Elevated plasma il-8 concentration is related to severity of systemic inflammation in chronic spontaneous urticaria.

Chronic spontaneous urticaria (CSU) is associated with activation of acute phase response. Questions arise regarding its association with other inflammatory mediators. To determine plasma IL-8 concentration in CSU patients and its association with C-reactive protein (CRP) concentration, a nonspecific inflammatory marker of the disease activity, concentrations of plasma IL-8 and serum CRP were measured in CSU patients and compared with healthy controls. IL-8 and CRP concentrations were significantly higher in CSU patients as compared with the healthy subjects. In addition, there were significant differences in IL-8 and CRP concentrations between CSU patients with moderate-severe symptoms and the healthy subjects. Plasma IL-8 and serum CRP concentrations showed a significant correlation with urticaria activity score (UAS). Additionally, a significant positive correlation was observed between IL-8 and CRP concentrations. Up-regulations of IL-8 and its association with the marker of clinical and inflammatory activity suggest a role of this cytokine in the pathogenesis of CSU.

Decreased plasma kallikrein 5 concentrations in patients with chronic spontaneous urticaria.

Little is known about the role of the kallikrein-kinin system in chronic spontaneous urticaria (CSU). Kallikrein 5 (KLK5), a trypsin-like enzyme, is the most abundant in the skin and plays a role in itching and inflammatory reaction. In this study, we determined plasma KLK5 concentration, and its associations with acute phase response in CSU patients. Concentrations of KLK5 in plasma and CRP in serum were measured in patients with CSU of varying severity and in the healthy subjects. Plasma KLK5 concentrations were significantly lower in CSU (all) and moderate-severe CSU patients, as compared with the controls. There were no significant differences in KLK5 concentration in mild CSU patients as compared with the healthy subjects and moderate-severe CSU patients. No correlation was observed between KLK5 and CRP concentrations in the patients. It may be considered that circulating kallikrein 5 is down-regulated in CSU patients, however its potential role and the possible underlying mechanism are unknown.

The relationship between circulating concentrations of interleukin 17 and C reactive protein in chronic spontaneous urticaria.

BACKGROUND: Up-regulation of interleukin 17 (IL-17) family cytokines and acute phase response have been observed in patients with chronic spontaneous urticaria (CSU). It has been demonstrated that IL-17 stimulates C-reactive protein (CRP) expression. AIM: To determine relationship between circulating concentrations of IL-17 and CRP in CSU. METHODS: Concentrations of IL-17 in plasma and CRP in serum were measured in patients with CSU of varying severity and in the healthy subjects. RESULTS: IL-17 and CRP concentrations were significantly higher in CSU patients as compared to the healthy subjects. In addition, there were significant differences in IL-17 and CRP concentrations between CSU patients with mild, moderate-severe symptoms and the healthy subjects. CRP did not correlate significantly with IL-17. CONCLUSIONS: Increased circulating IL-17 concentration may represent an independent index of systemic inflammatory response in CSU, which is not related to increased CRP concentration.

Quantification of image contrast of infarcts on computed tomography scans.

Introduction Accurate identification of infarcts in non-contrast computed tomography (NC-CT) scans of the brain is fundamental in the diagnosis and management of patients with stroke. Quantification of image contrast properties at the boundaries of ischemic infarct regions in NC-CT can contribute to a more precise manual or automatic delineation of these regions. Here we explore these properties quantitatively. Methods We retrospectively investigated 519 NC-CT studies of 425 patients with clinically confirmed ischemic strokes. The average and standard deviation (SD) of patients' age was 67.5 ± 12.4 years and the average(median)±SD time from symptoms onset to NC-CT examination was 27.4(12)±35.7 h. For every scan with an ischemic lesion identified by experts, the image contrast of the lesion vs. normal surrounding parenchyma was calculated as a difference of mean Hounsfield Unit (HU) of 1-5 consecutive voxels (the contrast window width) belonging to the lesion and to the parenchyma. This contrast was calculated at each single voxel of ischemic lesion boundaries (previously delineated by the experts) in horizontal and vertical directions in each image. The distributions of obtained horizontal, vertical and both contrasts combined were calculated among all 519 NC-CTs. Results The highest applicative contrast window width was identified as 5 voxels. The ischemic infarcts were found to be characterized by 6.60 HU, 8.28 HU and 7.55 HU mean values for distributions of horizontal, vertical and combined contrasts. Approximately 40-50% of the infarct boundary voxels were found to refer to the image contrast below 5 HU. Conclusion Low image contrast of ischemic lesions prevents accurate delineation of the infarcts in NC-CT.

