in lung recipients in the long-term period after lung transplant was higher than in waitlisted patients (p = 0.03). Analysis of the relationship between miRNA expression levels and external respiration function in the long-term post-transplant period showed that miR-142 expression level (r = 0.61; p = 0.04) positively correlates with the Tiffeneau-Pinelli index. This strong correlation, which exceeds 85%, indicates the presence of restrictive lung diseases. A year and more after transplantation, it was found that in the recipients, there were close positive correlations between miR-27, miR-142, miR-424 expression levels and blood leukocyte concentration, as well as between the miR-142 expression level and the sCD40L concentration during this period. Conclusion. A comparative study of the expression level of miRNAs (miR-27, miR-101, miR-142, miR-339 and miR-424) in the blood plasma of patients suffering from end-stage chronic lung diseases of various origin and in lung recipients enables us to conclude that further studies of the miRNA panels are needed in order to assess their effectiveness as potential molecular and genetic markers of post-transplant complications.

Trait-specific effects of exogenous triiodothyronine on cytokine and behavioral responses to simulated systemic infection in male Siberian hamsters

Seasonal changes in day length enhance and suppress immune function in a trait-specific manner. In Siberian hamsters (Phodopus sungorus) winter-like short days (SDs) increase blood leukocyte concentrations and adaptive T cell dependent immune responses, but attenuate innate inflammatory responses to simulated infections. Thyroid hormone (TH) signaling also changes seasonally and has been implicated in modulation of the reproductive axis by day length. Immunologically, TH administration in long days (LD) enhances adaptive immune responses in male Siberian hamsters, mimicking effects of SDs. This experiment tested the hypothesis that T3 is also sufficient to mimic the effects of SD on innate immune responses. Adult male hamsters housed in LDs were pretreated with triiodothyronine (T3; 1 μg, s.c.) or saline (VEH) daily for 6 weeks; additional positive controls were housed in SD and received VEH, after which cytokine, behavioral, and physiological responses to simulated bacterial infection (lipopolysaccharide; LPS) were evaluated. SD pretreatment inhibited proinflammatory cytokine mRNA expression (i.e. interleukin 1β, nuclear factor kappa-light-chain-enhancer of activated B cells). In addition, the magnitude and persistence of anorexic and cachectic responses to LPS were also lower in SD hamsters, and LPS-induced inhibition of nest building behavior was absent in SD. T3 treatments failed to affect behavioral (food intake, nest building) or somatic (body mass) responses to LPS in LD hamsters, but one CNS cytokine response to LPS (e.g., hypothalamic TNFα) was augmented by T3. Together these data implicate thyroid hormone signaling in select aspects of innate immune responses to seasonal changes in day length.

Blood Leukocyte Concentrations, FEV1 Decline, and Airflow Limitation. A 15-Year Longitudinal Study of World Trade Center-exposed Firefighters.

