MMP-19 deficiency causes aggravation of colitis due to defects in innate immune cell function.

Matrix metalloproteinases (MMPs) are potential biomarkers for disease activity in inflammatory bowel disease (IBD). However, clinical trials targeting MMPs have not succeeded, likely due to poor understanding of the biological functions of individual MMPs. Here, we explore the role of MMP-19 in IBD pathology. Using a DSS-induced model of colitis, we show evidence for increased susceptibility of Mmp-19-deficient (Mmp-19(-/-)) mice to colitis. Absence of MMP-19 leads to significant disease progression, with reduced survival rates, severe tissue destruction, and elevated levels of pro-inflammatory modulators in the colon and plasma, and failure to resolve inflammation. There was a striking delay in neutrophil infiltration into the colon of Mmp-19(-/-) mice during the acute colitis, leading to persistent inflammation and poor recovery; this was rescued by reconstitution of irradiated Mmp-19(-/-) mice with wild-type bone marrow. Additionally, Mmp-19-deficient macrophages exhibited decreased migration in vivo and in vitro and the mucosal barrier appeared compromised. Finally, chemokine fractalkine (CX3CL1) was identified as a novel substrate of MMP-19, suggesting a link between insufficient processing of CX3CL1 and cell recruitment in the Mmp-19(-/-) mice. MMP-19 proves to be a critical factor in balanced host response to colonic pathogens, and for orchestrating appropriate innate immune response in colitis.

Elevated intracranial pressure, low cerebral perfusion pressure, and impaired brain metabolism correlate with fatal outcome after severe brain injury.

BACKGROUND: New brain tissue monitoring techniques (tissue oxymetry, microdialysis) provide direct information about the state of brain oxygenation and brain metabolism in patients with severe traumatic brain injury (TBI). Despite this information being limited to a small region of the brain surrounding the probes, it could be associated with such global parameters as the clinical outcome. OBJECTIVE: To study the predictive value of monitoring brain oxygenation and metabolism on clinical outcome in patients in the acute phase of severe TBI. METHODS: An observational study of 20 patients with a severe TBI was undertaken, utilizing intracranial pressure (ICP), cerebral perfusion pressure (CPP), brain tissue oxygenation, and brain metabolism monitoring. We correlated the clinical outcome of the patients with the following parameters: ICP, CPP, brain tissue oxymetry (PbtO2), glucose and glycerol levels, and the lactate/pyruvate (LP) ratio. Further, we analyzed the relationship between ICP, CPP, PbtO2, and the metabolism parameters. RESULTS: We found a correlation of the mean ICP values (8.73 ± 1.18 in group A vs. 26.32 ± 5.01 mmHg in group B, p < 0.005), the mean CPP values (84.82 ± 2.02 in group A vs. 66.62 ± 4.64 mmHg, p < 0.005), the LP ratio (37.36 ± 3.44 vs. 199 ± 87.97, p < 0.05), and glycerol levels (62.07 ± 12.14 vs. 215 ± 46.52 µmol/l, p < 0.05) with the clinical outcome. High ICP correlated with both a high LP ratio (Spearman R = 0.61, p < 0.05), and elevated glycerol concentrations (Spearman R = 0.48, p < 0.05). A low CPP correlated with a high LP ratio (Spearman R = -0.57, p < 0.05), while a low PbtO2 correlated with a high LP ratio (Spearman R = -0.49, p < 0.05). CONCLUSIONS: High ICP, low CPP, an elevated mean LP ratio, and high glycerol concentrations in the acute phase predict fatal outcome 6 months after TBI. Further, high ICP, low CPP, and low PbtO2 correlate with impaired brain metabolism.

Xylanases of anaerobic fungus Anaeromyces mucronatus.