Lower serum B12 vitamin concentrations are not accompanied by hyperhomocysteinaemia in chronic spontaneous urticaria.

BACKGROUND: Lower serum vitamin B12 concentrations have been observed in patients with chronic spontaneous urticaria (CSU). It is known that vitamin B12 deficiency is closely related to hyperhomocysteinaemia, which is associated with a proinflammatory state. AIM: To assess the relationship between vitamin B12 status and concentrations of homocysteine (Hcy) with acute phase response in patients with CSU. METHODS: Circulating concentrations of vitamin B12, Hcy and C-reactive protein (CRP) were measured in 42 patients with CSU of varying severity, and compared with 19 healthy controls (HCs). RESULTS: Significantly lower concentrations of vitamin B12 and higher concentrations of CRP were observed in the serum of the patients with CSU compared with HCs (P < 0.01 and P < 0.001, respectively). However, there were no significant differences in plasma Hcy concentrations between the investigated groups. In addition, no correlations were found between the concentrations of vitamin B12, Hcy and CRP. CONCLUSIONS: Lower values of vitamin B12 concentration in patients with CSU were not associated with higher Hcy concentrations, suggesting that such patients do not have functional vitamin B12 deficiency.

Evaluation of circulating vascular endothelial growth factor and its soluble receptors in patients suffering from persistent allergic rhinitis.

BACKGROUND: Overexpression and enhanced release of vascular endothelial growth factor (VEGF) have been detected in various types of allergic inflammation, including asthma. AIM: To further evaluate the pattern of systemic release of VEGF in atopic allergy, free circulating VEGF was measured in patients with persistent allergic rhinitis (PAR). METHODS: The concentrations of VEGF and its soluble receptors (sVEGF-R1 and VEGF-R2) in plasma were measured in patients with PAR sensitized to house dust mites and the healthy subjects. RESULTS: No significant differences were found between PAR patients and healthy subjects with respect to plasma levels of VEGF and its receptors. CONCLUSIONS: It seems that free circulating VEGF may not be elevated in PAR patients. Moreover, on the basis of the present study as well as the earlier ones, it appears likely that systemic release of VEGF varies among patients with distinct clinical manifestation of atopy; may depend on severity/activity and the extent of inflammatory response.

Effective treatment of different phenotypes of chronic urticaria with omalizumab: Case reports and review of literature.

Despite the excellent efficacy and safety profile of omalizumab in chronic spontaneous urticaria (CSU), there are scarce data concerning its role in the treatment of refractory cases with different phenotypes of urticaria. We describe our experience with the therapy of nine patients with CSU co-existing with delayed pressure urticaria (DPU) or angioedema or both and refractory to treatment with high-dose antihistamines. The first patient, with severe CSU and recurrent angioedema, did not respond well to cyclosporine A or corticosteroids and suffered from numerous side effects of long-term corticosteroid therapy. The second patient presented with severe symptoms of DPU, which first of all prevented any daily activities of the professional routines. Both patients showed a complete remission of urticaria after the first injection of omalizumab. The third patient with CSU and severe DPU had been ineffectively treated for more than 20 years with various medications. Following the administration of omalizumab, the symptoms of CSU subsided but those of DPU intensified, and the drug was withdrawn after two cycles. In another four patients with refractory CSU and angioedema, the symptoms subsided after the first administration of omalizumab, and the patients have been in remission for about 5 weeks. In the remaining two patients, the symptoms did not resolve despite four 300 mg doses of omalizumab. It is important to establish a therapeutic regimen with omalizumab (150-300 mg; every 4-8 weeks) tailored to individual patient's needs and dependent on the type of urticaria; this may minimize unnecessary the medication exposure, adverse drug effects, and healthcare costs.