Rescue/recovery work at the World Trade Center disaster site (WTC) caused a proximate decline in lung function in Fire Department of the City of New York firefighters. A subset of this cohort experienced an accelerated rate of lung function decline over 15 years of post-September 11, 2001 (9/11) follow-up. To determine if early postexposure blood leukocyte concentrations are biomarkers for subsequent FEV1 decline and incident airflow limitation. Individual rates of forced expiratory volume in 1 second (FEV1) change were calculated for 9,434 firefighters using 88,709 spirometric measurements taken between September 11, 2001, and September 10, 2016. We categorized FEV1 change rates into three trajectories: accelerated FEV1 decline (FEV1 loss >64 ml/yr), expected FEV1 decline (FEV1 loss between 0 and 64 ml/yr), and improved FEV1 (positive rate of change >0 ml/yr). Occurrence of FEV1/FVC less than 0.70 after 9/11 defined incident airflow limitation. Using regression models, we assessed associations of post-9/11 blood eosinophil and neutrophil concentrations with subsequent FEV1 decline and airflow limitation, adjusted for age, race, smoking, height, WTC exposure level, weight change, and baseline lung function. Accelerated FEV1 decline occurred in 12.7% of participants (1,199 of 9,434), whereas post-9/11 FEV1 improvement occurred in 8.3% (780 of 9,434). Higher blood eosinophil and neutrophil concentrations were each associated with accelerated FEV1 decline after adjustment for covariates (odds ratio [OR], 1.10 per 100 eosinophils/μl; 95% confidence interval [CI], 1.05-1.15; and OR, 1.10 per 1,000 neutrophils/μl; 95% CI, 1.05-1.15, respectively). Multivariable-adjusted linear regression models showed that a higher blood neutrophil concentration was associated with a faster rate of FEV1 decline (1.14 ml/yr decline per 1,000 neutrophils/μl; 95% CI, 0.69-1.60 ml/yr; P < 0.001). Higher blood eosinophil concentrations were associated with a faster rate of FEV1 decline in ever-smokers (1.46 ml/yr decline per 100 eosinophils/μl; 95% CI, 0.65-2.26 ml/yr; P < 0.001) but not in never-smokers (P for interaction = 0.004). Higher eosinophil concentrations were also associated with incident airflow limitation (adjusted hazard ratio, 1.10 per 100 eosinophils/μl; 95% CI, 1.04-1.15). Compared with the expected FEV1 decline group, individuals experiencing accelerated FEV1 decline were more likely to have incident airflow limitation (adjusted OR, 4.12; 95% CI, 3.30-5.14). Higher post-9/11 blood neutrophil and eosinophil concentrations were associated with subsequent accelerated FEV1 decline in WTC-exposed firefighters. Both higher blood eosinophil concentrations and accelerated FEV1 decline were associated with incident airflow limitation in WTC-exposed firefighters.

P099 The impact of lymphocyte purity on flow cytometry crossmatch (FCXM) assay. It’s not purely theoretical

Aim Lymphocyte flow cytometric crossmatch (FCXM) is used in pre-transplant risk assessment to detect donor specific antibodies (DSA). Many labs use density medium centrifugation to isolate donor cells for FCXM, which yields variable lymphocyte purity (15–90%) depending on the original concentration of lymphocytes and other leukocytes in donor blood, as well as sample quality and age. However, it is unclear whether variation in lymphocyte purity can effect FCXM assay. The goal of this study was to assess the impact of donor lymphocyte purity on FCXM results. Methods Enriched lymphocytes (Ly; >97%), neutrophils (Nu; >98%) and monocytes (Mo; >92%) were isolated from 5 volunteer donors using subset specific EasySep Direct kits (STEMCELL). Whole leukocyte (WL) cell preps were obtained by mixing enriched Ly, Nu and Mo in equal proportion (approx. 33% each) to mimic poor quality cell preps. Ly and WL cell preps were treated with pronase (2.35 kunitz units/ml) or control (PBS) and used as targets in FCXM against negative control (NC) and several dilutions of positive control (PC) sera. Median channel fluorescence (MCF) shifts from NC obtained with Ly vs WL target cell preps were compared. Results T and B cell FCXM NC reactivity was similar with Ly and WL cells. Pronase treatment was equally effective at reducing B cell background in Ly and WL cell preps (191 vs 198 MCF shift). Interestingly, both T and B cells FCXM performed with WL targets showed significantly reduced PC reactivity compared to Ly cells (Fig. 1). This was true at all PC concentrations (1/50–1/400) and was seen with both pronase treated and untreated cells. Average MCF decrease in pronase treated Ly vs WL preps was 49 MCF for T and 68 MCF for B cell FCXM, and in untreated preps it was 41 MCF for T and 61 MCF for B cell FCXM. Conclusion Lymphocyte purity has significant impact on FCXM results. Weak DSA could be missed if low purity lymphocyte preps are used in FCXM assay. Use of highly enriched lymphocytes will improve detection of DSA and reduce variability of FCXM results. Download : Download high-res image (220KB) Download : Download full-size image

