Studies on the Authorship of Albumen Vintage Photographs: A Combined Experimental and Chemometric Approach.

The differences in albumen photographs from vintage photographic studios were identified by energy-dispersive X-ray fluorescence spectroscopy and Fourier transform infrared spectroscopy. The results inspired the concept of finding common features characteristic of a given photographic studio. The obtained measurement data (i.e., positions of vibrational bands for characteristic groups of albumen and the mass contents of chosen elements) were analyzed chemometrically by employing the Principal Component Analysis (PCA). The PCA technique allowed us to reduce the number of relevant experimental parameters characterizing the unique features of the photographic objects. The two major components were able to distinguish the photographic objects in terms of their authorship and the time to produce a photograph. The method developed was examined for a selected group of photographs consisting of albumen prints from three Polish photographic ateliers. To validate ED-XRF measurements and, consequently, the chemometric findings, reference albumen photo samples were designed and prepared. The empirical functional relationships between the content of photochemically reduced silver particles on the photographic paper and several physicochemical factors, including time of exposure to UV light, AgNO3 concentration in a fixed bath, and concentrations of other additives, were proposed. These results can be used for the prediction of the experimental conditions under which the investigated photographs were developed.

Increased risk of acute pancreatitis occurrence in smokers with rs5751901 polymorphisms in GGT1 gene.

Objectives: The study was aimed to assess γ­glutamyltransferase (GGT) activity and concentration as a marker of oxidative stress induced by exposure to tobacco smoke in acute pancreatitis (AP) course. Examination of the relationship between GGT activity/concentration and single-nucleotide polymorphism (SNP rs5751901 and rs2236626) in GGT1 gene was performed. Subjects and methods: We examined SNPs in 38 AP patients and 51 healthy subjects by PCR-RFLP methods. GGT concentration in blood was measured with the use of the ELISA method; GGT activity and GSH concentration were measured by the Szasz and Patterson methods, respectively. Results: In the non-AP smokers group with TC genotype for SNPrs5751901 an increased blood GGT activity compared to smokers with CC genotypes was shown. In the course of AP was observed an elevated GGT activity and the value of GGT activity/GGT concentration ratio in smokers compared to non-smokers, in AP patients with TC genotypes and CC genotypes, respectively, for both SNP: rs5751901 and rs2236626. In the group of smoking AP patients with the CC and TC genotypes in rs5751901 locus and CC and TT genotypes in rs2236626 locus a decreases in GSH concentration during hospitalization were noted. Conclusions: SNP rs5751901 and rs2236626 cause changes in GGT activity. Smoking in the AP course contributes to increased GGT activity and excessive GSH use up in patients with TC and CC genotypes for both SNPs. Exposure to smoke xenobiotics enhances (3-fold) the risk of AP occurrence in individuals with TC genotypes for SNP rs5751901.

Relationship between somatostatin and interleukin-6: A cross-sectional study in patients with acute pancreatitis.

OBJECTIVES: The aim of the analysis is to determine dynamic changes in somatostatin (SS) and interleukin-6 (IL-6) concentrations during in acute pancreatitis (AP). METHODS: The influence of tobacco smoking on IL-6 and SS levels in the serum of non-smoking (n = 10) and smoking (n = 27) patients with diagnosed AP and control group: non-smoking (n = 44), smoking (n = 42) and passive smoking (n = 29) healthy persons was proved. The concentration of IL-6 and SS was determined by means of ELISA. Differences between the groups analyzed were tested using the U Mann Whitney test. The Spearman rank correlation analysis was used to evaluate the correlations. RESULTS: The concentrations of IL-6 and SS were significantly higher in smoking patients with AP and healthy persons when compared with non-smoking population on every day (1 day: p = 0.0002, p = 0.015; 3 day: p = 0.005, p = 0.001 and 7 day: p = 0.025, p = 0.038). Dynamic changes in concentrations of IL-6 and SS in the serum of patients with AP were demonstrated in the ensuing days of the disease. In case of non-smoking and smoking patients, significant positive correlations between IL-6 and SS was observed. CONCLUSIONS: These findings suggest that some of the antiinflammatory effects of SS against acute pancreatitis may be mediated by reducing the local proinflammatory cytokine secretion in the pancreas.

