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Background: Motility disorders of upper gastrointestinal tract are common in critical illness and associated with significant clinical consequences. However, detailed quantitative and qualitative analyses of esophageal motor functions are lacking. Therefore, we aimed to characterize the key features of esophageal motility functions using high-resolution impedance manometry (HRIM) and to evaluate an objective link between esophageal motor patterns, gastric emptying, and gastroesophageal reflux. We also studied the prokinetic effects of metoclopramide., Methods: We prospectively performed HRIM for 16 critically ill hemodynamically stable patients. Patients were included if they had low gastric volume (LGV; < 100 mL/24 h, n = 8) or high gastric volume (HGV; > 500 mL/24 h, n = 8). The HRIM data were collected for 5 h with intravenous metoclopramide administration (10 mg) after the first 2 h., Results: The findings were grossly abnormal for all critically ill patients. The esophageal contraction vigor was markedly increased, indicating prevailing hypercontractile esophagus. Ineffective propulsive force was observed for 73% of esophageal activities. Panesophageal pressurization was the most common pressurization pattern (64%). Gastroesophageal reflux predominantly occurred with transient lower esophageal sphincter relaxation. The common features of the LGV group were a hyperreactive pattern, esophagogastric outflow obstruction, and frequent reflux. Ineffective motility with reduced lower esophageal sphincter tone, and paradoxically fewer reflux episodes, was common in the HGV group. Metoclopramide administration reduced the number of esophageal activities but did not affect the number of reflux episodes in either group., Conclusion: All critically ill patients had major esophageal motility abnormalities, and motility patterns varied according to gastric emptying status. Well-preserved gastric emptying and maintained esophagogastric barrier functions did not eliminate reflux. Metoclopramide failed to reduce the number of reflux episodes regardless of gastric emptying status. Trial registration ISRCTN, ISRCTN14399966. Registered 3.9.2020, retrospectively registered. https://www.isrctn.com/ISRCTN14399966 . more...

Objectives: Colorectal cancer (CRC) remains a major health burden. Although screening is recommended and considered beneficial, further data on its positive effects are needed for worldwide implementation., Study Design: The aim of our national multicentre prospective observational study was to reveal and document clinicopathological differences in CRC diagnosed by screening and presented by disease symptoms as well as assess the efficiency of the screening programme in the Czech Republic., Methods: Between March 2013 and September 2015, a total of 265 patients were enrolled in 12 gastroenterology centres across the Czech Republic. Patients were divided into screening and symptomatic groups and compared for pathology status and clinical characteristics. Screening was defined as a primary screening colonoscopy or a colonoscopy after a positive faecal occult blood test in an average-risk population., Results: The distribution of CRC stages was significantly (statistically and clinically) favourable in the screening group (predominance of stages 0, I and II) compared with the non-screening group (P < 0.001). The presence of distant and local metastases was significantly less frequent in the screening group than in the symptomatic group (P < 0.001). Patients in the screening group had a higher probability of radical surgery (R0) than those diagnosed based on symptoms (P < 0.001). Systemic palliative treatment was indicated in two patients in the screening group compared with 23 patients in the non-screening group (P = 0.018)., Conclusion: CRC diagnosed by screening disclosed less advanced clinicopathological characteristics and results in patients with a higher probability of radical surgery (R0) than diagnoses established based on symptoms, with subsequent management differing accordingly between both groups. These results advocate the implementation of a suitable worldwide screening programme., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.) more...

