Search

CD163 is a multi-ligand scavenger receptor exclusively expressed by monocytes and macrophages, which is released after their activation during sepsis (sCD163). The biological relevance of sCD163, however, is not yet clear. We now demonstrate that sCD163 exhibits direct antimicrobial effects by recognizing a specific subfragment ((6) F1(1) F2(2) F2(7) F1) of fibronectin (FN) bound to staphylococcal surface molecules. Moreover, contact with staphylococci promotes sCD163-shedding from monocyte surface via induction of metalloproteinases ADAM10 and ADAM17. sCD163 subsequently binds to Staphylococcus aureus via FN peptides and strongly amplifies phagocytosis as well as killing by monocytes and to a lesser extend by neutrophils. This mechanism exhibits additional paracrine effects because staphylococci additionally opsonized by sCD163 induce higher activation and more efficient killing activity of non-professional phagocytes like endothelial cells. Targeting pathogen-bound FN by sCD163 would be a very sophisticated strategy to attack S. aureus as any attempt of the pathogen to avoid this defence mechanism will automatically bring about loss of adherence to the host protein FN, which is a pivotal patho-mechanism of highly invasive staphylococcal strains. Thus, we report a novel function for sCD163 that is of particular importance for immune defence of the host against S. aureus infections.

Glucocorticoids (GC) are the most widely used immunosuppressive agents in clinical medicine. Recently we showed that GC enhance survival of human monocytes and induce a specific anti-inflammatory monocyte subtype which actively induces resolution of inflammation. We now investigated if cytokines IL-4, IL-6 and IL-10, which, like GC, have mostly anti-inflammatory effects on macrophages, would have GC-like effects also on monocytes. Human monocytes were stimulated with either cytokine, GC or combination thereof, and resulting effects on apoptosis, adherence, migration, phagocytosis, ROS production and cell surface phenotype were determined. We found that IL-4, IL-6, and IL-10 had either less or different effects on various anti-inflammatory functions of monocytes compared to GC. As such, IL-4 and IL-6 alone did not delay apoptosis while IL-10 even enhanced it. However, IL-6 or IL-10 increased GC-mediated protection from apoptosis when applied together with GC. Thus, the potential of GC to induce anti-inflammatory human monocytes is unique and not mimicked by the investigated cytokines. However, IL-6 and IL-10 amplify GC-induced anti-inflammatory and pro-resolution mechanisms by enhancing survival of GC-induced monocytes and thus sustaining their function. This combined effect of GC and cytokines could be important for the physiological switch from amplification towards resolution phase of inflammation.

BACKGROUND Treatment of breast cancer can lead to abnormally shaped anteroposterior spinal curvatures, which consequently leads to functional disturbances of trunk muscles. The aim of the paper was to investigate the correlation between trunk muscle function and type of body posture in women receiving treatment for breast cancer. MATERIAL AND METHODS Fifty women following treatment of breast cancer were included in the study. The average age of the women was 61 years (± 8.67), average body height was 157.5 cm (± 5.49) and average body weight was 74 kg (± 13.10). 84% of the women underwent a modified radical mastectomy, while 16% underwent a breast-sparing procedure. The average time since surgery was 6.4 years (± 5.15). Additionally 90% of women were subjected to adjuvant treatment. The force and velocity parameters of the flexor and extensor muscles of trunk were assessed in all participants at angular velocities of 60 degrees/s and 120 degrees/s in an isokinetic examination unit. A photogrammetric evaluation of the anteroposterior curvatures of the spine was also carried out. On the basis of the value of a compensation index (μ), the participants were divided into three body posture subgroups: kyphotic (K), lordotic (L) and balanced (R. RESULTS 40% of the women (20 patients) demonstrated a balanced body posture with equal thoracic and lumbar curvatures, 30% (15 patients) demonstrated a kyphotic body posture with a dominance of the kyphotic curvature over the lordotic one, and another 30% (15 patients) demonstrated a dominance of the lordotic over kyphotic curvature. Women with a lordotic body posture had the highest correlation between the activity of trunk muscles at loads of 60 degrees/s and 120 degrees/s and body posture in the sagittal plane. Minimal correlation, in turn, was observed in the women with a balanced body posture. CONCLUSIONS 1. The most marked involvement of trunk muscles was observed in women with a lordotic body posture. 2. Women with a balanced posture did not show any correlation between muscle activity and the parameters characterizing this posture.