The anaerobic fungus Anaeromyces mucronatus KF8 grown in batch culture on M10 medium with rumen fluid and microcrystalline cellulose as carbon source produced a broad range of enzymes requisite for degradation of plant structural and storage saccharides including cellulase, endoglucanase, xylanase, alpha-xylosidase, beta-xylosidase, alpha-glucosidase, beta-glucosidase, beta-galactosidase, mannosidase, cellobiohydrolase, amylase, laminarinase, pectinase and pectate lyase. These enzymes were detected in both the intra- and extracellular fractions, but production into the medium was prevalent with the exception of intracellular beta-xylosidase, chitinases, N-acetylglucosaminidase, and lipase. Xylanase activity was predominant among the polysaccharide hydrolases. Extracellular production of xylanase was stimulated by the presence of cellobiose and oat spelt xylan. Zymogram of xylanases of strain KF8 grown on different carbon sources revealed several isoforms of xylanases with approximate molar masses ranging from 26 to 130 kDa.

[Thrombolysis in central retinal artery occlusion using alteplase]. / Trombolýza okluze centrální retinální arterie alteplázou.

AIM: The systemic acute thrombolysis (fibrinolysis) is one of methods how to treat occlusion of the central retinal artery. We present our first results of this rarely used treatment. MATERIAL AND METHODS: The patients were treated by means of i.v. infusion of the fibrinolytic alteplasis (plasminogen tissue activator, Actilyse) with the dose 0.9 mg/kg of body weight, first as a bolus of 10% of the dose, the rest was slowly applied while monitoring vital functions and coagulations parameters. The contraindications of the thrombolysis are especially tumors, bleeding, any surgery in last three months, and the brain stroke in the medical history, and diseases of the liver and kidneys. From November 2006 to April 2007, we treated 5 patients by means of thrombolysis; two patients with the occlusion of the temporal branch retinal artery (BRAO) with characteristic quadrant scotoma of the visual field and decrease of the visual acuity, and three patients with central retinal artery occlusion or hemi-occlusion (CRAO). RESULTS: The central visual acuity improved in all patients the next day after the thrombolysis. Both patients with the branch retinal artery occlusion had visual acuity 1.0, the delay between the thrombosis and the treatment was more than 12 hours. In two out of three patients with the central occlusion the visual acuity improved from practical blindness to 0.66. The third patient came with the latency of more than 30 hours; he registered partial improvement shortly after the thrombolysis, but later, the visual acuity decreased again. CONCLUSION: The thrombolytic treatment is definitely an advantage comparing to the conservative treatment. According to many contraindications and possible complications, it is necessary to select the eligible patient carefully. The treatment can be done on specialized in-patient department with the possibility to monitor vital functions and coagulations factors during the treatment as well as after it. In four out of five of our patients, the treatment finished with fast recovery of visual acuity almost to the normal.The effect of the treatment was good up to 18 hours after the occlusion; after 30 hours it was bad.

[Decompressive craniectomy in the treatment of posttraumatic edema and the contribution of new diagnostic methods]. / Dekompresivní kraniektomie v lécbe posttraumatického edému mozku a prínos nových monitorovacích metod.

The authors present a case report of a young male who suffered a brain injury complicated with malignant posttraumatic edema managed with bilateral decompressive craniectomy after conservative treatment failure. They further discuss current surgical approach to posttraumatic brain edema and contribution of new diagnostic methods in secondary brain damage management.

Arteriovenous fistula as a complication of renal biopsy.

BACKGROUND: We evaluated the incidence and history of arteriovenous fistula (AVF) after kidney biopsy and assessed the use of superselective embolisation for treatment. METHODS: Case report of a 10-year-old boy with nephrotic syndrome. Renal biopsy (RB) in this patient was complicated with AVF. Immediately after RB was undertaken, microscopic haematuria was observed, within 48 hours after the biopsy life-threatening haematuria due to pseudoaneurysm started. Renal angiography was carried out, which demonstrated a hyperthrophic aberrant artery in the region of the bottom pole of the left kidney, from which blood was instantaneously flowing through a high-flow arteriovenous fistula (AVF). RESULTS: Embolization was carried out using small platinum coils (MWCE-18S-3/2, -18S-4/2, -18S-5/2TORNADO Embolization Microcoil) and the tissue adhesive Histoacryl. CONCLUSIONS: The technique of superselective embolisation using coaxial catheter is a safe method in the treatment of post biopsy AVFs and pseudoaneurysm (Fig. 3, Ref. 6).

