We studied absorption, distribution, metabolism, and excretion of polyamines (putrescine, spermidine, and spermine) in the gastrointestinal tract using 15N-labeled polyamines as tracers and ionspray ionization mass spectrometry (IS-MS). The relatively simple protocol using rats bearing solid tumors provided useful information. Three 15N-labeled polyamines that were simultaneously administered were absorbed equally from gastrointestinal tract, and distributed within tissues at various concentrations. The uptake of 15N-spermidine seemed preferential to that of 15N-spermine since the concentrations of 15N-spermidine in the liver and tumors were higher, whereas those of 15N-spermine were higher in the kidney, probably due to the excretion of excess extracellular spermine. Most of the absorbed 15N-putrescine seemed to be lost, suggesting blood and tissue diamine oxidase degradation. Concentrations of 15N-spermidine and 15N-spermine in the tumor were low. We also describe the findings from two rats that were administered with 15N-spermine. The tissue concentrations of 15N-spermine were unusually high, and significant levels of 15N-spermidine were derived from 15N-spermine in these animals.
Mice with juvenile visceral steatosis (JVS) develop remarkable cardiac hypertrophy and exhibit an increased number of mitochondria in their heart. However, the biochemical characteristics and physiological functions of these mitochondria cardiac are little known. Here we show that the respiratory activities at state 3 with glutamate plus malate or succinate in the heart mitochondria of JVS mice were greatly decreased to 47% or 77%, respectively, compared with those of control mice. The contents of cytochromes a+a3, b, and c+c1 in the heart mitochondria of these mice were also decreased, to 51%, 45%, and 79%, respectively, of those of the control mice. Oligomycin-sensitive ATPase activitiy in these mitochondria, however, was increased to about 2 times over that of the control mice. Surprisingly, the ATP-Pi exchange activity of the heart mitochondria of JVS mice was greatly decreased, to 35% of that of control mice. On the other hand, the expression levels of 2 subunits of H+-ATP synthase, i.e., coupling factor 6 and α subunit, in heart mitochondria from control and JVS mice were almost the same. These results indicate that the coordinate regulation of mitochondrial proliferation and gene expression for components of the oxidative phosphorylation system was markedly defective in the heart of JVS mice. Our current results also suggest the presence of a novel regulatory mechanisms of ATP synthase activities in the heart.
An acid sialidase partially purified from porcine liver was activated by incubation at 37 °C under acidic pH. This activation was dependent on pH, time and temperature, but not inhibited by amastatin, an inhibitor of aminopeptidase A, in contrast to the case of human placental sialidase. The effects of inorganic anions on the two sialidases from porcine liver and from human placenta were investigated. Among the anions tested, halide ions, especially chloride and bromide ions, markedly enhanced the activation of the two sialidases. However, nitrate, sulfate, sulfite and pyrosulfite ions rarely affected the activation of sialidase from porcine liver, while all of them enhanced the activation of human placental sialidase. The activation of the enzyme from porcine liver was depressed at concentrations of greater than 100 mM of sodium chloride, whereas the enzyme from human placenta was held at maximum activation until 1 M sodium chloride. These results suggest the possibility of the participation of enzyme functions different from that of human placental sialidase in the activation process of sialidase.
Papyriflavonol A, a new prenylated flavonol isolated from Broussonetia papyrifera, selectively inhibits recombinant human secretory phospholipase A2s (sPLA2s). Papyriflavonol A was found to inhibit human group IIA and V sPLA2s potently and irreversibly in a dose-dependent manner, with respective IC50 values of 3.9 and 4.5 μM. The inhibitory effects of papyriflavonol A against bovine group IB (IC50 of 76.9 μM) and the human group X (IC50 of 225 μM) sPLA2s were weaker than those against human group IIA and V sPLA2s, and human group IIF sPLA2 was not inhibited. In addition, papyriflavonol A potently inhibited the stimulus-induced production of leukotriene C4 with an IC50 value of approximately 0.64 μM in mouse bone marrow-derived mast cells. In addition, papyriflavonol A significantly reduced IgE-dependent passive cutaneous anaphylaxis in rats. These results indicate that papyriflavonol A provides a basis for novel types of antiinflammatory drugs.
