Gelatinases A and B and Antioxidant Enzyme Activity in the Early Phase of Acute Myocardial Infarction.

Oxidative stress plays important roles in the pathophysiology of acute myocardial infarction (AMI). The aim of this study was to investigate the correlation of the oxidative stress status and matrix metalloproteinase activity in AMI patients in comparison to controls. This study included 136 subjects: 68 patients with AMI (42 males/26 females; mean age 58.5 ± 10.5 years) and 68 controls (37 males/29 females; mean age 60.2 ± 12.4 years). Gelatinases A and B were assayed using gelatin zymography, enzyme activities were obtained spectrophotometrically. Gelatinase A and B activities were increased in the AMI patients' group compared to the control. Activities of serum superoxide dismutase (SOD) and xanthine oxidase (XO) were significantly higher in AMI patients (106.53 ± 23.45 U/l, P < 0.001 and 158.18 ± 29.59 U/l, P < 0.001) than in the control group (55.99 ± 10.79 U/l and 79.81 ± 7.93 U/l). The activity of catalase (CAT) in the sera of AMI patients was lower (271.31 ± 7.53 U/l, P < 0.005) than in the control group (305.94 ± 97.28 U/l). Plasma glutathione peroxidase (GPx) and glutathione reductase (GR) in AMI patients were significantly higher (582.47 ± 184.81 U/l, P < 0.001 and 59.64 ± 21.88 U/l, P < 0.001) than in the control group (275.32 ± 104.69 U/l and 47.71 ± 20.05 U/l). The present findings demonstrate activation of gelatinases A and B and oxidative stress markers in the early stage of AMI. Gelatinases, detected at high levels in AMI patients only, indicate their noticeable predisposition for becoming additional biomarkers of the early phase of AMI.

The Validation Study of Neurofilament Heavy Chain and 8-hydroxy-2'-deoxyguanosine as Plasma Biomarkers of Clinical/Paraclinical Activity in First and Relapsing-Remitting Demyelination Acute Attacks.

Although current evidence mainly suggests immunopathogenesis of demyelination and neurodegeneration in multiple sclerosis (MS), there are results which document the importance of other factors, such as oxidative stress and its mediated injuries. The oxidative stress intensity in axonal damage during acute demyelination is little known. We performed this study as a cross-sectional biomarker validation study in order to evaluate the parameters of axonal damage (phosphorylated neurofilaments heavy chain (pNF-H)) and oxidative stress (8-hydroxy-2'-deoxyguanosine (8-OHdG)) in plasma of patients with initial and relapsing-remitting demyelination attacks, defined as clinically isolated syndrome (CIS) and relapsing-remitting multiple sclerosis (RRMS); and the correlations between these parameters and biological (index of blood brain barrier (BBB) permeability), clinical (index of disease progression), and radiological (T1-Gd-enhancing lesion volume) activities of disease. Both parameters were increased in CIS and RRMS compared to control subjects (p < 0.05). The positive correlations were observed between 8-OHdG values and index of BBB permeability, clinical severity of disease, and demyelinated brain lesion volume, in CIS group (r > 0.50; p < 0.05). Similar correlations were obtained between pNF-H values and the above parameters, as well as the index of disease progression, in RRMS group (r > 0.30; p < 0.05). There was a significant correlation between values of 8-OHdG and pNF-H only in CIS group, r = 0.52, p < 0.05. While the plasma values of 8-OHdG reflect the degree of acute demyelination in CIS, pNF-H values reflect that in RRMS. The obtained results must be reevaluated in similar prospective studies related to their prognostic values.

[Magnetic Resonance Imaging, Arthroscopy and Near-Infrared Spectroscopy (NIRS) for Grading Osteoarthritis in the Knee]. / Methodenvergleich von Kernspintomografie, Arthroskopie und Nah-Infrarot-Spektroskopie (NIRS) bei der Bestimmung des Arthrosegrads des Kniegelenks.