[Diagnostics and treatment of hepatocellular carcinoma. Recommendations of the Portal Hypertension Working Group of the Czech Hepatology Society and the J.E. Purkinje Czech Medical Society ]. / Diagnostika a lécba hepatocelulárního karcinomu.

Hepatocellular carcionma (HCC) is almost exclusively associated with liver cirrhosis as a significant HCC risk marker in advanced countries. Applicable therapy depends on early diagnosis, and risk patients should be screened for the presence of HCC on a regular basis. Liver ultrasound and determination of alpha-fetoprotein serum levels (AFP) are the screening methods used. Spiral CT is the most often used method for HCC staging. Non-invasive methods may under certain circumstances replace aimed biopsy. There are 3 basic curative therapies for the early stage of HCC: liver transplantation, surgical resection and different methods of local destruction of tumour (i.e., ethanolisation, thermoablation, etc.). Patients at medium stage of HCC may profit from chemoembolisation. Current available systemic chemotherapy is ineffective. Patients with advanced HCC are treated symptomatically. Patient survival prognosis after the application of one of the above treatment methods may be similar with that for HCC free cirrhosis patients, however, prognosis for advanced HCC patients is bad, with survival period from one to nine months.

[C-reactive protein in diagnosis of complications in renal insufficiency and failure]. / C-reaktivní protein v diagnostice komplikací pri ledvinové nedostatecnosti a selhání.

C-reactive protein (CRP) is one of the positive proteins in an acute phase. It is produced in hepatocytes in response to cytokines activity, especially to IL-6. Its increase is the second biggest after significant bacterial and cardiovascular insults. It reaches its peak between 24 and 48 hours. CRP monitoring makes possible monitoring of the intensity of the pathologic process and to control efficiency of treatment measures according to fluctuation of its level. According to its serum values it can reflect a place of inflammation, e.g. in upper or lower airways, urinary tract etc. It helps to distinguish between bacterial and viral inflammations and to identify size of vascular lesions such as acute myocardial infarction, cerebral infarction, decompensation of atherosclerosis. Because of its easy detection and quick elevation CRP has not only a diagnostic importance but also a prognostic one and is a predictor of a risk of atherosclerosis. Although long lasting renal insufficiency (LLRI), renal failure (RF) and regular dialysis treatment (RDT) are indicated to elevate CRP level, authors present proves that adequately treated patient compensated with an adequate dialysis treatment has normal CRP values for a long time in spite of long lasting comorbidities including atherosclerosis. There has been done a long term monitoring of 10 patients with LLRI and 22 patients with RDT. Their CRP was monitored via a turbidimetric method using sets K-Assay made by company Kamya Bio Comp. Elevated CRP in the samples reflects an acute insult such as infection, cardiovascular disease, diabetes decompensation and last but not least quality of a dialysis treatment.

[Ambulatory monitoring of blood pressure in regular dialysis therapy]. / Ambulantní monitorování krevního tlaku v pravidelném dialyzacním lécení.

Ambulatory monitoring of the blood pressure (AMBP) makes it possible to diagnose in hypertensive patients the so-called dipper phenomenon, i.e. a drop of the BP during the night provided that the patient is asleep. The absence of this phenomenon implies as a rule serious damage of the cardiovascular apparatus, brain or kidneys. By means of an apparatus ABP monitoring type 90207 of Space Labs. Inc. a group of 16 patients in regular dialysis treatment (RDT) was examined and the blood pressures were evaluated before and after dialysis. Patients with the dipper profile reacted more adequately during dialysis i.e. by a drop of the blood pressure due to the loss of excessive fluid which they retained during the interdialysis period, as compared with the group with a non-dipper profile which may be exposed to a greater risk of cardiovascular complications. The authors conclude that detection of the absence of the non-dipper phenomenon can reveal risk patients. AMBP can explain so-called paradoxical hypertension at the end of haemodialysis despite major removal of fluids by ultrafiltration, and that moxonidine participates in a significant way in the elimination of the non-dipper phenomenon.

