Tolvaptan reduces the required amount of albumin infusion in patients with decompensated cirrhosis with uncontrolled ascites : a multicenter retrospective propensity score-matched cohort study.

Background: The aim of this retrospective study was to determine whether tolvaptan treatment reduces the amount of albumin administered, volume of ascites removed, and frequency of paracentesis procedures in patients with decompensated cirrhosis with uncontrolled ascites with conventional diuretics. Patients and methods: The control (C) group included patients treated with conventional diuretics. The tolvaptan (T) group included patients treated with both tolvaptan and conventional diuretics. Both groups were matched according to baseline parameters. The amount of albumin administered, volume of ascites removed, and frequency of paracentesis within 30 days of onset of uncontrolled ascites were compared between the two groups. Results: After matching, 74 patients (C=37, T=37) were included. Baseline parameters (C vs. T group) were as follows: age, 69.5 ± 9.3 vs. 70.4 ± 11.0 years (p = 0.702) ; males, 24 (64.9%) vs. 25 (67.6%) (p = 0.999) ; patients with hepatocellular carcinoma, 17 (45.9%) vs. 18 (48.6%) (p = 0.999) ; serum albumin levels at treatment initiation, 2.76 ± 0.48 vs. 2.73 ± 0.49 g/dL (p = 0.773), and serum creatinine levels at treatment initiation, 1.18 ± 1.23 vs. 1.09 ± 0.48 g/dL (p = 0.679). In the C vs. T groups, respectively, mean amount of albumin administered was 51.0 ± 31.4 vs. 33.4 ± 29.8 g/month (p = 0.016) ; mean volume of ascites removed was 2,905 ± 4,921 vs. 1,824 ± 3,185 mL/month (p = 0.266) ; and mean frequency of paracentesis was 0.92 ± 1.46 vs. 0.89 ± 1.45 procedures (p = 0.937). Conclusions: Tolvaptan reduced the use of albumin infusion in patients with decompensated cirrhosis and was effective and acceptable for uncontrolled ascites.

A change in the timing for starting systemic therapies for hepatocellular carcinoma: the comparison of sorafenib and lenvatinib as the first-line treatment.

Aim: The aim of this retrospective multicenter study was to evaluate the differences in the timing for starting systemic therapies as the first-line treatment for hepatocellular carcinoma (HCC). Methods: A total of 375 patients with HCC treated with sorafenib from May 2009 to March 2018 and 56 patients treated with lenvatinib from March 2018 to November 2018 at our affiliated hospitals were included in this study. Results: The median ages of the sorafenib and lenvatinib groups were 71.0 (interquartile range [IQR]: 64.0-77.0) and 73.5 (IQR: 68.0 -80.0) years old, and 300 (80.0%) and 42 (75.0%) patients were men, respectively. The Barcelona Clinic Liver Cancer stage was early, intermediate and advanced in 39 patients (10.4%), 133 patients (35.5%) and 203 patients (54.1%) in the sorafenib group and 1 patient (1.8%), 17 patients (30.4%) and 38 patients (67.9%) in the lenvatinib group, respectively. In the analysis of intermediate HCC, patients who satisfied the criteria of TACE failure/refractoriness (P=0.017), those with ALBI grade 1 (P=0.040), and those with a serum AFP level < 200 ng/ml (P=0.027) were found more frequently in the lenvatinib group than in the sorafenib group, with statistical significance. The objective response rate (ORR) of lenvatinib was 34.8% in the overall patients and 46.7% in the intermediate-stage HCC patients, which was significantly higher than sorafenib (P=0.001, P=0.017). Conclusions: The emergence of lenvatinib has encouraged physicians to start systemic chemotherapy earlier in intermediatestage HCC patients.