Decreases in Paraoxonase-1 Activities Promote a Pro-inflammatory Effect of Lipids Peroxidation Products in Non-smoking and Smoking Patients with Acute Pancreatitis.

Aim: The study investigated the extent to which tobacco smoke exposure causes changes in lipids biochemistry through measurement blood concentrations of: paraoxonase-1 (PON-1) activities as lipid-bound enzyme into cell membrane, concentration of malonyldialdehyde (MDA), protein adducts of 4-hydroxynonenal (HNE-adducts), oxidized low density lipoproteins (oxLDL), total cholesterol (CH) and high-density lipoprotein cholesterol (HDL). Additionally, the activity of P isoform of glutathione S-transferase (GST-π) was measured. Methods: Investigations were performed in the blood of patients with acute pancreatitis (AP) on the 1st, 3rd and 7th day of hospitalization and in healthy volunteers. The activities of PON-1 forms, GST-π were determined spectrophotometrically. Concentrations of PON-1, MDA, HNE-adducts, oxLDL, HDL, CH were measured using commercial tests. Results: Near 2-fold higher concentrations of MDA, HNE-adducts, oxLDL, correlating with inflammatory markers in AP patients compared to healthy subjects were demonstrated, which were accompanied by gradually increasing CH/HDL ratio during hospitalization. During hospital treatment, decreased activities of all PON-1 subtypes were observed in AP patients compared to healthy subjects, more pronounced in tobacco smokers. A decreased PON-1 phosphotriesterase activity in non-AP control group smokers compared to non-smokers was noted. In non-smoking AP patients GST-π activity normalized during hospitalization in contrast to smokers. Conclusions: GST-π and PON-1 phosphotriesterase activities seem to be a sensitive marker of pro/antioxidative imbalance in smokers. Lipids peroxidation products generated during AP can intensify preexisting inflammation. Increasing stay in the hospital was associated with worsening of lipids peroxidation markers and the parameters of lipid profile, in both non-smoking and smoking AP patients, what can indicate that the oxidative-inflammatory process are not extinguished.

Resistin as a Prooxidant Factor and Predictor of Endothelium Damage in Patients with Mild Acute Pancreatitis Exposed to Tobacco Smoke Xenobiotics.

OBJECTIVES: The study was aimed to assess the influence of tobacco smoke exposure on the intensity of inflammation measured by IL-6, α1-antitripsin (AAT) and α1-acid glycoprotein (AGP) concentrations, and Cd level and oxidative stress intensity measured by advanced oxidation protein product (AOPP) concentration in the blood of healthy subjects and AP patients during hospitalization. Endothelin-1 (ET-1) and resistin concentrations, markers of endothelium injury, were determined. RESULTS: An increased IL-6 concentration in healthy smokers compared to nonsmokers and AP patients compared to controls was shown. An increased AAT and AGP concentrations during hospitalization of AP patients were noted, in both smokers (AAT, AGP) and nonsmokers (AAT). In comparison to control groups, in AP patients, a 2-fold increased resistin concentration correlating with ET-1 concentration and decreased albumin concentration accompanied by increased AOPP concentration were demonstrated. AOPP concentration was higher in smokers with AP compared to nonsmokers and gradually enhanced during their hospitalization. CONCLUSIONS: Tobacco smoke exposure can have a proinflammatory effect in both healthy subjects and AP patients. Increased resistin concentration in AP patients negatively correlating with albumin concentration has prooxidative effect on this protein resulting in enhanced AOPP level. Increased resistin concentration can intensify AAT and AGP production during AP.

The Cause and Effect Relationship of Diabetes after Acute Pancreatitis.