PURPOSE: This study was aimed to evaluate the meaningfulness of the MRI Score WORMS (Whole Organ Magnetic Resonance Imaging), the arthroscopic WOAKS (Whole Organ Arthroscopic Knee Score) and the result of NIRS (near-infrared spectroscopy) measurements. MATERIALS AND METHODS: A total of 49 patients with knee pain (> 3 months) underwent MRI with a standardised protocol. In the results the WORMS was calculated. The WOAKS was calculated from the results of an arthroscopic evaluation. In the same procedure, NIRS measurements were performed in the identical 14 regions of interest. From these measurements, the WOAKS_NIRS was calculated. RESULTS: The highest grade of degeneration in all evaluations was found in the patella. The medial compartment showed moderate lesions compared with the lateral compartment. The relative WORMS was only 3.7 % (95 % CI 2.8-4.6; 0-15.6 %). During arthroscopy, we calculated a mean WOAKS of 15.2 % (95 % CI 13.2-17.2; 5-39 %). The degree of joint degeneration was highest in NIRS measurements. The mean WOAKS_NIRS was 50.9 % (95 % CI 48.1-53.7 %). These differences are significant (p < 0.001). CONCLUSION: The methods to detect early cartilage degenerations in MRI are flawed. Thus in our patients, we detected a full grade of degeneration in only 3.7 % of the patients. Arthroscopy mostly gives higher damage within the knee joint. The initial stages of cartilage lesion are usually undetectable. Spectroscopy has the best sensitivity for the evaluation of early degeneration within the hyaline cartilage. The clinical relevance of our results is still unclear. Further outcome studies are needed.

[Characteristics of Focal Degenerative Cartilage Lesions in the Knee Joint. A Radiologic, Spectroscopic, Histological and Biochemical Study]. / Eigenschaften fokaler degenerativer Knorpelschäden am Kniegelenk. Eine radiologische, spektroskopische histologische und biochemische Untersuchung.

OBJECTIVE: The aim of this study was to perform a macroscopic, spectroscopic and biochemical/histological examination of the defect margins of grade IIIb cartilage lesions in the patella, the medial femoral condyle, the corresponding articular surface and the remaining cartilage surfaces of the knee joint. Our null hypothesis was that there would be no difference in characteristics between the cartilage surrounding the defect, the corresponding articular surface and the remaining articular surfaces of the knee joint on the one hand and the cartilage within the defect on the other. METHOD: The study included ten patients treated for focal cartilage lesions (ICRS classification grade IIIb) by autologous cartilage transplantation (ACT). All patients underwent a preoperative magnetic resonance imaging scan (1, 5 T). The articular cartilage lesions were classified pursuant to the recommendations of the International Cartilage Repair Society (ICRS). During the arthroscopic procedure, spectroscopic examinations were performed to measure the degree of cartilage degeneration in a total of 14 defined areas including the defect itself and the region of the defect margins. Biopsies for a histological and biochemical examination (collagen II, glycosaminoglycan, DNA) were taken from the centre of the defect and the defect margins that seemed to be intact on macroscopic examination. RESULTS: All knee joints had focal grade IIIb cartilage lesions with an intact margin and an intact corresponding articular surface. The readings obtained on spectroscopic examination both in the defect, the apparently intact margins, the corresponding articular surface and all other examined areas of the knee suggested that severe degenerative changes had already occurred in the cartilage. The histological and biochemical examinations of the residual cartilage in the centre of the defect and the apparently intact margins revealed no significant differences. CONCLUSIONS: Focal cartilage lesions frequently occur in the main weight-bearing zones of the patella and the medial femoral condyle. If they are the result of degenerative changes in the knee joint, the residual cartilage in the defect does not differ from the cartilage of the defect margins, the corresponding articular surface and the other cartilage surfaces. This leads to the conclusion that focal cartilage defects seen in degenerative joint damage are only one aspect of general joint degeneration.

The Role of Matrix Metalloproteinase 3 and 9 in the Pathogenesis of Acute Neuroinflammation. Implications for Disease Modifying Therapy.

Matrix metalloproteinases (MMPs) are proteolytic enzymes that are involved in a variety of physiological and pathological processes, including those in CNS. In this study, plasma values of MMP-3 and MMP-9 have been compared in clinically isolated syndrome (CIS) and relapsing-remitting multiple sclerosis (RRMS) patients during their acute attacks, in relation to the biological activity of disease. Therefore, we compared the MMPs plasma values regarding Expanded Disability Status Scale (EDSS), progression index of disease (PID), acute brain lesion volume seen on magnetic resonance imaging (MRI) and index of blood-brain barrier (BBB) permeability destruction. The obtained results demonstrated higher plasma values of MMPs in both study groups than control values (p < 0.05). No statistical significances have been detected comparing the obtained values of both enzymes between CIS and RRMS group (p > 0.05). In both CIS and RRMS groups, the patients with higher EDSS showed higher MMPs plasma values (p < 0.05). The MMPs values were also significantly higher in both study patients with higher total number comparing to those with lower number of MRI brain lesion (p < 0.05) (beyond MMP-3 in RRMS). All obtained correlations, between MMPs and EDSS, PID, volume of MRI Gd-enhancement brain lesions, and index of BBB permeability, were positive (p < 0.05.) This study demonstrates alterations of both tested MMPs with closed correlation with the disease biological activity. Although MMPs are being implicated in the pathogenesis of acute neuroinflammation, the MMPs modulation might be useful in the future design of disease modifying therapy with the specific target profile.