Objective: Playing an instrument often requires a certain posture and asymmetric position that may affect the anteroposterior spinal curvatures and may lead to postural asymmetry. The aim of the study was to evaluate the spinal curvatures in the sagittal plane and the magnitude of asymmetries in the trunk in the frontal plane in a group of music students in comparison with a control group. Methods: The group of 67 students aged 20 to 26 years was made up of 2 subgroups: the musicians (violin playing students of the Academy of Music in Wroclaw) and the control group (physical therapy students who played no instruments). The examination included an interview, measuring of somatic characteristics, and evaluation of body posture by means of the photogrammetric method. Results: The spinal curvatures of the instrumentalists in the sagittal plane differ from the control group mainly in terms of length and depth parameters. Compared with the control group, the musicians were characterized by statistically more significantly longer and deeper thoracic kyphosis (P b .01) and more shallow lumbar lordosis (P b .05), a greater angle of thoracic kyphosis (P b .005), and a smaller inclination angle of the thoracolumbar and lumbosacral section of the spine (P b .01). Conclusion: In the group of musicians, the asymmetries in the area of shoulders and waist triangles as well as the distance of the spinous processes from the C7 to S1 line were more frequent. (J Manipulative Physiol Ther 2012;35:319-326) Key Indexing Terms: Music; Posture; Moire Topography

Human body posture changes in the course of ontogenesis. The changes are brought about by both internal factors (illness) and external factors (injury). The negative consequences of a medical condition such as breast cancer, together with the treatment process, undoubtedly contribute to disturbance of body posture. The aim of this paper was to evaluate the type of body posture in women after treatment of breast cancer on the basis of anteroposterior spinal curves.The study involved a group of 51 (Group 1) women following treatment of breast cancer and a group of 37 healthy women (Group 2). The average age of the women in Group 1 was 61 years, and the average age of the healthy women was 58 years. All participants underwent a photogrammetric examination of body posture. Postural types were defined on the basis of the value of a compensation index (μ) as kyphotic, balanced, and lordotic. The following subtypes were distinguished within these three categories, depending on the shape of the spinal curves: kyphotic subtype I, II, III; balanced subtype I, II, III; and lordotic subtype I, II, III.The post-mastectomy group and the control group did not differ with regard to age (p=0.09), making it possible to continue the analysis of postural differences, which revealed significant differences at p=0.00008. In the group of women after treatment of breast cancer, 82.3% demonstrated a faulty body posture, compared to only 35.1% of the controls. There was no significant relationship between the quality of body posture and oncological treatment.A significantly higher incidence of faulty body postures was observed among women after treatment of breast cancer.

Active resolution of inflammation is a previously unrecognized process essential for tissue homeostasis. Monocytes play a pivotal role in the generation as well as resolution of inflammation. Glucocorticoids (GCs) are widely used anti-inflammatory agents. We demonstrate that GCs exhibit antiapoptotic effects in monocytes resulting in differentiation to an anti-inflammatory phenotype. The molecular basis of this novel antiapoptotic effect is a prolonged activation of the extracellular signal regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway resulting in inhibition of caspase activities and expression of antiapoptotic genes via activation of c-Myc. We identified up-regulation and activation of A3 adenosine receptor (A3AR) as the initial trigger of this antiapoptotic pathway. In summary, we deciphered a novel molecular pathway promoting survival of anti-inflammatory monocytes. Specific activation of A3AR or its downstream signaling pathways may thus be a novel strategy to modulate inflammation in autoimmune disorders with fewer side effects via induction of inflammatory resolution rather than immunosuppression.

Objective The etiology and pathology of pain in the lumbosacral region and pelvis of pregnant women during and after pregnancy have not been fully determined. This study evaluated if lower back pain during pregnancy and after childbirth is connected with static alterations in the alignment of the pelvis, dysfunction of sacroiliac joints, and irritation of the pelvic ligaments and spine in the lumbosacral region. Methods This study was carried out on a group of 30 women in their eighth month of pregnancy and through 3 months after childbirth. Techniques of manual examination were used to determine the strain. Static alteration of the pelvis was evaluated in both the sitting and standing positions on the basis of alignment of the posterior superior iliac spines. Irritation of the iliolumbar ligaments, sacrotuberous, sacroiliac, and interspinous ligaments was evaluated by means of pressure palpation. Disorders of sacroiliac joint function were evaluated with the Patrick FABERE test, the standing Gillet test, and the standing and sitting flexion tests. Results The most frequently irritated ligaments in the lumbar region are interspinous (60%), iliolumbar (40%), and sacroiliac (36%). Conclusions In women, in their eighth month of pregnancy and after childbirth, disorders of static alterations in pelvis alignment and sacroiliac joint dysfunction may occur. The state of pregnancy may result in strain symptoms in the lumbosacral region and pelvis with variable pain intensifying in various static positions.