The Editorial Committee of this Journal wishes to acknowledge with regret that "Trypsinogen hL expressed in the human lung is a new member of the trypsinogen family" by Imamura Y., Katsu M., Sakai K., Okumura Y., Ariga H. and Kido H. which was published in Vol. 26 (2003), p. 361 of this Journal, has been found that the sequence published in BPB is not the human sequence but the mouse one, according to proposal by the authors.
Thus it should be noted that the above-mentioned paper by Imamura Y., Katsu M., Sakai K., Okumura Y., Ariga H. and Kido H. should be deleted.
For this reason, as of May 28, 2004, this article (including its abstract and references) has been removed from this site.
In the liver, the multidrug resistance (MDR) protein P-glycoprotein (P-gp) is physiologically expressed at the bile canalicular membrane, where it participates in the biliary excretion of various lipophilic drugs and xenobiotics. Previous studies showed that the immunosuppressive agent cyclosporine A (CsA) modulates P-gp and exerts a hepatotrophic influence in the regenerating liver. Hepatocytes isolated from regenerating rat liver, after 2/3 partial hepatectomy (PH 2/3), were used as an in vivo experimental model of cells with high proliferating activity in order to investigate whether CsA influences cellular levels of P-gp in those cells. Male Wistar rats were treated with CsA (20 mg/kg body weight) for 4 d preoperatively and 1 d postoperatively, and regenerating hepatocytes were isolated by collagenase perfusion 12, 24 and 48 h after PH 2/3. Flow cytometry and Western blotting studies with the monoclonal antibodies C494 and C219 showed that after PH 2/3, cellular levels of P-gp were initially suppressed, 12 h after PH 2/3, by 23%, but were significantly elevated thereafter, 24 and 48 h after PH 2/3 by 28% and 73%, respectively. In CsA pretreated animals, P-gp levels were increased even in normal hepatocytes by 34%, and an additional augmentation was seen in hepatocytes from 24 and 48 h regenerating livers (60% and 56%, respectively). In summary, we demonstrate for the first time that CsA has an additive effect on the expression of P-glycoprotein during liver regeneration in the rat. Therefore, induction of P-gp might also be considered in patients receiving CsA after liver transplantation for hepatocellular carcinoma and chemotherapy as an adjuvant treatment for the prevention of tumor recurrence.
The Panax ginseng has been used as traditional medicine for past several years among oriental people. The present investigation has been made to assess the radioprotective efficacy of ginseng root extract in the testicular enzymes of Swiss albino mice. The Swiss albino mice were divided into different groups. (i) Ginseng treated group: The animals were administered 10 mg/kg body weight ginseng root extract i.p. (ii) Radiation treated group: The animals were exposed to 8 Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 cm. (iii) Combination group: Animals were administered ginseng extract continuously for 4 d and on 4th day they were irradiated to 8 Gy gamma radiation after 30 min of extract administration. The animals from above groups were autopsied on day 1, 3, 7, 14 and 30. Biochemical estimations of acid and alkaline phosphatases and Lipid peroxidation (LPO) in testes were done. In ginseng treated group acid and alkaline phosphatases activity and LPO level did not show any significant alteration. In irradiated animals there was a significant increase in acid phosphatase activity and LPO level. However, significant decline in alkaline phosphatase activity was observed. The treatment of ginseng before irradiation causes significant decrease in acid phosphatase and LPO level and significant increase in alkaline phosphatase activity. One of the cause of radiation damage is lipid peroxidation. Due to lipid peroxidation, lysosomal membrane permeability alters and thus results in release of hydrolytic enzymes. So, an increase in acid phosphatase was noticed after radiation treatment. The alkaline phosphatase activity is associated with membrane permeability and different stages of spermatogenesis. Due to membrane damage and depletion of germ cells of testes after irradiation the enzyme activity was decreased. Ginseng markedly inhibits lipid peroxidation. It acts in indirect fashion to protect radical processes by inhibition of initiation of free radical processes and thus reduces the radiation damages in testes of Swiss albino mice.
