The localization of mycobacterial 17beta-hydroxysteroid dehydrogenase (17beta-OH SDH) was studied using cell fractionation and cytochemical investigation. Mycobacterium sp. Et1 mutant strain derived from Mycobacterium sp. VKM Ac-1815D and characterized by increased 17beta-OH SDH activity was used as a model organism. Subcellular distribution study showed both soluble and membrane-bound forms of mycobacterial 17beta-hydroxysteroid dehydrogenase. The cytochemical method based on a copper ferrocyanide procedure followed by electron microscopic visualization was applied in order to investigate the intracellular localization of bacterial 17beta-OH SDH in more detail. The enzyme was found to be located in the peripheral cytoplasmic zone adjoining the cytoplasmic membrane (CM). 17beta-OH SDH was loosely membrane bound and easily released into the environment under the cell integrity failure.

Segmental renal scarring occurs in experimental obstructive uropathy in the multipapillary porcine kidney, and segmental abnormalities in renal perfusion are likely to be responsible. This preliminary study examines the urinary excretion of the potent locally active vasoconstrictor endothelin 1 (ET 1) in a pig model of renal obstruction and subsequent relief. Significant urinary excretion of ET 1 from the postobstructive kidney was found to occur after longstanding obstruction. Preglomerular arteriolar stenosis may be the cause of the renal ischaemia in obstruction that is at first reversible but later becomes irreversible if the stimulus persists. ET 1 may be implicated in the pathogenesis of this injury.

In the present study we characterized a "crosstalk" mechanism between transforming growth factor beta-1 (TGF beta-1) and endothelin-1 (ET1) signaling pathways in neonatal cardiac myocytes. A 5 minute pretreatment with 1 ng/ml concentrations of TGF beta-1 attenuated ET1-induced negative chronotropic effects and translocation of the alpha, delta and varepsilonPKC isozymes to the particulate cell fraction. We found no effect of TGF beta-1 on responses induced by the P(2) purinergic agonist ATP or phorbol ester. Treatment of cardiac myocytes with acidic fibroblast growth factor (aFGF) did not alter ET1- or ATP-mediated effects on contraction rate or translocation of PKC isozymes to the particulate fraction. Our studies suggest that TGF beta-1 may act as a negative modulator of ET1- but not ATP- or phorbol ester-induced PKC isozyme signaling events in neonatal cardiac myocytes. A better understanding of the complex ET1 and TGF beta-1 signaling mechanisms in neonatal heart cells should enhance our knowledge regarding the interplay between these pathways.

To assess if cold-induced vasoconstriction may persist during exercise and contribute to the development of myocardial ischaemia, we studied 11 patients with exertional angina and angiographically proven coronary artery disease, in all cases involving the proximal portion of the left anterior descending artery. Great cardiac vein flow (GCVF) was measured by the thermodilution technique and the coronary resistance of the abnormally perfused anterior region (ARCR) was calculated as the quotient of mean arterial pressure and GCVF. All patients performed a supine bicycle exercise test (ET1) until angina occurred. After recovery, they underwent a cold pressor test (CPT) and then performed a second exercise test (ET2) while cold stimulation was continued. During ET1, ARCR decreased (from 1.53 +/- 0.43 to 1.04 +/- 0.35 mmHg ml-1 min-1, P less than 0.001) as a result of the metabolic vasodilation, while it rose, although non significantly, during CPT despite the increase in double product (P less than 0.001), reflecting the augmented myocardial oxygen consumption. However, such abnormal response to CPT did not persist during ET2, because ARCR decreased to a value non significantly different from that achieved at peak ET1. In five patients, who showed a reduced exercise tolerance during ET2, ARCR dropped by 22% during ET2 compared with 34% decrease during ET1. However, such a difference was unlikely to account for the reduction in exercise tolerance, because the double product at peak ET2 was never lower than the corresponding value at peak ET1. Our data show that the inappropriate vasoconstriction induced by CPT in an abnormally perfused myocardial region does not persist during exercise, when metabolic vasodilation occurs.(ABSTRACT TRUNCATED AT 250 WORDS)