There has been a growing interest in studies examining the importance of the aquatic environment in non-invasive treatment of scoliosis. The water environment is an important element in the correction of body posture abnormalities. The aim of this paper is to evaluate the influence of corrective exercises in a water environment on the shape of the antero-posterior curves of the spine and on the functional status of the locomotor system of children with Io scoliosis.The study involved a group of 94 children aged 8-13 years with Cobb Io scoliosis. Computer-aided photogrammetry was used to analyse the shape of the antero-posterior curves of the spine. The functional evaluation of the children's musculoskeletal system involved several functional tests: the Dega wall test, Thomas test, Lasegue test, and Kraus-Weber test. The tests were carried out twice: before a 6-month programme of corrective exercises in a water environment, and immediately on completion of the rehabilitation programme. The programme included swimming and corrective exercises in water.A statistical analysis of the dimensions of the anterio-posterior curves of the spine revealed a significant increase in the total length of the spine and the length of thoracic kyphosis. Besides, the bending angle of the trunk, the angle of thoracic kyphosis and the angle of lumbar lordosis were reduced. The analysis also showed an increase in the bending angle of the upper spine. Shoulder mobility also increased significantly following the programme of corrective aquatic exercises.1. The programme of corrective exercises had an influence on the length and angle of thoracic kyphosis. 2. The muscle strength of the lower part of the erector spinae also increased, as did shoulder mobility measured by the Dega test.

The aim of this study was to assess the changes of the bioelectric activity of the selected muscles and their impact on the functioning of the shoulder joint in people with impingement syndrome. The study covered 58 subjects aged between 24 and 85, who were treated for impingement syndrome in the years 2004-2006. The average duration of the disease was 40 months. The following muscles were tested for bioelectric activity using surface myography: deltoid, supraspinatus, infraspinatus, latissimus dorsi, greater pectoral and biceps brachii on the healthy and the diseased sides. A significant drop in activity of the deltoid and the infraspinatus muscles on the diseased side was observed. The following muscles showed comparable activity on both sides: the supraspinatus, latissimus dorsi and the greater pectoral muscle. The activity of the biceps brachii muscles grew during resisted movements. The drop in the activity of the deltoid and the infraspinatus muscles on the affected side is an important factor responsible for changes of the active mobility of the shoulder and for the development of instability of the shoulder joint. A similar activity of the latissimus dorsi, greater pectoral and biceps brachii muscles on both sides indicates a development of the compensatory mechanisms and the role of those muscles in the dynamic stabilisation of the shoulder joint.

Impingement syndrome (IS) is one of the most common causes of progressive dysfunctions of the shoulder joint. The aim of the study was to evaluate the changes in the joint's mobility in patients with IS and to find correlations between the angular values of particular motion ranges.The study involved 58 people aged 24-85 years, treated for IS from 2004 to 2006. The average duration of the disease was 40 months. A comparative examination of active mobility on the healthy and affected side was carried out according to the SFTR method.A comparison of mean values of the motion ranges indicated significant limitations of active mobility in the affected shoulder. The greatest mobility deficit was found in the external rotation range. Moreover, there was a statistically significant correlation between the angular values of internal rotation and the values of the remaining motion ranges.Impingement syndrome.

The study aimed to assess changes in shoulder joint mobility and symmetry as well as changes in the bioelectrical activity of selected muscles important for the functioning of the shoulder complex in order to determine optimal physiotherapeutic management in patients with shoulder impingement syndrome.The study involved 58 persons aged 24-85 years treated for shoulder impingement syndrome from 2004 to 2006. Symptoms had been present for 40 months on average. The examination of patients included in each case measurement of active mobility ranges using the SFTR method, photogrammetry-based assessment of shoulder symmetry and an examination of the bioelectrical activity of muscles using surface electromyography.The results demonstrated that the degree of limitation of external rotation, which considerably affects the other motion ranges, is the most important index of shoulder joint dysfunction. The decrease in the bioelectrical activity of the deltoid muscle is caused by pain avoidance, with high activity of the pectoralis major and latissimus dorsi muscles indicating the development of compensatory mechanisms. A spatial displacement of the scapula referred to as a wing-like position impairs the scapulobrachial rhythm.Therapeutic management should firstly aim to eliminate pain, then to restore shoulder joint stability by instructing the patient on how to properly centre the humeral head, and, subsequently, to restore normal movement patterns and the scapulobrachial rhythm, and finally to gradually increase the range of active mobility, in particular of external rotation.