Arteriovenous malformations of the orofacial area.

Vascular lesions are pathological residues of the embryonic vascular system and can be divided into two main groups. The first group comprises haemangiomas, which are typical of childhood and involute spontaneously. The second group is formed by lesions without active proliferation, which include, among others, arteriovenous malformations that are congenital and grow proportionately with the subject. The authors present two cases of arteriovenous malformations of the orofacial area and discuss possibilities for modern diagnosis and treatment. Precise diagnosis and effective treatment of vascular lesions should be ensured by a diagnostic and therapeutic team of specialists from several disciplines (maxillofacial, ENT, plastic and general surgeon, paediatrician, haematologist, anaesthesiologist and possibly a neurosurgeon), headed by an intervention radiologist.

Metabolic aspects of myocardial disease and a role for L-carnitine in the treatment of childhood cardiomyopathy.

OBJECTIVES: A multicenter retrospective study was conducted to investigate the possible metabolic causes of pediatric cardiomyopathy and evaluate the outcome of patients treated with L-carnitine. METHODS: Seventy-six patients diagnosed with cardiomyopathy were treated with L-carnitine in addition to conventional cardiac treatment, and 145 patients were treated with conventional treatment only. There were 101 males and 120 females between 1 day and 18 years old. Cardiomyopathy diagnoses included dilated (148 patients), hypertrophic (42 patients), restrictive (16 patients), mixed diagnosis (11 patients), and 4 with an unknown type. Of 76 L-carnitine-treated patients, 29 (38%) had evidence to suggest a disorder of metabolism, and of 145 control patients, 15 (10%) were suspected to have a disorder of metabolism. These metabolic disorders were thought to be the cause for the cardiomyopathy of the patients. The duration of L-carnitine treatment ranged from 2 weeks to >1 year. Information was collected on length of survival (time-to-event), clinical outcome, echocardiogram parameters, and clinical assessments. Data were collected at intervals from baseline to study endpoint, death, transplant, or last known follow-up visit. RESULTS: L-Carnitine-treated patients were younger than control patients and had poorer clinical functioning at baseline, yet they demonstrated lower mortality and a level of clinical functioning and clinical severity comparable to control patients on conventional therapy by the end of the study. An analysis of the interaction between clinical outcome and concomitant medications unexpectedly revealed that the population of patients treated with angiotensin-converting enzyme (ACE) inhibitors (40% of patients) had significantly poorer survival (although their greater likelihood for poor survival may possibly have made them more likely to receive ACE inhibitors). CONCLUSION: Results suggest that L-carnitine provides clinical benefit in treating pediatric cardiomyopathy. There is a need for further exploration of potential explanatory factors for the higher mortality observed in the population of patients treated with ACE inhibitors.

[Hemodynamic changes after sclerotherapy of esophageal varices in children]. / Hemodynamické zmeny po sklerotizacích jícnových varixu u detí.

The authors present an account on favourable late results of sclerotization of oesophageal varices in children. After sclerotization not only an immediate effect can be observed such as arrest of haemorrhage but also long-term favourable effects are found in the venous circulation of the splanchnic area.

Regulation of cathepsin B activity by cysteine and related thiols.