Kampo medicine, Stephania tetrandra Radix (Stephania) in Boi-ogi-to increases the blood insulin level and falls the blood glucose level in streptozotocin (STZ)-diabetic ddY mice. These actions of Stephania are potentiated by Astragalus membranaceus Bunge Radix (Astragali) in Boi-ogi-to (Liu et al., J. Traditional Med., 17, 253—260, 2000). In the present study, actions of bis-benzylisoquinoline alkaloids isolated from Stephania were investigated in the hyperglycemia of STZ-diabetic mice. A main bis-benzylisoquinoline alkaloid, fangchinoline (0.3—3 mg/kg) significantly fell the blood glucose level of the diabetic mice in a dose-dependent manner. The effect of fangchinoline was 3.9-fold greater than that of water extract of Stephania. However, another main compound, tetrandrine (1—100 mg/kg) did not have any effect. The water extract of Astragali did not affect singly but potentiated the anti-hyperglycemic action of fangchinoline (0.3 mg/kg). Out of used compounds (1 mg/kg) isolated from Stephania, fangchinoline, fangchinoline 2′-N-α-oxide and 2′-N-norfangchinoline, which are substituted with 7-hydroxy side chain for 7-O-methyl side chain, decreased to near 50% of high blood glucose level. In addition, tetrandrine 2′-N-β-oxide, tetrandrine 2′-N-α-oxide, tetrandrine 2-N-β-oxide, fangchinoline 2′-N-α-oxide, which are added to 2- or 2′-N-oxide side chain, also decreased to near 50% of the high blood glucose level. In conclusion, fangchinoline but not tetrandrine from Stephania shows the anti-hyperglycemic action in the STZ-diabetic mice. The demethylation of 7-O-position and/or addition of 2- or 2′-N-oxide side chain in bis-benzylisoquinoline compounds in Stephania have a role for the induction of the anti-hyperglycemic actions.
The effects of nefiracetam on the cerebral adenylyl cyclase (AC) activity of animals with microsphere embolism-induced memory dysfunction were examined. Sustained cerebral ischemia in the right cerebral hemisphere was induced by an injection of microspheres into the right internal carotid artery of rats. To examine learning and memory function, the water maze test was performed from day 7 to day 10 after the operation. The escape latency of the microsphere-embolized (ME) rat in the water maze task was longer than that of the sham-operated (Sham) rat, suggesting that spatial memory dysfunction occurred in the ME rat. Gsα and Gi1/2α protein levels in the cerebral cortex, striatum and hippocampus of the ME rat, when determined on day 11, were similar to those of the Sham rats. The basal AC activity in the striatum, but not in the other two regions, of the ME rat decreased. The AC activity in the presence of 10 μM colforsin daropate (Col), a direct stimulator of AC, was increased by approximately 20-fold in sham animals and 7- to 10-fold in the ME rat. Treatment of the ME rat with 10 mg/kg/d nefiracetam p.o. from day 1 to day 10 after the operation shortened the escape latency, restored the basal AC activity in the striatum, and reversed the Col-induced increases in AC in these three regions without any changes in the cerebral Gsα and Gi1/2α protein levels. These results suggest that nefiracetam-mediated activation of AC activity may contribute to the improvement of memory and learning function in sustained cerebral ischemia.