Background In a previous study, we demonstrated excessive activation of neutrophils leading to high concentrations of S100A12, a member of the damage-associated molecular pattern molecules (DAMP), in systemic-onset juvenile idiopathic arthritis (SOJIA) which is not found in other inflammatory diseases. Binding to the Receptor for Advanced Glycation Endproducts (RAGE) has been shown but exact molecular mechanisms of S100A12 leading to pro-inflammatory responses in immune cells are still lacking.

Lipopolysaccharide (LPS) as major constituent of Gram-negative bacteria is known to elicit a rapid pro-inflammatory primary response in immune cells. Multiple findings indicate that monocytes next to its crucial function in the early stage of LPS signaling might additionally play an essential role in the resolution of systemic inflammatory responses. We therefore compared gene expression profiles of short-term and long-term LPS-stimulated monocytes. Surprisingly, we revealed an anti-inflammatory gene expression program after long-term LPS treatment. Blocking different signaling pathways revealed p38 MAP kinase to be significantly involved in the regulation of this anti-inflammatory program. Functional clustering analysis showed that chromatin remodeling genes are significantly overrepresented in the group of p38 MAP kinase-dependent genes. To test whether epigenetic modifications cause transcriptional silencing in long-term LPS-treated monocytes and whether this process is p38 MAP kinase-dependent we performed chromatin-immunoprecipitations. We confirmed for pro-inflammatory genes an increased trimethylation of histone 3 (H3K4me3) and increased acetylation of histone 4 (AcH4) after 4 h LPS treatment indicating increased transcription whereas after 16 h of LPS treatment AcH4 decreased. Blocking p38 MAP kinase reversed the long-term LPS effect causing a strong increase of AcH4 and methylation of K79 both indicators of high transcriptional activity. Our results show that p38 MAP kinase contributes significantly to an anti-inflammatory gene expression program in monocytes after long-term LPS treatment. The application of p38 MAP kinase inhibitors currently proposed as novel anti-inflammatory agents might result in deleterious effects on innate immune-regulatory mechanisms.

Glucocorticoids (GCs) represent a mainstay of treatment in most chronic inflammatory diseases. However, many forms of vasculitis relapse after cessation of GC medication or remain primarily unresponsive. Assuming the vasculature and especially endothelial cells (ECs) as origin of the inflammatory process we investigated the effects of GCs on ECs in comparison to monocytes. We treated human micro- and macrovascular ECs as well as human monocytes for 4 h and 16 h with GC or GC + TNF or pretreated ECs with GC followed by TNF stimulation. Testing several genes by quantitative RT-PCR as well as by microarray analysis considering more than 17,000 genes we found no significant gene expression changes in ECs neither with GC alone nor for GC on TNF-induced genes which was in strict contrast to the impressive GC-response of monocytes. We checked that all known GC receptor (GR) subtypes as well as potential cofactors of GC were readily expressed in untreated and inflammatory stimulated ECs. However, in immunofluorescence stainings we surprisingly found that GC treatment failed to induce a nuclear shift of the GR in ECs in contrast to a substantial shift in steroid-sensitive monocytes. Summarizing our results indicate that the molecular prerequisites for the implementation of GC effects on gene regulation are not given in ECs in contrast to myeloid cells. The anti-inflammatory effects of GCs in vasculitis might primarily act on infiltrating leucocytes without targeting ECs which may be the molecular basis for symptomatic effects but lack of long-lasting success of GC-treatment in many forms of vasculitis.

Results We identified GM-CSF-dependent regulation of 402 genes, including important immune-regulatory molecules. Functional clustering of GM-CSF-regulated genes indicated induction of monocytic properties such as lymphocyte activation and tumour necrosis factor (TNF) receptor binding as well as repression of apoptosis, oxidative burst, cell motility and migration. Expression data show an up-regulation of M2-phenotype genes (CCL 13, 23 and 17) and down-regulation of M1-phenotype genes (CXCL10, 11 and 13) indicating a GM-CSF induced alternative macrophage activation. Independent functional analysis revealed suppression of apoptosis by GM-CSF. However GM-CSF induces apoptosis in monocytes activated by TNFα which may contribute to anti-inflammatory actions of GM-CSF treatment in vivo.

The aim of this work is the evaluation of changes at the shoulder complex in (STS) and the selection of physiotherapeutic techniques which allow for the global changes and let streamline results of more effi cient treatment. Fifty eight people at age 24-85 took part in the researches. All of them were cured because of the sydrome of the subacromial tightness between 2004 and 2006. The average time of the incidence was 40 months. Measurements of the range of active mobility were performed at all people. The SFTR method was used. The symmetry of shoulders was evaluated by photogrammetric method and the bioelectric activity of muscles was investigated using superfi cial electromyography. The effi cient treatment should start when sharp aching symptoms have disappeared. It should include the reconstruction of the shoulder joint’s stability through learning how properly the head of the shoulder joint should be focused by using PNF method. The effi cient treatment should also restore the correct movement and the scapulohumeral rhythm thorough scapular bone’s mobility and gradually increase the range of the active mobility. The range of the active mobility should allow for the external rotation which is needed to improve other ranges of movement and fi nally to rebuild the correct power of muscles.