Chemokines are chemotactic cytokines produced by leukocytes and other types of cells including tumor cells. Their action is determined by the expression of cognate receptors and subsequent signaling in target cells, followed by the modulation of cytoskeletal proteins and the induction of other responses. In tumors, chemokines produced by neoplastic/stroma cells control the leukocyte infiltrate influencing tumor growth and progression. Tumor cells also express functional chemokine receptors responding to chemokine signals, promoting cell survival, proliferation and metastasis formation. Chemokines may be detected in serum of cancer patients, but due to the paracrine nature of these molecules, more significant concentrations are found in the tumor adjacent, non-vascular fluids, collectively called tumor proximal fluids. This review summarizes the expression of CC and CXC chemokines in these fluids, namely in interstitial fluid, pleural, ascitic, and cyst fluids, but also in urine, saliva, cerebrospinal fluid, cervical secretions and bronchoalveolar lavage fluid. Most comparative clinical studies reveal increased chemokine levels in high-grade tumor proximal fluids rather than in low-grade tumors and benign conditions, indicating shorter survival periods. The data confirm peritumoral fluid chemokines as sensitive diagnostic and prognostic markers, as well as offer support for chemokines and their receptors as potential targets for antitumor therapy. more...

Aim: The aim of our prospective study was to evaluate the development of postpartum anal incontinence in patients with inflammatory bowel disease (IBD) compared to healthy women., Material and Methods: Patients with IBD and healthy controls enrolled in the study from January 1st 2013 to November 30th 2016 and filled in the anal incontinence questionnaire in the beginning of pregnancy and after vaginal delivery. The results were statistically processed using suitable tests., Results: A total of 57 women were enrolled, 17 (29.8 %) with ulcerative colitis, 23 (40.4 %) with Crohn's disease, and 17 (29.8 %) healthy controls. Incidence of postpartum anal incontinence is comparable across all groups; there was no statistically significant difference between the IBD and control groups (Kruskal-Wallis test by ranks with Dunn correction, non-significant). Postpartum anal incontinence was strongly correlated with the extent of perineal injury (r = 0.80; p < 0.0001; Pearson's linear correlation)., Conclusions: Women with inflammatory bowel disease in remission do not exhibit higher incidence of postpartum anal incontinence (PPAI) compared to healthy controls; the key correlate of PPAI appears to be the extent of obstetric injury, consistently across all study groups. These results suggest that concerns about postpartum anal incontinence development should not be an indication for Caesarean section in IBD patients (Tab. 6, Fig. 1, Ref. 34). more...

Introduction: Percutaneous endoscopic gastrostomy (PEG) is a minimally invasive routine procedure used to provide long-term enteral nutrition in selected patients with impaired oral intake. The knowledge of clinical, technical and safety features of PEG is an important issue in clinical practice.The aim of this study was to evaluate the popu-lation of patients selected for PEG insertion, describe certain aspects of the insertion procedure, assess the service life of PEG, as well as account for the number of complications and patient mortality in the six-month period following the procedure., Methods: We used a retrospective analysis of data from medical database. The data were collected in a single endoscopic tertiary-referral center for a period of seven and a half years., Results: We evaluated 326 PEG insertions performed on 292 patients with a median age of 63 years (IQR 55-70). Mortality in the six-month period following PEG insertion was 26 %. Prevailing indications for PEG insertion were oncological (53 %) and neurological (40 %) diseases, with certain fluctuation of these numbers during the observed period according to changing demands of the two medical specialties. Local anesthesia alone was applied in 56 % of patients, 38 % underwent analgosedation and 6 % required general anesthesia. Median duration of the procedure (from insertion of endoscope to its final extraction) in 68 consecutive procedures was 6 minutes (IQR 5-8). Median interval between PEG introduction and its first replacement in 21 patients was 22 months (IQR 14-31, range 4-76). 61 patients underwent PEG extraction during the observed period, 66 % of whom had oncological disease. Periprocedural complications were seen in 5.8 % of patients, of these one patient (0.3 %) suffered a serious complication. The buried bumper syndrome was observed in four patients (1.2 %), all of whom had neurological disease., Conclusions: PEG is a relatively safe procedure and can be performed in a short time using local anesthesia or analgosedation in a majority of patients. The population of patients indicated for PEG insertion reflects primarily the current needs of neurological and oncological departments. Most patients within the observed group benefit from PEG insertion for more than six months., Key Words: analgosedation - complications - mortality - percutaneous endoscopic gastrostomy. more...