The application of EHF therapy for the combined treatment of severe poisoning with the scolding liquids

The objective of the present study was to compare the results of the treatment and changes in the measurements of the laboratory characteristics in the patients suffering from severe poisoning with the scolding liquids under effect of the combined treatment including the application of extremely high frequency electromagnetic radiation (EHF therapy). The study included ten patients given a course of EHF therapy and 10 ones treated without it. The patients of the two groups developed the oppositely directed alterations in the rheological properties of blood and hemostasis including the elevation of the hemorheological viscoelastic constituent, erythrocyte and platelet aggregation, the increase in blood viscosity at a high shear velocity and, conversely, its decrease at the low and moderate shear velocities and hematocrit. Moderate leukocytosis was observed during this period along with the 3-4 rise in the concentration of lymphocytes involved in the apoptotic process and the substantial increase (1,4 - 8,1 times) in the values of parameters of endotoxicosis (the leukocytic index of intoxication and the erythrocyte sedimentation rate). The use of EHF therapy for the management of this category of the patients at the stage of their medical rehabilitation produced a modulating hemorheological effect manifested as the reduction of the elevated hemorheological characteristics, with the especially pronounced increase in the parameters below the respective normal values. In the group comprising the control patients, the undesirable changes occurred more frequently than in the previous one; specifically, deviations of the parameters of interest from their normal values were more pronounced. EHF therapy resulted in the reduction of the concentration of dead leukocytes in blood and a two-fold decrease in the amount of lymphocytes at the later stages of apoptosis whereas the concentration of the lymphocytes just entering apoptosis was elevated; this situation reflected the process of sanogenesis. In contrast, this parameter decreased in the patients of the group of comparison. The reduction in the level of endogenous intoxication under effect of EHF therapy manifested itself as a more significant decrease of leukocytic index of intoxication and erythrocyte sedimentation rate in the patients of the main group in comparison with those of the group of comparison. It is concluded that the inclusion of EHF therapy in the combined treatment of severe poisoning with the scolding liquids reduces the time of epithelization of gastrointestinal mucosa by 4.7 days on the average. Accordingly, the duration of the hospital stay can be decreased by 3.8 days.

Fluctuation of neutrophil counts around parturition in Holstein dairy cows with and without retained placenta

Retained placenta (RP) is often diagnosed in high-yielding dairy cows and can negatively affect reproductive performances. The objective of the present study was to investigate the hematological and biochemical profile of cows with RP before and immediately after parturition, with particular emphasis on neutrophil counts, since a previous study demonstrated the presence of peripheral neutropenia in dairy cows with RP sampled a few days after parturition. Results from 12 Holstein cows affected by RP and from 17 clinically healthy controls sampled one week pre-partum, within 12h after calving and between 48 and 72h after parturition were compared between groups and over time. Compared with controls, cows with RP had lower lymphocyte counts before parturition, lower leukocyte and neutrophil counts at parturition, lower monocyte counts at all times, and higher β-hydroxybutyrate before and after parturition. Erythroid and biochemical parameters were similar over time in both groups, whereas RP cows did not show the increase of neutrophil counts that occurs in controls at parturition. Hence, the finding of a lower neutrophil count in a routinely hemogram performed at parturition could be used as an alarm signal suggesting to monitor the affected animals. Moreover, although the underlying pathogenetic mechanism should be better investigated, the present study describes for the first time the association between altered blood leukocyte concentrations at parturition in RP compared to control cows.