The posture of people with shoulder impingement syndrome (SIS) is a result of adaptive defensive posturing to decrease the intensity of pain in the affected joint. The aim of this work is to characterise trunk and shoulder girdle positioning in patients with SIS.The study involved 58 patients treated for SIS in the years 2004-2006. Symptoms had been present for 40 months on average. A photogrammetric study was performed with the use of a MORA 4G system. It consisted in measuring lordosis and kyphosis, as well as the symmetry of some selected anthropometric points in the frontal plane.Changes in posture presenting as an increased angle of trunk inclination in the sagittal plane and in the frontal plane were observed in all patients. There was asymmetry of bony points as regards the position of the scapula and the waist triangles.The impingement syndrome is associated with displacement of all bony points analysed. Changes in posture are a result of adaptive mechanisms. Trunk asymmetry is secondary to changes in the spatial position of the scapula.

Background. One of the key elements in the examination and evaluation of body posture is defining the shape and size of anterior-posterior curvature of the spine and the location of symmetrical bone points in the trunk area. The aim of our study was to determine which somatic features and parameters of spinal curvature in the sagittal plane show statistically significant differences among children with given types of body posture. Material and methods. We examined 298 children (146 boys and 152 girls) attending four primary schools in Wroclaw, Poland. The examinations were carried out in specially prepared darkened rooms, using computer equipment to evaluate body posture. Results. The most significant similarities regardless of gender were observed in the inclination of the lumbosacral spine (a) and the angle of thoracic kyphosis, as well as in the size and index of thoracic kyphosis. The highest values in these parameters were found in children with lordotic posture, in whom the lowest values were found in the inclination of the upper thoracic segment (g) and the angle and size of lumbar lordosis. However, in children with kyphotic posture the most significant differences were observed in the length of thoracic kyphosis, and the least significant in the actual length of lumbar lordosis. Conclusions. The size parameters and indexes of anterior-posterior spinal curvature appeared to be the least differentiating factors among posture types. The strongest similarity of posture types was found in somatic features and weight-height ratios.

Background. The goal of our study was to assess the outcome of the surgical treatment of scoliosis using the Cotrel- Dubousset method. Material and methods. The study population consisted of 30 girls, aged 13-19 years, with right-sided thoracic scoliosis, grades II, III and IV. All these subjects were operated using the Cotrel-Dubousset method. The research program included measurements of the primary scoliosis angle (Cobb's angle), the vital capacity (VC) of the lungs, chest mobility, and body height. Examinations were performed before surgery, immediately after surgery, and one year after surgery. Results. Comparison of the Cobb's angle before, directly after, and a year after surgery showed that the Cotrel-Dubousset method caused significant correction of scoliosis. The mean VC was below normal before surgery, and despite some improvement remained below physiological values one year later. Chest mobility examinations found that respiratory mobility was adequate before surgery. The mean body height increase one year after surgery was statistically significant. Based on the Risser test performed before surgery we found that ossification was still in progress in the majority of patients one year later. Conclusions. Surgical treatment using the Cotrel-Dubousset method had a significant impact on the correction of scoliosis, as measured by Cobb's angle. The operation did not, however, cause a significant increase of VC in girls with lateral, primarily thoracic scoliosis. A large increase in height was observed in the operated patients. This may have been evoked by significant correction of the scoliosis.

Background. The aim of our study was to assess self-correction of body posture depending on the position assumed. We made observations of the body posture of subjects in the habitual-unrestrained position and the forced erect position, and on that basis we assessed the capacity for self-correction in girls with lateral idiopathic scoliosis. Material and method. The research group consisted of 20 girls, aged 11-18 years (ave. 13.8), divided into two groups depending on the location of the primary scoliosis. The quality of the body posture in habitual and erect position was assessed by computerized photogrammetry, using the moiré technique. Results. In both groups, the mean values in habitual position indicate moderate (indicated by angular indexes) or large asymmetry (indicated by linear indexes). After the subjects had assumed upright position, linear indexes indicated moderate scoliosis and angular indexes indicated slight scoliosis. There were statistically significant differences in asymmetry of the trunk between the habitual and right position in the group of primarily thoracal and primarily thoracolumbar scolioses. A high correlation coefficient may be useful for detecting these types of scolioses. In both groups the asymmetry of the bone points was primarily large or moderate. The differences between the two types of scoliosis were not statistically significant. The only statistically significant correlation was observed between the differences in the distance of the lower scapular angles from the spine (OL).