2017 International Consensus on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care Science With Treatment Recommendations Summary

The International Liaison Committee on Resuscitation has initiated a near-continuous review of cardiopulmonary resuscitation science that replaces the previous 5-year cyclic batch-and-queue approach process. This is the first of an annual series of International Consensus on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care Science With Treatment Recommendations summary articles that will include the cardiopulmonary resuscitation science reviewed by the International Liaison Committee on Resuscitation in the previous year. The review this year includes 5 basic life support and 1 pediatric Consensuses on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care Science With Treatment Recommendations. Each of these includes a summary of the science and its quality based on Grading of Recommendations, Assessment, Development, and Evaluation criteria and treatment recommendations. Insights into the deliberations of the International Liaison Committee on Resuscitation task force members are provided in Values and Preferences sections. Finally, the task force members have prioritized and listed the top 3 knowledge gaps for each population, intervention, comparator, and outcome question

Percutaneous radiofrequency ablation for hepatocellular carcinoma located in the caudate lobe of the liver.

AIM: This study aimed to evaluate the effectiveness and safety of radiofrequency ablation (RFA) for hepatocellular carcinoma (HCC) located in the caudate lobe of the liver. PATIENTS AND METHODS: Between 2012 April and 2014 February, 142 patients with HCC meeting the Milan criteria were enrolled in this study. Of these patients, nine patients had HCC located in the caudate lobe (caudate group). Six of the nine cases were located in the Spiegel lobe, two cases were located in the paracaval portion and one case was located in the caudate process. We evaluated the local recurrence rate and RFA-related complications in the caudate group and non-caudate group. RESULTS: The local recurrence rate in the caudate group was 12.5% at 1 year and 12.5% at 2 years, while the local recurrence rate in the non-caudate group was 14.9% at 1 year and 29.0% at 2 years; there were no significant differences between the groups. No complications were observed in the caudate group, and minor complications were observed in six patients (4.5%) in the non-caudate group. No major complications or mortalities were observed in either group, and the complication rates were not significantly different between the groups (P = 1). CONCLUSIONS: RFA for HCC in the caudate lobe and the non-caudate lobe has equivalent effectiveness and safety. RFA is a promising treatment option for HCC arising in the caudate lobe.

Biocompatibility of cross-linked hyaluronate (Gel-200) for the treatment of knee osteoarthritis.

OBJECTIVE: To compare the biocompatibility and immunogenicity of two intra-articular hyaluronan formulations, Gel-200 (Gel-One(®)) and hylan G-F 20 (Synvisc(®) series). EXPERIMENTAL DESIGN: A comparison of the biocompatibility of Gel-200 and hylan G-F 20 was made using a rat subcutaneous air pouch model and the knee joint of normal rabbits. Immunogenicity was evaluated using a homologous passive cutaneous anaphylaxis (PCA) assay in guinea pigs. RESULTS: In the air pouch model in rats, characteristic fibrous belts formed in the subcutaneous tissue. Injection of hylan G-F 20 into the air pouch induced granulomatous nodules primarily composed of macrophages, multinucleated giant cells, and eosinophils accompanied with the test material in the center of the nodules in the fibrous belt. Furthermore, the thickness of the fibrous belt in the hylan G-F 20 group increased significantly compared to the saline group. Injection of Gel-200 into the air pouch induced neither granulomatous inflammation nor significant thickening of fibrous belt, while foamy macrophages containing the test material were observed. Intra-articular injection of hylan G-F 20 into the rabbit knee joints induced granulomatous inflammation, eosinophil infiltration, and significant increase in the number of cells in the synovial fluid, while these findings were absent in the Gel-200 group. In the immunogenicity assay, hylan G-F 20 induced a positive PCA reaction, but the Gel-200 did not. CONCLUSION: Gel-200 showed more favorable biocompatibility and less immunogenicity compared to hylan G-F 20. Gel-200 is expected to be a single injection hyaluronan product with less safety concerns for the treatment of knee osteoarthritis (OA) pain.

Contribution of conservation genetics in assessing neotropical freshwater fish biodiversity

Human activities have a considerable impact on hydrographic systems and fish fauna. The present review on conservation genetics of neotropical freshwater fish reveals that DNA analyses have been promoting increased knowledge on the genetic structure of fish species and their response to environmental changes. This knowledge is fundamental to the management of wild fish populations and the establishment of Evolutionary Significant Units capable of conserving genetic integrity. While population structuring can occur even in long-distance migratory fish, isolated populations can show reduced genetic variation and be at greater risk of extinction. Phylogeography and phylogeny have been powerful tools in understanding the evolution of fish populations, species and communities in distinct neotropic environments. Captive fish can be used to introduce new individuals and genes into the wild and their benefits and disadvantages can be monitored through genetic analysis. Understanding how fish biodiversity in neotropical freshwaters is generated and maintained is highly important, as these habitats are transformed by human development and fish communities are increasingly exploited as food sources to sustain a growing human population.