We studied the mode of regulation of the activity of mature cathepsin B (CB) by L-cysteine and some related thiols. The activity of CB with Z-Arg-Arg-NHMec as substrate was gradually inhibited over a range of increasing concentration of Cys, Cys methyl ester (CysOMe), Cys ethyl ester (CysOEt), N-acetyl-Cys (N-AcCys) and 3-mercaptopropionic acid. However, the inhibition of CB peaked at a definite value of [Cys], [CysOMe], [CysOEt] and [N-AcCys] and was gradually reversed over a range of higher concentrations of Cys and its esters. The maximum inhibitory concentrations of Cys, CysOME, CysOEt and N-AcCys showed a positive relationship to the pKa(RSH) values of the thiols and those of CysOEt and Cys decreased with increasing pH. The capability of the thiols to overcome their own inhibitory effect on CB was dependent on the concentration of their thiolate anion (RS-). However, the preincubation-dilution experiments showed that Cys and N-AcCys did not interact with active CB via a covalent mode. The inhibition of CB by N-AcCys was competitive and could be reversed by CysOMe. This activity-recovering effect of CysOMe was concentration-dependent and obeyed the Michaelis-Menten saturation kinetics over a profound increase of [RS-]. CB reacting in an environment of concurrently decreasing [RS-] and increasing [RSH], which was achieved by means of carboxylesterase-catalyzed deesterification of CysOEt to Cys, was progressively inhibited. Cys and N-AcCys also inhibited the fragmentation of histone H4 by CB and their concentration-dependent inhibitory profiles were qualitatively similar to those observed with Z-Arg-Arg-NHMec. Taken together, the results indicate that the RSH form of Cys and related thiols inhibits the activity of CB while the RS- form of these thiols counteracts or reverses the inhibitory action of the RSH form. This previously unrecognized thiol-thiolate anion regulation mechanism might be involved in a dynamic regulation of CB activity in endosomes and lysosomes and at the sites of lysosome-driven pericellular proteolysis.

Cysteine proteases and cysteine protease inhibitors in non-small cell lung cancer.

In this study we investigated the levels of two lysosomal cysteine protease proteins cathepsin B (CB) and cathepsin L (CL) and the levels of three cysteine protease inhibitor proteins stefin A (SFA), stefin B (SFB) and cystatin C (CNC) in squamous-cell lung carcinoma (SQCLC) and matched lung parenchyma specimens and examined the inhibition of CB and cathepsin C (CC) activities by endogenous inhibitors in extracts from SQCLC, lung adenocarcinoma (LAC) and lung parenchyma specimens. We found that Stage I SQCLCs contained significantly increased levels of CB protein, CB activity and SFA protein as compared to matched lungs. Neither the levels of CL protein nor the levels of SFB protein nor the levels of CNC protein in Stage I SQCLCs and the lungs were significantly different, but the levels of CB and CL proteins as well as the levels of SFA and SFB proteins showed significant positive correlation in SQCLCs. In SQCLCs as well as in the lungs the level of SFB protein was significantly higher than the level of SFA protein or the level of CNC protein. In the lungs the levels of SFA protein and CNC protein revealed a weak negative correlation trend. In extracts from SQCLCs the level of SFA protein showed a weak negative correlation with the residual CB activity (i.e. the activity remaining after extract preincubation) whereas in extracts from the lungs the level of CNC protein displayed a weak negative correlation trend with the residual CB activity and with the residual CC activity. We observed that SQCLCs and LACs contained not only a significantly increased activity of CB but also a significantly higher inhibitory potential against the activity of endogenous CB as compared to matched lungs. Leupeptin, a small inhibitor of CB, was capable to protect CB in lung carcinoma and lung parenchyma extracts from preincubation-induced inhibition, revealing an active-site directed and competitive nature of CB inhibition by endogenous cystatins. Ultrafiltration passaged protein preparations of nominal Mr < or = 30,000 obtained from extracts of SQCLCs inhibited significantly higher quantities of activity of purified bovine spleen CC than did such protein preparations from matched lungs. Reaction courses of purified bovine spleen CC that had been preincubated with such protein preparations resembled those of endogenous CC from SQCLC and lung extracts showing a slow steady-state approach. These observations and the relaxation kinetics of CC from SQCLC and lung extracts suggest that CC in the extracts may be complexed with some cystatins. In conclusion, our results indicate that quantitatively different combinations of cystatins are the major constituents of the inhibitory potential against CB and CC in SQCLCs and the lungs.

Cathepsin B, thiols and cysteine protease inhibitors in squamous-cell lung cancer.