Endothelin (ET) acts at selected brain loci to elicit a pressor response and vasopressin (AVP) secretion. The pressor action of centrally acting ET is mediated via enhanced efferent sympathetic nerve activity. ET-induced VP secretion depends upon the ET receptor subtype and the brain region involved. ET(A)R activation at the subfornical organ (SFO) increases mean arterial pressure and renal sympathetic nerve activity (RSNA) as well as AVP secretion in awake rats. These effects are only partly mediated by glutamatergic receptors in paraventricular nucleus (PVN). Recent data indicate dendritic release of AVP may act as a neurotransmitter. We therefore hypothesized that dendritic release of AVP from magnocellular PVN neurons contributes to the increase in arterial pressure and RSNA due to ET(A) receptor activation at SFO. Male Sprague Dawley rats equipped with vascular catheters, renal nerve electrodes, and intracerebral cannulae directed into SFO and magnocellular PVN bilaterally were studied 48hr after recovery in the awake state. Hemodynamic and neural parameters were monitored continuously. Microinjection of 5 pmol ET1 into SFO increased mean arterial pressure by 15.8 +/- 4.2 mmHg accompanied by reflex decreases in heart rate and RSNA. Microinjection of 100 ng of the V(1a) receptor antagonist alone bilaterally into the PVN did not change baseline parameters; however, the pressor response to ET1 was significantly attenuated with mean arterial pressure increasing only by 6.1 +/- 3.0 mmHg (P<0.05). Reflex changes in heart rate and RSNA did not change. These findings support the concept that dendritic release of VP from magnocellular neurons within the PVN mediates, at least in part, the pressor response to ET(A) receptor activation at the SFO.

BACKGROUND

The need for transvenous lead extraction procedures of coronary sinus (CS) leads is increasing due to rising numbers of implanted cardiac resynchronization therapy devices during the past decade.

METHODS

From January 2009 to June 2013, 27 CS leads were scheduled for extraction in 27 patients (mean age (63.1 ± 14.6) years). Indications for lead extraction were infection in 13 and lead dysfunction in 14 cases. Isolated extraction of CS leads was performed in eight, extraction of multiple leads in 19 cases. Among leads with an implant time of ≥ 12 months (n = 19) mean implant duration (MID) was (46.4 ± 15.2) (12-76) months. Groups were formed depending on infectious or non-infectious indications (INF vs. Non-INF), and the use or non-use of extraction tools (ET1 vs. ET0).

RESULTS

Among patients with an implant duration of ≥ 12 months, complete procedural success was 94.7% and clinical success 100%. Operative mortality was zero. In the INF versus NON-INF groups complete procedural success (100% vs. 91.7%, P = 0.43), mean number of required extraction tools (0.7 (0-2) vs. 0.9 (0-3), P = 0.65) and MID (49.1 ± 15.0 vs. 44.7 ± 15.8, P = 0.83) did not differ significantly. Comparing the groups ET1 and ET0 showed no significant differences in complications (n = 1 vs. n = 1, P = 0.81) and MID (47.0 ± 17.5 vs. 45.5 ± 12.6, P = 0.71).

CONCLUSIONS

In specialized centers transvenous lead extraction of coronary sinus leads with a mean implant duration of almost four years can be performed safely and effectively. Neither non-infectious indications nor the use of extraction tools negatively affected the outcome of the procedure.

Amino-terminal fragments of PTHrP were previously shown to increase regional blood flow in laboratory animals. Since PTHrP is produced in the lactating mammary gland and associated nutrient vessels, we examined the effects of peptide fragments of PTHrP on the hemodynamics of the mammary gland of dried sheep. The left arterial mammary blood flow measured using ultrasonic flow probes in four dried Lacaune ewes was 233 +/- 11 ml/minute. It was significantly increased when synthetic human PTHrP-(1-34) or (1-86) fragments were injected into the mammary artery. The effect was dose dependent for PTHrP-(1-34), varying between 0.0075 and 0.3 nmol/kg body weight. PTHrP-(140-173) fragment lacked any vasorelaxant activity. Synthetic human endothelin (ET1) decreased arterial blood flow in a dose-dependent manner. This decrease was inhibited by PTHrP-(1-34), and this inhibition was PTHrP dose related. When ET1 (10 pmol/kg body weight) was injected together with PTHrP-(1-86) (100 pmol/kg body weight), only a significant increase in mammary blood flow was observed. Thus, PTHrP produced by the lactating mammary gland may be involved in the regulation of mammary blood flow.

