Bioenergetic effect of liposomal coenzyme Q10 on myocardial ischemia reperfusion injury.

The antioxidant and bioenergetic effects of CoQ10 are well known but its clinical utility is limited by the requirement for enteral administration. A newly developed liposomal CoQ10 (CoQ) is water soluble and capable of intravenous administration. The purpose of this study is to determine the mechanism by which acute administration CoQ protects myocardium from reperfusion (Rp) injury. Rats were pretreated with CoQ 10 mg/kg i.v. 30 min prior to the experiment. Control rats were pretreated with liposome only. Hearts were excised and subjected to equilibration, 25 min of normothermic ischemia and 40 min of Rp on a Langendorff apparatus. At end Rp, CoQ hearts recovered 74 +/- 5% of their DP vs. 50 +/- 9% in control (p < 0.05). Aerobic efficiency was maintained (0.66 +/- 0.02 vs. control, 0.5 +/- 0.04, p < 0.003) and CoQ hearts lost less CK activity vs. control (p < 0.02). PCr and ATP were higher than control (p < 0.05, 0.02, respectively). Results show that i.v. CoQ improves recovery of function, aerobic efficiency, CK activity, and recovery of PCr and ATP after Rp. This suggests that acute administration of liposomal CoQ improves myocardial tolerance to I/R via its role as an antioxidant as well as improving oxygen utilization and high energy phosphate production.

Anti-MRSA activity of sophoraflavanone G and synergism with other antibacterial agents.

Anti-MRSA activity of sophoraflavanone G (SFG) and synergism between SFG and antibacterial agents against MRSA (methicillin-resistant Staphylococcus aureus) were evaluated by means of Minimal Inhibitory Concentrations (MIC). The MICs of SFG against 27 strains of MRSA ranged from 3.13 to 6.25 micrograms ml-1. Synergism between SFG and vancomycin hydrochloride (VCM) or fosfomycin (FOM) was observed (the fraction inhibitory concentration (FIC) indices were 0.16 and 0.48), while partial synergism was admitted between SFG and other antibacterial agents such as methicillin (DMPPC), cefzonam (CZON), gentamicin (GM), minocycline (MINO) and levofloxacin (LVFX) (the FIC indices were 0.71, 0.73, 0.69, 0.65 and 0.58, respectively). These findings suggest that SFG in combination with VCM or FOM may be useful in controlling MRSA infections.

Effects of preparations of Chinese medicinal prescriptions on digestive enzymes in vitro and in vivo.

The effects of preparations of Chinese medicinal prescriptions on the activities of digestive enzymes were investigated. The starch dextrinizing activity of pancreatin was inhibited by Keishi-bukuryo-gan, Sho-seiryu-to, Hachimi-jio-gan, Unsei-in and Keishi-ka-shakuyaku-to to below 40% of the control activity. Hachimi-jio-gan and Sho-seiryu-to, in particular, lowered the activity to 4% and 24% of the control activity, respectively. The protein digestive activity of pancreatin was lowered by Keishi-bukuryo-gan, Oren-gedoku-to, Ryutan-shakan-to, Tokaku-joki-to, Yokuinin-to and Hange-shashin-to, to 56 to 70% of the control activity. To investigate the effects of the preparations of Chinese medicinal prescriptions on digestive enzymes in vivo, 600 mg (185 kBq) of 125I-egg white albumin were administered orally to rats 30 min before the administration of 100 mg of Tokaku-joki-to or Oren-gedoku-to. The radioactivity which transferred to the blood was less in the animals pre-fed these prescriptions than in the control animals, indicating that the digestion of egg white albumin was delayed in the presence of the prescriptions.

[The cause of polyurethane catheter cracking during constant infusion of etoposide (VP-16) injection].

We studied the cause of cracking of a clinically used polyurethane (PU) catheter during the constant infusion of etoposide (VP-16) injection (Lastet), administered without dilution to patients as a part of combination high-dose chemotherapy. After VP-16 injection was infused into the PU catheter at a constant infusion rate (30 ml/h) for 24 h, a decrease in the elasticity (36% of untreated) and on increase in the length of the catheter (3.7%) were observed. These changes were significantly higher than those treated with the control saline. The similar changes of the PU catheter were observed after treatment with a basal solution containing polyethylene glycol 400 (PEG 400), polysorbate 80 and ethanol, which is the vehicle of the VP-16 injection, and with ethanol alone. Moreover, obvious degeneration of the internal wall (occurrence of spots like melting) and cutting face (micro-cracking) of the catheter was observed with an electron microscope after treatment with the vehicle. On the other hand, the elasticity or extension of the PU catheter were not changed after treatment with saline or PEG 400. From these findings, it was suggested that the degeneration and subsequent cracking of the PU catheter during the infusion of VP-16 injection was caused by ethanol contained in its injection solution. No cracking or morphological changes of polyvinyl chloride (PVC) and silicone catheters were found after treatment with the vehicle solution. However, since it has been reported in previous reports that di(2-ethylhexyl)phthalate was leached from PVC bags, the high dose chemotherapy with the dilution-free VP-16 injection should be achieved safely and effectively using a silicon catheter, rather than the PU catheter.