Compound A, a selective glucocorticoid receptor agonist, inhibits immunoinflammatory diabetes, induced by multiple low doses of streptozotocin in mice.

BACKGROUND AND PURPOSE: Type 1 diabetes is a multifactorial inflammatory disease that develops as a result of deregulated immune responses, causing progressive autoimmune destruction of insulin-producing beta cells of pancreas. 2-((4-acetoxyphenyl)-2-chloro-N-methyl) ethylammonium chloride, compound A (CpdA), is a selective glucocorticoid receptor (GR) agonist that displays strong anti-inflammatory and immunomodulatory activities. We investigated the therapeutic effectiveness of CpdA in a pharmacological model of type 1 diabetes in mice. EXPERIMENTAL APPROACH: The utility of CpdA in diabetes prevention was evaluated in vivo through its prophylactic administration to male C57BL/6 mice that received multiple low doses of streptozotocin for immunoinflammatory diabetes induction. The effect of CpdA on disease development was studied by measuring blood glucose and insulin level, histopathological examination, determination of the nature of infiltrating cells, pro- and anti-inflammatory cytokine production, and signalling pathways. KEY RESULTS: Prophylactic in vivo therapy with CpdA conferred protection against development of immunoinflammatory diabetes in mice by dampening the M1/Th1/Th17 immune response and switching it towards an anti-inflammatory M2/Th2/Treg profile, thus preserving beta cell function. CONCLUSIONS AND IMPLICATIONS: Anti-diabetic properties of CpdA are mediated through modulation of immune cell-mediated pathways, but without triggering adverse events. These findings provide basic information for the therapeutic use of selective GR agonists in the amelioration of islet-directed autoimmunity.

Carbon monoxide-releasing molecule-A1 inhibits Th1/Th17 and stimulates Th2 differentiation in vitro.

Carbon monoxide (CO) is endogenously produced by haeme oxygenase-1 and has profound effects on intracellular signalling processes, generating anti-inflammatory, antiproliferative and antiapoptotic effects. A boron-containing compound CORM-A1 is capable of releasing CO in such a way to mimic physiological functions of haeme oxygenase-1. Considering the importance of Th1/Th17 versus Th2 balance in the final outcome of immune and inflammatory responses in this study we focused on immune-modulatory effects of CORM-A1 on murine lymph node-derived T cells in vitro and its influence on T-cell proliferation, activation and differentiation. Anti-CD3/CD28 antibody-triggered lymph node cells proliferation remained unaffected after 24-hour CORM-A1 treatment, as well as the expression of the early activation marker CD25. However, CORM-A1 successfully reduced the secretion of the two representative pro-inflammatory cytokines, IFN-γ and IL-17, while the secretion of anti-inflammatory cytokine IL-4 remained unchanged. Furthermore, CORM-A1 efficiently reduced the percentage of CD4(+) IFN-γ(+) and CD4(+) IL-17(+) cells, whereas CD4(+) IL-4(+) cell population increased after treatment. Also, CORM-A1 significantly reduced expression of transcription factor RORγT, necessary for Th17 development, but the expression of Th1-related and Th2-related transcription factors (T-bet and GATA-3, respectively) remained unchanged. In conclusion, our findings indicate that CO has anti-inflammatory role through the regulation of balance between pro-inflammatory Th1/Th17 and anti-inflammatory Th2 cells. Observed immunomodulatory effects of CORM-A1 could be useful for developing novel therapeutic approaches in managing Th1/Th17-mediated immune disorders.

Effects of depsidones from Hypogymnia physodes on HeLa cell viability and growth.