OBJECTIVE

To use multilocus enzyme electrophoresis to determine the genetic structure of Staphylococcus intermedius from normal skin of dogs and those isolated from a variety of disease conditions and to distinguish clinically important strains in dogs.

METHODS

The diversity amongst 129 isolates of S intermedius from the skin and mucosa of 32 healthy dogs and 120 isolates from diseased sites in 120 individual dogs was examined using multilocus enzyme electrophoresis. Associations among ETs were examined to determine the diversity of isolates.

RESULTS

Twenty two ETs were distinguished comprising 21 containing isolates from diseased sites and 11 containing isolates from normal dogs. The majority of isolates (171 of 249; 69% were located in two ETs (ET1 and ET 4), that were not distinguishable phenotypically. ET 1 contained 94 isolates (54 isolates from healthy dogs and 40 isolates from diseased sites) and ET 4 contained 77 isolates (46 from healthy dogs and 31 isolates from diseased sites). Further, 77.5% of isolates from healthy dogs were present in ET 1 and ET 4 and 59% of isolates from diseased dogs belonged to the same two ETs. There was only a small difference in genetic diversity among isolates taken from healthy dogs (11 ETs; H = 0.182) and those isolates taken from clinical specimens from diseased dogs (21 ETs; H = 0.218). Of the 21 ETs from diseased sites, ET 16 contained all six isolates from Staphylococcal Scalded Skin Syndrome in racing Greyhounds.

CONCLUSIONS

The small difference in genetic diversity between isolates from the skin and mucosa of healthy dogs and isolates from various diseases, as well as the presence of the majority of isolates in two ETs, is consistent with the role of S intermedius as an opportunistic pathogen. The confinement of all Staphylococcal Scalded Skin Syndrome isolates within one ET is confirmation of this entity as a distinct disease of dogs.

The vasoconstrictor peptide endothelin-1 (ET1) has only recently been characterized and its effects are at present largely speculative. It has been hypothesized that ET1 acts on mesangial cells to cause vasoactive changes which might ultimately contribute to the development of glomerulosclerosis. Opposite to ET1, nitric oxide (NO) inhibits mesangial cell contraction and proliferation. NO activates soluble guanylic acid cyclase and the final product, cyclic GMP (cGMP), has been recently used as a marker of NO action. Urinary levels of ET1 and cGMP were detected in 58 patients with biopsy-proven glomerulonephritis (GN), including 36 IgA nephropathy (IgAGN), 30 with normal and 6 with impaired renal function, 10 patients with non-IgA mesangial GN and 12 pts with membranous GN (MGN) with normal renal function. Compared to normal controls (0.019 +/- 0.006 ng/min), urine ET1 levels were significantly higher in patients with normal renal function having IgAGN (0.035 +/- 0.017, p < 0.01), MGN (0.028 +/- 0.013, p < 0.05), non-IgA mesangial GN (0.027 +/- 0.012, p < 0.05) and those with IgAGN and renal failure (0.032 +/- 0.011, p < 0.01). However no difference was found between MGN patients and normals by deleting MGN cases with mild to moderate mesangial proliferation. The mean value of urinary cGMP in IgAGN patients with renal failure (0.186 +/- 0.117 nmol/min) was lower (p < 0.05) than that of each group with normal renal function (IgAGN: 0.378 +/- 0.010 nM/min; MGN: 0.338 +/- 0.064 nmol/min, non-IgAGN: 0.436 +/- 0.168 nmol/min). The same significant differences were obtained by correcting cGMP values for creatinine urinary excretion. Urinary ET/cGMP ratio (assumed as an index of the relative balance between vasoconstrictor and vasorelaxing factors) was found to be higher than normal (0.570 +/- 0.010 ng/nmol) both in IgAGN patients with normal renal function (0.103 +/- 0.064 ng/mol, p < 0.05), and in those with renal failure (0.203 +/- 0.108 ng/nmol, p < 0.02). Urinary cGMP values were not related to plasma levels of atrial natriuretic peptide (ANP). These data show that hyperexcretion of ET1 occurs in a number of patients with mesangial proliferative GN. In some of them, mainly those with established glomerular damage, the local production of ET1 is not counter-balanced by adequate cGMP biosynthesis.