Allometric and Phylogenic Comparisons of Circulating Leukocyte Concentrations between and within Birds and Mammals

The blood concentrations of leukocytes and the major classes of leukocytes were compared across over 200 species of birds and over 300 species of mammals based on phylogenic relations, body weight and diet. The concentration of leukocytes was markedly higher in birds than mammals reflecting elevated concentrations of all major classes but particularly those of lymphocytes, monocytes, eosinophils and basophils. Overall, there were no differences between blood leukocyte concentrations in placental and marsupial mammals. There were differences between leukocytes concentrations within the placental mammals, for instance being lower in mammalian species within the taxa Glires than in Laurasiatheria and Primatomorpha which in turn are lower than in Afrotheria and Xenarthra. There were also differences in leukocyte concentrations in birds being higher in species with the taxa Afroaves than in Australaves. There were phylogenic differences in the percentages of the leukocyte classes. For instance, in placental compared marsupial mammals, the percentage neutrophils is elevated, the percentage lymphocytes is depressed and neutrophil to lymphocyte (N:L) ratio was much higher. Moreover, the percentage heterophils is high, the percentage lymphocytes is low and heterophil to lymphocyte (H:L) ratio is greater in birds from the taxa Paleognathae than the Neognathae. There was a positive relationship between concentrations of leukocytes or neutrophils and log body weight.

Total Knee Arthroplasty Infected by Brucella Melitensis

Objectives:Brucella infection associated with arthroplasty is a rare event. We present the long-term results of two-stage revision knee arthroplasty due to TKA infection by Brucella melitensis.Methods:A 62-year-old man presented with a 4-month history of night sweats, high body temperature, and right knee pain in 2003. Two years previously, he had undergone a right TKA for treatment of gonarthrosis. Upon examination, the knee appeared swollen, warm, and sore, and a sinus tract formation had developed at the incision area. Bacteriologic cultures taken from the sinus tract discharge revealed B. melitensis. The patient’s erythrocyte sedimentation rate (ESR) was 83 mm/h, C-reactive protein (CRP) level was 12.38 mg/L, and blood leukocyte concentration was 9425/mm3. Plain radiography confirmed septic loosening. A two-stage revision TKA was planned. The first stage involved removal of the infected prosthesis and debridement of the remaining cement and necrotic tissue. Removal of the prosthetic components and cement and application of an antibiotic-loaded spacer (teicoplanin-impregnated) was also performed in the first stage. Oral rifampicin (900 mg) and doxycycline (300 mg) were administered daily for 12 weeks postoperatively. At the end of the antibiotic therapy, the patient was seen as an outpatient, and his ESR, CRP level, and antibody titer had decreased to normal values. Finally, prosthetic revision was performed in the second stage of the TKA.Results:Ten years following surgery, the patient had returned to full activities of daily living, described no knee pain, and was very satisfied with the outcome. No abnormalities associated with the femoral, tibial, or patellar components were observed on plain radiography 10 years following surgery. The results of a Brucella agglutination test at that time were negative.Conclusion:Osteoarticular disease related to Brucella remains a problem in endemic regions. Brucellosis should be kept in mind in the differential diagnosis of prosthetic joint infections, especially in patients with a history of brucellosis and exposure to possible sources of the bacteria. We believe that two-stage revision arthroplasty is the first-line treatment of a loose TKA infected with B. melitensis. In addition, if a systemic Brucella infection is identified, the patient should be managed with two-stage revision surgery.

Effects of experimental tail shortening on the phenotypic condition of barn swallowsHirundo rustica: implications for tail‐length evolution

