Renal tissue angiotensins during converting enzyme inhibition of angiotensin I in spontaneously hypertensive rat

To compare the effects of an angiotensin-converting enzyme inhibitor on circulating and tissue renin-angiotensin system, we measured different renin-angiotensin system parameters during the first day of treatment (Day 1) as well as after two weeks of treatment (Day 14). Ramipril was given orally once daily to adult male spontaneously hypertensive rats. Renin activity, angiotensin-converting enzyme activity and levels of angiotensin I and angiotensin II in the plasma, renal cortex and renal medullar were assessed at Day 1 and Day 14 of the treatment. In the plasma, both renin activity and angiotensin I increased 10 to 15 fold one to four hours after acute as well as at Day 14 of ramipril treatment and then returned to basal values within 24 hours. Plasma angiotensin II levels were not significantly decreased at Day 1 or Day 14. The decrease in the angiotensin II/angiotensin I ratio suggested a sustained inhibition of plasma angiotensin-converting enzyme at Day 14. In the renal cortex and medulla, a clearly different pattern was observed: in ramipril treated rats, renin activity in the renal cortex and medulla did not change at Day 1 but at Day 14 we observed a slight and sustained increase in renin activity. Despite very high basal levels of renin activity, angiotensin I levels in the renal cortex were comparable to those in the plasma. The angiotensin I level increased only one-fold one hour after ramipril intake at Day 1 and Day 14. This suggests that angiotensinogen may have a limiting role in the synthesis of angiotensin I in the kidney. Angiotensin II levels were slightly higher in the renal cortex and medulla than in the plasma suggesting local synthesis of the peptide. In the kidney, angiotensin II levels decreased one and four hours after the acute or prolonged ramipril treatment and the angiotensin II/angiotensin I ratio was reduced at the same time. Our results show that the responses of the plasma and kidney components of the renin-angiotensin system to angiotensin-converting enzyme inhibition are different in the plasma and the kidney suggesting that the circulating and tissue renin-angiotensin system are at least in part independent.     

DNA-based immunization against hepatitis B surface antigen (HBsAg) in normal and HBsAg-transgenic mice

Hepatitis B virus (HBV) remains a serious worldwide health problem and the possibility to control it will depend on the availability of safe, effective and affordable vaccines. Recombinant protein or plasma-derived vaccines containing HBV surface antigen (HBsAg) are safe and generally efficacious, however, they are too expensive for widespread use in areas of HBV endemicity and are only partially effective for treatment of HBV chronic carriers. Immunization of mice by injection of HBsAg-expressing plasmid DNA results in rapid induction of strong and long-lasting humoral and cell-mediated immune responses. Here we report optimization of the humoral response with the use of necrotizing agents, co-expression of cytokines or co-stimulatory molecules and formulation of the DNA with cationic liposomes. DNA-based immunization of HBsAg-transgenic mice can also overcome non-response to HBsAg. Thus, DNA vaccines against HBV may be useful for both prophylactic and therapeutic purposes.     

Effects of hydroxyl radicals on purified angiotensin I converting enzyme

Somatic angiotensin-converting enzyme (ACE) is a protein which contains two similar domains (N and C), each possessing a functional active site. The relationship between ACE, its natural substrates and oxygen free radicals is starting to be explored. On one hand, superoxide anions production is induced by angiotensin II and on the other hand, activated polynuclear neutrophils, through free radicals generation, alter endothelial ACE activity. In this study, we examined the impact of hydroxyl radicals (.OH) on purified ACE. .OH were produced using a generator: 2,2'-azo-bis 2-amidinopropane (GRH) provided by Lara-Spiral (Fr). GRH (3 mM), in a time-dependent fashion, inhibited ACE activity. When ACE was co-incubated for 4 h with GRH, its activity decreased by 70%. Addition of dimethylthiourea (DMTU: 0.03 to 1 mM) or mannitol + methionine (20/10 mM), two sets of .OH scavengers, produced a dose-dependent protection on ACE activity. To examine whether oxidation of thiol groups in the ACE molecule could be involved in the action of GRH, the effects of thiol reducing agents: mercaptoethanol and dithiotreitol (DTT) were investigated. These compounds produced a dose-dependent and significant protection; with 100% protection at 0.2 and 0.3 mM for mercaptoethanol and at 0.1 mM for DTT. The hydrolysis of two natural and domain-specific substrates were also explored. The hydrolysis of angiotensin I preferentially cleaved by the C domain was significantly (p < 0.01) inhibited by 57, 58 and 69% in contact with 0.3, 1 and 3 mM GRH [in nmol angio II formed/min/nmol of ACE, n = 4; 35.9 +/- 0.6 (control), 15.5 +/- 2.8 (GRH : 0.3 mM), 15.1 +/- 0.5 (1), 10.9 +/- 0.6 (3)]. The hydrolysis of the hemoregulatory peptide (hp), preferential substrate for the N domain was not affected by GRH at 0.3 mM and inhibited by 28% (not significant) by 1 mM GRH [in nmol ph hydrolized/min/nmol ACE, n = 4; 12.6 +/- 1.9 (control), 14.9 (GRH : 0.3 mM), 8.3 +/- 4.0 (1). These results demonstrated that .OH affect ACE activity and could suggest a privileged impact of GRH on the C domain. The precise sites of action of .OH remain unknown. The Cys residues near the active centers, by forming disulphide bridges during the oxidation could be of critical importance. Further studies will be needed to determine whether oxidative stress again ACE can be involved in the genesis of inflammatory vascular pathologies.     