The anti-proliferative activitiy of Hypogymnia physodes methanol extracts (ME) and its main constituents, physodalic acid (P1), physodic acid (P2), and 3-hydroxy physodic acid (P3), was tested on human cancer HeLa cell lines. Three lichen depsidones, P1, P2 and P3, were isolated from H. physodes ME using column chromatography and their structures were determined by UV, ESI TOF MS, 1H and 13C NMR. The content of P1, P2 and P3 in ME was determined using reversed-phase highperformance liquid chromatography with photodiode array detection. P1-3 represented even 70 % of the studied extract. The HeLa cells were incubated during 24 and 72 h in the presence of ME and depsidones P1, P2 and P3, at concentrations of 10-1000 µg/ml. Compounds P2 and P3 showed higher activity than compound P1. Half maximal inhibitory concentrations (IC50, µg/ml) of P1, P2, P3 and ME for 24-h incubation were 964, 171, 97 and 254 µg/ml, respectively, while for 72-h incubation they were 283, 66, 63 and 68 µg/ml. As far as we know, this is the first report on the effect of H. physodes ME and their depsidones on HeLa cells.

IL-17 and glutamate excitotoxicity in the pathogenesis of multiple sclerosis.

Immunoinflammatory-mediated demyelination, the main pathological feature of multiple sclerosis (MS), is regularly accompanied by neurodegenerative processes, mostly in the form of axonal degeneration, which could be initiated by glutamate excitotoxicity. In the current study, the relationship between Th17-mediated inflammatory and excitotoxic events was investigated during an active phase of MS. Cerebrospinal fluid (CSF) of patients with MS and control subjects was collected, and IL-17A and glutamate levels were determined. IL-17A level was significantly higher in patients with MS; whereas no statistically significant changes in glutamate concentrations were found. There was a direct correlation between IL-17A and glutamate levels; IL-17A levels were also associated with the neutrophil expansion in CSF and blood-brain barrier disruption. However, IL-17A level and the number of neutrophils tended to fall with disease duration. The results suggest that Th17 cells might enhance and use glutamate excitotoxicity as an effector mechanism in the MS pathogenesis. Furthermore, Th17 immune response, as well as neutrophils, could be more important for MS onset rather than further disease development and progression, what could explain why some MS clinical trials, targeting Th17 cells in the later stage of the disease, failed to provide any clinical benefit.

Unusual suicide with a chainsaw.

A 39-year-old male, with a history of multiple suicidal attempts and psychiatric pathology, a professional lumberjack, was found dead at the meadow with his throat cut and a chainsaw beside him. Autopsy revealed that all physical injuries were confined to the head, neck and left shoulder. Two major (long and wide) wounds were found and documented on both sides of the neck and head. A wound on the posterior and right lateral side of his neck and head was noted. Medical examiner noted an irregular rupture on the posterior-right side of the atlanto-occipital joint with impaired bone, but without any damage on the spinal cord. Another gaping cut was noted in the lower part on the left lateral side of his neck. Medical examiner noted that muscles of the left side of the neck, left common carotid artery, left internal jugular vein and left vagus nerve were completely cut off. The body of the C5 and C6 vertebra, with the spinal cord at that level, was completely cut. Also, there were multiple linear and striped parallel abrasions on the outer side of the left shoulder and one abrasion on the left lateral side of the neck. The conclusion of inquiries was "suicide by chainsaw".

The immunobiology of apotransferrin in type 1 diabetes.

The transferrin (Tf) family of iron binding proteins includes important endogenous modulators of the immune function that may modulate autoimmune diseases. To define more clearly the role of apotransferrin (apoTf) in type 1 diabetes we determined the impact of this protein on type 1 diabetes as investigated in islet cells, animal models and patient sera. First, we demonstrated that recombinant apoTf counteracts the cytokine-induced death of murine pancreatic islet cells. Secondly, human apoTf administration favourably influences the course of type 1 diabetes in animal models, resulting in protection against disease development that was associated with reduction of insulitis and reduced levels of proinflammatory cytokines. Finally, we confirmed that patients with newly diagnosed type 1 diabetes manifest significantly lower apoTf serum levels compared to healthy controls and patients with long-lasting disease. In conclusion, our data suggest the apoTf pivotal role in the perpetuation of type 1 diabetes pathology.

Macrophage migration inhibitory factor deficiency protects pancreatic islets from cytokine-induced apoptosis in vitro.