Some studies have suggested that tail streamers in the barn swallowHirundo rusticamay have been elongated 10–12 mm by sexual selection, but according to other studies, the length of these feathers is at the aerodynamic optimum or very close to it. To shed light on this issue, outermost tail feathers were experimentally shortened in male and female barn swallows by 1, 11 or 21 mm. Changes in four physiological parameters commonly used to estimate phenotypic condition in birds (weight, erythrocyte sedimentation rate, blood leukocyte concentration and heterophil/lymphocyte ratio) were checked one month later. Health improved (blood leukocyte concentration decreased) in the group of birds with tails shortened by 11 mm (both males and females), but body condition deteriorated (weight decreased) compared to the other two experimental groups. There was no significant effect of tail‐length manipulation on the other two physiological parameters. These contradictory results suggest trade‐offs between components of phenotypic condition. Possible negative relationships between condition‐related traits imply that using one or very few physiological parameters to estimate phenotypic condition might not be appropriate. The most plausible explanation for the turning point in phenotypic condition when streamers were shortened by 11 mm is that these feathers are 7–15 mm longer than the aerodynamic optimum in both sexes. Therefore, our results are consistent with the hypothesis that tail streamers have been elongated 10–12 mm by sexual selection. This conclusion disagrees with a previous study on the effect of experimental tail shortening on haematocrit, but the complexity of interpreting changes in haematocrit might account for this discrepancy.

Cell-autonomous iodothyronine deiodinase expression mediates seasonal plasticity in immune function

Annual rhythms in morbidity and mortality are well-documented, and host defense mechanisms undergo marked seasonal phenotypic change. Siberian hamsters (Phodopus sungorus) exhibit striking immunological plasticity following adaptation to short winter day lengths (SD), including increases in blood leukocytes and in the magnitude of T cell-mediated immune responses. Thyroid hormone (TH) signaling is rate-limited by tissue-level expression of iodothyronine deiodinase types II and III (dio2, dio3), and dio2/dio3 expression in the central nervous system gate TH-dependent transduction of photoperiod information into the neuroendocrine system. THs are also potent immunomodulators, but their role in seasonal immunobiology remains unexamined. Here we report that photoperiod-driven changes in triiodothyronine (T3) signaling mediate seasonal changes in multiple aspects of immune function. Transfer from long days (LD) to SD inhibited leukocyte dio3 expression, which increased cellular T4→T3 catabolism. T3 was preferentially localized in the lymphocyte cytoplasm, consistent with a non-nuclear role of T3 in lymphoid cell differentiation and maturation. Exposure to SD upregulated leukocyte DNA methyltransferase expression and markedly increased DNA methylation in the dio3 proximal promoter region. Lastly, to bypass low endogenous T3 biosynthesis in LD lymphocytes, LD hamsters were treated with T3, which enhanced T cell-dependent delayed-type hypersensitivity inflammatory responses and blood leukocyte concentrations in a dose-dependent manner, mimicking effects of SD on these immunophenotypes. T3 signaling represents a novel mechanism by which environmental day length cues impact the immune system: changes in day length alter lymphoid cell T3-signaling via epigenetic transcriptional control of dio3 expression.

Thoracic and abdominal lymphatic pump techniques inhibit the growth of S. pneumoniae bacteria in the lungs of rats.

Osteopathic physicians utilize manual medicine techniques called lymphatic pump techniques (LPT) to improve lymphatic flow and enhance immunity. Clinical studies report that LPT enhances antibody responses to bacterial vaccines, shortens duration of cough in patients with respiratory disease, and shortens the duration of intravenous antibiotic therapy and hospital stay in patients with pneumonia. The purpose of this study was to identify if thoracic LPT (Th-LPT) or abdominal LPT (Ab-LPT) would reduce Streptococcus pneumoniae colony-forming units (CFU) in the lungs of rats with acute pneumonia. Rats were nasally infected with S. pneumoniae and received either control, sham, Ab-LPT, or Th-LPT once daily for 3 consecutive days. On day 4 post-infection, lungs were removed and bacteria were enumerated. Three daily applications of either Ab-LPT or Th-LPT were able to significantly (p<0.05) reduce the numbers of pulmonary bacteria compared to control and sham. There were no significant differences in the percentage or concentration of leukocytes in blood between groups, suggesting neither Ab-LPT nor Th-LPT release leukocytes into blood circulation. Our data demonstrate that LPT may protect against pneumonia by inhibiting bacterial growth in the lung; however, the mechanism of protection is unclear. Once these mechanisms are understood, LPT can be optimally applied to patients with pneumonia, which may substantially reduce morbidity, mortality, and frequency of hospitalization.