Acute pancreatitis (AP) is an acute inflammation of the pancreas associated with high morbidity and mortality. Endocrine pancreatic insufficiency secondary to AP has drawn increasing attention in recent years. The aim of this paper is to analyze the available clinical and experimental literature to determine the cause and effect relationship of diabetes type 3c (T3cDM; pancreatogenic diabetes) after acute pancreatitis. The clinico-pathological features and management challenges of pancreatogenic diabetes overlap with other secondary causes of diabetes. A complex pathogenesis involving pancreatic exocrine insufficiency, dysfunction of insulin secretion, and insulin resistance is likely the cause of T3cDM after AP. To obtain an improved understanding of the pathophysiology of diabetes after AP, more research is now needed to understand the risk of complications related to the pancreas and diabetes in these patients.

Association of Genetic Variants in IL6 Gene (rs1800795) with the Concentration of Inflammatory Markers (IL-6, hs-CRP) and Superoxide Dismutase in the Blood of Patients with Acute Pancreatitis-Preliminary Findings.

In the course of acute pancreatitis, interleukin-6 plays an important role as a mediator in the inflammatory response. The course of inflammatory disease is associated with intensive oxidative stress, which may activate transcription factors leading to gene-expression changes. Isoenzymes of superoxide dismutase are involved in the defense against free radicals. This study aimed to evaluate changes in IL-6 concentration and the concentration/activity of superoxide dismutase isoenzymes (SOD1, SOD2, and SOD3) in the blood of patients with acute pancreatitis (AP) in terms of rs1800795 polymorphism in the IL6 gene. In the smoking AP patients group with the GC and GG genotypes, the plasma SOD1 concentration was significantly higher (p = 0.0146 and p = 0.0250, respectively) than in patients with CC genotype for SNP rs1800795 in the IL6 gene. An increase in SOD1 concentration in erythrocytes of AP patients with GC genotypes was also demonstrated compared to the individuals from the group with GG genotype (p = 0.0408). Furthermore, a positive correlation between IL-6 and SOD1 concentrations in the plasma of AP patients with GC genotype for SNP rs1800795 was shown. These results indicate that SOD1 may play a protective role against oxidative damage induced by inflammation in the group of AP patients with GC genotype.

Architecture of the thrombi removed from coronary artery bypass grafts in a patient with acute myocardial infarction.

Using scanning electron microscopy we analyzed thrombotic material removed from coronary bypass grafts in a 57-year-old woman with multilevel atherosclerosis presenting with acute myocardial infarction (AMI). A white thrombotic material removed from the marginal branch bypass that contained large amounts of activated platelets displaying pseudopodia clearly visible at a higher magnification with a relatively low amount of fibrin. The other thrombus obtained from the right posterior descendent branch (RPD) bypass showed a highly organized fibrin structure composed of thicker fibers with low amounts of cellular components. Our findings indicate that the thrombus structure is different in AMI patients in whom the infarct-related vessel is vein anastomosis compared to those with a native coronary artery occluded. These findings help explain resistance of such thrombi to fibrinolysis and faster plaque growth related to fibrin accumulation.

Changes in the Activity and Concentration of Superoxide Dismutase Isoenzymes (Cu/Zn SOD, MnSOD) in the Blood of Healthy Subjects and Patients with Acute Pancreatitis.

