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Glutamate-cysteine ligase (GCL) is a heterodimer enzyme composed of a catalytic subunit (GCLC) and a modifier subunit (GCLM). This enzyme catalyses the synthesis of γ-glutamylcysteine, a precursor of glutathione. cDNAs of the putative glutamate-cysteine ligase catalytic (Ace-GCLC) and modifier subunits (Ace-GCLM) of Ancylostoma ceylanicum were cloned using the RACE-PCR amplification method. The Ace-gclc and Ace-gclm cDNAs encode proteins with 655 and 254 amino acids and calculated molecular masses of 74.76 and 28.51kDa, respectively. The Ace-GCLC amino acid sequence shares about 70% identity and 80% sequence similarity with orthologs in Loa loa, Onchocerca volvulus, Brugia malayi, and Ascaris suum, whereas the Ace-GCLM amino acid sequence has only about 30% sequence identity and 50% similarity to homologous proteins in those species. Real-time PCR analysis of mRNA expression in L3, serum stimulated L3 and adult stages of A. ceylanicum showed the highest level of Ace-GCLC and Ace-GCLM expression occurred in adult worms. No differences were detected among adult hookworms harvested 21 and 35dpi indicating expression of Ace-gclc and Ace-gclm in adult worms is constant during the course of infection. Positive interaction between two subunits of glutamate-cysteine ligase was detected using the yeast two-hybrid system, and by specific enzymatic reaction. Ace-GCL is an intracellular enzyme and is not exposed to the host immune system. Thus, as expected, we did not detect IgG antibodies against Ace-GCLC or Ace-GCLM on days 21, 60 and 120 of A. ceylanicum infection in hamsters. Furthermore, vaccination with one or both antigens did not reduce worm burdens, and resulted in no improvement of clinical parameters (hematocrit and hemoglobin) of infected hamsters. Therefore, due to the significant role of the enzyme in parasite metabolism, our analyses raises hope for the development of a successful new drug against ancylostomiasis based on the specific GCL inhibitor., (Copyright © 2014 Elsevier B.V. All rights reserved.)

In a previous small series of surgically treated non-small cell lung cancer patients (NSCLC), we found that higher apoptotic index (AI) negatively influenced survival (Dworakowska D, Jassem E, Jassem J, Karmolinski A, Dworakowski R, Wirth T, et al. Clinical significance of apoptotic index in non-small cell lung cancer: correlation with p53, mdm2, pRb and p21WAF1/CIP1 protein expression. J Cancer Res Clin Oncol 2005; 131:617-623.). In this study we attempted to verify our previous finding in larger group of 170 NSCLC cases, additionally correlating AI to selected cell cycle regulators as well as a proliferation marker. Apoptosis was assessed with the use of the TUNEL technique, whereas the expression of p53, pRb, mdm2, p21(WAF1/CIP1), cyclin D1 and PCNA were assessed immunohistochemically. The mean and the median AI was 12 and 8, respectively. The expression of p53, pRb, mdm2, p21(WAF1/CIP1) proteins and cyclin D1 was found in 47%, 71%, 37%, 65% and 40% of cases, respectively. The mean and the median PCNA labeling index (PCNA LI) was 34 and 35, respectively. AI was not correlated with any patient characteristic or other tumor markers. In uni- and multivariate analysis AI, analysed separately or jointly with cell cycle regulators and PCNA LI, did not influence disease-free or over-all survival. However, patients with "very high AI/very high PCNA LI" had a particularly poor prognosis (P=0.001). Patients with "very low AI/negative pRb" phenotype survived for a shorter time in comparison to others (P=0.04). In addition, patients with the highest PCNA LI had a worse outcome in comparison to patients with the lowest PCNA LI (P=0.04), especially those with concomitant p53 protein expression (P=0.026) or lacking pRb protein expression (P=0.04). This study demonstrates that joint analysis of several factors involved in apoptosis, proliferation and cell cycle regulation, but not AI alone, might provide additional prognostic information in NSCLC patients.