BACKGROUND

Precise control of transgene expression is essential for a variety of applications ranging from gene-function analysis, biopharmaceutical manufacturing to next-generation molecular interventions in gene therapy and tissue engineering. The regulation of gene expression is currently a key issue for clinical implementation of gene-therapy-based treatments since desired transgene expression may need to be maintained within a narrow therapeutic window for successful treatment of a particular human disease.

METHODS

We have designed a novel bidirectional expression module that enables adjustable coregulation of two different transgenes in response to clinical doses of macrolide antibiotics. A bidirectional macrolide-responsive promoter consisting of a central operator module (ETR) specific for the macrolide-dependent transactivator (ET1) is flanked by two minimal promoters (P(hCMVmin); P(hsp70min)) which drive expression of two divergently oriented transgenes. Macrolide antibiotics modulate the binding affinity of ET1 to ETR and adjust expression of both transgenes to desired levels.

RESULTS

Bidirectional expression configurations enabled excellent macrolide-adjustable coregulation profiles of two secreted reporter genes or one-vector-based autoregulated fine-tuning of a single transgene in various transgenic rodent and human cell lines. Following implantation of microencapsulated CHO-K1 cell derivatives transgenic for macrolide-controlled bidirectional expression of erythropoietin (EPO) and the human secreted alkaline phosphatase (SEAP) intraperitoneally into mice, serum EPO and SEAP levels could be coadjusted to desired levels by administration of different erythromycin doses.

CONCLUSIONS

Based on their in vivo compatibility, the versatile bidirectional and macrolide-responsive expression modules represent an important advancement on the way to implementing targeted and conditional molecular interventions into a clinical reality.

It has been shown in vitro that endothelin 1 (ET1) differentially affects the human myometrial contractility according to the hormonal profile of women. Our purpose was to test the hypothesis that ovarian steroids influence the ET1 induced myometrial contractility. We performed three types of experiments. Myometrial tissues were exposed to 17beta-oestradiol (E), progesterone (P), E plus P (E+P) in concentrations 10(-10)M, 10(-8)M, 10(-7)M, 10(-6)M and 10(-4)M (Type I), ET1 in concentrations 10(-10)M, 10(-9)M, 10(-8)M, 10(-7)M and 10(-6)M (Type II) and E+ET1, P+ET1 and E+P+ET1 in concentrations ranging from 10(-10)M to 10(-6)M (Type III). Tissue exposure to E, P and E+P did not significantly alter the pattern of spontaneous myometrial motility. ET1 (10(-6)M) induced a sustained long-lasting contraction, the initial part of which lasted 34 +/- 4 min, elevating the initial baseline by 190 +/- 20%. This was followed by ripples of gradually increasing amplitude with no regular contractions up to the end of the period of observation (120 min). Addition of P or E+P to ET1 markedly restricted (p<0.05) the elevation of initial baseline (P+ET1: 68 +/- 8%, P+E+ET1: 67 +/- 8%), and significantly shortened (p<0.01) the duration of the alterations (P+ET1: 21 +/- 3 min, P+E+ET1: 26 +/- 3 min). These results demonstrate the lack of any significant effect of E and P or their combinations on the pattern of spontaneous myometrial motility in post-menopausal women. However, P alone or in combination with E exerted an inhibitory action on ET1 -induced contractility on human post-menopausal myometrium. The physiological significance of these findings remains to be clarified.

Endothelin-1 (Et1), like angiotensin II, is implicated in postnatal maturation and development. The present study was designed to identify Et1 receptors and subtype Et1 receptors present in rat kidney between 1 and 30 days of postnatal life. On day 1, high-affinity and high-density Et1 binding sites were identified in rat kidney. The dissociation constant and maximum binding for ET1 to membranes from whole kidney were 0.073 +/- 0.05 nM and 1,345.9 +/- 73 fmol/mg protein, respectively. On day 30, affinity and receptor density were markedly decreased. The dissociation constant and maximum binding were 0.147 +/- 0.021 nM (P < 0.01) and 633.2 +/- 56.4 fmol/mg protein (P < 0.001), respectively. Using BQ 123 (EtA-selective antagonist) and sarafotoxin S6c (EtB-selective agonist), the two Et1 receptor subtypes EtA and EtB were identified in 1- and 30-day-old rat kidney. BQ 123 selectively recognized EtA receptors with high affinity (2.9 +/- 0.44 on day 1 and 4.0 +/- 0.5 nM on day 30) and sarafotoxin S6c bound with higher affinity EtB receptors (0.871 +/- 0.14 on day 1 and 0.717 +/- 0.12 nM on day 30). Between birth and day 30, the EtA binding capacity was decreased (304 +/- 27 vs. 752 +/- 202 fmol/mg protein, P < 0.05), whereas EtB binding was not affected (514 +/- 87 vs. 656 +/- 171 fmol/mg protein, NS). The decrease in the total number of Et1 receptors during the 1st month of life may be due to the concomitant decrease in the number of EtA receptors. Increased Et1 receptor density in early postnatal life suggests an influence of Et1 on immature kidney circulation and/or kidney growth.