Polysaccharide of Astragali radix enhances IgM antibody production in aged mice.

The effect of Astragali Radix (AR) on IgM antibody production in mice of various ages (10 weeks, 36 weeks and 60 weeks) was examined. The antibody production levels in the 36- and 60-week-old mice were significantly decreased to about 70 and 60% of that in the 10-week-old mice. The enhancement effect of a crude polysaccharide AR fraction on the antibody production was nil in the 10-week-old mice, but significant enhancement effects were observed in the 36- and 60-week-old mice, compared to the age-matched control. Two polysaccharides active in the enhancement of the IgM antibody production in the aged mice were isolated from the high molecular weight fraction of AR by cetavlon precipitation, ion-exchange and gel permeation chromatography. The molecular masses of these polysaccharides were calculated by HPLC in salt solution. Only one major peak was observed for each, and their molecular masses were estimated to be 1.2 x 10(4) and 2.2 x 10(4). The major components of these polysaccharides were neutral carbohydrates (89.3 and 95.5%), followed by uronic acid and protein; glucose was the predominant sugar component.

Predictors of the progression of diabetic nephropathy and the beneficial effect of angiotensin-converting enzyme inhibitors in NIDDM patients.

A progressive decline in glomerular function occurs in diabetic nephropathy. The predictive effects of progression promoters were examined in 182 non-insulin-dependent diabetic patients from a baseline serum creatinine concentration of 133 mumol/l. During a total of 605 person-years follow-up, 107 patients developed end-stage renal failure requiring dialysis. The rate of decline of renal function was highly variable. Urinary protein excretion was the strongest predictor correlated to the rate of decline, followed by diastolic and systolic blood pressure, total cholesterol and platelet count, while the protective effects were seen in serum albumin and haematocrit. Adjustment for urinary protein excretion revealed that diastolic blood pressure, familial predisposition to hypertension, serum albumin, and smoking were independent significant predictors. Angiotensin converting enzyme inhibitors (ACE-I) significantly retarded the development of end-stage renal failure compared to antihypertensives other than ACE-I (mostly nifedipine), and the effect was evident particularly in patients with proteinuria below the median (2.5 g/24 h) (presumably those who responded to ACE-I). A complex effect of proteinuria in association with blood pressure elevation, familial predisposition to hypertension, hypoalbuminaemia, and smoking may play an important role in the progression of nephropathy.

Passive protection of neonatal calves against bovine coronavirus-induced diarrhea by administration of egg yolk or colostrum antibody powder.

The protective effect of egg yolk and colostrum powders prepared from hens and cows vaccinated with inactivated bovine coronavirus (BCV) antigen was evaluated in a challenge model with a virulent BCV strain. Twenty three calves from BCV-free herds were randomly divided into control and several treatment groups. All calves were orally challenged with 1 x 10(9) TCID50 of the virulent Kakegawa strain of BCV at 24 to 36 h after birth. Calves in treatment groups received either egg yolk powder or cow colostrum containing BCV specific antibodies. Daily treatment with these antibody preparations started 6 h until 7 days post-challenge. Control calves which received no antibody had severe diarrhea and all died within 6 days after infection. In contrast, calves fed milk containing egg yolk or colostrum with neutralization titers of 1:2560 or 1:10,240 respectively all survived and had positive weight gain unlike the other treatment groups. These results indicate that the orally administered egg yolk and colostrum powders protected against BCV-induced diarrhea in neonatal calves and that the egg yolk used provided a higher degree of protection compared to colostrum powder on a titer basis. Treatment with whole egg yolk from immunized hens therefore provides a more efficacious alternative to the existing methods of specific passive protection against BCV.

Protective effect of astragali radix by oral administration against Japanese encephalitis virus infection in mice.