Interleukin-8 (IL-8, CXCL8) was originally discovered as a powerful attractor and activator of polymorphonuclear leukocytes. It was soon recognized that IL-8 also affects proliferation and migration of cancer cells, tumor angiogenesis and metastasis, and that it is expressed in many cancerous cell types. IL-8 protein expression is usually increased in serum of cancer patients, but markedly higher concentrations are found in cancer-associated non-vascular extracellular fluids, such as pleural effusion, ascites and cyst fluid. Elevated concentrations of IL-8 are indicative of malignant processes also in cerebrospinal fluid, urine, saliva, interstitial fluid and cervicovaginal secretions. Higher IL-8 levels are typically found in high-grade peritumoral fluids rather than low-grade tumors and benign conditions, with the exception of inflammatory processes. In line with recent molecular biology investigations, it appears that the local IL-8 production is related to its malignant origin and to tumor progression. Hence, IL-8 in peritumoral fluid is to be taken into consideration while assessing tumor character and monitoring the tumor progression/remission status. Besides, the data here collected justify the attempts to find an IL-8-targeted inhibitory therapy more...

A soil-plant transfer study was performed using soil from a former uranium ore processing factory in South Bohemia. We present the results from greenhouse experiments which include estimates of the time required for phytoremediation. The accumulation of (226)Ra by different plant species from a mixture of garden soil and contaminated substrate was extremely variable, ranging from 0.03 to 2.20 Bq (226)Ra/g DW. We found differences in accumulation of (226)Ra between plants from the same genus and between cultivars of the same plant species. The results of (226)Ra accumulation showed a linear relation between concentration of (226)Ra in plants and concentration of (226)Ra in soil mixtures. On the basis of these results we estimated the time required for phytoremediation, but this appears to be too long for practical purposes., (Copyright (c) 2008 Elsevier Ltd. All rights reserved.) more...

Phytoremediation of selected pharmaceuticals (diclofenac, ibuprofen, and acetaminophen) using Armoracia rusticana and Linum usitatissimum cell cultures and by hydroponically cultivated Lupinus albus, Hordeum vulgaris, and Phragmites australis plants in laboratory conditions is described. During in vitro experiments, the best results for acetaminophen were achieved using Armoracia rusticana hairy root cultures, where 100% of the starting amount was removed from the media during eight days. Total removal of ibuprofen and diclofenac was achieved using a Linum usitatissimum suspension culture after one and six days, respectively. In the hydroponic arrangement, the best results were achieved for Lupinus, where acetaminophen was totally removed from media during two or four days in concentrations of 0.1 or 0.2 mM, respectively. The best effectiveness of ibuprofen removal (50% of starting amount) was found in case of Phragmites. Effectiveness of all tested plants for diclofenac removal was low. The best removal was achieved using Phragmites in the case of 0.2 mM concentration-67% of the starting amount and Hordeum for 0.1 mM starting concentration, 56%. more...

Besides massive expression in inflammatory pleural effusions, inflammatory markers are also present in cancerinduced pleural effusions. Recent advances in cancer biology point to a role of inflammatory signaling in cancer and encourage reconsidering the diagnostic and prognostic value of inflammatory markers. Here an attempt was made to relate protein levels of inflammatory markers to underlying malignant processes in the pleural space. Pleural effusions from lung cancer patients (n=116) were subjected to a multifactorial analysis covering 13 inflammatory markers. The composition of tumor-associated effusions was compared with that of parainflammatory pleural effusions (n=30), transudates (n=18), and serum values, and evaluated in relation to cancer origin, histology, cytology, pleural involvement, treatment history, and survival time. Inflammatory markers were significantly expressed in pleural effusions of paraneoplastic origin when compared to transudates and most serum levels. Values in pleura-invading and metastatic tumor-associated effusions were typically higher than those of other tumors. Many markers correlated negatively with survival, most prominently IL-8 (r=-0.36, p=0.001) and VEGF (r=-0.35, p=0.001). It appears that most inflammatory markers are highly expressed in tumor-associated pleural effusions, reflecting to some extent tumor origin and localization. Despite the lower efficacy of inflammatory markers in the differentiation between exudative pleural effusions, some inflammatory markers may represent potential prognostic markers of malignant processes in the pleural space. more...