In this paper we evaluated the bronchodilator effects of SPFF [2-(4-amino-3-chloro-5-trifluomethyl-phenyl)-2-tert-butylamino-ethanol chloride], a newly synthesized β2 adrenergic agonist in guinea pigs and rabbits, in comparison with other β2 adrenergic agonists, isoprenaline or salbutamol. We studied in vitro the bronchodilator effects of SPFF and isoprenaline on isolated guinea pig trachea strips with or without the precontraction of bronchocontractors (acetylcholine and histamie). The positive chronotropic effects of SPFF and isoprenaline on isolated guinea pig left atria were also tested in vitro. Potency values (pD2, pA2 or ED50) were determined from the cumulative concentration–response curves. The results showed that SPFF and isoprenaline dose-dependently relaxed the isolated guinea pig trachea strips and the pD2 values of both drugs were 7.66±0.68 and 8.79±0.19, respectively. Moreover, we confirmed that the bronchodilator effect of SPFF was due to the activation of β2 adrenoceptor because this effect was easily antagonized by ICI-118551 (pA2 8.90±0.01), a specific β2 adrenoceptor antagonist. SPFF also dose-dependently relaxed the isolated guinea pig trachea strip precontraction with acetylcholine or histamine with ED50 values of 10.2±0.7 μM and 550±38.2 nM, respectively. Furthermore, the positive chronotropic effect of SPFF on isolated guinea pig left atria (pD2 5.41±0.38) was much weaker than that of isoprenaline (pD2 8.75±0.24), which implied that SPFF was more selective to airway β2 adrenoceptor than isoprenaline; the β1/β2 selectivity assay also showed that SPFF was about 162 times more selective to β2 adrenoceptor than isoprenaline. A radioligand binding experiment using guinea pig lung and cardiac ventricle as β2 and β1 adrenoceptor sources, respectively, also demonstrated that SPFF possesses high affinity (27.3 nM) and selectivity (4.6 fold) to β2 adrenoceptors. The protective effects of SPFF and salbutamol on bronchospasm induced by bronchoconstrictor aerosol in guinea pigs in vivo were investigated, and the Konzett and Rössler experiment in rabbits in vivo was also carried out. SPFF significantly prolonged the latency time of histamine and acetylcholine induced asphyxiation collapse in guinea pigs: the ED50 value of SPFF i.g. was 0.32±0.05 mg·kg−1 in this experiment. Meanwhile, the ED50 values of salbutamol was 2.37±0.22, which meant that the bronchorelaxation effect of salbutamol was about 6 times less potent than that of SPFF. The Konzett and Rössler experiment performed in anesthetized rabbit showed that intraduodenal administration of SPFF exerted action of longer duration than salbutamol. From the results above we suggested that SPFF was a potent, long-acting bronchodilator with relatively higher β2 adrenoceptor selectivity.
Many people suffer from intractable bedsores, which sometimes develop because of chronic metabolic failure in patients. An extract of the root of Lithospermun erythrorhison (SK) has been reported to have an effect on wound healing. However, the effects of SK have not been studied in chronic wounds, such as bedsores. The healing-impaired diabetic (db/db) mouse is a good model for the investigation of clinical healing therapies. Therefore, we examined whether SK accelerates wound healing in db/db mice. Full-thickness round wounds of 6-mm diameter were created on the backs of mice. After applying SK, we covered the wound with a film dressing to keep it moist. At three weeks, wound closure was complete in SK-treated mice but not in controls. Capillary vessel number and collagen synthesis increased early in wound healing in SK-treated wounds. At this time, vascular endothelial growth factor (VEGF)-positive neutrophils had infiltrated the wound and the appearance of apoptotic fibroblasts and endothelial cells in the granulation tissue was more advanced than in the controls. Where the wound was covered with epithelium, there tended to be less infiltration of VEGF-positive cells and apoptotic cells. These results suggest that the inflammatory phase was shortened, and the proliferative and maturation phases were advanced by SK. It is known that SK also has antibacterial activity. Therefore, we conclude that SK is useful for wound healing in db/db mice, and could potentially help patients with intractable bedsores.
Oleic acid-induced hypoxemia is an animal model of acute respiratory distress syndrome (ARDS). Increased capillary permeability is a cause of hypoxemia in lung injury. Endothelial cells form a major capillary barrier, and disruption of the barrier appears to involve a decreased level of ATP in the cells. Phosphoenolpyruvate (PEP) is an endogenous substance that is one of the ATP precursors and can cross some cell membranes via anion exchanger. We examined the effect of PEP on oleic acid-induced lung injury in guinea pigs. An intravenous injection of oleic acid (15 μl/kg) caused severe hypoxemia. Pretreatment with PEP at a dose of 2, 20, or 200 μmol/kg attenuated the oleic acid-induced decrease in the arterial partial pressure of oxygen in a dose-dependent manner. Furthermore, PEP attenuated the oleic acid-induced increase in vascular permeability in the proximal and distal bronchi, as indicated by the extravascular leakage of Evans Blue dye. The combination of PEP with ATP (4 μmol/kg) showed no additional inhibitory effect on oleic acid-induced lung injury, compared with PEP alone. We suggest that PEP is a promising candidate to prevent hypoxemia in acute lung injuries associated with increased vascular permeability, such as ARDS.