This study was aimed at evaluating the changes in the concentration and activity of all superoxide dismutase isoenzymes (SOD1, SOD2, SOD3) in the blood of patients with acute pancreatitis (AP) and healthy subjects, taking into account the extracellular (plasma) and intracellular (erythrocyte lysate) compartment. The relationships between the activity/concentration of SODs, metal concentration and the markers of inflammation were evaluated. To assess the pro/antioxidative imbalance, the malonyldialdehyde (MDA) concentration and the value of total antioxidant capacity (TAC) were measured. The impact of single-nucleotide polymorphism (SNP) in the SOD1 gene (rs2070424) on the activity/concentration of SOD1 as the main isoenzyme of the SOD family was also analyzed in this study. The SOD2 activity in erythrocytes was increased compared to plasma: 10-fold in the AP patient group and 5-fold in healthy subjects. The plasma of AP patients showed an increased SOD1 concentration and decreased SOD2 and SOD3 concentrations compared to healthy subjects. The Cu/Zn SOD (SOD1 + SOD3) concentration in plasma of AP patients was elevated compared to healthy subjects, but changes in plasma Cu/Zn SOD (SOD1 + SOD3) activity in the examined groups were not observed. An influence of SNP rs2070424 in the SOD1 gene on the total activity of SOD in AP patients (with AG genotype), accompanied by an increased IL-6 concentration, was observed. In oxidative stress conditions induced by inflammation, the participation of individual forms of plasma SOD isoenzymes in total antioxidative activity of SOD changed. A significant increase in the intracellular SOD1 concentration in plasma of AP patients proves the important role of this isoenzyme in the neutralization of oxidative stress induced by impaired Cu and Zn homeostasis. The presence of increased concentration of SOD2 in erythrocytes of healthy subjects and AP patients confirms the important function of this isoenzyme in the antioxidative defense.

A comparative study of osteopontin expression, Ki67 index and prognosis in squamous cell carcinoma and cysts of the oral cavity.

BACKGROUND: Recent studies have shown a strong relationship between the expression of osteopontin and oral carcinogenesis. Osteopontin (OPN) has been shown to play a major role in regulating the aggressiveness of cancer cells and promote tumor growth. Odontogenic cysts are an essential aspect of oral and maxillofacial pathology. They are relatively frequent lesions with different clinical behavior. Some of them may have a proliferative pattern of growth and neoplastic nature. Evaluation of osteopontin expression with Ki-67 index may help examine clinical behavior and recurrence of oral squamous cell cancer and radicular cyst patients. METHODS: A total of 44 oral cavity cancer cases and 21 cysts samples were analyzed by immunohistochemical staining. Data used for analysis were derived from medical records. The following information was obtained from all patients' medical records: survival, age, sex, lymph node status, tumor size, and location, as well as grade and histologic type of tumor. Expression status of OPN and Ki-67 was statistically assessed. RESULTS: Our data demonstrated that for summary immunoreactive scores of OPN and Ki-67 expressions in OSCC vs. RC patients statistical significance was found for both markers' between OSCC and RC groups. Moreover, osteopontin is significantly higher expressed in larger OSCC tumors. CONCLUSIONS: In conclusion, the role of OPN expression both in oral squamous cancer cells and radicular cyst and possible correlation with demographic and clinicopathological features remain undetermined in some aspects, further high-powered studies to develop a more standardized assessment of Ki-67 and osteopontin expression in OSCC and are needed.

The role of pancreatic polypeptide in pancreatic diseases.

The aim of this study was to review the diagnostic significance of pancreatic polypeptide (PP) in pancreatic diseases. PP may play a significant role in monitoring the development of the disease and the patient's healing process, particularly after the removal of a portion of the pancreas. Determining PP in acute pancreatitis is quite controversial. At the 1st stage of severe pancreatic damage, there is excessive PP release followed by its fall. In patients with chronic pancreatitis, a significant decrease in PP secretion was found in the presence of a food stimulant. In this case, PP could be a good marker for determining the stage of pancreatitis. Pancreatic polypeptide also functions as a hepatic glucose regulator. PP increases hepatic insulin sensitivity, resulting in reduced hepatic glucose production. Therefore, impaired hepatic insulin sensitivity in chronic pancreatitis is abrogated after the PP administration. Endocrine pancreatic tumors initially grow without specific symptoms. In contrast, they are almost always correlated with elevated serum pancreatic polypeptide. Therefore, the level of PP may be a good diagnostic parameter confirming the presence of pancreatic cancer. Depending on the type of disease, the polypeptide concentration can be increased or decreased, evidencing the disease progress or regression.