During pathogenesis of diabetes, pancreatic islets are exposed to high levels of cytokines and other inflammatory mediators that induce deterioration of insulin-producing beta cells. Macrophage migration inhibitory factor (MIF) plays a key role in the onset and development of several immunoinflammatory diseases and also controls apoptotic cell death. Because the occurrence of apoptosis plays a pathogenetic role in beta cell death during type 1 diabetes development and MIF is expressed in beta cells, we explored the influence of MIF deficiency on cytokine-induced apoptosis in pancreatic islets. The results indicated clearly that elevated MIF secretion preceded C57BL/6 pancreatic islets death induced by interferon (IFN)-γ + tumour necrosis factor (TNF)-α + interleukin (IL)-1ß. Consequently, MIF-deficient [MIF-knock-out (KO)] pancreatic islets or islet cells showed significant resistance to cytokine-induced death than those isolated from C57BL/6 mice. Furthermore, upon exposure to cytokines pancreatic islets from MIF-KO mice maintained normal insulin expression and produced less cyclooxygenase-2 (COX-2) than those from wild-type C57BL6 mice. The final outcome of cytokine-induced islet apoptosis in islets from wild-type mice was the activation of mitochondrial membrane pore-forming protein Bcl-2-associated X protein and effector caspase 3. In contrast, these apoptotic mediators remained at normal levels in islets from MIF-KO mice suggesting that MIF absence prevented initiation of the mitochondrial apoptotic pathway. Additionally, the protection from apoptosis was also mediated by up-regulation of prosurvival kinase extracellular-regulated kinase 1/2 in MIF-KO islets. These data indicate that MIF is involved in the propagation of pancreatic islets apoptosis probably via nuclear factor-κB and mitochondria-related proteins.

The reduced glutathione and S-nitrosothiols levels in acute phase of experimental demyelination--pathophysiological approach and possible clinical relevancy.

UNLABELLED: Multiple sclerosis (MS) is characterized by inflammatory process associated with nitric oxide (NO) and the related species production in CNS, which can nitrosylate protein thiols and modulate their structure and functions, also reducing the CNS content of redox active compounds, such as glutathione (GSH). We have evaluated the relationships between S-nitrosothiols (RSNO) and GSH in the experimental model of MS - experimental autoimmune encephalomyelitis (EAE), during the treatment with inducible NO synthase inhibitor - aminoguanidine (AG) and thiol donor molecule - N-acetyl-L-cysteine (NAC). MATERIAL AND METHODS: EAE was induced by myelin basic protein, dissolved in phosphate-buffered saline (PBS), emulsified in the complete Freund's adjuvant (CFA) followed by injections of Pertussis toxin. Animals assigned to the control (PBS), EAE, CFA, EAE+AG, AG, EAE+NAC and NAC groups were scored daily for the clinical signs of EAE. RSNO and GSH were evaluated in whole encephalitic mass and cerebellum. RESULTS: RSNO concentration was increased in EAE-untreated animals compared to the AG and NAC-treated EAE animals (p<0.05). Also, during the treatment with AG and NAC, GSH concentration was increased compared to the untreated animals (p<0.05). The EAE clinical signs were reduced in EAE-treated animals compared to the other groups (p<0.05). CONCLUSION: The findings of our work suggest a potential role of RSNO and GSH in early clinical presentation of experimental MS, that might be also useful as predictive parameters for MS treatment directed to increased GSH and thiol pool in CNS.

Is insulin-dependent diabetes and obesity a predisposition for endometrial and pancreatic carcinoma?

Among 178 patients operated for endometrial carcinoma during a five-year period, 17 were re-operated at the Institute of Surgery (9.5%) because of pancreatic head carcinoma. The frequency of insulin-dependent diabetes was pointed out in patients-- 28% of those who were first diagnosed with endometrial carcinoma. Moreover in the same group diagnosed with endometrial carcinoma, we found 17 to have pancreatic carcinoma, and among those there were 12 cases that had diabetes (70.58%).

Bone metastasis arising from a polyp of the cervix as the first symptom in generalized multi-organ adenocarcinoma.