It has been shown that ascorbate (AsA) and its stable derivative, ascorbic acid 2-O-α-glucoside (AA-2G), do not elicit neurite outgrowth in PC12 cells. However, these ascorbates are synergistically enhanced by both dibutyryl cyclic AMP (Bt2cAMP)- and nerve growth factor (NGF)-induced neurite outgrowth in this model. In the present study, the effects of a series of novel lipophilic ascorbate derivatives, 6-acylated ascorbic acid 2-O-α-glucosides (6-Acyl-AA-2G), on neurite outgrowth induced by Bt2cAMP and NGF were examined in PC12 cells. We found that all the tested acylated ascorbate derivatives enhanced neurite formation induced by both agents in a dose-dependent manner. Of the 6-Acyl-AA-2G derivatives, 6-octanoyl ascorbic acid 2-O-α-glucoside (6-Octa-AA-2G) enhanced the Bt2cAMP-induced phosphorylated MAPK p44 and p42 expression. A α-glucosidase inhibitor, castanospermine, completely abrogated the promotion of neurite outgrowth and MAPK expression by 6-Octa-AA-2G. Addition of 6-Octa-AA-2G (0.5 mM) to PC12 cells caused a rapid and significant increase in intracellular AsA content, which reached a maximum and was maintained from 12 to 24 h after the culture. These findings suggest that 6-Acyl-AA-2G is rapidly hydrolyzed to AsA within the cell and enhances neurite differentiation through the interaction with the inducer-activated MAPK pathway.
Tryptanthrin, a biologically active compound found in the medicinal plant Polygonum tinctorium, reportedly has several biological activities. We investigated the effects of tryptanthrin on cytokine production by lymphocytes in response to staphylococcal enterotoxin B (SEB), which causes a variety of disorders in humans based on its induction of large amounts of immunostimulatory cytokines. Tryptanthrin dose-dependently inhibited interferon-γ (IFN-γ) and interleukin-2 production by mouse spleen cells and Peyer's patch (PP) lymphocytes in vitro. The efficacy of tryptanthrin was further studied in a mouse model in vivo. Tryptanthrin was administered orally 2 h after an oral challenge with SEB. Nineteen hours after SEB administration, PP lymphocytes were prepared, and IFN-γ production by PP lymphocytes was examined. The production of IFN-γ increased after SEB administration, and the elevated IFN-γ production was significantly inhibited by tryptanthrin treatment. These results suggest that tryptanthrin may be effective in the treatment of disorders of the intestines, such as food poisoning, that are associated with activated lymphocytes.
We investigated the effects of restraint and footshock stress and norepinephrine treatment on gastric emptying. The gastric emptying was significantly inhibited by restraint stress. β3-adrenergic antagonist canceled the inhibition of gastric emptying caused by restraint stress, while β1-, β2-, α1-, and α2-adrenergic antagonists did not affect the inhibition. Norepinephrine treatment also inhibited gastric emptying. The inhibition by norepinephrine treatment was canceled by β1-, β2-, and β3-adrenergic antagonists, but not by α1- and α2-adrenergic antagonists. On the other hand, footshock stress did not affect gastric emptying. These results suggest that β3-adrenoceptors play an important role in the inhibition of gastric emptying caused by restraint stress, while nonspecific β-adrenoceptors are involved in that caused by norepinephrine treatment.
The inhibitory effects of (−)-matairesinol (MTS) isolated from Thujopsis dolabrata var. hondai on the activities of four distinct Ser/Thr-protein kinases [two casein kinases (CK-I and CK-II), A-kinase and C-kinase] were determined in vitro. It was found that (i) MTS inhibits the activities of CK-I and C-kinase α (ID50=approx. 10 μM) in a dose-depedent manner, but high doses are required to inhibit A-kinase activity (ID50=approx. 90 μM); (ii) the autophosphorylation of CK-I is more sensitive to MTS (ID50=approx. 0.2 μM); (iii) MTS inhibits CK-I activity in a manner similar to that observed with CK-I-7 (a CK-I inhibitor); and (iv) the compound inhibits CK-I activity by affecting ATP binding in a mixed type manner. These results indicate that MTS is a potent CK-I inhibitor in vitro.