A series of 25 rye (Secale cereale L.) inbred lines was tested with respect to three mitochondrial sequence-characterized amplified region (SCAR) polymorphisms. The analysis revealed a close association between the marker-determined mitotypes and plasmotypes (cytoplasm types known from breeding data) represented by the inbreds. The mitochondrial markers also confirmed normal (N-) cytoplasmic character of three wild rye species: Secale montanum, S. vavilovii and S. kuprijanovii. For 186 plants from open-pollinated cultivars of Turkish and South American origin, cytoplasm identification was performed with the use of crossing with double non-restoring tester. In 77 plants the normal (N) cytoplasm was detected, and for 63 of these the PCR analysis was performed producing results which were consistent with the genetic data based on testcrossing and phenotype assessment. The mitochondrial markers also confirmed a presence of sterility-inducing cytoplasm in the remaining 109 plants. Moreover, the markers allowed for differentiation between Pampa (P-) and Vavilovii (V-) cytoplasmic individuals. For 60 plants the latter results were verified using crosses with a line maintaining P-cytoplasmic sterility and acting as a restorer of the V-cytoplasm. For two of these plants contradicting results were produced with the applied methods of cytoplasm identification and the basis of this discrepancy is discussed. Regardless of the identification method, widespread occurrence of a sterility-inducing cytoplasm was revealed, especially in South American populations.

Neoadjuvant chemotherapy before resection is being the standard of care for stage IIIA non-small cell lung cancer in many institutions. The risk of complications in patients undergoing thoracotomy after induction chemotherapy remain controversial. We reviewed our experience. From 1998 to 2003, 29 patients underwent pulmonary resection after induction chemotherapy for advanced non-small cell lung cancer. Pneumonectomies were performed for 16 (55.2%) patients (2 right sleeve pneumonectomy and 1 pneumonectomy with wedge excision of tracheal carina), lobectomies for 11 (37.9%) patients (3 right upper sleeve lobectomy), segmentectomies for 1 (3.45%) patient and explorative thoracotomy for 1 (3.45%) patient. There were 3 (10.3%) postoperative deaths, all after right pneumonectomy; 2 caused by pneumonia of the left lung, 1 caused by pulmonary embolism in patient after re-thoracotomy for hemothorax. The postoperative complications included pneumonia in 2 patients, postoperative bleeding in 2, hemothorax in 1, prolonged intubation in 1, vocal cord paralysis in 2, cardiac arrhythmia in 2, atelectasis in 1 and residual air space in 1, resulting in 41,4% morbidity. Most of complications occurred after right pneumonectomy (45.5%). The mortality of patients who had received induction chemotherapy was higher than that of a comparative group of 1529 who underwent lung resection or only exploration without induction chemotherapy during the same period, and the difference was significant (10.3% vs 4.1%; p = 0.01). Morbidity differences were. not significant (p = 0.94).

Hemorrheological and humoral abnormalities and excessive platelet activity can predict the development of cardiovascular complications in patients with essential hypertension. A study was conducted to assess the influence of gender on these factors and the interrelations between changes in hemorrheology and the sympatho-adrenal system in 54 patients (18 women, 36 men) with essential hypertension (aged 39.6 +/- 9.7 years) and 25 healthy volunteers (10 women, 15 men; aged 36.0 +/- 7.2 years). A decrease in erythrocyte deformability (p < 0.01) was found in the hypertensive men compared with the hypertensive women. Hematocrit (p < 0.01), blood viscosity at the shear rates of 0.3 s-1 (p < 0.01) and 6 s-1 (p < 0.01), plasma viscosity (p < 0.01), erythrocyte aggregation (p < 0.01), and neuropeptide Y (p < 0.02) concentrations were higher in the hypertensive men than in the hypertensive women. A positive correlation between blood fibrinogen and serotonin was found in the pooled hypertensive group and in the hypertensive men (p < 0.01) and between blood viscosity (shear rate 6 s-1) and neuropeptide Y in the pooled hypertensive group (p < 0.01). Neuropeptide Y correlated with filtration time of 1 mL blood in the hypertensive men (p < 0.05) and in the pooled normotensive group (p < 0.01) and with beta-thromboglobulin in the hypertensive women (p < 0.001). A positive correlation was also found in the hypertensive men between erythrocyte and platelet aggregation (p < 0.01) and between beta-thromboglobulin and adrenaline (p < 0.01). Hemorrheological and humoral abnormalities are more pronounced in men than in women with essential hypertension and may contribute to the increased incidence of cardiovascular events in men.