Our patient was a 40-year-old female with a positive familial history for malignancies but no chronic diseases. After two vaginal deliveries without any reported difficulties, the patient had no intermenstrual bleeding, postcoital bleeding, leucorrhea or hypermenorrhea, abnormal vaginal bleeding, or postmenstrual bleeding, except during the past five-year period when a polyp-like change in the cervix was found. There was no indication for polypectomy, considering the fact that the patient had no symptoms, had an iodine positive Schiller test, as well as regular cytological smears on Papanicolaou testing. It is noteworthy that the patient had no symptoms until changes in the stool and painful sensation in the hip area. The patient was subjected to extensive surgery by a team composed of a gynecologist, surgeon and orthopedist. During Werthaim-Meigs surgery, four positive glandules and cervical adenocarcinoma Stage II were found. The colon was removed, as a right hemicolectomy, as well as the iliac bone upper segment. Unfortunately, considering the changes in the tissue of the colon and cervix, we considered the condition to be "generalized" adenocarcinoma.

Spermidine influence on the nitric oxide synthase and arginase activity relationship during experimentally induced seizures.

Nitric oxide (NO), a potential candidate for a modulator of convulsive activity, is a mediator in several pathological events in the central nervous system. The polyamines, spermidine (Spd) and spermine, are neuromodulators influencing the metabolism of L-arginine and NO production. Here we examined the effects of Spd on NO production and arginase activity during convulsions induced by pentylenetetrazol (PTZ). Male Wistar rats were allocated into four experimental groups of 8 animals each and received the following treatments: I (control)--saline, intraperitoneally (i.p.); II (PTZ)--seizures induced by pentylenetetrazol (100mg/kg bw i.p); III (Spd)--Spd (1 mg/kg bw i.p.) 50 min before PTZ application; IV (Mid)--antiepileptic Midazolam (100 mg/kg bw) 45 min before PTZ. In brain cortex, striatum, hippocampus, cerebellum, and brainstem homogenates, nitrite + nitrate levels and arginase activity were determined. Spermidine showed proepileptic effects. shortening seizure latency and inducing a more profound increase of NO production than PTZ in all brain structures. PTZ reduced arginase activity, whereas Spd pretreatment increased enzyme activity, with the most profound effects in cerebellum and brainstem. The results point out the importance of polyamine and arginine metabolism in the brain during seizures, suggesting a regulatory role for polyamines and arginase in NO production.

Does polyamine oxidase activity influence the oxidative metabolism of children who suffer of diabetes mellitus?

Diabetes mellitus is a metabolic disease characterized by inadequate secretion of insulin. Polyamine oxidase (PAO), a FAD-containing enzyme is involved in the biodegradation of Sp and Spd, catalyzing the oxidative deamination of Sp and Spd, resulting in production of ammonia (NH(3)), corresponding amino aldehydes and H(2)O(2). Malondialdehyde (MDA) and acrolein (CH2=CHCHO), potentially toxic agents, which induce oxidative stress in mammalian cells, are then spontaneously formed from aminoaldehydes. The main signs of oxidative stress in diabetic children were the values of HbA1c and MDA levels. Polyamines have an insulin-like action. Antiglycation property of spermine and spermidine has been recently confirmed. There are no data in the literature about plasma polyamine oxidase (PAO) activities in children with type 1 diabetes. The idea of this study was to evaluate the polyamine metabolism through the estimation of polyamine oxidase activity. We have study children with newly diagnosed type 1 diabetes mellitus (n = 35, age group of 5-16 years, as well as age-matched healthy control subjects (n = 25). The biochemical investigations were done on diabetic children who have the pathological values of glucose (9.11-17.33 mmol/l) and glycosylated Hb (7.57-14.49% HbA(1c)). The children in the control group have referent values of glucose and glycated hemoglobin (4.11-5.84 mmol/L and HbA(1c) 4.22-6.81% of the total Hb. Glucose levels in blood plasma and glycosylated hemoglobin in erythrocythes hemolysates (HbA1c) were measured by using standard laboratory methods. PAO activity in venous blood plasma and the amount of malondialdehyde (MDA) were measured by the spectrophotometric methods. PAO activity, glycemia, HbA1c and MDA were significantly increased in diabetic children compared to the control subjects. PAO activity in children with type 1 diabetes mellitus was very high. The findings of higher blood HbA(1C) and MDA levels confirm the presence of oxidant stress in children with type 1 diabetes mellitus and demonstrate that PAO activity may participate in these circumstances.

Metabolic correlations of glucocorticoids and polyamines in inflammation and apoptosis.