BACKGROUND

Prolonged incubation of porcine isolated coronary artery (PCA) to lipopolysaccharide (LPS) causes a moderate reduction in vessel constrictive responsiveness. This has been attributed mainly to the induction of nitric oxide synthase (NOS). We aimed to investigate the role of induction of cyclo-oxygenase (COX) and expression of endothelin receptor 1-A (ET1(A)) in modulating the vascular responses of PCA in vitro.

METHODS

Segments of PCA were exposed to 100 microg ml(-1) LPS overnight. L-Arginine 0.4 mM was included in the medium in some preparations to examine the influence of intracellular nitric oxide, and the influence of extracellular donor sodium nitroprusside (SNP) was also examined in separate experiments. After overnight incubation, the contractile function of the artery was evaluated by the isometric tension recording test. The non-selective NOS inhibitor (L-NAME), non-selective COX inhibitor (indomethacin), COX-1 inhibitor (FR 122047), COX-2 inhibitor (NS 398), and ET1(A) receptor antagonist (FR 139317) were added into the organ bath 30 min before eliciting contractile responses to KCl or U46619 separately or in combinations. Vascular relaxations to 10 nM Substance P (SP) were also assessed.

RESULTS

L-Arginine did not potentiate the effects of LPS. SNP caused a quantitatively larger reduction in the responsiveness to KCl and U46619 compared with 100 microg ml(-1) LPS. Post exposure to a combination of indomethacin and FR 139317, indomethacin or NS 398 alone enhanced the inhibitory effects of LPS, but FR 122047 or FR 139317 alone failed to modify the responses to LPS. L-NAME fully reversed the changes induced by LPS combined with indomethacin and NS398. In terms of the relaxation by SP, LPS failed to change the magnitude; none of the agents used affected the response except L-NAME which abolished it.

CONCLUSIONS

NOS and COX-2 are both activated by overnight exposure to LPS in vascular smooth muscle from PCA in vitro. The prostanoid produced by COX-2 functionally antagonizes the effects of induction of NOS.

This study was conducted to evaluate the responsiveness of human nonpregnant myometrium to endothelin 1 (ET1) (10(-10) M-10(-6 )M) and KCl (80 mM) in relation to the hormonal profile of the women, who were allocated into three groups: group 1, premenopausal follicular phase, n=14, group 2, premenopausal luteal phase, n=20, and group 3, postmenopausal women, n=12. At a concentration of 10(-6 )M, ET1 in both groups 1 and 2 induced very low ripples of high frequency (group 1: 80+/-14%, n=5, group 2: 314+/-63%, n=11; P<0.05 compared with the pretreatment frequency) which lasted significantly longer in group 2 (29+/-2 min, n=10, P<0.05) than in group 1 (20+/-2 min, n=5), increasing the basal tone (group 1: 57.9+/-6%, n=5, group 2: 64.4+/-4%, n=6), the amplitude of myometrial contractility (group 1: 1.2+/-0.07 g, n=5, group 2: 1.6+/-0.1 g, n=7, P<0.05) and the area under the contractility curve (AUC; group 1: 8.4+/-1.1 gxmin, n=6, group 2: 11.9+/-1.6 g x min, n=11). In group 3, ET1 (10(-6 )M) created a sustained long-lasting contraction (initial phase: 43+/-6 min, n=6) characterized by the complete obliteration of spontaneous contractility with no ripples at all, and increasing significantly (P<0.05) the amplitude of myometrial contractility (2.8+/-0.5 g, n=6), the AUC (24.7+/-3.3 g x min, n=6), as well as the basal tone (183.6+/-21%, n=6) compared with the two premenopausal groups. In all three groups KCl exposure induced an initial rise (mean amplitude value: 1.1 g) followed by a relaxation phase to the primal baseline level (mean duration value: 12 min). Addition of ET1 (10(-6 )M) to KCl (80 mM) induced a similar pattern of contractility to that evoked by ET1 alone which, compared with KCl alone lasted significantly longer (P<0.05) in all three groups (group 1: 20+/-2 min, n=6; group 2: 23+/-2 min, n=6; group 3: 35+/-3 min, n=5). In group 3, the percentage change in basal tone was significantly smaller following KCl than after the combination of KCl plus ET1 (149+/-16%, n=5; P<0.01), indicating a different mechanism of contractility between KCl and ET1. These results demonstrate for the first time differences in myometrial response to ET1 between pre- and postmenopausal women. It is suggested that KCl and ET1 affect uterine contractility through different mechanisms and that ovarian steroids may play a regulatory role in human uterine responsiveness to ET1.