Circulating soluble receptor for advanced glycation end products is decreased and inversely associated with acute phase response in chronic spontaneous urticaria.

BACKGROUND: Activation of receptor for advanced glycation end products (RAGE) leads to the proinflammatory response and the release of its soluble form (sRAGE) which appears to function as an anti-inflammatory feedback mechanism. AIM: To determine serum sRAGE concentration in CSU patients and its association with C-reactive protein (CRP) concentration, a nonspecific inflammatory marker of the disease activity. METHODS: Concentrations of sRAGE and CRP were measured in serum of CSU patients and compared with the healthy controls. RESULTS: Serum sRAGE concentrations were significantly decreased in CSU patients, especially those more severely affected. In addition, significant inverse correlations were observed between sRAGE and CRP concentrations. CONCLUSIONS: Down-regulation of sRAGE and its association with acute phase response suggest a role for RAGE activation in the pathogenesis of CSU. It seems that lower serum sRAGE concentration may enhance the urticarial processes.

Pentraxin-3 as a local inflammatory marker in chronic spontaneous urticaria.

BACKGROUND: The pentraxin family plays an important role in the acute phase response to immune-inflammatory processes. The short pentraxin, C-reactive protein (CRP) is a marker of chronic spontaneous urticaria (CSU) activity, reflecting the systemic effects of inflammatory mediators associated with the disease. It is known, that the long pentraxin, pentraxin 3 (PTX3) is produced at the sites of inflammation, therefore may better reflect activity of the local inflammatory processes. To assess the relevance of PTX3 in CSU patients and its association with CRP. METHODS: Plasma PTX3 and serum CRP concentrations were measured in patients with CSU of varying severity as well as in the healthy subjects. RESULTS: The concentrations of PTX3 and CRP were significantly increased in more severe CSU patients, when compared to mild CSU and the healthy controls. There was a significant correlation between concentrations of PTX3 and CRP. CONCLUSIONS: In contrast to CRP, PTX3 is produced at the sites of inflammation, therefore it seems that elevated PTX3 may result from activation of cells involved in local urticarial processes. Finally, the correlation between these two pentraxins suggests that they may be upregulated by the same mechanisms associated with acute phase response in CSU.

How to isolate urothelial cells? Comparison of four different methods and literature review.

The aim of this study is to present the comparison of four different methods for urothelial cell isolation and culture and compare them to methods cited in the literature. Four different techniques were examined for urothelium isolation from rat bladders. Isolation effectiveness was calculated using trypan blue assay. Confirmation of isolated cell phenotype and comparison with native bladder tissue was confirmed using immunohistochemical (IHC), immunocytochemical (ICC) and immunofluorescence (IF) analysis. The method with bladder inversion and collagenase P digestion resulted in the highest number of isolated cells. These cells showed positive expression of cytokeratin 7, 8, 18, α6-integrin and p63. Our results and the literature review showed that the best method for urothelium bladder isolation is dissection of the epithelium layer from other bladder parts and digestion of mechanically prepared tissue in a collagenase solution.

Is delayed pressure urticaria associated with increased systemic release of sCD40L?

BACKGROUND: Elevated levels of soluble CD40 Ligand (sCD40L) were found in serum but not in plasma of patients with chronic spontaneous urticaria (CU). What is important is that sCD40L has proinflammatory properties, and its elevated plasma level may indicate increased risk of cardiovascular events. These observations should stimulate further evaluation of sCD40L in different forms of urticaria. AIM: In the present study, sCD40L plasma level was investigated in delayed pressure urticaria (DPU). METHODS: As platelets are predominant and variable sources of sCD40L, we investigated sCD40L concentration in platelet-poor plasma (PPP), which seems the best way to minimize the potential contribution of these cells to the ligand level. RESULTS: Plasma sCD40L concentration was significantly increased in the DPU group compared to the healthy controls. CONCLUSIONS: It seems that DPU is associated with increased systemic release of sCD40L, which is believed to derive predominantly from activated platelets. The present study as well as the earlier contributions suggest that distinct cells activity, including platelets, may be identified in different types of urticaria.