Atividades humanas têm um grande impacto sobre os sistemas hidrográficos e a fauna de peixes. A presente revisão em genética da conservação dos peixes de água doce neotropicais revela que as análises de DNA têm promovido um crescimento no conhecimento da estrutura genética das espécies e suas respostas às alterações ambientais. Este conhecimento é fundamental para o manejo das populações de peixes nativos e para o estabelecimento de Unidades Evolutivas Significantes capazes de conservar a integridade genética dessas populações. Enquanto o fenômeno de estruturação de populações pode ocorrer mesmo em peixes migradores de longas distâncias, populações isoladas podem mostrar reduzida variação genética e apresentar grande risco de extinção. Em adição, a filogeografia e a filogenia têm dado importantes contribuições para o entendimento da evolução das populações, espécies e comunidades de peixes em distintos ambientes neotropicais. Populações de cativeiro podem ser utilizadas como fonte de introdução de novos indivíduos e genes no ambiente natural e seus benefícios e desvantagens podem ser monitorados por meio de análises genéticas. É altamente importante, portanto, entender como é gerada e mantida a biodiversidade de peixes de água doce neotropicais, uma vez que os habitats são transformados pelo desenvolvimento humano e as comunidades de peixes têm sido intensamente exploradas como alimento de sustentação desse crescimento das populações humanas.

Contribution of conservation genetics in assessing neotropical freshwater fish biodiversity / Contribuição da Genética da Conservação para o conhecimento da biodiversidade dos peixes de água doce neotropicais

Human activities have a considerable impact on hydrographic systems and fish fauna. The present review on conservation genetics of neotropical freshwater fish reveals that DNA analyses have been promoting increased knowledge on the genetic structure of fish species and their response to environmental changes. This knowledge is fundamental to the management of wild fish populations and the establishment of Evolutionary Significant Units capable of conserving genetic integrity. While population structuring can occur even in long-distance migratory fish, isolated populations can show reduced genetic variation and be at greater risk of extinction. Phylogeography and phylogeny have been powerful tools in understanding the evolution of fish populations, species and communities in distinct neotropic environments. Captive fish can be used to introduce new individuals and genes into the wild and their benefits and disadvantages can be monitored through genetic analysis. Understanding how fish biodiversity in neotropical freshwaters is generated and maintained is highly important, as these habitats are transformed by human development and fish communities are increasingly exploited as food sources to sustain a growing human population.

Atividades humanas têm um grande impacto sobre os sistemas hidrográficos e a fauna de peixes. A presente revisão em genética da conservação dos peixes de água doce neotropicais revela que as análises de DNA têm promovido um crescimento no conhecimento da estrutura genética das espécies e suas respostas às alterações ambientais. Este conhecimento é fundamental para o manejo das populações de peixes nativos e para o estabelecimento de Unidades Evolutivas Significantes capazes de conservar a integridade genética dessas populações. Enquanto o fenômeno de estruturação de populações pode ocorrer mesmo em peixes migradores de longas distâncias, populações isoladas podem mostrar reduzida variação genética e apresentar grande risco de extinção. Em adição, a filogeografia e a filogenia têm dado importantes contribuições para o entendimento da evolução das populações, espécies e comunidades de peixes em distintos ambientes neotropicais. Populações de cativeiro podem ser utilizadas como fonte de introdução de novos indivíduos e genes no ambiente natural e seus benefícios e desvantagens podem ser monitorados por meio de análises genéticas. É altamente importante, portanto, entender como é gerada e mantida a biodiversidade de peixes de água doce neotropicais, uma vez que os habitats são transformados pelo desenvolvimento humano e as comunidades de peixes têm sido intensamente exploradas como alimento de sustentação desse crescimento das populações humanas.