Objective: An anaerobic type of glycolysis exemplified by hyperproduction of the lactate dehydrogenase (LDH) subunit M has been detected in lung tumours, while a similar pattern has been found in concomitant pleural effusions (PE). The aim of this study was to verify the presence of the LDH subunit M in PEs of different aetiology and to compare its expression with markers of inflammation., Material and Methods: LDH isoenzymes were estimated and the LDH5/LDH1 coefficient was calculated in paraneoplastic PEs (n = 99), including subgroups with a different tumour ultrastructure, origin and pleural involvement. The expression pattern was compared with parainflammatory PEs (n = 21), transudates (n = 16) and with the expression of 13 inflammatory markers in PEs., Results: The LDH5/LDH1 coefficient was higher in PEs associated with non-small-cell lung cancer (NSCLC) and with pleura-invading tumours, and lower in PEs of small-cell lung cancer and tumours without a confirmed pleural involvement. The LDH5/LDH1 coefficient positively correlated with uPA, IL-8, IL-10, sICAM, sVCAM, MPO and MMP-9., Conclusions: In accordance with inflammatory markers, it appears that the expression of LDH and its isoenzymes in PEs reflects the host reaction in pleural space and, in NSCLC, may also feature the anaerobic phenotype of cancer cells. more...

Cathepsins S and H are present in immune cells and tissues and may play a role in the activation of an adoptive immune response. Our goal was to assess their protein levels in pleural fluids from 82 patients who underwent thoracentesis or thoracoscopy for therapeutic or diagnostic reasons and to relate them to an inflammatory, neoplastic or hemodynamic origin. Pleural effusions were also analyzed for a panel of 13 inflammatory or proliferative markers to test possible links to a nonspecific host reaction. Increased levels of cathepsin S were found in parainflammatory and cancer-related effusions compared to transudates. Cathepsin H levels were elevated only in parainflammatory effusions, whereas the levels in cancer-related effusions were comparable to transudates. Cathepsin S values significantly correlated with LDH, alpha-1-AT, VEGF, sICAM, sVCAM, MPO, uPA, MMP-9/TIMP-1, IL-8 and MCP-1, but not with CRP, IL-10 or cathepsin H. In contrast to cathepsin S, cathepsin H values did not correlate with markers of inflammation, indicating a specific role for cathepsin H in the pleural host response. In conclusion, the estimation of cathepsin S and cathepsin H may help to distinguish between effusions of different etiology. more...

Objective: Progelatinase B/proMMP-9 has recently been identified as an indicator of pleural inflammation, presumably originating from granulocytes. The aim of this study was to verify the origin of progelatinase B by simultaneous estimation of specific markers of neutrophil recruitment and activation in pleural effusions following induced pleurisy and pleural injury., Material and Methods: Sixty-three samples of pleural fluid from patients undergoing therapeutic talc pleurodesis (n = 8) and explorative thoracoscopy (n = 3) collected before and at different time intervals after the intervention were analyzed for progelatinase B and neutrophil gelatinase-associated lipocalin (NGAL)-gelatinase complex by substrate electrophoresis, for myeloperoxidase (MPO) and interleukin-8 (IL-8) by immunoadsorbent sandwich assay, as well as for leukocyte count, C-reactive protein (CRP) and total protein (TP)., Results: A significant increase in free and NGAL-complexed progelatinase B, MPO and IL-8 was recorded within 48 h following treatment in all subjects. Progelatinase B was strongly correlated with NGAL-gelatinase complex (r = 0.88, p = 0.001), MPO (r = 0.81, p = 0.001), neutrophil count (r = 0.75, p = 0.01) and IL-8 (r = 0.71, p = 0.001), but not with CRP and TP., Conclusions: The results support the neutrophil origin of the proenzyme, which confirms progelatinase B as an indicator of a local inflammatory reaction. Quantifying the inflammatory reaction may be helpful in the evaluation of both the technical variants of therapeutic pleurodesis and finer discrimination of paraneoplastic effusions. more...