Analysis of procalcitonin and CRP concentrations in serum of patients with chronic spontaneous urticaria.

BACKGROUND: Our previous findings showed the importance of analysing the peripheral markers of acute phase response (APR) activation, C-reactive protein (CRP) and IL-6 in the context of urticaria activity and severity. However, these biomarkers do not reliably differentiate between APR to infectious and the disease severity. AIM: In order to investigate a possible association between the immune-inflammatory activation markers CRP and procalcitonin (PCT). METHODS: Serum PCT and CRP concentrations were measured in patients with CU of varying severity as well as in healthy subjects. RESULTS: Serum PCT and CRP concentrations were significantly increased in more severe CU patients when compared to healthy controls and mild CU, and within the CU population there was a significant correlation between concentrations of PCT and CRP. Serum PCT concentrations remained within normal ranges in most CU patients and were only slightly elevated in some severe CU cases. CONCLUSIONS: PCT serum concentration may be only slightly elevated in some cases of severe CU. Upregulation of PCT synthesis accompanied by parallel changes in CRP concentration reflects a low-grade systemic inflammatory response in CU. PCT should be considered as a better marker than CRP to distinguish between APR to infection and an active non-specific urticarial inflammation.

Acute phase inflammatory markers in patients with non-steroidal anti-inflammatory drugs (NSAIDs)-induced acute urticaria/angioedema and after aspirin challenge.

BACKGROUND: Active chronic urticaria, identified as a mast cell- and basophil-dependent inflammatory disorder of the skin is able to elicit acute phase response (APR). However, systemic inflammatory response in different types of urticaria is poorly characterized. AIM: To determine APR pattern in a clearly defined group of patients with acute urticaria and/or angioedema - induced by NSAIDs. METHODS: Plasma IL-6 and serum C-reactive protein (CRP) concentrations were studied in 17 patients with NSAIDs-induced acute urticaria/angioedema (NSAIDsAU) and in 20 healthy controls. Eleven patients who used NSAIDs were presented at the emergency room with acute urticaria/angioedema while the remaining six manifested the symptoms during the aspirin challenge test. Patients were examined in a dynamic manner: during the acute phase, and next, after subsidence of the symptoms. RESULTS: CRP and IL-6 concentrations increased significantly in patients with NSAIDsAU as compared with their asymptomatic period and the healthy subjects. In addition, NSAIDsAU patients showed elevated concentration of the biomarkers following aspirin provocation with the baseline values recovered in the asymptomatic period. CONCLUSION: These results indicate that an acute systemic inflammatory response is activated in patients with NSAIDs-induced urticaria and/or angioedema. The study supports the evidence proving that up-regulation of CRP and IL-6 in urticaria/angioedema does not necessarily reflect any concomitant infection or other inflammatory processes, but may be due to the disease itself.

Vitamin D analogs decrease in vitro secretion of RANTES and enhance the effect of budesonide.

PURPOSE: Eosinophils appear to be central inflammatory cells in the pathogenesis of rhinosinusitis with nasal polyps (NP). One of the most predominantly recognized eosinophil chemoattractants is RANTES. The aim of this study was to assess the influence of vitamin D (VD) derivates on RANTES expression in the culture of nasal polyp fibroblasts. MATERIAL AND METHODS: NP fibroblast cell cultures derived from 16 patients with NP were first stimulated with bacterial LPS and than incubated in increasing concentrations (from 10(-7)M to 10(-4)M) of calcitriol, tacalcitol or budesonide and in combination with one of VD derivate with budesonide in 1:1, 1:3 and 3:1 ratios. Quantitative analysis of RANTES level was conducted in culture supernatants using an ELISA method. RESULTS: The highest calcitriol concentration (10(-4)M) as well as tacalcitol at 10(-5)M and 10(-4)M reduced RANTES production significantly compared to the control (201.1pg/ml, 338.7pg/ml, 211.3pg/ml v 571.78pg/ml; p<0.05). Budesonide and calcitriol administered in 1:3 ratio and budesonide and tacalcitol in 1:1 and 1:3 reduced RANTES concentration significantly better than each of the drug used in monotherapy (p<0.05). Budesonide and tacalcitol in 1:1 and 1:3 ratios suppressed RANTES production to the lowest level (171.8±97.6pg/ml and 178.7±105.22pg/ml, respectively). CONCLUSION: Active VD compounds via downregulation of RANTES production exert a potential role as a complementary element in the therapy of chronic rhinosinusitis with NP. Compounds consisting of budesonide and VD derivate have an advantage over both drugs used in monotherapy.