The goal is to describe occurrences of functional motion system disorders in patients who exhibit the lumbar spinal disc disease. It is assumed that the lower back pain- radicular and pseudoradicular is caused by the lumbar spinal disc disease.Studies were performed in 19 patients who exhibited MRI determined hernia disc in the lumbar spine. Patients were 40-50 years old. Diagnosis of dysfunctions of motion system was determined by utilizing the following manual diagnostic tests: Kibler skin fold test, palpation of spine and pelvis ligaments exams, examination of painfulness processus spinosus, areas of active pain points in muscle tissue; and further examined the state of nerve irritation in accordance to Laseque test and cross Laseque test.The study results indicate symptoms of syndrom pseudoradiatic pain occurred in the majority of examined patients (40-60%). Examination of a localized irritation in the area of dermantom lead to the conclusion that skin pain occurred at L3-S1 levels (60-80% of examined patients) and Th5-Th7 (30-35% of examined patients). The analysis of pain occurrences in the area of pelvis ligaments , lead to the conclusion that all examined ligaments are susceptible for irritations.This particular group of examined patients experienced lower back pain as a result of radicular or/and pseudoradicular pain . Subsequently, any physical therapy treatments in patients who suffer from the lumbars spinal disc disease should be performed individually in each patient because of radicular or pseudoradicular symptoms.

Monocytes and macrophages may either promote or down-regulate inflammatory reactions depending on their state of activation. The effects of glucocorticoids (GCs), the most widely used immunosuppressive drugs, on monocytes are currently not well defined. By analyzing the GC-induced expression pattern in human monocytes by microarray technology, we identified for the first time GC-dependent regulation of 133 genes, including anti-inflammatory molecules such as adenosine A3 receptor, CD1d, and IL-1 receptor II. The results were independently confirmed by real-time polymerase chain reaction (PCR) and flow cytometry. Functional clustering of GC-regulated genes indicated induction of monocytic properties such as phagocytosis and motility as well as repression of adhesion, apoptosis, and oxidative burst. These predictions were confirmed by independent functional assays. GCs up-regulate fMLP receptors and specifically promote chemotaxis to this chemoattractant. Furthermore, GCs promote survival of an anti-inflammatory monocytic phenotype in inflammatory reactions, probably by inhibition of apoptosis because of oxidative stress. GCs limit tissue damage because of induction of antioxidative properties and high capacity for phagocytosis of proinflammatory agents. Thus, GC treatment did not cause a global suppression of monocytic effector functions but results in differentiation of a specific anti-inflammatory phenotype which seems to be actively involved in resolution of inflammatory reactions.

S100A8 and S100A9 are two proinflammatory molecules belonging to the S100 family of calcium-binding proteins. Common to all S100 proteins S100A8 and S100A9 form non-covalently associated complexes which have been shown to exhibit different functional properties. Besides dimerization, recent research is focused on the importance of higher oligomeric structures of S100 proteins induced by bivalent cations. While S100A8/S100A9-heterodimers are formed in the absence of calcium, tetramerization is strictly calcium-dependent. Heterodimer formation is not a simple process and our biophysical analyses (FRET, ESI-MS) demonstrate that simply mixing both subunits is not sufficient to induce complex formation. Steps of denaturation/renaturation are necessary for the recombinant complex to show identical biophysical properties as S100A8/S100A9 obtained from granulocytes. In addition to calcium both proteins are able to bind zinc with high affinity. Here we demonstrate for the first time by different biophysical methods (MALDI-MS, ESI-MS, fluorescence spectroscopy) that zinc-binding, like calcium, induces (S100A8/S100A9) 2 -tetramers. Using mass spectrometric investigations we demonstrate that zinc triggers the formation of (S100A8/S100A9) 2 -tetramers by zinc-specific binding sites rather than by interactions with calcium-specific EF-hands. The zinc-induced tetramer is structurally very similar to the calcium-induced tetramer. Thus, like calcium, zinc acts as a regulatory factor in S100A8/S100A9-dependent signaling pathways.