We studied altered gene expressions in BALB/3T3 cells treated by different tumor promoters in the promotion phase of a transformation assay, an in vitro model of a two-stage carcinogenicity test, using fluorescent mRNA differential display analysis. Expression of the NP95 gene, which was previously found to be the gene of a murine nuclear protein associated with cell proliferation, was increased in the cultures treated by 12-O-tetradecanoylphorbol-13-acetate (TPA), okadaic acid, and orthovanadate. The upregulation of NP95 mRNA was confirmed by reverse transcription-PCR, and Northern blot. TPA, okadaic acid, and orthovanadate enhanced cell proliferation as measured by a 5-bromo-2′-deoxyuridine incorporation assay. The expression level of NP95 mRNA was not affected by the treatment with typical carcinogens benzo[a]pyrene and 3-methylcholanthrene at concentrations at which they act as initiators of cell transformation. These facts may imply that the enhancement of cell transformation by these tumor promoters is due, at least in part, to the acceleration of cell proliferation. NP95 mRNA was also increased in the transformed BALB/3T3 cells. Overexpression of NP95 may also participate in the maintenance of the transformed phenotype.
N-Acyloctahydropyrido[3,2,1-ij][1,6]naphthyridines were synthesized as derivatives of matrine-type alkaloids, and the structure–activity relations were examined by the acetic acid-induced abdominal contraction test. The antinociceptive potencies of N-acyloctahydropyrido[3,2,1-ij][1,6]naphthyridines were significantly lower than those of (+)-matrine. The antinociceptive effects of N-benzyloctahydropyrido[3,2,1-ij][1,6]naphthyridines are approximately 5.6 to 6.5 times less than those of N-benzoyloctahydropyrido[3,2,1-ij][1,6]naphthyridine. These findings suggest that the amide group of matrine-type alkaloids is an essential functional group that influences antinociceptive potency. The antinociceptive effect of 4c was markedly antagonized by pretreatment with Naloxone, and that of 3c partially so.
Enzyme inhibitory activities of 14 iridoids previously obtained from two Malaysian medicinal plants, Saprosma scortechinii and Rothmannia macrophylla, were evaluated in vitro using soybean lipoxygenase and bovine testis hyaluronidase. Most of the iridoids, including asperulosidic acid, paederosidic acid, and an epimeric mixture of gardenogenins A and B, did not show any effect on the enzyme activities, except for the bis-iridoids, which inhibited the lipoxygenase activity with their IC50 values of approximately 1.3 times that of a known inhibitor, fisetin. Structural modification of asperulosidic acid and paederosidic acid through enzymatic hydrolysis by β-glucosidase resulted in their inhibition towards the enzyme activities, and these activities were enhanced by the presence of some amino acids (lysine, leucine or glutamic acid) or ammonium acetate. Mixtures of gardenogenins A and B; isomers of non-glucosidic iridoids, incubated with amino acid or ammonium acetate did not show any inhibitory effect on the enzyme activities during the 6 h incubation period, except for lysine where spontaneous reaction between the iridoids and amino acid resulted in the inhibition of lipoxygenase activity. The results from these biomimetic reactions suggested that the iridoid aglycons and the intermediates formed by these reactive species could inhibit the enzyme activities, and thus substantiate previous reports that the formation of iridoidal aglycons is a prerequisite for the iridoid glycosides to demonstrate some of the biological activities. In addition, the results also indicated that it is worthwhile to further explore these intermediates as potential anti-inflammatory agents.
An ethanol (EtOH) extract of the soft coral (Sinularia sp.), collected in Okinawa, demonstrated a potent inhibitory effect on lipopolysaccharide (LPS)-induced tumor necrosis factor-α (TNF-α) production by murine macrophage-like cells (RAW264.7). The activity-guided purification of the EtOH extract resulted in the isolation of two norditerpenes, norcembrenolide (1) and sinuleptolide (2). These structures were identified from the spectroscopic data. Norcembrenolide (1) and sinuleptolide (2) inhibited TNF-α production in a dose-dependent manner, and showed a more potent effect than prednisolone at the concentration of 33 μg/ml. They also exhibited an inhibitory effect on nitric oxide (NO) production not influenced by cytotoxicity.