Markers of pulmonary hypertension in potential candidates to heart transplantation.

BACKGROUND: Pulmonary hypertension (PH) with pulmonary vascular resistance (PVR) 320-480 ARU resistant to therapy is a contraindication for orthotopic heart transplantation (HTX). AIM OF THE STUDY: evaluation of pulmonary hypertension in candidates for heart transplantation. STUDY POPULATION AND METHODS: 44 patients (pts) with dilated cardiomyopathy (Group I) and 34 patients with ischemic heart disease (Group II) were assessed. Evaluation of PH was done according to the following protocol: 1st measurement (cardiac catheterization): PVR (pulmonary vascular resistance) > or = 320 ARU--infusion of NTG (nitroglycerine) 1 microg/kg/min. (during 10 min.), 2nd measurement: PVR > or = 320 ARU--infusion of NTG 2 microg/kg/min. (during 10 min.), 3rd measurement: final PVR evaluation. If PVR was less than 320 ARU in 2nd or 3rd measurement, the reversibility of PH was diagnosed, and when PVR > or = 320 ARU was observed in 3rd measurement, fixed PH was diagnosed. RESULTS: No significant difference in baseline PVR between the groups was observed (Group I--332,7 ARU and Group II--327.6 ARU). In 23 patients with PVR > or = 320 ARU reduction of mean values of PVR, MPAP (mean pulmonary artery pressure) and TPG (transpulmonary gradient) was seen (p < 0.001), significantly more prominent in 11 pts from Group I. There was no correlation between duration of the disease and degree of PH. HTX was performed in 25 patients. PVR at 72 hours after HTX was similar in 7 pts with reversible PH and 18 pts with PVR < 320 ARU in 1st measurement. CONCLUSIONS: 1. Nitroglycerine used according to the protocol is sufficient in determining PH. 2. The reversibility of PH could depend on type of cardiomyopathy but not directly on symptom duration.

Presence of B cells within aortic valves in patients with aortic stenosis: Relation to severity of the disease.

BACKGROUND: Aortic stenosis (AS) shares several similarities with atherosclerosis. Recent reports showed that B cells are implicated in atherosclerosis progression through macrophage-B cells bidirectional interaction. We aimed to study the in loco presence of B cells within aortic valves and to determine its modulators. METHODS: Thirty-seven patients with severe AS were studied. Immunohistochemistry was performed on valve leaflets using antibodies against CD20, B cell-activating factor of the tumor necrosis factor family receptor (BAFF-R) and CD68. Plasma inflammatory markers were also determined. RESULTS: The B cells were detected within aortic leaflets from 5 to 31/mm(2) (17.9±11.6/mm(2)). Double-staining showed that 27±13.5% of B cells express BAFF-R. There were positive correlations between the number of B cells and macrophages (r=0.45, p=0.018), and between macrophages and B cell-associated BAFF-R expression (r=0.66, p=0.002). The number of B cells was associated with the valve calcification (r=0.41, p=0.039), and with the maximum transvalvular gradient (r=0.63, p=0.02). The BAFF-R expression was positively correlated with maximum transvalvular gradient (r=0.39, p=0.031) and negatively with aortic valve area (r=-0.41, p=0.048). There were no correlations between the number of B cells and plasma markers. CONCLUSIONS: It might be hypothesized that, like in atherosclerosis, increasing number of B cells within aortic valves may accelerate inflammation and thus potentiate the progression of AS.

Second-Line Chemotherapy of Advanced Colorectal Cancer: Predictive and Prognostic Factors.