Late Dysphagia and dyspnea as complications of esophagogastroduodenoscopy in delayed pressure urticaria: case report.

Delayed pressure urticaria (DPU) is characterized by swelling in the area of sustained pressure on the skin. The reported case was a potentially life-threatening complication due to mucosal edema following esophagogastroduodenoscopy (EGD). A 37-year-old man, suffering from severe DPU and chronic spontaneous urticaria, had undergone EGD due to dyspeptic symptoms. A few hours after the EGD procedure, the patient showed both dysphagia and dyspnea. A physical examination indicated massive tongue base and pharynx edema. We suggest that these symptoms were most likely due to the pressure exerted by EGD. No other apparent origins such as angioedema or late-phase allergic reaction to drugs were identified. One should be aware of the increased risk of developing airway and gastrointestinal obstruction during medical procedures associated with compression, such as EGD or endotracheal intubation, in DPU patients.

Doxorubicin-induced F-actin reorganization in cofilin-1 (nonmuscle) down-regulated CHO AA8 cells.

The actin cytoskeleton plays an important role in many cellular processes, including cell mortality, mitosis, cytokinesis, intracellular transport, endocytosis and secretion but also is involved in gene transcription. The dynamics of the actin cytoskeleton is controlled by different classes of actin-binding proteins (ABPs) which regulate the polymerization of actin filaments. In this report we used siRNA against cofilin-1 (nonmuscle) to demonstrate the effect of cofilin on the nuclear and cytoplasmic actin pools in CHO AA8 cells after exposition to various concentrations of doxorubicin. The immunofluorescence studies showed doxorubicin dose dependent tendency to formation the multinucleated giant cells, but also the increase of fluorescence intensity of cofilin in nuclei of untransfected cells. Induction of cell death with doxorubicin treatment in untransfected cells revealed both mitotic catastrophe (in both lower and higher doxorubicin doses) and apoptosis (mostly in higher doxorubicin doses), whereas among cofilin-1 down-regulated cells we observed only mitotic catastrophe. The results suggest that cofilin has apoptosis-inducing ability and that mitotic catastrophe is independent from F-actin content in cell nucleus. In this point of view we conclude that different mechanisms of chromatin reorganization are involved in these two processes. Moreover, we suppose that apoptosis and mitotic catastrophe are independent from each other.

Actin reorganization in CHO AA8 cells undergoing mitotic catastrophe and apoptosis induced by doxorubicin.

Doxorubicin (DOX) is a drug widely used in cancer chemotherapy. Although it has been proven that DOX kills tumor cells, the triggered modes of cell death are not fully understood. There is some evidence that, depending on the dose of DOX, the treated cells undergo senescence, mitotic catastrophe, apoptosis or necrosis. The aim of this study was to assess the type of CHO AA8 cell death induced with different DOX doses. In this context, we also assessed organization and distribution of F-actin, which integrity was suggested to be indispensable for apoptosis. Following treatment with 0.5 and 1 microM DOX, the giant multinucleated cells with extended network of fine microfilaments appeared. Notably, in the nuclei of the enlarged cells microscopy and cytometric analysis showed the presence of F-actin. DOX (2.5 microM) caused the appearance of the giant cells and with apoptotic features and signs of autophagy vacuolization. Flow cytometric studies indicated a dose-dependent increase in the number of TUNEL-positive cells and cells stained with both Annexin V and PI. Cell cycle analysis revealed the increase in the hyperploid DNA content. Our results suggest that treatment of CHO AA8 cells with different DOX doses caused mitotic catastrophe that was followed by apoptosis with signs of autophagy. The increase in F-actin content in the nuclei of the dying cells was evident. We hypothesize that in CHO AA8 cells F-actin may be involved in chromatin reorganization undergoing cell death.