Cells can die either by apoptosis, necrosis or by a form of cell death that displays morphologic characteristics of both forms (Los et al., 2002; Proskuryakov et al , 2002; Sreedhar & Csermely, 2004). Furthermore, both necrosis (where the cell membrane loses its integrity and the cell contents are released causing an inflammatory response) and apoptosis (where the cell content remains "well-packed" in the apoptotic bodies and inflammation does not occur): (/) can be caused by related (patho)-physiological stimuli; (//) can be partially prevented by anti-apoptotic mechanisms; and (///) can shift from one form to the other (Los et al., 2002). Thus, depending on which form of cell death prevails, an appropriate detection method should be chosen that accurately detects and quantifies the process. Furthermore, some methods will detect only one form of cell death, whereas other methods are not able to distinguish between different forms. Anticancer drugs may not necessarily induce classical apoptosis in cancer cells. In addition, some components of the apoptotic machinery may participate in processes

Despite significant progress in cancer therapy, the outcome of the treatment is often unfavorable. Better treatment monitoring would not only allow an individual more effective, patient-adjusted therapy, but also it would eliminate some of the side effects. Using a cytochrome c ELISA that was modified to increase sensitivity, we demonstrate that serum cytochrome c is a sensitive apoptotic marker in vivo reflecting therapy-induced cell death burden. Furthermore, increased serum cytochrome c level is a negative prognostic marker. Cancer patients whose serum cytochrome c level was normal 3 years ago have a twice as high probability to be still alive, as judged from sera samples collected for years, analyzed recently and matched with survival data. Moreover, we show that serum cytochrome c and serum LDH-activity reflect different stages and different forms of cell death. Cellular cytochrome c release is specific for apoptosis, whereas increased LDH activity is an indicator of (secondary) necrosis. Whereas serum LDH activity reflects the "global" degree of cell death over a period of time, the sensitive cytochrome c-based method allows confirmation of the individual cancer therapy-induced and spontaneous cell death events. The combination of cytochrome c with tissue-specific markers may provide the foundation for precise monitoring of apoptosis in vivo, by "lab-on-the-chip" technology. (c) 2005 Wiley-Liss, Inc.

During acute inflammation, leukocyte infiltration is mostly neutrophilic, but later monocytes prevail. The majority of inflammatory cells, particularly neutrophilic polymorphonuclear leukocytes (PMNs), become apoptotic at later stages of inflammation and are phagocytosed by neighboring cells, mostly by macrophages. Recently, it has been found that human peripheral blood monocytes also recognize apoptotic cells, which primes them to increased production of interleukin (IL)-10--a cytokine known to reduce phagocytes' ability to engulf and kill pathogens. Based on the above, we studied monocytes' ability to phagocytose and kill Staphylococcus aureus while in contact with apoptotic cells.Monocytes isolated by elutriation were co-cultured with apoptotic PMNs or Jurkat cells and exposed to viable, human serum-opsonized S. aureus. To induce apoptosis PMNs were cultured overnight while Jurkat cells were UV-treated. Apoptosis, phagocytosis of bacteria and intracellular superoxide production were measured by flow cytometry. Production of reactive oxygen species was also followed by measurement of chemiluminescence. The bactericidal effect was determined by standard colony forming units method.Data presented show that contact of monocytes with apoptotic neutrophils and Jurkat cells had no influence on monocyte phagocytosis of S. aureus, the generation of reactive oxygen species, or the killing of bacteria.The data obtained suggest that monocytes attracted to the inflammatory site are not deficient in their ability to cope with pathogens after contact with apoptotic cells despite increased production of IL-10.

Monocytes/macrophages undergo apoptosis and are in contact with apoptotic cells both in vitro and in vivo. The data show that monocytes undergoing spontaneous apoptosis in vitro change their cytokine production profile. We demonstrate that the lipopolysaccharide (LPS)-induced production of interleukin-10 (IL-10) is up-regulated, while production of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) is either not affected or reduced. These differences seen both at the protein and mRNA level directly correlate with the appearance of apoptotic cells in the culture. Flow cytometry analysis using double staining, surface with annexin V and intracellular with anti-IL-10, suggested that annexin V-negative monocytes are the predominant source of IL-10. Analysis of sorted populations of monocytes indicated that the increase in IL-10 synthesis appears to result from direct interactions between non-apoptotic and apoptotic cells at the time of stimulation. Also non-apoptotic, freshly isolated monocytes produced more IL-10 upon stimulation with LPS, Staphylococcus aureus or zymosan when apoptotic neutrophils were added to the culture. In contrast, monocyte-derived macrophages did not produce more IL-10 in the presence of apoptotic neutrophils. Finally, we found that the presence of apoptotic monocytes in the culture may influence specific immune responses. The data show that in the presence of annexin V-positive monocytes CD4-positive memory T cells produce less IFN-gamma upon stimulation with purified protein derivative of tuberculin, which could be partially reversed by anti-IL-10 neutralizing antibodies. We conclude that these findings might illustrate the mechanisms operating within an inflammatory site and play an important immunoregulatory role during the resolution of inflammation and specific immune responses.