BACKGROUND: Colorectal cancer progression presents a significant clinical problem. After its dissemination, the foundation of its treatment comprises of palliative chemotherapy. OBJECTIVES: The aim of this study was to assess the predictive and prognostic value of clinical response to second line treatment (with capecitabine or with a two-drug regimen including irinotecan) and to analyze its relation to selected clinical and pathological variables with respect to time to disease progression. MATERIAL AND METHODS: The retrospective analysis of 164 patients with advanced colorectal cancer treated in 2001- -2008 included chosen clinical, pathological and follow-up data. RESULTS: Response to second-line chemotherapy was observed in 34 out of 164 patients: In 18/82 in the irinotecan group (22%) and in 16/82 in the capecitabine group (19.5%). The mean survival time to progression following the second line of treatment amounted to 5.85 and 6.2 months respectively. Statistically, a higher number of patients in good condition of 0 to 1 was documented in the group responding to treatment. Significant correlation was documented between primary stage of the disease and time to progression in patients treated with capecitabine (p = 0.0258). The recurrence of the disease was observed in 44/45 patients following operation with radical intention but with an insufficient number of excised lymph nodes. A significantly longer time to progression was observed in women treated with capecitabine. In logistic regression, lack of treatment response was found to be an independent factor affecting the time to disease progression. Patients who did not respond to the second line of treatment demonstrated a significantly shorter time to disease progression than patients who responded to it and they showed a significantly higher number of patients with leucopenia during treatment. CONCLUSIONS: Clinical response to treatment in both treated groups is of significant importance for the probability of local recurrence of the disease, preservation of a good patient's condition and the higher level of leukocytes during treatment.

Indications for reoperation for valvular prosthesis dysfunction in urgent and elective patients.

OBJECTIVES: Comparison analysis of indications for operation, early clinical outcomes and hospital mortality between patients reoperated for valvular prosthesis dysfunction urgently and electively. MATERIAL AND METHODS: Retrospective data analysis of 94 patients (pts) reoperated for valve prosthesis dysfunction at Department of Cardiovascular Surgery and Transplantology in Kraków, Poland between January 1999 and December 2002. RESULTS: Valvular reoperation was elective procedure in 69 pts (73.4%)--group A: 46 males (M), 23 females (F), age 23-71 (av. 51.31 +/- 10.2). Urgent operation was performed in 25 pts--group B: 17 M, 8 F, age 29-77 years (av. 5.41 +/- 13.26). In group B: 9 patients underwent reoperation for dysfunction of aortic valve (2 allogenic, 7 mechanical), 13 pts for mitral valve dysfunction and 3 pts for both aortic and mitral valve dysfunction. In group A: 24 pts for mitral valve dysfunction, 37 pts for aortic valve dysfunction (25 allogenic, 12 mechanical) and 8 pts for both aortic and mitral valve dysfunction. In group B: 16 pts (64%) were operated in NYHA class IV; 8 in cardiogenic shock; on which 8 pts with active endocarditis, 3 pts with sepsis and 1 patient in septic shock. Average operative risk calculated as Euroscore was 30.86 in group B and 13.15 in group A (p<0.05). Overall hospital mortality was 12.6% (12 pts). In group B: 7 pts died (28%), 3 pts due to multiorgan failure, 1 patient due to sepsis, 2 pts due to aorta rupture and 1 due to left ventricle rupture during cardiopulmonary rescuscitation. In group B 5 pts died (7.2%). CONCLUSION: Patients reoperated urgently for valve prosthesis dysfunction were in worse preoperative state, had significantly higher preoperative risk and in hospital mortality than patients reoperated electively.

Dysfunction of primary aortic valve prosthesis--allogenic versus mechanical--preoperative differences.