We investigated the protective effect of Astragali Radix (AR) by oral administration against Japanese encephalitis virus (JEV) infection in mice, the pharmacological effects of AR extracts (AE) in different origin, and the chemical composition of the AEs. A protective effect was demonstrated in all four AEs used, however, the effective grade for each one was different. In the control group, an increase of hemagglutination inhibition (HI) antibody titer was observed in all mice surviving 25 d after JEV inoculation. However, the increase of HI antibody titer was not observed in some animals administered an AE. In the control group, the rate of HI antibody positive mice was 90% 3 d after JEV inoculation, while the four groups which received the AE had a 30-60% positive rate. In mice which received the AE, the peritoneal exudate cell (PEC) numbers increased significantly compared to the control group. The predominant cell population of PECs in mice receiving the AE was macrophages, and in the PEC, the active oxygen (AO) production was high. From these results, we propose that the protective effect of AE by oral administration is based on a non-specific mechanism during the early stage of infection, before shifting to antibody production, and that macrophages play an important role in this resistance to JEV infection, e.g., by inducing the production of AO. In the chemical composition of each AE, carbohydrate was the major component.

Protective effect of Astragali Radix by intraperitoneal injection against Japanese encephalitis virus infection in mice.

We examined the protective effect of Astragali Radix extracts (AE) by intraperitoneal injection against Japanese encephalitis virus (JEV) infection in mice. A protective effect was observed by all four samples of AE used. However, the degree of effectiveness for each AE was different. The observed survival rates of the groups injected with sample A (from Shanhsi, Japanese name Sansei-syo) and sample D (from Hokkaido) extracts were higher than 80% at 21 d after JEV inoculation. The groups injected with sample B (from Hopei, Japanese name Kahoku-syo) and sample C (from Hsiahsi, Japanese name Sensei-syo) extracts had a 60% survival rate. The increase in hemagglutination inhibition antibody titer was negligible in mice that survived 21 d after JEV inoculation. The antiviral effect of AE was examined by plaque assay in vitro, but no antiviral effect was shown. In mice injected with AE, the peritoneal exudate cell (PEC) numbers increased significantly, compared to the control. In these PEC, active oxygen production was also high. Also the group as a whole displayed a high survival rate against JEV infection, these were so strong. From these results, we propose that the protective effect of AE is dependent on a non-specific mechanism during the early stage of infection, before it shifts to antibody production, and that PEC plays an important role.

Passage of chicken egg yolk antibody treated with hydroxypropyl methylcellulose phthalate in the gastrointestinal tract of calves.

Two types of chicken egg yolk antibody samples for oral passage trials in calves were prepared: (1) hydroxypropyl methylcellulose phthalate (HPMCP) antibody powder (HAP)--a powder produced by spray-drying a supernatant obtained after precipitation of lipids from egg yolk with HPMCP and (2) control antibody power (CAP)--a powder produced from an antibody solution with HPMCP. Antibody activity and pattern of distribution of both antibody preparations in the gastrointestinal tract of calves were compared by enzyme-linked immunosorbent assay. At 2 hr post administration, anti-K99 fimbrial antibodies from both the CAP and the HAP were detected in the abomasum of calves with titers of 1:128 and 1:256, respectively. However, at 4 hr, anti-K99 fimbrial titers of the CAP and the HAP were reduced to 1:2 and 1:64, respectively, due to digestion in the abomasum. These results indicated that the egg yolk antibody powder with HPMCP was more resistant against gastric juice in the stomach, thereby, ensuring a transfer of functional antibodies to the small intestine of calves after oral administration.

Effect of TJ-960 (a Japanese herbal medicine) on free radical changes within an iron-induced focal epileptogenic region in rat brain measured by in vivo L-band electron spin resonance.

In vivo L-band electron spin resonance (ESR) system was used to measure the decay of nitroxide radicals in the living rat cerebrum containing a cortical area with an iron-induced epileptogenic region. Carbamoyl-PROXYL (3-carbamoyl-2,2,5,5-tetramethyl-1-pyrrolinyloxy) was injected intraperitoneally. The decay of nitroxide radicals in the iron-injected regions at the subacute stage of epileptogenesis was prolonged. The Japanese herbal medicine, sho-saiko-to-go-keishi-ka- shakuyakuto (TJ-960), a compound known to have free radical scavenging activity, inhibited the delay of nitroxide radical decay in the rat cerebrum with an iron-induced epileptic focus following injection of carbamoyl-PROXYL. TJ-960 did not affect the decay of nitroxide radicals in the control rats' brain. These data show that TJ-960 has the scavenging activity for free radicals generated within an iron-induced epileptogenic regions of rat brain.