Pulmonary arterial hypertension (PAH) is a progressive and a life-threatening disease with its high morbidity and mortality ratios. On searching for new shining targets in pathogenesis, we noticed, in our previous studies, urotensin-II (UII) in systemic sclerosis with potent angiogenic and pro-fibrotic features. Owing to the mimicking properties of UII with endothelin-1 (ET1), we attempted to investigate the effect of palosuran in a PAH rat model. Thirty rats were randomly divided into three groups, with each group comprising 10 rats: group 1 (control group) received the vehicle subcutaneously, instead of monocrotaline (MCT) and vehicle; group 2 (MCT group) received subcutaneous MCT and vehicle; and group 3 (MCT + palosuran group) received subcutaneous MCT and palosuran. Serum UII, ET1, transforming growth factor-β1 (TGF-β1) levels, pulmonary arteriolar pathology of different diameter vessels, and cardiac indices were evaluated. The ET1, TGF-β1, and UII levels were significantly diminished in the treatment group, similar to the controls (p < 0.001). Right ventricular hypertrophy index and mean pulmonary arterial pressure scores were also significantly reduced in the treatment group (p = 0.001). Finally, in the 50-125-μm diameter arterioles, in contrast to Groups 3 and 1, there was a statistically significant thickness (p < 0.01) in the arteriolar walls of rats in Group 2. The treatment effect on arteries of more than 125-μm diameters was found to be valuable but not significant. Owing to its healing effect on hemodynamic, histological, and biochemical parameters of MCT-induced PAH, palosuran as an antagonist of UII might be an optional treatment alternative for PAH.

A model is proposed to describe the electrical activity and intracellular calcium dynamics of vascular smooth muscle cells (SMC) induced by endothelin (ET1). The conductance of the nonselective channels (NSCs), proportional to the ET1-receptor complex (ET . R), is intracellular calcium dependent. Inositol (1,4,5)-trisphosphate (IP3) produced by ET1 releases Ca2+ from the IP3-sensitive Ca2+ store. The transient increase of intracellular Ca2+ triggers the release of Ca2+ from the Ca(2+)-sensitive store by a Ca(2+)-induced Ca2+ (CICR) mechanism and activates the Ca(2+)-activated K+ current (IK,Ca). The inward current (Iin) via the NSC can depolarize the cell to a level at which the L-type Ca2+ current becomes activated (ICa). The level of depolarization is determined by the relative amplitude of (Iin + ICa + IK,Ca) and the voltage- and time-dependent K+ current. The model simulations show that (a) in cells without a CICR mechanism, short-lasting stimulation by ET1 elicits higher membrane potential and Ca2+ than long-lasting stimulation; (b) in cells with or without a CICR mechanism, a reduction of normal membrane capacitance (1 muf/cm2) results in either significant and sustaining or oscillatory membrane potential and intracellular calcium concentration. The applicability of the model to the study of electrical activity and calcium dynamics associated with hypercholesterolemia is discussed.