Contribution of conservation genetics in assessing neotropical freshwater fish biodiversity.

Human activities have a considerable impact on hydrographic systems and fish fauna. The present review on conservation genetics of neotropical freshwater fish reveals that DNA analyses have been promoting increased knowledge on the genetic structure of fish species and their response to environmental changes. This knowledge is fundamental to the management of wild fish populations and the establishment of Evolutionary Significant Units capable of conserving genetic integrity. While population structuring can occur even in long-distance migratory fish, isolated populations can show reduced genetic variation and be at greater risk of extinction. Phylogeography and phylogeny have been powerful tools in understanding the evolution of fish populations, species and communities in distinct neotropic environments. Captive fish can be used to introduce new individuals and genes into the wild and their benefits and disadvantages can be monitored through genetic analysis. Understanding how fish biodiversity in neotropical freshwaters is generated and maintained is highly important, as these habitats are transformed by human development and fish communities are increasingly exploited as food sources to sustain a growing human population.

Comparison of the percutaneous absorption of hydrophilic and lipophilic compounds in shed snake skin and human skin.

The in vitro transdermal permeation of eight hydrophilic drugs (antipyrine, L-dopa, dopamine hydrochloride, diclofenac sodium, 5-fluorouracil, isoprenaline hydrochloride, nicorandil and morphine hydrochloride) and eight lipophilic drugs (aminopyrine, cyclobarbital, ibuprofen, indomethacin, isosorbide dinitrate, flurbiprofen, ketoprofen and lignocaine) was determined using shed snake skin of Elaphae obsoleta and human skin. The permeation parameters and physiological characteristics of the skin, e.g. the water and lipid content, and the thickness of shed snake skin and human skin were evaluated and compared. In shed snake skin, the permeability coefficients (P) of lipophilic drugs were in the same range as those through the human skin (0.9 to 1.8-times); whereas those of hydrophilic drugs were remarkably lower (3.3 to 6.1-times). The thickness and lipid content of shed snake skin and human stratum corneum were not significantly different (P > 0.05), whereas the water content of shed snake skin was significantly lower than that of human stratum corneum (P < 0.05). The lower permeability of shed snake skin for hydrophilic compounds might be caused by the lower porosity of skin strata. The results suggested a potential use of shed snake skin as barrier membrane for lipophilic compounds percutaneous absorption studies in vitro.

cDNA cloning and functional characterization of Xenopus laevis DNase gamma.

We report here the cDNA cloning and functional analysis of Xenopus DNase gamma (xDNase gamma). Two forms of cDNAs are isolated from adult spleen: one composing a 933 bp open reading frame for the enzymatically active xDNase gamma protein, and the other encoding an inactive short alternative form. Northern blot analysis revealed that the xDNase gamma mRNA is expressed in spleen, liver, testis, and ovary. xDNase gamma expression is scarcely detected in the tail muscle of tadpoles; however, it increases during metamorphosis and reaches a maximum during the late metamorphic climax. The ectopic expression of xDNase gamma results in the appearance of extensive DNA fragmentation in C2C12 myoblasts after the induction of apoptosis. In contrast, Xenopus DNase I fails to induce apoptotic DNA ladder formation under the same conditions. Our results suggest a possible involvement of xDNase gamma in apoptosis during amphibian metamorphosis.

Molecular paramagnetic semiconductor: crystal structures and magnetic and conducting properties of the Ni(dmit)(2) salts of 6-oxoverdazyl radical cations (dmit = 1,3-dithiol-2-thione-4,5-dithiolate).