Glucocorticoid hormones (GC) are essential in all aspects of human health and disease. Their anti-inflammatory and immunosuppressive properties are reasons for therapeutic application in several diseases. GC suppress immune activation and uncontrolled overproduction and release of cytokines. GC inhibit the release of pro-inflammatory cytokines and stimulate the production of anti-inflammatory cytokines. Investigation of GC's mechanism of action, suggested that polyamines (PA) may act as mediators or messengers of their effects. Beside glucocorticoids, spermine (Spm) is one of endogenous inhibitors of cytokine production. There are many similarities in the metabolic actions of GC and PA. The major mechanism of GC effects involves the regulation of gene expression. PA are essential for maintaining higher order organization of chromatin in vivo. Spermidine and Spm stabilize chromatin and nuclear enzymes, due to their ability to form complexes with negatively charged groups on DNA, RNA and proteins. Also, there is an increasing body of evidence that GC and PA change the chromatin structure especially through acetylation and deacetylation of histones. GC display potent immunomodulatory activities, including the ability to induce T and B lymphocyte apoptosis, mediated via production of reactive oxygen species (ROS) in the mitochondrial pathway. The by-products of PA catabolic pathways (hydrogen peroxide, amino aldehydes, acrolein) produce ROS, well-known cytotoxic agents involved in programmed cell death (PCD) or apoptosis. This review is an attempt in the better understanding of relation between GC and PA, naturally occurring compounds of all eukaryotic cells, anti-inflammatory and apoptotic agents in physiological and pathological conditions connected to oxidative stress or PCD.

Arginase activity and magnesium levels in blood of children with diabetes mellitus.

Under physiological conditions insulin controls the metabolism of carbohydrates, lipids and proteins. Diabetes mellitus is a metabolic disease characterized by a disturbance in the intermediary metabolism of glucose and glucose-induced insulin release. Arginase (L-arginine amidinohydrolase, EC 3.5.3.1) modulates nitric oxide synthase activity by regulating intracellular L-arginine availability. In diabetes mellitus, a decrease in nitric oxide bioavailability is a central mechanism for endothelial dysfunction. The aim of our study was to assess arginase activity in the blood of children with diabetes mellitus. Blood arginase activity, serum glucose (14.155 +/- 4.197 mmol/L; p < .001) and blood HbA1c (11.222 +/- 3.186 %; p < .001), were significantly higher in diabetic children than in healthy controls, whereas the magnesium (Mg2+) level, a cofactor of many enzymes, was significantly lower (0.681 +/- 0.104 micromol; p < .001). In diabetic children, arginase activity, hyperglycemia (r = 0.143), and the HbA1, level (r = 0.381) showed a positive correlation between but a negative correlation between Mg2+ and arginase activity (r= -0.206). The higher arginase activity and the lower Mg2+' levels in diabetic children could be a consequence of reduced insulin action and increased protein catabolic processes in these pathophysiological conditions. The inverse directions of arginase activity and serum Mg2+ levels are in agreement with this concept.

Anti-tumor effect of Coriolus versicolor methanol extract against mouse B16 melanoma cells: in vitro and in vivo study.

Numerous studies have shown immunostimulatory and anti-tumor effects of water and standardized aqueous ethanol extracts derived from the medicinal mushroom, Coriolus versicolor, but the biological activity of methanol extracts has not been examined so far. In the present study we investigated the anti-tumor effect of C. versicolor methanol extract (which contains terpenoids and polyphenols) on B16 mouse melanoma cells both in vitro and in vivo. In vitro treatment of the cells with the methanol extract (25-1600 microg/ml) reduced melanoma cell viability in a dose-dependent manner. Furthermore, in the presence of the methanol extract (200 microg/ml, concentration IC(50)) the proliferation of B16 cells was arrested in the G(0)/G(1) phase of the cell cycle, followed by both apoptotic and secondary necrotic cell death. In vivo methanol extract treatment (i.p. 50 mg/kg, for 14 days) inhibited tumor growth in C57BL/6 mice inoculated with syngeneic B16 tumor cells. Moreover, peritoneal macrophages collected 21 days after tumor implantation from methanol extract-treated animals exerted stronger tumoristatic activity ex vivo than macrophages from control melanoma-bearing mice. Taken together, our results demonstrate that C. versicolor methanol extract exerts pronounced anti-melanoma activity, both directly through antiproliferative and cytotoxic effects on tumor cells and indirectly through promotion of macrophage anti-tumor activity.