A case of pheochromocytoma in 81 year old woman hospitalized due to choledocholithiasis is reported. During premedication pronounced blood pressure elevation was observed. Operation was discontinued and diagnostic of pheochromocytoma was carried out. USG and CT scanning revealed right sided adrenal tumor. Elevated urine catecholamines and methoxycatecholamines were found. The patient underwent adrenalectomy and cholecystectomy. Histological examination confirmed diagnosis of pheochromocytoma. The patients done uneventful recovery.

Objective of the study was assessment of the usefulness of determination of noradrenaline (NA) and adrenaline (A) in urine and blood as well as the total methoxycatecholamines (MNA +MA), vanillylmandelic acid (VMA), DOPA and dopamine (DA) urinary excretion in diagnosis of pheochromocytoma. The experience based on 155 patients with pheochromocytoma (105F, 50M, age 18-82 yrs) diagnosed in the Department of Hypertension and Angiology Academy of Medicine in Warsaw will be discussed. In all patients excluding 2 cases pheochromocytoma has been proven histopathologically. The most considerable diagnostic usefulness of MNA + MA indication was proven. MNA + MA was increased in 96.6 patients. In 89.6% patients an increased excretion of NA and A or one of this catecholamines was demonstrated. An increased excretion of VMA was demonstrated in 75%. The excretion of DOPA and dopamine was tested in 120 cases. An increased excretion of DA was shown in 31% and DOPA in 16%.

The purpose of our study was to assess the potential role of spin-echo (SE), chemical shift, and gadolinium-enhanced magnetic resonance imaging (MRI) in the differentiation of adrenal masses. Seventy-two adrenal masses (26 nonhyperfunctioning adenomas, 16 aldosterone-secreting adenomas and 6 other different benign cortical masses, 18 pheochromocytomas, and 6 malignant masses) in 63 patients were evaluated with spin-echo sequences, chemical shift imaging (CSI) and gadolinium diethylenetriamine penta-acetic acid (Gd-DTPA) dynamic studies. Ratios and indices of signal intensity for all examined MRI methods were calculated and examined for significance of difference between different types of adrenal masses. Quantitative magnetic resonance evaluation of adrenal masses showed significant differences (at least alpha < 0.01) between nonhyperfunctioning adenomas vs. pheochromocytomas or vs. malignant lesions or vs. aldosterone-secreting adenomas and between pheochromocytomas vs. malignant lesions. The most specific indicators of adrenal mass character proved to be the CSI ratio based on opposed-phase and in-phase two-dimensional fast low-angle shot (FLASH) images, reflecting lipid content in the lesion, and Gd-DTPA dynamic studies ratios reflecting contrast agent inflow and washout in the lesion: WoMAX/LAST and Dyn1.2-3.2. There was no overlap of CSI ratio between adenomas and pheochromocytomas. The overlap of ranges of CSI ratio between nonhyperfunctioning adenomas and aldosterone-secreting adenomas was only 18.5%. There was no overlap of WoMAX/LAST ratio between adenomas and pheochromocytomas, or adenomas and malignant lesions. The overlap of ranges of Dyn1.2-3.2 ratio between pheochromocytomas and malignant lesions was only 17.6%. MRI enables good visualization and specific characterization of adrenal masses. The optimal MRI protocol for the adrenal region is presented.

A case of a pheochromocytoma with haemorrhagic necrosis resulting in signs of acute abdomen with shock is reported. Diagnosis of a ruptured adrenal tumor was made by CT scanning. Elevated urine and plasma catecholamines as well as histological examination of the removed tumor confirmed the clinical suspicion of pheochromocytoma. The patient made uneventful recovery and is asymptomatic 2 years after surgery. Ruptured pheochromocytoma as a cause of abdominal emergency is discussed in view of the existing literature.