OBJECTIVES: Comparison of preoperative characteristics between patients reoperated for allogenic and mechanical aortic valve prosthesis dysfunction. METHODS: Retrospective data analysis of 47 patients reoperated for aortic valve prosthesis dysfunction at Department of Cardiovascular Surgery and Transplantology in Kraków, Poland between January 1999 and December 2002. RESULTS: In analyzed group 27 patients were operated for dysfunction of allogenic valve (Gr. A), 20 pts for mechanical valve (Gr. B). Group A: 21 males (M), 6 females (F), age 23-68 years old (av. 47.10 +/- 11.71); Group B: 18 M, 2 F, age 38-77 (av. 54.73 +/- 11.03), p<0.05. Time between operations in group A was 69-258 months (av. 164.34 +/- 66.02) months and in group B 0.5-348 months (av. 77.27 +/- 98.29) p<0.01. In group A, 5 pts and in group B, 7 pts were in NYHA group IV preoperatively. Echocardiography revealed that diastolic left ventricular dimension (LVDD) was larger in group A: av. 6.7 cm and 5.36 cm in group B, p<0.05. 2 pts from group A and 7 pts from group B were operated urgently (p<0.05), including 3 pts with aortic dissection. Definite infective endocarditis was diagnosed in 6 pts from group A and 11 pts from group B, p<0.05. In group A, 1 patient was urgently operated for infective endocarditis with vegetations, he died due to multiorgan failure. In group B, 6 pts died including all 3 with aortic dissection. CONCLUSIONS: Patients reoperated for allogenic aortic valve prosthesis dysfunction are: younger, reoperated after a much longer period of time with slower valve dysfunction progress (larger LVDD), most frequently reoperated electively, less frequently with endocarditis as the cause of valve dysfunction than in patients reoperated for mechanical aortic valve dysfunction.

Does diabetes accelerate the progression of aortic stenosis through enhanced inflammatory response within aortic valves?

Diabetes predisposes to aortic stenosis (AS). We aimed to investigate if diabetes affects the expression of selected coagulation proteins and inflammatory markers in AS valves. Twenty patients with severe AS and concomitant type 2 diabetes mellitus (DM) and 40 well-matched patients without DM scheduled for valve replacement were recruited. Valvular tissue factor (TF), TF pathway inhibitor (TFPI), prothrombin, C-reactive protein (CRP) expression were evaluated by immunostaining and TF, prothrombin, and CRP transcripts were analyzed by real-time PCR. DM patients had elevated plasma CRP (9.2 [0.74-51.9] mg/l vs. 4.7 [0.59-23.14] mg/l, p = 0.009) and TF (293.06 [192.32-386.12] pg/ml vs. 140 [104.17-177.76] pg/ml, p = 0.003) compared to non-DM patients. In DM group, TF-, TFPI-, and prothrombin expression within valves was not related to demographics, body mass index, and concomitant diseases, whereas increased expression related to DM was found for CRP on both protein (2.87 [0.5-9]% vs. 0.94 [0-4]%, p = 0.01) and transcript levels (1.3 ± 0.61 vs. 0.22 ± 0.43, p = 0.009). CRP-positive areas were positively correlated with mRNA TF (r = 0.84, p = 0.036). Diabetes mellitus is associated with enhanced inflammation within AS valves, measured by CRP expression, which may contribute to faster AS progression.

Fibrin presence within aortic valves in patients with aortic stenosis: association with in vivo thrombin generation and fibrin clot properties.

A role of coagulation in the pathogenesis of aortic stenosis (AS) is unknown. The aim of this study was to investigate the fibrin (Fn) presence and its determinants in calcified stenotic aortic valve leaflets. Twenty-one patients with dominant AS and 17 well-matched patients with dominant aortic insufficiency (AI) undergoing aortic valve replacement were studied. Immunofluorescence analysis was performed on decalcified leaflets using antibodies against human Fn and tissue factor (TF). Fn-positive (41.4%) and TF-positive (25.3%) areas were increased in AS valves compared with AI valves (7.9% and 5.9%, respectively, both p<0.001). Patients with AS had elevated plasma D-dimer (236.4 ± 28 ng/ml, p=0.002) and prothrombin fragment 1+2 (F1.2) (261.7 ± 27.1 pM, p=0.005) compared to AI subjects (142.8 ± 10 ng/ml and 131.2 ± 1.3 pM, respectively). In AS patients Fn-positive areas correlated with TF-positive areas (r=0.68, p=0.0005), D-dimer (r=0.45, p=0.018), F1.2 (r=0.64, p=0.002), the time required for plasma fibrin clot formation (r=0.44, p=0.015) and maximum absorbance of fibrin clots (r=-0.38, p<0.0001), but not with clot permeability or lysis time. Thickness of Fn layer within AS valves was associated with maximum transvalvular gradient (r =0.41, p=0.048). Patients with maximal gradient above 75 mmHg (n=11) showed significant associations between Fn-positive area and both maximal (r =0.63) and mean (r =0.67) transvalvular gradients. Large fibrin amounts, mostly co-localised with TF, are present within the valve leaflets of patients with advanced AS. In vivo thrombin generation and fibrin clot formation are associated with the extent of Fn presence within leaflets, which might contribute to the AS progression.