Four kinds of 1:1 and 1:3 salts of 3-[4-(trimethylammonio)phenyl]-1,5-diphenyl-6-oxoverdazyl radical cation ([1](+)) and its mono- and dimethyl derivatives ([2](+) and [3](+)) with Ni(dmit)(2) anions (dmit = 1,3-dithiol-2-thione-4,5-dithiolate) ([1](+)[Ni(dmit)(2)](-) (4), [2](+)[Ni(dmit)(2)](-) (5), [3](+)[Ni(dmit)(2)](-) (6), and [1](+)[Ni(dmit)(2)](3)(-) (7)) have been prepared, and the magnetic susceptibilities (chi(M)) have been measured between 1.8 and 300 K. The chi(M) values of salts 5 and 7 can be well reproduced by the sum of the contributions from (i). a Curie-Weiss system with a Curie constant of 0.376 (K emu)/mol and negative Weiss constants (THETAV;) of -0.4 and -1.7 K and (ii). a dimer system with strong negative exchange interactions of 2J/k(B) = -354 and -258 K, respectively. The dimer formations in Ni(dmit)(2) anions have been ascertained by the crystal structure analyses of salts 4-6. In salts 4 and 6, Ni(dmit)(2) dimer molecules are sandwiched between two verdazyl cations, indicating the formation of a linear tetramer in 4 and 6. The magnetic susceptibility data for salts 4 and 6 have been fitted to a linear tetramer model using an end exchange interaction of 2J(1)/k(B) = -600 K and a central interaction of 2J(2)/k(B) = -280 K for 4 and 2J(1)/k(B) = -30 K and 2J(2)/k(B) = -580 K for 6, respectively. The results of the temperature dependence of the g(T) value in salts 4-6 obtained by ESR measurement also support the above analyses. The 1:1 salts 4-6 are insulators. On the other hand, the conductivity of the 1:3 salt 7 at 20 degrees C was sigma = 0.10 S cm(-)(1) with an activation energy E(A) = 0.099 eV, showing the semiconductor property. Salt 7 is a new molecular paramagnetic semiconductor.

Magnetic semiconductor: structural, magnetic, and conducting properties of the salts of the 6-oxoverdazyl radical cation with M(dmit)(2) anions (M = Ni, Zn, Pd, and Pt, dmit = 1,3-dithiol-2-thione-4,5-dithiolate).

Five kinds of (1:1), (1:3), and (2:1) salts of 3-[4-(diethylmethylammonio)phenyl]-1,5-diphenyl-6-oxoverdazyl radical cation [V](+) with M(dmit)(2) anions (M = Ni, Zn, Pd, and Pt, dmit = 1,3-dithiol-2-thione-4,5-dithiolate) ([V](+)[Ni(dmit)(2)](-) (1), [V](+)[Ni(dmit)(2)](3)(-) (2), [V](+)(2)[Zn(dmit)(2)](2-) (3), [V](+)(2)[Pd(dmit)(2)](2-) (4), and [V](+)(2)[Pt(dmit)(2)](2-) (5)) and an iodide salt of [V](+) ([V](+)[I](-) (6)) have been prepared, and the magnetic susceptibilities (chi(M) values) have been measured between 1.8 and 300 K. The chi(M) of the (1:1) Ni salt (1) can be well reproduced by the sum of the contributions from (i) a Curie-Weiss system with a Curie constant (C) of 0.376 K emu/mol and a negative Weiss constant (theta) of -1.5 K and (ii) the one-dimensional Heisenberg antiferromagnetic alternating chain system with 2J(A-B)/k(B) = -274 K (alternation parameter alpha = J(A-C)/J(A-B) = 0.2). The chi(M) of the (1:3) Ni salt (2) can be well explained by the two-term contributions from (i) the Curie-Weiss system with C = 0.376 K emu/mol and theta = -5.0 K and (ii) the dimer system with 2J/k(B) = -258 K. The magnetic properties of 1 and 2 were discussed based on the results obtained by crystal structure analysis and ESR measurements of 1 and 2. The chi(M) values of the (2:1) Zn, Pd, Pt salts 3, 4, and 5 and [V](+)[I](-) salt 6 follow the Curie-Weiss law with C = 0.723, 0.713, 0.712, and 0.342 K emu/mol and theta = -2.8, -3.1, -2.6, and +0.02 K, respectively, indicating that only the spins of the verdazyl radical cation contribute to the magnetic property of these salts. The salts 1, 3, and 5 are insulators. On the other hand, the conductivity (sigma) of the Ni salt 2 and Pd salt 4 at 20 degrees C was sigma = 8.9 x 10(-2) and 1.3 x 10(-4) S cm(-)(1) with an activation energy E(A) = 0.11 and 0.40 eV, respectively. The salts 2 and 4 are new molecular magnetic semiconductors.