Investigated group consisted of 28 patients (21 female and 7 male) with pheochromocytoma in mean age of 39.9 +/- 13 years, while control group comprised 25 healthy volunteers in mean age 36.0 +/- 7.2 years. Following parameters were determined: plasma and blood viscosity, erythrocyte and platelets aggregation, erythrocyte deformability, hematocrit, beta-thromboglobulin plasma concentration, plasma fibrinogen concentration, lipids concentration, noradrenaline, adrenaline, serotonin, neuropeptide Y plasma concentrations. Plasma fibrinogen and beta-thromboglobulin were increased in pheochromocytoma before surgery after pharmacological treatment and also after excision of the tumor. Whereas altered erythrocyte deformability before any treatment, only. Plasma neuropeptide Y concentration was higher in pheochromocytoma before surgery and after pharmacological treatment. There was no correlation between plasma catecholamines, plasma fibrinogen, beta-thromboglobulin, and erythrocyte deformability. Our results suggest that rheological abnormalities observed in patients with pheochromocytoma may contribute to vascular complications in this subjects. Sympathetic nervous system seems to influence indirectly observed rheological abnormalities.

The aim of the study was to investigate endothelin-1 (ET-1) and neuropeptide Y (NPY) plasma concentrations in renal venous blood of hypertensive patients with unilateral renal artery stenosis (URAS). The study was performed in 22 patients with URAS and 18 patients diagnosed as essentially hypertensive. In each subject renal arteriography and renal vein catheterisation was performed. Blood samples for ET-1, NPY and plasma renin activity (PRA) were withdrawn from renal veins and vena cava inferior, and for ET-1 and NPY from the aorta. Patients with URAS were divided in two subgroups according to the renal vein renin ratio. Both in nine patients with URAS and ratio > 1.5 and in 13 patients with URAS and ratio < 1.5, ET-1 and NPY plasma concentrations evaluated in renal venous blood of the ischaemic kidney were not different from those assessed in the contralateral side, in vena cava inferior and in the aorta. In essential hypertension, the mean ET-1 and NPY plasma concentrations of both renal veins were not different from the ET-1 and NPY plasma values assessed in renal vein of stenosed and contralateral side, vena cava and aorta of patients with URAS with and without activation of the renin system. Our study indicates that chronic ischaemia does not affect ET-1 and NPY plasma concentrations in renal venous blood of hypertensive URAS patients both with and without activation of the renin system.

Background: Atrial natriuretic peptide (ANP) has been extensively studied in cardiovascular disorders in recent years. Particular attention has been paid to the role of ANP in the pathogenesis of hypertension and congestive heart failure and in the neurohormonal response to myocardial infarction. However, no published data are available on the significance of ANP in hypertensive patients with acute myocardial infarction., Methods: We studied the relationship between plasma ANP concentration and systolic blood pressure, diastolic blood pressure, left atrial dimension, left ventricular diastolic dimension and left ventricular mass index in patients with essential hypertension in the acute phase of myocardial infarction. Plasma ANP concentrations were determined at 0, 4, 8, 16, 24, 48 and 72 h after admission in 40 patients with a first myocardial infarction (18 hypertensive patients, group 1; 22 normotensive patients, group 2). Left atrial dimension and left ventricular diastolic dimension were assessed echocardiographically within the first 48 h of acute myocardial infarction., Results: Maximum and mean plasma ANP values at 0, 4, 8, 16, 24, 48, and 72 h as well as mean ANP concentrations within the 72 h period were higher in group 1 than in group 2 (P < 0.001). Plasma ANP concentration, left atrial dimension (r = 0.59) and left ventricular diastolic dimension (r = 0.56) were positively correlated in both groups., Conclusion: Acute myocardial infarction is characterized by a more pronounced rise in plasma ANP concentration in hypertensive patients than in those without a history of hypertension. Plasma ANP concentration correlates with left heart chamber sizes.