Gelatinase B/MMP-9 is a member of matrix metalloproteinases with a major role in extracellular matrix degradation, cell proliferation and migration. Its proenzyme form has also been reported in pleural fluids as an inducible species, but its relation to pleural pathology has not yet been fully clarified. The primary goal of this study was to evaluate proMMP-9 as a potential marker for differentiating pleural effusions of both malignant and non-malignant origin. Pleural fluid samples were studied from 194 patients, including tumor etiology in 133 cases, inflammatory disorders in 33, transudates in 12, and unspecified disorders in 16 patients. The concentrations of proMMP-9 were estimated by means of immunoassays and/or by scanning zymography. Samples were also examined for C-reactive protein (CRP). The analysis of proMMP-9 showed significant differences among the etiological groups with the highest concentrations in para-inflammatory exudates, intermediate in para-neoplastic exudates, and the lowest in transudates. However, the analysis of the para-neoplastic group revealed a distinct heterogeneity with a minor portion of fluids reaching values typical for para-inflammatory effusions. A subsequent sorting based on tumor histology showed increased levels particularly in exudates associated with metastatic tumors. Interestingly, proMMP-9 values in general correlated with CRP, a systemic marker of inflammation. Thus, MMP-9 proenzyme appears to complement traditional markers distinguishing pleural fluids of different origin. Yet, the differentiation between paraneoplastic and para-inflammatory exudates must be regarded with caution due to the presence of a high-expressive paraneoplastic sub-population, including effusions associated with metastatic tumors. more...

Previous analyses of pleural effusions of different origin revealed high levels of matrix metalloproteinase-9 proenzyme (proMMP-9), predominantly in parainfectious exudates. To confirm the etiological association with inflammation and to explore time course of release of proMMP-9 under conditions of a local inflammatory reaction, the authors estimated proMMP-9 in pleural fluids of patients undergoing therapeutic talc pleurodesis (n = 20) and explorative thoracoscopy (n = 4), before, and at different time intervals after the intervention. The authors also compared proMMP-9 values with proinflammatory markers, such as interleukin-6 (IL-6), C-reactive protein (CRP), and leukocyte count. As revealed through scanning substrate electrophoresis, proMMP-9 levels increased within the first hours a!nd culminated 16 to 48 hours after intervention, amounting to 1.5- to 102 -fold of initial concentration (mean = 14.8 -fold). MMP-9 proenzyme values significantly correlated with the levels of proinflammatory cytokine IL-6 (r =.79, n = 47, p <.01) and with neutrophil counts (r =.75, n = 45, p <.01). In conclusion, the results present proMMP-9 as a novel early indicator of acute host-response in pleural space that complements traditional proinflammatory markers and might be useful in monitoring pleural inflammatory processes. more...

Both degradative and limited proteolysis are involved in physiological processes. Once activated, proteinases are controlled by endogenous inhibitors (antiproteinases). A variety of genetic disorders and exogenous microbial proteinases disturb the balance between proteinases and cognate inhibitors. alpha 1-Antitrypsin deficiency is a model disorder resulting in an insufficient control of leukocyte elastase with a subsequent alveolar tissue damage. It appears that the manifold molar excess of inhibitor required to compensate the deficiency, is the consequence of both the local relative inaccessibility of the serpin, and the mode of proteinase-serpin interaction, in accord with the "branched pathway" mechanism. Slow-binding kinetics and the leak of native proteinase from the complex is illustrated by the peptidolytic action of porcine elastase in the presence of human alpha 1-antitrypsin and by gelatin zymography, respectively. (Tab. 2, Fig. 5, Ref. 44.) more...