We investigated activities of the cysteine protease cathepsin B (CB; EC 3.4.22.1), the levels of reduced glutathione (GSH) and cysteine and the activity of gamma-glutamyltransferase (gamma-GT; EC 2.3.2.2) in squamous-cell lung carcinoma (SQCLC) and the lung parenchyma specimens from surgically treated patients. The basal CB activity, assayed in tissue extracts in the absence of exogenous activators, was significantly higher in SQCLC compared to the lung. The residual CB activity, remaining in tissue extracts after preincubation at 37 degrees C, was not any longer significantly different in SQCLC and the lungs. The inhibited CB activity, calculated as the difference between the basal and residual CB activities, was significantly higher in SQCLC compared to the lung. In the case of the cysteine protease cathepsin C (CC; EC 3.4.14.1), neither the basal nor the residual nor the inhibited CC activities in SQCLC and the lung were significantly different. Compared to CC, the powerfulness of endogenous cysteine protease inhibitors to inhibit CB was much higher in both SQCLC and the lung. The cysteine protease inhibitors from SQCLC and the lung which effectively inhibited CB could be related to the inhibitors with an apparent M(r) ranging from 10,000 to 30,000. Isoelectric focusing studies indicated significant differences in the progress of inhibition of the activity of CB isoforms in SQCLC and lung parenchyma extracts. The levels of both GSH and Cys were significantly higher in SQCLC compared to the lung and the level of GSH was significantly higher in Stage III tumors compared to Stage I tumors. The activity of gamma-GT was not significantly different in SQCLC and the lung but it was significantly higher in Stage I tumors compared to Stage III tumors and showed a significant negative correlation with GSH level in SQCLC. Dithiothreitol did not increase the basal activity of CB from SQCLC and the lung which indicates that reversibly oxidized forms of CB do not accumulate in the tumors and the lungs. The basal activity of CB from SQCLC and the lung was competitively inhibited by Cys. Moreover, increasing Cys concentrations had a modulatory effect on the basal activity of CB from SQCLC and the lung which was featured by Cys-induced inhibition of CB activity and by subsequent Cys-effected recovery of CB activity from its previous inhibition by Cys.

Lysosomal dipeptidyl-peptidases I and II in human squamous cell lung carcinoma and lung parenchyma.

In the present work we studied the levels of activities of dipeptidyl-peptidase I (or cathepsin C, DPP-I) and dipeptidyl-peptidase II (DPP-II) and examined their isoelectric focusing profiles in matched pairs of human squamous cell lung carcinoma (SQCLC) and the lung from surgically treated patients (n = 33). The mean specific activities of DPP-I and DPP-II were higher in SQCLC (Stages I and II) than in the lung, but only the activity of DPP-II in Stage I SQCLC was significantly higher compared to the lung. The activities of both enzymes were higher in the tumor than in the lung in 10 of 20 Stage I SQCLC patients, but only in 3 of 13 Stage II SQCLC patients. The specific activities of DPP-I and DPP-II in the lungs showed a good correlation while the correlation of both enzyme activities in SQCLCs was poor. We observed only a small and mutually comparable activation of DPP-I in extracts from SQCLCs and from the lungs by dithiothreitol. The isoelectric focusing profile of several DPP-II forms in SQCLCs and the lungs was similar and the single major DPP-II isoform revealed in the tumors and lungs showed a pIapp of 5.3-5.2. The isoelectric focusing profile of DPP-I showed multiple enzyme forms in SQCLCs (pIapp 6.3-4.5) as well as in the lungs (pIapp 6.4-4.8). In SQCLCs, as well as in the lungs, the activities of the DPP-I forms with pIapp values < or = 5.6 were shifted by neuraminidase treatment to the site of the major DPP-I isoform with pIapp of about 6.0 and the zymograms then showed an another DPP-I with pIapp of 5.7, which was less discernible in the lung. In some patients, the DPP-I forms with pIapp values < or = 5.6 from SQCLC retained a greater percentage of activity distribution than did the DPP-I pIapp-counterparts from the lung.