A polymorphic, telomeric-like sequence microsatellite in the Neotropical fish Prochilodus.

A microsatellite locus from the Neotropical fish genus Prochilodus was isolated using PCR-based isolation of microsatellite arrays. Of 470 positive clones, 15 were sequenced, and 10 of them showed an (AATTT)(n) repeat. Primers were designed, and analysis of polymorphism revealed 11 alleles in three Prochilodus species. Fluorescence in situ hybridization analysis showed signals predominantly in the telomeric regions of several chromosomes. The description of this microsatellite may contribute to studies of the population structure of this fish group.

Cloning and functional characterization of a new subtype of the amino acid transport system N.

We have cloned a new subtype of the amino acid transport system N2 (SN2 or second subtype of system N) from rat brain. Rat SN2 consists of 471 amino acids and belongs to the recently identified glutamine transporter gene family that consists of system N and system A. Rat SN2 exhibits 63% identity with rat SN1. It also shows considerable sequence identity (50-56%) with the members of the amino acid transporter A subfamily. In the rat, SN2 mRNA is most abundant in the liver but is detectable in the brain, lung, stomach, kidney, testis, and spleen. When expressed in Xenopus laevis oocytes and in mammalian cells, rat SN2 mediates Na(+)-dependent transport of several neutral amino acids, including glycine, asparagine, alanine, serine, glutamine, and histidine. The transport process is electrogenic, Li(+) tolerant, and pH sensitive. The transport mechanism involves the influx of Na(+) and amino acids coupled to the efflux of H(+), resulting in intracellular alkalization. Proline, alpha-(methylamino)isobutyric acid, and anionic and cationic amino acids are not recognized by rat SN2.

Differential influence of cAMP on the expression of the three subtypes (ATA1, ATA2, and ATA3) of the amino acid transport system A.

Treatment of HepG2 cells with forskolin led to 60-100% stimulation of system A activity, measured as the Na+-dependent uptake of alpha-(methylamino)isobutyric acid. The stimulation was reproducible with cholera toxin and dibutyryl cAMP, and inhibitable by H7, a non-specific protein kinase inhibitor. The stimulatory effect was eliminated by cycloheximide and actinomycin D. The forskolin effect was associated with an increase in the maximal velocity of the transport system, with no change in substrate affinity. These cells express three different subtypes of system A (ATA1, ATA2, and ATA3). Treatment with forskolin increased the steady-state levels of ATA1 and ATA2 mRNAs, but decreased that of ATA3 mRNA.

Effect of pH on skin permeation enhancement of acidic drugs by l-menthol-ethanol system.

The effect of pH on the skin permeation enhancement of three acidic drugs by the l-menthol-ethanol system was investigated. The total flux of acidic drugs from the system remarkably varied over the pH range 3.0-8.0, and the permeation enhancement factor depended on the system pH and drug. A skin permeation model, which consists of two permeant (unionized and ionized) species, two system (oily and aqueous) phases, and two permeation (lipid and pore) pathways, was developed. The assumptions were made that only the unionized species can distribute to the oily phase and transport via the lipid pathway. The model explained the relationship between the concentration of drug in the aqueous phase and system pH. The skin permeability data were also described by the model and permeability coefficients corresponding to the physicochemical properties of permeant were calculated for the lipid and pore pathways. The model simulation showed that the permeation of acidic drugs occurred from the aqueous phase and the oily phase acted as a reservoir. Whether the total flux increased with increase of pH was dependent on the lipophilicity of drug. These results suggest that the pH of l-menthol-ethanol system should be given attention to elicit the maximum permeation enhancement.

Characterization of loops of the yeast mitochondrial ADP/ATP carrier facing the cytosol by site-directed mutagenesis.