In a previous study, kinetic assays showed that pNiXa, an Ni(II)-binding serpin of Xenopus oocytes and embryos, strongly inhibits bovine chymotrypsin, weakly inhibits porcine elastase, and does not inhibit bovine trypsin. In this study, analyses by SDS-PAGE and gelatin zymography showed that an SDS-resistant complex is formed upon the interaction of pNiXa with bovine chymotrypsin. No such pNiXa-enzyme complex was detected after pNiXa interactions with porcine elastase, bovine trypsin, or human cathepsin G. The major products of pNiXa cleavage by the four proteinases were partially sequenced by Edman degradation. The cleavage products were also tested by immunoblotting with an antibody to the His-cluster of pNiXa, and by radio-blotting with 63Ni(II). These assays showed that chymotrypsin and elastase cleave pNiXa at the P1-P1 (Thr-Lys) peptide bond near the C-terminus, while trypsin and cathepsin G cleave pNiXa at specific peptide bonds near the N-terminus, within an interesting 26-residue segment, rich in Lys and Gln, that separates the His-cluster of pNiXa from the rest of the molecule. The segment lacks homology to other serpins, but resembles a domain of Xenopus POU3 transcription factor. This study identifies the specific sites for interactions of four serine proteinases with pNiXa, indicates that pNiXa inhibition of chymotrypsin involves a serpin-like mechanism, and shows that 63Ni(II)-binds to the His-cluster of pNiXa. more...

A Ni(II)-binding serpin, pNiXa, is abundant in Xenopus oocytes and embryos. Kinetic assays show that purified pNiXa strongly inhibits bovine alpha-chymotrypsin (Ki = 3 mM), weakly inhibits porcine elastase (K1 = 0.5 microM), and does not inhibit bovine trypsin. The reversible, slow-binding inhibition of alpha-chymotrypsin by pNiXa is unaffected by Ni(II). Ovochymase in egg exudates is inhibited by pNiXa, but to a limited extent, even at high pNiXa concentrations. An octadecapeptide that models the His-rich domain (-HRHRHEQQGHHDSAKHGH-) of pNiXa forms six-coordinate, octahedral Ni(II)-complexes when the N-terminus is acetylated, and a square-planar Ni(II)-complex when the N-terminus is unblocked. Spectroscopy reveals two distinct types of octahedral Ni(II)-coordination to the N-acetylated octadecapeptide, involving, respectively, 3-4 and 5-6 imidazole nitrogens; the octadecapeptide undergoes partial, reversible precipitation in pH- and Ni(II)-dependent fashion, suggesting an insoluble, Ni(II)-coupled (Hx)n-dimer. Such (Hx)n-peptide interaction is confirmed by an enzyme-linked biotinavidin assay with N-biotin-KHRHRHE-amide and N-acetyl-KHRHRHE-resin beads, which become coupled after adding Ni(II) or Zn(II). H2O2 oxidation of 2'-deoxyguanosine to mutagenic 8-hydroxy-2'-deoxyguanosine is enhanced by the octahedral Ni(II)-octadecapeptide complex, although the effect is more intense with the square-planar Ni(II)-octadecapeptide complex. Immunoperoxidase staining of whole mounts with pNiXa antibody shows that pNiXa is distributed throughout gastrula-stage embryos and is localized during organogenesis in the brain, eye, spinal cord, myotomes, craniofacial tissues, and other sites of Ni(II)-induced anomalies. Patterns of pNiXa staining are similar in controls and Ni(II)-exposed embryos. Binding of Ni(II) to pNiXa may cause embryotoxicity by enhancing oxidative reactions that produce tissue injury and genotoxicity. Although the natural target proteinases for pNiXa inhibition have not been established, pNiXa may be an important regulator of proteolysis during embryonic development. more...

The enzymic activity of secreted aspartic proteinase (SAP) in 117 clinical isolates of Candida albicans strains ranges from 0.0 do 0.12 mumol Tyr/min/ml of culture supernatants following 24 h incubation in a YCB-BSA-glucose medium. SAP was purified by the modification of a combined chromatographic method (Banerjee et al., 1991) and a final gel filtration. The diagnostic value of the SAP activity estimation and immunochemical use of purified enzyme are discussed. more...