Maternal plasma ET-1 levels and the immunolocalisation of ET1 in the fetal membranes of pre-eclamptic primigravidae at>>/=28 weeks, gestation were studied. The levels of maternal plasma ET1 and immunoreactive ET-1 were increased in pre-eclampsia. Immunoreactive ET-1 was localised in the amnion, chorion and decidua of normal pregnant women as well as those with preeclampsia-eclampsia. Intense labelling was observed in moderate pre-eclampsia (BP 140/90 - 170/110 mmHg) with very intense labelling in severe pre-eclampsia (BP >170/110 mmHg), especially in the amniotic epithelium, chorionic villi, maternal blood vessels, cytotrophoblasts and giant cells of the decidua. The increased ET-1 levels demonstrated in fetal membranes of pre-eclamptic women are probably produced in a paracrine and/or autocrine manner, contributing to the hypertension, vasospasm and fetal growth restriction characteristic of the syndrome. A larger study would be required to show significant change in endothelin production in pre-eclampsia.

Specific endothelin-1 (ET1) binding sites have been demonstrated in membranes derived form normal (NP) and benign hyperplasic (BPH) human prostate using an 125I-ET1 binding assay. 125I saturation experiments and Scatchard analysis demonstrated the existence of a homogeneous population of binding sites with high affinity (Kd(app)) and density (B(max)), respectively, 106 +/- 15 pM and 1086 +/- 399 fmol/mg protein for NP (n = 5) and 168 +/- 26 pM and 964 +/- 445 fmol/mg protein for BPH (n = 5). We demonstrated the presence of two subtypes of ET1 receptors, ETA and ETB, by means of the following ET1 competitors: ET2, ET3, and BQ123 (which is selective for the ETA receptor), and IRL1620 and sarafotoxine c (S6c) (which are selective for the ETB receptor). The displacement curves allowed us to conclude that the large majority (85%) of the ET1 receptors in normal and hyperplasic human prostate are of the A subtype.

Endothelin (ET1) and vasoactive intestinal contractor (VIC) stimulate quiescent Swiss 3T3 cells to resume DNA synthesis acting synergistically with epidermal growth factors (EGF) and other mitogens. The peptide [D-Arg1,D-Phe5,D-Trp7,9,Leu11] substance P has been identified as a broad spectrum neuropeptide antagonist which blocks the binding and biological effects of the Ca2(+)-mobilizing neuropeptides bombesin, vasopressin, and bradykinin. In the present study we show that [D-Arg1,D-Phe5,D-Trp7,9,Leu11] substance P also acts as an ET1/VIC antagonist as judged by the following criteria: a) inhibition of specific 125I-labelled ET1 binding to a ET1/VIC receptor in a competitive and dose-dependent manner; b) blocking of the rapid increase in the cytosolic Ca2+ concentration promoted by ET1 or VIC; and c) inhibition of DNA synthesis stimulated by VIC in the presence of EGF. The inhibitory effects of [D-Arg1,D-Phe5,D-Trp7,9,Leu 11] substance P on Ca2+ mobilization and DNA synthesis were reversed by increasing the concentration of VIC. This is the first time that a peptide structurally unrelated to ET1 or VIC is shown to block the binding and mitogenic effects of peptides of the endothelin family.

Whole cells and crude extract of Mycobacterium sp. VKM Ac-1815D mutant strain Et1 were shown to carry out 17beta-reduction, 17beta-dehydrogenation and 1(2)-reduction of 3-keto-C(19)-steroids. Two 17-hydroxy steroid dehydrogenases (17-OH SDH) were partially purified from the strain by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-sephacel and gel-filtration on Bio-Gel A. The enzymes differed in chromatographic properties and specific activities. One enzyme--17-OH SDH (2) (tetramer, M(r) approximately 210,000) was found to be responsible for bi-directional reduction-oxidation of steroids at C 17, whereas the other one--17-OH SDH (1) (monomer, M(r) approximately 68,000) specifically catalysed 17beta-dehydrogenation of 17-hydroxysteroids (testosterone and 1(2)-dehydro testosterone). The 17beta-reduction of 1-ene-17-ketosteroids was accompanied by 1(2)-reduction. A role of 1-ene-reductase as a steroid-binding protein associated with 17-OH SDH (2) in Mycobacterium sp. is discussed.