Background: Endothelin-1 (ET-1) may play an important role in the development and maintenance of hypertensive states. In patients with essential hypertension, ET-1 plasma concentration increases or remains unchanged. The aim of the present study was to investigate ET-1 plasma concentration in patients with mild-to-moderate essential hypertension and its interrelationship with catecholamines, neuropeptide Y and atrial natriuretic peptide (ANP)., Methods: The study included 37 patients (mean age 38.2 +/- 1.6 years) with mild-to-moderate essential hypertension. The control group consisted of 21 healthy volunteers (mean age 35.6 +/- 1.4 years). ET-1, neuropeptide Y and ANP were determined by radioimmunoassay methods and catecholamine plasma concentration was determined radioenzymatically., Results: Our study shows that plasma ET-1 and neuropeptide Y levels are elevated in patients with essential hypertension compared with a control group. No correlation was found in either of the groups between plasma ET-1 level and plasma neuropeptide Y, catecholamine or ANP concentrations., Conclusion: Our results suggest that ET-1 is relevant in the development and maintenance of elevated blood pressure.

The aim of the study was to investigate to what extent inhibition of nitric oxide (NO) formation, cyclooxygenase and converting enzyme activities and vasopressin V1 receptors blockade affects the cardiovascular system in conscious, freely moving normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats. The experiments were performed on 33 WKY and on 33 SHR. In series 1 (8 WKY and 8 SHR) animals received bolus injection of N omega-nitro-L-arginine - NLA (10 mg/kg), in series 2 (6 WKY and 6 SHR) bolus injection of indomethacin (10 mg/kg). In series 3 (8 WKY and 8 SHR) the animals received captopril as initial bolus (1 mg/kg) followed by constant infusion (1 mg/kg/min), in series 4 (11 WKY and 11 SHR) vasopressin V1 receptor antagonist 1-(1-mercapto-4-methylcyclohexaneacetic acid)-8-arginine-vasopressin (MeCAAVP) was infused (1.52 micrograms/kg/min). In series 1 in WKY NLA elicited a long-lasting, significant increase in mean blood pressure (max 46 +/- 3 mmHg at 40 min). In SHR mean blood pressure raises were not significant (max 22 +/- 6 mmHg). In series 2, both in WKY and SHR, indomethacin elicited only transient, nonsignificant increases in mean blood pressure. In series 3, the mean blood pressure fall during the captopril infusion was more pronounced in SHR than in WKY (45 +/- 2 mmHg vs 13 +/- 2 mmHg respectively). In series 4 vasopressin V1 receptor blockade caused a nonsignificant fall in mean blood pressure, both in WKY and SHR.(ABSTRACT TRUNCATED AT 250 WORDS)

Three patients with giant cell arteritis (GCA) observed recently in our department are described. One of them demonstrated clinical picture of polymyalgia rheumatica, the other two presented with clinical symptoms of GCA. This presentation seems to be very helpful for correct diagnosis in cases with difficult recognition.

The aim of the study was to investigate blood pressure responses and changes in heart rate after bolus administration of endothelin-3 [ET-3] in conscious, freely moving SHR and WKY rats and DOCA-salt hypertensive and normotensive Wistar rats. The effect of ET-3 on blood pressure and heart rats was investigated for four doses equal to 250 ng, 500 ng, 1000 ng and 2000 ng of ET-3. Our study shows that in experimental models of hypertension changes in blood pressure were predominantly characterized by the pronounced hypotensive phase with no significant raises in blood pressure. In normotensive animals cardiovascular responses to ET-3 were biphasic, with initial depressor phase followed by long-lasting, significant pressor effect.