Stimulation of alpha1A-adrenoceptors in Rat-1 cells inhibits extracellular signal-regulated kinase by activating p38 mitogen-activated protein kinase

We report a patient with cholangiocellular carcinoma with tumor thrombi in the main portal trunk who has survived for 9.5 years after hepatic resection. A 57-year-old woman underwent an extended left lobectomy, and resection of the caudate lobe plus the main portal trunk for a liver tumor that had a portal tumor thrombus in the main portal trunk. The portal vein was reconstructed with an autologous vein graft obtained from the external iliac vein. Histological examination of the resected specimen revealed moderately differentiated tubular adenocarcinoma compatible with cholangiocellular carcinoma. Factors contributing to the patient's long-term survival are discussed. Aggressive surgical resection can be effective even for such an advanced case of cholangiocellular carcinoma.     

Characterization of the human gonadotropin-releasing hormone receptor heterologously produced using the baculovirus/insect cell and the Semliki Forest virus systems

1. Two eukaryotic viral systems, the baculovirus/insect cell and the Semliki Forest virus systems, were tested for heterologous expression of human gonadotropin-releasing hormone receptor (GnRHR) cDNA. 2. An unmodified as well as a c-myc epitope-tagged human GnRH receptor was produced in two insect cell lines (Spodoptera frugiperda, Trichoplusia ni) after infection with the respective recombinant baculoviruses. In both insect cell lines, the receptor was identified by immunoblot analysis as a triplet of bands between 35 and 40 kDa. After deglycosylation of the receptor the molecular mass decreased to 35 kDa. The GnRH receptor was localized in membrane compartments within the infected insect cells. However, only in membranes of infected Trichoplusia ni insect cells could approximately 2000 receptors per cell be detected. 3. Production of the GnRH receptor in BHK cells using the Semliki Forest virus system resulted in approximately 50,000 receptors per cell. A maximal yield of 0.42 pmol/mg membrane protein was obtained 24 hr after electroporation of BHK cells with in vitro synthesized RNA. Binding of the antagonist [125I]Cetrorelix was saturable with a KD of 1.3 nM. The receptor produced in the BHK cells was further characterized by ligand displacement studies. The rank order of agonist and antagonist affinities was Cetrorelix > Triptorelin > Antide > GnRH.     

Synchronous tabu search parallelization strategies for multicommodity location-allocation with balancing requirements

We study and compare synchronous parallelization strategies for tabu search. We identify the most promising parallelization approaches, and evaluate the impact on performance and solution quality of some important algorithmic design parameters: length of the synchronization steps, number of processors, handling of exchanged information, etc. Parallelization approaches are implemented and compared by using a tabu search algorithm for multicommodity location-allocation problems with balancing requirements.     

Cognitive Operations in the Generation Effect on a Recall Test: Role of Aging and Divided Attention.

Generation effect (generated words are better memorized than read words) of anagrams, rhymes, and associates of target words was examined in young, elderly, and very old subjects. Experiments 1 and 2 showed that only young subjects benefit from the generation effect in a free-recall test when the rule is of a phonological nature. Experiments 3, 4, and 5 showed that the generation effect of rhymes was due to a resources-dependent self-initiated process. Experiments 4 and 5 showed that in a divided-attention situation, generation effect of rhymes is not significant in young subjects, but that the generation effect of semantic associates remains significant for both groups (Experiment 5). The results are discussed within the environmental support framework and the transfer-appropriate processing framework. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Wastewater treatment in a hybrid activated sludge baffled reactor

A novel hybrid activated sludge baffled reactor (HASBR), which contained both suspended and attached-growth biomass perfect mixing cells in series, was developed by installing standing and hanging baffles and introducing plastic brushes into a conventional activated sludge (CAS) reactor. It was used for the treatment of domestic wastewater. The effects on the operational performance of developing the suspended and attached-growth biomass and reactor configuration were investigated. The change of the flow regime from complete-mix to plug-flow, and the addition of plastic brushes as a support for biofilm, resulted in considerable improvements in the COD, nitrogen removal efficiency of domestic wastewater and sludge settling properties. In steady state, approximately 98+/-2% of the total COD and 98+/-2% of the ammonia of the influent were removed in the HASBR, when the influent wastewater concentration was 593+/-11 mg COD/L and 43+/-5 mg N/L, respectively, at a HRT of 10 h. These results were 93+/-3 and 6+/-3% for the CAS reactor, respectively. Approximately 90+/-7% of the total COD was removed in the HASBR, when the influent wastewater concentration was 654+/-16 mg COD/L at a 3h HRT, and in the organic loading rate (OLR) of 5.36kgCOD m(-3) day(-1). The result for the CAS reactor was 60+/-3%. Existing CAS plants can be upgraded by changing the reactor configuration and introducing biofilm support media into the aeration tank.     

New Raman spectrometer using a digital micromirror device and a photomultiplier tube detector for rapid on-line industrial analysis. Part I: description of the prototype and preliminary results

In this paper, a prototype of a new generation of Raman spectrometers, based on the use of a monochromator, a digital micromirror device as light modulator, and a photomultiplier tube as detector of the Raman light, is described. This spectrometer, containing no moving parts, is inexpensive, robust, and very precise. New in concept, this spectrometer makes it possible to record, in addition to classical Raman spectra, the intensity at several selected points of the spectrum and/or the total intensity in several selected intervals at the same time with great accuracy, thus giving new possibilities for analytical applications. Also, the work presented demonstrates the possibilities of this very simple prototype for rapid on-line industrial analysis, with an example of quantitative analysis of binary and ternary mixtures of xylene isomers. The precision obtained is satisfactory (errors of prediction approximately 3% in 5-6 seconds per sample).