Increased thrombin generation and platelet activation are associated with deficiency in high molecular weight multimers of von Willebrand factor in patients with moderate-to-severe aortic stenosis.

BACKGROUND: High molecular weight von Willebrand factor (vWF) multimers (HMWM) are often deficient in patients with severe aortic stenosis (AS) owing to shear stress-enhanced proteolysis of vWF. It has also been reported that AS is associated with increased activation of blood coagulation. OBJECTIVE: To investigate whether patients with AS with a deficiency in vWF HMWM have enhanced thrombin generation and platelet activation in vivo. DESIGN: Based on the analysis of vWF HMWM performed using immunolocalisation, 11 subjects with vWF HMWM deficiency (low %HMWM group) were identified and compared with 42 patients with AS with a normal distribution of vWF HMWM (normal %HMWM group). Plasma thrombin markers thrombin-antithrombin complexes (TAT) and prothrombin factor 1+2 (F1.2) plus platelet activation markers soluble CD40 ligand (sCD40L), ß-thromboglobulin and P-selectin were also measured. PATIENTS: 48 consecutive patients with severe AS and five with moderate AS, free of angiographically-proven coronary artery disease and clinically overt bleeding, were studied. RESULTS: Patients in the low %HMWM group had 34.8% higher maximal transvalvular gradient (p = 0.0003) and 44.8% higher mean gradient (p = 0.0002) than those in the normal %HMWM group. Thrombin formation was enhanced in the low %HMWM group (F1.2, 284.5 ± 63.7 vs 216.9 ± 62.5 pmol/l, p = 0.004; thrombin-antithrombin, 4.89 ± 1.3 vs 4.06 ± 0.9 µg/l, p = 0.02) and both markers showed inverse correlations with the percentage of vWF HMWM (r = -0.59, p = 0.002; r = -0.42, p = 0.03, respectively). In the low %HMWM group sCD40L (279.4 ± 60.7 vs 221.4 ± 41.7 pmol/l, p = 0.003) and ß-thromboglobulin (73.1 ± 9.2 vs 64.5 ± 8.5 IU/ml, p = 0.04), but not P-selectin, were also higher than in the remaining patients with AS. CONCLUSION: Patients with advanced AS deficient in vWF HMWM are characterised by enhanced thrombin formation and platelet activation. This observation indicates the ambivalent impact of high shear stress in AS on haemostasis and might help explain two aspects of AS-Heyde syndrome and increased risk of thromboembolism.

Cysteine peptidases and their inhibitors in breast and genital cancer.

Cysteine proteinases and their inhibitors probably play the main role in carcinogenesis and metastasis. The metastasis process need external proteolytic activities that pass several barriers which are membranous structures of the connective tissue which includes, the basement membrane of blood vessels. Activities of the proteinases are regulated by endogenous inhibitors and activators. The imbalance between cysteine proteinases and cystatins seems to be associated with an increase in metastatic potential in some tumors. It has also been reported that proteinase inhibitors, specific antibodies for these enzymes and inhibition of the urokinase receptor may prevent cancer cell invasion. Some proteinase inhibitor could serve as agents for cancer treatment.