The interaction between catecholamines (CA) and ANP is not clearly established. The effects of excess endogenous CA on ANP secretion can be investigated in patients with pheochromocytoma. We studied 27 patients with surgically and histologically proven pheochromocytoma (P) aged 19-70 years. In 16 of these patients plasma ANP study was repeated after surgical removal of the tumour. The control group (C) consisted of 20 healthy volunteers aged 21-48 years. Moreover, 42 patients with uncomplicated mild to moderate essential hypertension (EH) aged 18-48 years were also studied. In P higher plasma ANP concentration versus C, EH was found (51.9 +/- 8.1; 25.5 +/- 1.5; 19.3 +/- 1.5 fmol/ml, respectively). In 16 patients with P, increased plasma ANP level (mean 63.3 +/- 12.6 fmol/ml) declined after surgical removal of the tumour (mean 22.4 +/- 2.9 fmol/ml). In the P patients no relationship was found between plasma ANP and hormonal patterns of the tumour or between plasma ANP and plasma catecholamines, whereas significant positive correlations between plasma ANP and both systolic and diastolic blood pressure and heart rate were demonstrated. These results suggest that excess CA produced by the chromaffin tumour induce ANP secretion via stimulation of adrenergic receptors. However, influence of the haemodynamic changes evoked by CA cannot be excluded. It is suggested that increased secretion of ANP may be of some importance in maintaining blood pressure homeostasis in patients with pheochromocytoma.

We evaluated the effect of captopril treatment on left ventricular mass (LVM) and function in sustained mild-to-moderate primary hypertension. In 16 men aged 34.5 +/- 6.4 yrs. M-mode echo and pulsed wave Doppler recordings were obtained prior to and after 2 months captopril treatment (50-150 mg daily). No significant changes in heart rate, systolic blood pressure, ejection fraction, fractional shortening, cardiac output, LVM and Doppler-derived indices of mitral inflow were observed. Diastolic blood pressure measured during examination fell from 102.2 +/- 13.3 to 90.3 +/- 9 mmHg (p less than 0.026). No consistent correlations between catecholamines and indexes of left ventricular function were found. Two-month captopril therapy in mild-to-moderate hypertension results in diastolic blood pressure lowering with no effect on LVM or left ventricular function.

The aim of the study was to find out whether vasopressin (AVP) modifies hypotensive and heart rate accelerating effects of atrial natriuretic peptide (ANP) in normotensive (WKY) and spontaneously hypertensive (SHR) conscious rats. The effect of i.v. administration of 1; 2 and 4 micrograms of ANP on blood pressure (MP) and heart rate (HR) was compared during i.v. infusion of 0.9% NaCl (NaCl), NaCl+AVP (1.2 ng kg-1 min-1) and NaCl+dEt2AVP (V1 receptors antagonist, 0.5 microgram kg-1 min-1). AVP increased MP in SHR and WKY and decreased HR in SHR. V1 antagonist decreased MP and increased HR only in SHR. In SHR ANP decreased MP and increased HR during NaCl, AVP and V1 antagonist infusion. In WKY these effects were observed only during AVP administration. In each experimental situation hypotension and tachycardia induced by ANP were greater in SHR than in WKY. In both strains ANP induced changes in MP and HR were enhanced during AVP in comparison to NaCl infusion. V1 antagonist did not modify effects of ANP in WKY and SHR. The results indicate that ANP abolishes hypertensive response induced by blood AVP elevation and that the basal levels of endogenous vasopressin acting through V1 receptors does not interfere with hypotensive action of ANP neither in WKY nor in SHR.

The aim of the present study was to compare effects of intravenous infusion of vasopressin AVP and V1 receptors blockade on blood pressure and heart rate in normotensive (WKY) and spontaneously hypertensive (SHR) rats. A 20 min vasopressin infusion (1.2 ng/kg/min) elicited significantly greater increase in mean blood pressure (MP) in SHR than in WKY. Heart rate was significantly reduced in SHR while nonsignificantly in WKY. A 20 min dEt2 AVP (V1 antagonist) infusion (0.5 microgram/kg/min) elicited significant decrease in MP and increase in heart rate (HR) in SHR, but produced no effect in WKY. The data indicate that SHR are more susceptible to pressor and hypotensive effects of sustained elevation of AVP and AVP antagonist. The results support the hypothesis that AVP may contribute to pathogenesis of hypertension.