Dietary fatty acid modulates actions of nucleotides on humoral immune responses.

Polynucleotides enhance T cell-dependent antibody production in culture. Impaired antibody production in mice fed a nucleotide-free diet can be easily restored by in vivo supplementation of both a mononucleotide-nucleoside mixture (OG-VI) and polynucleotides. Polynucleotides appear to act partly by modulating antigen presentation processes mediated by cell surface molecules. We examined whether dietary fatty acid manipulation alters nucleotides' actions on humoral immunity. Antibody production was studied in C57B1/6 mice fed I) a nucleotide-free diet high in saturated fatty acid (SFA diet), 2) a nucleotide-free diet high in polyunsaturated fatty acid (PUFA diet), and 3) a regular nucleotide-free diet (control). In vivo and in vitro T cell-dependent antibody production decreased in all groups, but mice fed the SFA diet produced more antibody in vivo than did mice in the other diet groups. Spleen cells from mice fed the SFA diet also produced more interferon-alpha when stimulated with mitogens than did those from mice fed the control diet. In contrast, polynucleotides enhanced in vitro antibody production much less efficiently in mice fed the SFA diet than in the other mice and in vivo supplementation of OG-VI was also less effective in restoring impaired antibody production in these mice. A diet with a high content of SFA may alter nucleotides' action on humoral immune responses, in addition to its direct effects on immune functions.

Molecular cloning and characterization of a plant serine acetyltransferase playing a regulatory role in cysteine biosynthesis from watermelon.

Serine acetyltransferase (SATase; EC 2.3.1.30), which catalyzes the reaction connecting serine and cysteine/methionine metabolism, plays a regulatory role in cysteine biosynthesis in plants. We have isolated a cDNA clone encoding SATase by direct genetic complementation of a Cys- mutation in Escherichia coli using an expression library of Citrullus vulgaris (watermelon) cDNA. The cDNA encodes a polypeptide of 294 amino acids (31,536 Da) exhibiting 51% homology with that of E. coli SATase. DNA-blot analysis indicated the presence of a single copy of the SATase gene (sat) in watermelon. RNA hybridization analysis suggested the relatively ubiquitous and preferential expression in the hypocotyls of etiolated seedlings. Immunoblot analysis indicated the accumulation of SATase predominantly in etiolated plants. L-Cysteine, an end product of the cysteine biosynthetic pathway, inhibited the SATase in an allosteric manner, indicating the regulatory function of SATase in this metabolic pathway, whereas beta-(pyrazole-1-yl)-L-alanine, a secondary metabolite formed partly through the cysteine biosynthetic pathway, showed no inhibitory effect. A multi-enzyme complex was formed from recombinant proteins of SATase and cysteine synthase (O-acetylserine(thiol)-lyase) from watermelon, suggesting efficient metabolic channeling from serine to cysteine, preventing the diffusion of intermediary O-acetyl-L-serine.

A convenient screening test for hypoxic cell radiosensitizers/cytotoxins.

A convenient in vitro screening test using E. coli B/r for evaluating a variety of hypoxic cell radiosensitizers/hypoxic cell cytotoxins has been developed for the initial selection of candidates in medicinal/organic chemistry laboratories. E. coli cells were used for convenience since: (1) the bacterium is grown using commercially available broths, where it multiplies rapidly, and requires little specialized equipment for growth and handling. (2) More is known about the genetics and biochemistry of the radiation damage to these cells and their repair than any other organism.

Research note: avidity of chicken yolk antibodies to enterotoxigenic Escherichia coli fimbriae.

The yolk antibodies from chickens and the serum and colostrum antibodies from cows were obtained after immunization of these animals with inactivated bacterin or purified K99 fimbriae from enterotoxigenic Escherichia coli (ETEC). The avidity of anti-K99 fimbriae antibodies produced from either chickens or cows was measured by competitive binding assay of ELISA. The yolk antibodies competed strongly with the serum and colostrum antibodies from immunized cows and inhibited 40 to 80% of the binding of these antibodies. Results demonstrate that the avidity of antibodies obtained from immunized chickens compares with that obtained from immunized cows. Thus, the yolk antibody from immunized chickens, aside from its use for prophylaxis against some infectious diseases, may also serve as effective ligand for purification of biologically active substances such as fimbrial antigens by affinity chromatographic procedures.