Immunoelectron microscopy in mycosis fungoides and benign dermatoses. Expression of CD3, CD4 and CD7 receptors.

UNLABELLED: Mycosis fungoides is an epidermotropic cutaneous T-cell lymphoma (CTCL).Specimens for presented study were taken from sixteen patients with MF confirmed by immunohistochemical methods and PCR and from nine patients with benign dermatoses. To demonstrate CD3, CD4 and CD7 antigens immunogold method was used. We saw morphological differences between lymphocytes from MF and benign dermatoses. In MF, CD3 and CD4 were present rather in form of clusters placed on the surface of cell. On the contrary -CD3 to CD7 distribution analysis showed that these antigens were present rather individually, however there were seen clusters as well. In MF tumor stage labelling decreased in following order: CD7, CD3 and CD4. We also found internalisation of studied antigens via the coated structures of the cell membrane -especially in tumor stage. In benign dermatoses the majority of all receptors was present on the cell membrane. Our work showed differences in localization of studied antigens, between MF stages, what can suggest their possible translocation in cells. We also found loss of CD3 and CD7 antigens in tumor stage what might be use as adiagnosis tool for this disease. KEYWORDS: mycosis fungoides, benign dermatoses, CD3, CD4 and CD7 receptors, immumogold labelling.

Expression of cyclin A in intestinal biopsies from children with celiac disease.

The aim of this study was to determine the expression of cyclin A and describe its distribution in biopsy samples taken from children with suspected and confirmed celiac disease as well as in control samples. Investigated material consisted of 37 intestinal biopsies: 19 taken from patients with confirmed celiac disease, 9 from patients with its suspicion and 9 from healthy patients, who served as control. Immunohistochemical and immunogold methods were used to estimate cyclin A expression. In celiac disease samples morphological changes in epithelial cells, typical for disease, were shown. We observed weaker cyclin A expression, however there were also some cells with strong labeling in cytoplasm, near the nucleus. In control and suspected celiac disease groups cyclin A was present in the brush border, nucleus and whole cytoplasm, especially in proximity to the nucleus. In conclusion, these studies enabled us visualized pattern of distribution of cyclin A but let us also to presume that observed decrease of expression and its distribution might function as additional factor which could be taken under consideration to establish terminal diagnosis. We are aware of the fact that these are very first observations and that this subject needs to be further investigated with the use of additional methods and samples.

Hyperthermia-induced reorganization of microtubules and microfilaments and cell killing in CHO AA8 cell line.

The aim of this study was to elucidate the effects of hyperthermic treatment on cell morphology and the cytoskeleton in CHO AA8 cell line. The effects of exposure to elevated temperature were analyzed in CHO AA8 cell line by fluorescence microscopy and flow cytometry. The 30 min, at 44.5 degrees C heat shock treatment resulted in the collapse of microtubules (MTs) and microfilaments (MFs) around the nucleus followed by their recovery 24 h after heating. The initial collapse of these cytoskeletal systems, observed 15 min after treatment, was accompanied by the appearance of cells with reduction of volume, shrunken cytoplasm and condensed chromatin. 24 h afterwards, there was the increase in the number of cells with restored and extended MT and MF cytoskeletons. Most of them were larger in size compared to the control cells and had multiple nuclei. 48 h after heat shock the highest number of the giant cells with alternation in nuclear morphology was seen. Flow cytometry analysis revealed the increase in the number of cells with externalized phosphatidylserine 24 h and 48 h after hyperthermic treatment. These results suggest that following heat shock, CHO AA8 cells undergo mitotic catastrophe that presumably represents one of the events resulting in apoptosis.