Administration of anesthetic and analgesic prevent the suppression of many leukocyte responses following surgical castration and physical dehorning

The objectives of the current research were to determine the physiological effects and responses of many leukocytes following surgical castration and/(or) physical dehorning and the influence of anesthetics and analgesics in 3-month-old calves. Eighty 3-month-old Holstein bull calves were completely randomized to treatments in a 2×2×2 factorial arrangement with castration, dehorning, and anesthetic/analgesic as the main effects. Peripheral blood samples were collected just before (0) and 0.5, 1.5, 2.5, 4, 6, 24, and 72h after the respective procedure(s) and analyzed for total leukocyte and differential counts, as well as plasma cortisol and haptoglobin concentrations. Blood from the 0, 0.5 and 24h collections were analyzed for many ex vivo leukocyte responses. Data were analyzed using a repeated measures analysis of variance with the fixed effects of treatment, time, and the interaction of treatment×time. Pre-planned contrasts were performed to determine the effect of (1) management procedure (castration and/(or) dehorning), (2) anesthetic/analgesic, and (3) were the management procedures additive. There were treatment×time interactions (P<0.05) on plasma cortisol and haptoglobin concentrations as well as for total leukocyte and neutrophil concentrations in blood. Castration and dehorning increased cortisol concentrations and the effect of the procedures was additive (P<0.02). Dehorning alone elicited a greater (P<0.05) cortisol response than castration alone. In contrast, the leukocytosis and neutrophilia was greater (P<0.01) among castrated calves. In addition, haptoglobin concentrations at 24h after castration were elevated (P<0.01) in calves that were castrated. Both castration and dehorning suppressed (P=0.04) many leukocyte responses including the secretion of tumor necrosis factor-α when whole blood cultures were stimulated with lipopolysaccharide, surface expression of l-selectin on peripheral blood neutrophils, and the oxidative burst intensity of peripheral blood neutrophils when co-cultured with an Escherichia coli. The effects of castration and dehorning on blood leukocyte counts or any of the leukocyte responses were not additive (P>0.23). Castration and dehorning effects of plasma haptoglobin concentrations tended (P=0.10) to be additive at 72h after the procedure(s). Prior administration of local anesthetic and a systemic analgesic attenuated (P<0.001) the cortisol response and prevented (P=0.03) the observed leukocytosis, neutrophilia, and leukocyte suppression. These data suggest that calves should be castrated and dehorned on the same day rather than spreading them out across two days and calves should be administered pain relief prior to performing either procedure.

Serum myeloperoxidase is independent of the risk factors of atherosclerosis

The main hypothesis of the study was that as serum myeloperoxidase (MPO) concentration is known to indicate the progression of the atherosclerotic process, MPO may be associated with common risk factors of atherosclerosis. Therefore, the presence of these risk factors (especially elevated glucose and lipid concentrations) should predict an increased MPO level during the subsequent months. We also hypothesized an association of MPO with markers of other chronic diseases involving inflammation. Fifty-three patients with ischemic heart disease were followed for 24 weeks by biweekly visits, during which the basic MPO level was measured (500 measurements in total, 2-12 per patient). The association of the patients' typical MPO with the risk factors of atherosclerosis and other personal determinants was examined by trend analysis and analysis of variance. MPO was statistically significantly associated with blood leukocyte, neutrophil, and lymphocyte concentrations of the patients (P=0.001-0.003). MPO was also associated with high-sensitivity C-reactive protein (P=0.02). MPO was not associated with markers of lipid and glucose metabolism, of atherosclerosis, or of other chronic diseases. Contradictory to our hypotheses, the results indicate that the serum MPO level is independent of the commonly measured risk factors of atherosclerosis and markers of other chronic diseases. Consequently, the findings suggest that MPO-related acute pathologic events (such as plaque destabilization) are not associated with the preceding glucose or lipid values. However, the results support the third hypothesis and previously reported view that MPO is a marker of inflammation in patients of ischemic heart disease.