To characterize structural features of the regions of the yeast type 2 ADP/ATP carrier (yAAC2) facing the cytosol, we prepared its Cys-less mutant, in which all four cysteine residues were replaced by alanine residues. The Cys-less mutant functioned like native yAAC2, showing that the cysteine residues are not essential. We then prepared cysteine mutants by substituting Ser(21) in the putative N-terminal region, Ala(124) and Ser(222) in the first and second loops facing cytosol, respectively, and Leu(312) in the C-terminal region of the Cys-less mutant for cysteine and examined the labeling of the substituted cysteine residues of the mutants with the membrane-impermeable SH reagent eosin-5-maleimide (EMA) from the cytosol. EMA labeled all the mutants, showing that all regions containing mutated residues faced the cytosolic side. The effects of transport inhibitors on EMA labeling were also examined. From the results, the location and conformation of the region around mutated residues were discussed.

Significant expression of functional human type 1 mitochondrial ADP/ATP carrier in yeast mitochondria.

As a first step to characterize the unknown functional properties of the human mitochondrial ADP/ATP carrier (AAC), we tried to express human type 1 AAC (hAAC1) in Saccharomyces cerevisiae. Expression of hAAC1 in yeast mitochondrial membrane was very low, although its transcript level was high. Its expression was improved greatly by replacement of its N-terminal region with the corresponding region of yeast type 2 AAC (yAAC2), as observed with the bovine type 1 AAC (bAAC1). This chimeric hAAC1 showed similar ADP transport activity to that of chimeric bAAC1, corresponding to the transport activity of bAAC1 in bovine heart mitochondria. These results suggested that the N-terminal region of yAAC2 is important for expression of the mammalian carriers in yeast mitochondria. Using the present expression system, studies on the functional properties of the human AAC isoforms in relation to their structures are now possible.

Significant effect of the N-terminal region of the mitochondrial ADP/ATP carrier on its efficient expression in yeast mitochondria.

The low-level expression of the bovine heart mitochondrial ADP/ATP carrier (bovine type 1 ADP/ATP carrier (bAAC1)) in the yeast mitochondrial membrane is significantly improved by replacement of its N-terminal region with corresponding regions of the yeast type 1 and 2 carriers (yAAC1 and yAAC2) (Hashimoto, M., Shinohara, Y., Majima, E., Hatanaka, T., Yamazaki, N., and Terada, H. (1999) Biochim. Biophys. Acta 1409, 113--124). To understand why the bAAC1 chimeras were highly expressed in yeast mitochondria, we examined the effects of the length and sequence of the N-terminal region extending into the cytosol on the expression of bAAC1 and yAAC2 derivatives in yeast mitochondria. For this, their N-terminal regions were replaced with peptide fragments of various lengths and sequences derived from those of bAAC1, yAAC1, and yAAC2. We found that a specific amino acid sequence and a definite length of the N-terminal region of yAAC2 were required for high expression of bAAC1 and yAAC2 in yeast mitochondria. We also examined the steady-state transcript levels and expression of these derivatives in whole yeast cells. Based on our results, we discuss the role of the N-terminal region in efficient expression of bAAC1 and yAAC2 in yeast mitochondria.

Stereoselective pharmacokinetics and pharmacodynamics of organic nitrates in rats.

Plasma concentration and vasodilating effect after i.v. bolus injection of stereoisomeric organic nitrates were evaluated. Pharmacokinetics of mononitrates was analyzed with a linear one-compartment model. The apparent volumes of distribution were almost identical, but systemic clearances were different among stereoisomers. The concentration data after dinitrate administration could be described based on a two-compartment model with elimination only from the central compartment via metabolism to mononitrate, and then mononitrate-dependent metabolic clearance was estimated. In the vasodilation by mononitrate administered intravenously, the maximum effect was not observed. The reduction of mean arterial pressure from baseline level was related to plasma concentration with a log-linear model. The pharmacological effect following dinitrate dosing was analyzed by a sigmoidal Emax model assuming a simple additive effect of dinitrate and mononitrate. Although almost the same Hill's constant and maximum effect (Emax) values were estimated, the concentrations required to produce 50% of Emax (EC50) differed among stereoisomers. The clearance and EC50 values of stereoisomers with nitrate group at the exo position were generally higher than those with the same group at the endo position. This suggests that the stereostructure of organic nitrates controls the vasodilator potency and duration of action.