Introduction Monocytes are circulating cells with high plasticity. They respond to various stimuli with distinct activation and differentiation patterns, are able to secrete several humoral factors and they contribute to inflammation in the immune system, either by governing host defense response to invading pathogens or driving reactions to selfmolecules in conditions of tissue-damage. Control of these mechanisms is necessary to ensure the self-limitation of inflammatory reactions and avoid perpetuated autoinflammation or autoimmunity. This aspect of immunoregulation is crucial and has been mainly associated with adaptive immunity. To date it is unclear how activated monocytes can regulate early cytokine signals promoting their survival or cell death.

Background Active resolution of inflammation is a previously unrecognized process essential for tissue homeostasis 1 . Monocytes and macrophages are the central component of the immune system critical not only for initiation and maintenance of the immune response but also for the resolution of inflammation. Glucocorticoids (GC) are still the most widely used anti-inflammatory agents acting on many targets in a cell-specific manner, including monocytes and macrophages 2 . We previously demonstrated that the treatment of monocytes with GC does not suppress monocytes functions, but rather induces specific differentiation of cells with anti-inflammatory phenotype 3,4 . Nevertheless the mechanism of GC action on monocytes during inflammation is currently not well defined. Objectives The aim of our studies was to investigate the effects of GC on pro-inflammatory monocytes. Methods GC-, LPS- and GC-LPS-induced gene expression patterns in monocytes were analysed by microarray technology. The results were independently confirmed by real-time PCR. Protein expression and activation was analysed by Western Blot and Flow Cytometry. Monocyte functions pivotal for innate immunity – migration, chemotaxis, phagocytosis, killing, oxidative burst – were assessed. Results As expected, treatment of LPS-stimulated monocytes with GC resulted in inhibited expression of many of pro-inflammatory factors. Nevertheless, many of them have not been described to be regulated by GC in monocytes so far. Surprisingly, we have found that GC-LPS treatment of monocytes led also to synergistical up-regulation of many genes, which are not induced by GC or LPS alone. Moreover, we observed additive effects of GC and LPS on expression of many anti-inflammatory genes which was much more pronounced compared to monocytes stimulated with GC alone. Analysis of specific functions of monocytes have shown that GC-LPS treatment inhibited specifically monocyte adherence, but enhanced spontaneous migration, chemotaxis, phagocytosis and killing of pathogens, and engulfment of apoptotic cells as well as the ability to produce anti-inflammatory lipid mediators. Conclusions Our results demonstrate that GC do not simply suppress LPS-mediated activation of monocytes, but rather induce their re-programming toward a specific anti-inflammatory phenotype involved in resolution of inflammation. Interestingly, this anti-inflammatory phenotype is not associated with reduced but rather with enhanced anti-microbial activity. Unravelling specific GC-actions on monocytes may thus define novel pathways allowing anti-inflammatory therapy without enhanced risk for severe infections. References Serhan et al. FASEB J. 2007, 21:325-332. Buttgereit et al. Arthritis Rheum. 2004, 50:3408-3417. Ehrchen et al. Blood 2007, 109:1265-1274. Barczyk et al. Blood 2010, 116: 446-455. Disclosure of Interest None Declared

BACKGROUND: In recent years, we have been investigating the circulating as well as the mucosal expression levels of CD25HighCD4+ regulatory T-cells and the CD4+ T-cells lacking CD28 (CD28-CD4+), which reflect immune dysfunction, with the hope of finding an insight into the abnormalities of immune profile in patients with ulcerative colitis (UC). In the present study, we have looked at the peripheral expression profiles of the CD25HighCD4+ T-cell and the CD28-CD4+ T-cell phenotypes in patients with UC, in active stage, in quiescent and after undergoing colectomy. METHODS: We studied 19 patients with active UC (aUC), 19 with UC in remission (rUC), and 15 healthy volunteers as a control group (HC). The mean Lichtiger's clinical activity index was 13.7±3.0 in the aUC group and 3.4±1.3 in the rUC group. Further, 8 of 19 patients in the aUC group who had undergone colectomy due to insufficient response to medications and persisting severe disease were studied as a colectomy group (cUC). Peripheral blood samples from aUC patients were analyzed in active stage (before starting their treatment), and from rUC and cUC groups at 12 months after achieving remission or colectomy. Lymphocytes were stained with anti CD4, CD25 and CD28 antibodies for analyses by fluorescence-activated cell sorter (FACS) in flow cytometry. The expressions (%) of CD25HighCD4+ and CD28-CD4+ T cells were compared between the three UC and theHC groups. RESULTS: The%of CD25HighCD4+ T cells was significantly higher in the rUC group than in the aUC group (P