Endothelin (Et) and nitric oxide (NO) may serve as chemical mediators of hypoxia-induced pulmonary hypertension. Plasma levels of Et1 were elevated 2-fold while levels of nitrate, a NO metabolite, decreased in rats exposed to 10 days of hypoxia (10% O2). Administration of l-arginine, the precursor for NO, decreased Et, increased nitrate, and decreased right ventricular hypertrophy in hypoxic animals. By increasing plasma NO levels, the right ventricular hypertrophy and right heart failure seen in hypoxia-induced pulmonary hypertension in human patients may be prevented.

Endothelins; (ET1, ET2 and ET3) are a family of peptides which acts on different smooth muscle preparations inducing a slow long lasting contraction. We investigated the effects of ET 1 modulatory action on adrenergic, cholinergic and serotoninergic transmission on an isolated mouse's stomach with gastric nerves. The endothelin 1 stimulation of the mouse stomach tone was abolished by the specific serotonin antagonist methizergid. This study suggests that endothelin 1 plays a role in the regulation of nonvascular smooth muscle tone. The endothelin effect was dependent on free intracellular Ca++ which can be recruited from an extracellular solution as well as from intracellular stores. Complete reduction of Ca++ from the extracellular solution with a simultaneous depletion of calcium stores abolished endothelin 1 depolarization of BC3H1 cells.

OBJECTIVE

This study aimed to compare the effects of mother's milk, sucrose and pacifier use to overcome pain during painful interventions to the newborns on the crying time and pain.

BACKGROUND

Various non-pharmacological methods are used to overcome the pain associated with painful interventions with newborns.

METHODS

A prospective, randomised, controlled study involved 120 newborns in Turkey.

METHODS

The population consisted of healthy newborns hospitalised in the gynaecology clinics of Trabzon Delivery and Children's Diseases hospital between February 2007-January 2008. The newborns who had blood sampling by heel stick were divided into four groups: mother's milk, sucrose, pacifier and control groups with 30 newborns in each. Data collection was performed using an information form on the newborn characteristics, which was developed by the researchers in the light of literature, clinical IR ear thermometer ET1 for temperature measurement, OXIMAX N-65 Pulse oxymeter for oxygen saturation and heart rate and neonatal infant pain scale for the measurement of the behavioural responses of newborns. Results. No differences were determined between the groups for heart rate and oxygen saturation in the newborns during painful interventions (p>> 0·05). Sucrose followed by pacifier was the most effective method of reducing the crying time in the newborns.

CONCLUSIONS

The results indicate that all three practices reduce the behavioural responses to pain at a higher rate than in the control group.

CONCLUSIONS

Health care personnel should perform painful interventions to the newborns while the babies are held by their mothers and during the procedure use of sucrose should be the primary choice.

Introduction: Protein Tyrosine Phosphatase 1B (PTP1B) and endoplasmic reticulum stress (ERS) are involved in the septic inflammatory response. Their inhibition is associated with improved survival in murine models of sepsis. The objective was to describe PTP1B and ERS expression during septic shock in human. Material and Methods: Prospective study including patients admitted to intensive care unit (ICU) for septic shock. Blood samples were collected on days 1 (D1), 3 and 5 (D5). Quantitative PCR (performed from whole blood) evaluated the expression of genes coding for PTP1B (PTPN1) and key elements of ERS (GRP78, ATF6, CHOP) or for endothelial dysfunction-related markers (ICAM1 and ET1). We analyzed gene variation between D5 and D1, collected glycemic parameters, insulin resistance and organ failure was evaluated by Sequential Organ Failure Assessment (SOFA) score. Results: We included 44 patients with a mean SAPS II 50 ± 16 and a mortality rate of 13.6%. Between D1 and D5, there was a significant decrease of PTPN1 (p < 0.001) and ATF6 (p < 0.001) expressions. Their variations of expression were correlated with SOFA variation (PTPN1, r = 0.35, CI 95% [0.05; 0.54], p = 0.03 and ATF6, r = 0.45 CI 95% [0.20; 0.65], p < 0.001). We did not find any correlation between PTPN1 expression and insulin resistance or glycemic parameters. Between D1 and D5, ATF6 and PTPN1 expressions were correlated with that of ET1. Conclusions: Our study has evaluated for the first time the expression of PTP1B and ERS in patients with septic shock, revealing that gene expression variation of PTPN1 and ATF6 are partly correlated with the evolution of septic organ failure and with endothelial dysfunction markers expression.