CD40 agonist antibody mediated improvement of chronic Cryptosporidium infection in patients with X-linked hyper IgM syndrome

X-linked hyper-IgM syndrome (XHM) is a combined immune deficiency disorder caused by mutations in CD40 ligand. We tested CP-870,893, a human CD40 agonist monoclonal antibody, in the treatment of two XHM patients with biliary Cryptosporidiosis. CP-870,893 activated B cells and APCs in vitro, restoring class switch recombination in XHM B cells and inducing cytokine secretion by monocytes. CP-870,893 infusions were well tolerated and showed significant activity in vivo, decreasing leukocyte concentration in peripheral blood. Although specific antibody responses were lacking, frequent dosing in one subject primed T cells to secrete IFN-g and suppressed oocyst shedding in the stool. Nevertheless, relapse occurred after discontinuation of therapy. The CD40 receptor was rapidly internalized following binding with CP-870,893, potentially explaining the limited capacity of CP-870,893 to mediate immune reconstitution. This study demonstrates that CP-870,893 suppressed oocysts shedding in XHM patients with biliary cryptosporidiosis. The continued study of CD40 agonists in XHM is warranted.

Effects of bacterial lipopolysaccharide injection on white blood cell counts, hematological variables, and serum glucose, insulin, and cortisol concentrations in ewes fed low- or high-protein diets1

Bacterial lipopolysaccharide endotoxins (LPS) elicit inflammatory responses reflective of acute bacterial infection. We determined if feeding ewes high-CP (15.5%) or low-CP (8.5%) diets for 10 d altered inflammatory responses to an intravenous bolus of 0 (control), 0.75 (L75), or 1.50 (L150) μg of LPS/kg of BW in a 2 × 3 factorial arrangement of treatments (n = 5/treatment). Rectal temperatures, heart and respiratory rates, blood leukocyte concentrations, and serum cortisol, insulin, and glucose concentrations were measured for 24 h after an LPS bolus (bolus = 0 h). In general, rectal temperatures were greater (P ≤ 0.05) in control ewes fed high CP, but LPS increased (P ≤ 0.05) rectal temperatures in a dose-dependent manner at most times between 2 and 24 h after the bolus. Peak rectal temperatures in L75 and L150 occurred 4 h after the bolus. A monophasic, dose-independent increase (P ≤ 0.023) in serum cortisol occurred from 0.5 to 24 h after the bolus, with peak cortisol at 4 h. Serum insulin was increased (P ≤ 0.016) by LPS in a dose-dependent manner from 4 to 24 h after the bolus. Insulin did not differ between control ewes fed high- and low-CP diets but was greater (P < 0.001) in L75 ewes fed low CP compared with high CP and in L150 ewes fed high CP compared with low CP. Increased insulin was not preceded by increased serum glucose. Total white blood cell concentrations were not affected (P ≥ 0.135) by LPS, but the neutrophil and monocyte fractions of white blood cells were increased (P ≤ 0.047) by LPS at 12 and 24 h and at 24 h after the bolus, respectively, and the lymphocyte fraction was increased (P = 0.037) at 2 h and decreased (P ≤ 0.006) at 12 and 24 h after the bolus. Red blood cell and hemoglobin concentrations and hematocrit (%) were increased (P ≤ 0.022) by LPS at 2 and 4 h after the bolus. Rectal temperatures and serum glucose were greater (P ≤ 0.033) in ewes fed a high-CP diet before LPS injection, but these effects were lost at and within 2.5 h of the bolus, respectively. Feeding high-CP diets for 10 d did not reduce inflammation in ewes during the first 24 h after LPS exposure but may benefit livestock by preventing acute insulin resistance when endotoxin exposure is mild.