We investigated the inhibitory effect of Cyclocarya paliurus (Batal.) Iljinskaja (C. paliurus) extract on postprandial hyperlipemia in mice. A single oral administration of C. paliurus extract (250 mg/kg) suppressed an increase in plasma triacylgycerol (TG) levels when fed with 5 ml/kg of lard and olive oil. The inhibition rates were 28.6% and 24.1%, respectively, but free fatty acid (FFA) levels in plasma were not significantly affected as compared with control group mice. In addition, C. paliurus extract showed inhibitory activity toward pancreatic lipase, a key enzyme of dietary TG absorption, with an IC50 of 9.1 μg/ml in vitro. Our results suggested that the hypolipemic action of C. paliurus extract was probably interrelated with suppression of the activity of digestive lipase, and as a result, the blood lipid level was reduced.
Specific gene delivery into hepatoma cells by liposomes and specific gene expression under the control of the cyclin A promoter were examined in HepG2 cells, a hepatoblastoma cell line that overexpresses cyclin A. A plasmid carrying the luciferase gene under the cyclin A promoter sequence was condensed with poly-L-lysine and encapsulated into anionic asialofetuin-labeled liposomes (AF-liposomes), which were preferentially taken up by hepatocytes through the action of the asialoglycoprotein receptor (AgpR). AF-liposomes delivered plasmids to the hepatoma cells by receptor-mediated endocytosis through the AgpR, and transgene expression could be achieved under the control of the cyclin A promoter. Furthermore, a fusogenic lipid, DOPE, as a liposomal component was required for the enhancement of transfection efficiency of AF-liposomes.
A series of clinical studies on the cytochrome P450 2C19 (CYP2C19) genotype and the pharmacokinetics and pharmacodynamics of three proton pump inhibitors (PPIs), omeprazole, lansoprazole and rabeprazole, have been conducted to establish the individualized pharmacotherapy based on the CYP2C19 genotyping, and in the present study, the CYP2C19 genotype-dependency was more pronounced for omeprazole than the other two. Herein, to validate further the difference among 3 PPIs in CYP2C19 genotype-dependency on the phenotype, a comparative in vitro study was conducted using the human liver microsomes and newly developed anti-human CYP antibodies. The residual concentrations of omeprazole and lansoprazole in 5 lots of human liver microsomes were dependent on the CYP2C19 activities, however, for rabeprazole, there was no correlation. The hydroxylation of omeprazole was more inhibited by anti-CYP2C19 antibody than lansoprazole, whereas anti-CYP3A4 antibody showed similar inhibition. In conclusion, the relative contribution of CYP2C19 on total metabolism of 3 PPIs elucidated herein coincided with the CYP2C19 genotype-dependent pharmacokinetics.
Six clinically isolated strains of Serratia marcescens were tested for their drug resistance. All showed fairly high resistance to many antimicrobial agents tested including norfloxacin, streptomycin, ampicillin, erythromycin, tetracycline, chloramphenicol, and antimicrobial dyes. Using the drug-hypersensitive strain of Escherichia coli KAM32 as the host, we cloned the genes responsible for multidrug resistance from chromosomal DNA of one of the strains of S. marcescens, NUSM8906. We obtained 28 hybrid plasmids that made host cells resistant to several antimicrobial agents. Many of the transformants harboring each of the plasmids showed multidrug resistance, and some showed resistance to specific drugs. The hybrid plasmids were classified into several groups based on their drug specificity. It appears that each class of plasmid carries different types of drug resistance genes. Analysis of such genes will reveal the multiple mechanisms involved in multidrug resistance in S. marcescens.
The compounds in diesel exhaust particles (DEP) that are responsible for vasodilatation were isolated and characterized for the first time. From benzene extract of DEP, 2-methyl-4-nitrophenol, 3-methyl-4-nitrophenl and 4-nitrophenol were isolated, and their vasodilatation activities were confirmed. 3-methyl-4-nitrophenol caused dilatation of rat thoracic artery, and the other two nitrophenols, also showed vasodilatation activities.