The effect of phosphatidylserine administration on memory and symptoms of attention-deficit hyperactivity disorder: a randomised, double-blind, placebo-controlled clinical trial.

BACKGROUND: Attention-deficit hyperactivity disorder (ADHD) is the most commonly diagnosed behavioural disorder of childhood, affecting 3-5% of school-age children. The present study investigated whether the supplementation of soy-derived phosphatidylserine (PS), a naturally occurring phospholipid, improves ADHD symptoms in children. METHODS: Thirty six children, aged 4-14 years, who had not previously received any drug treatment related to ADHD, received placebo (n = 17) or 200 mg day(-1) PS (n = 19) for 2 months in a randomised, double-blind manner. Main outcome measures included: (i) ADHD symptoms based on DSM-IV-TR; (ii) short-term auditory memory and working memory using the Digit Span Test of the Wechsler Intelligence Scale for Children; and (iii) mental performance to visual stimuli (GO/NO GO task). RESULTS: PS supplementation resulted in significant improvements in: (i) ADHD (P < 0.01), AD (P < 0.01) and HD (P < 0.01); (ii) short-term auditory memory (P < 0.05); and (iii) inattention (differentiation and reverse differentiation, P < 0.05) and inattention and impulsivity (P < 0.05). No significant differences were observed in other measurements and in the placebo group. PS was well-tolerated and showed no adverse effects. CONCLUSIONS: PS significantly improved ADHD symptoms and short-term auditory memory in children. PS supplementation might be a safe and natural nutritional strategy for improving mental performance in young children suffering from ADHD.

Lentivirus IL-10 gene therapy down-regulates IL-17 and attenuates mouse orthotopic lung allograft rejection.

The purpose of the study was to examine the effect of lentivirus-mediated IL-10 gene therapy to target lung allograft rejection in a mouse orthotopic left lung transplantation model. IL-10 may regulate posttransplant immunity mediated by IL-17. Lentivirus-mediated trans-airway luciferase gene transfer to the donor lung resulted in persistent luciferase activity up to 6 months posttransplant in the isograft (B6 to B6); luciferase activity decreased in minor-mismatched allograft lungs (B10 to B6) in association with moderate rejection. Fully MHC-mismatched allograft transplantation (BALB/c to B6) resulted in severe rejection and complete loss of luciferase activity. In minor-mismatched allografts, IL-10-encoding lentivirus gene therapy reduced the acute rejection score compared with the lentivirus-luciferase control at posttransplant day 28 (3.0 ± 0.6 vs. 2.0 ± 0.6 (mean ± SD); p = 0.025; n = 6/group). IL-10 gene therapy also significantly reduced gene expression of IL-17, IL-23, and retinoic acid-related orphan receptor (ROR)-γt without affecting levels of IL-12 and interferon-γ (IFN-γ). Cells expressing IL-17 were dramatically reduced in the allograft lung. In conclusion, lentivirus-mediated IL-10 gene therapy significantly reduced expression of IL-17 and other associated genes in the transplanted allograft lung and attenuated posttransplant immune responses after orthotopic lung transplantation.

MMP-dependent migration of extrapulmonary myofibroblast progenitors contributing to posttransplant airway fibrosis in the lung.

Myofibroblasts play a central role in fibroproliferative airway remodeling in obliterative bronchiolitis (OB) after lung transplantation. The purpose of the study is to elucidate the mechanisms whereby matrix metalloproteinases (MMPs) contribute to myofibroblast-mediated allograft airway fibrosis. In an intrapulmonary tracheal transplant model of OB, broad-spectrum MMP inhibitors, SC080 and MMI270 reduced the number of myofibroblasts at day 28 without changing differentiation, proliferation or apoptosis of myofibroblasts or fibroblasts. Next, myofibroblasts in allograft airway fibrosis were demonstrated to be almost exclusively of extrapulmonary origin by analyzing RT1A(n) positive myofibroblasts in an animal model combining orthotopic lung transplantation (from Lewis (RT1A(l)) to F1 (Brown-Norway (RT1A(n)) x Lewis)) and intrapulmonary tracheal transplantation (from a Wister-Furth rat (RT1A(u)) into the transplanted Lewis-derived lung). Using peripheral blood mononuclear cells (PBMCs) that can differentiate into alpha-SMA positive myofibroblasts in vitro, we demonstrated their contribution to the myofibroblast population of allograft airway fibrosis in vivo using a fluorescence-labeling cell tracking system. Moreover, PBMC-derived fibroblast-like cells expressed high levels of MMP-9 and MMP-12 and their migration was inhibited by MMP inhibitors in a wound healing assay. In conclusion, MMP-dependent migration of PBMC-derived myofibroblast precursors is an important contributing mechanism to the development of allograft airway fibrosis.

Normothermic ex vivo perfusion prevents lung injury compared to extended cold preservation for transplantation.

Treatment of injured donor lungs ex vivo to accelerate organ recovery and ameliorate reperfusion injury could have a major impact in lung transplantation. We have recently demonstrated a feasible technique for prolonged (12 h) normothermic ex vivo lung perfusion (EVLP). This study was performed to examine the impact of prolonged EVLP on ischemic injury. Pig donor lungs were cold preserved in Perfadex for 12 h and subsequently divided into two groups: cold static preservation (CSP) or EVLP at 37 degrees C with Steen solution for a further 12 h (total 24 h preservation). Lungs were then transplanted and reperfused for 4 h. EVLP preservation resulted in significantly better lung oxygenation (PaO(2) 531 +/- 43 vs. 244 +/- 49 mmHg, p < 0.01) and lower edema formation rates after transplantation. Alveolar epithelial cell tight junction integrity, evaluated by zona occludens-1 protein staining, was disrupted in the cell membranes after prolonged CSP but not after EVLP. The maintenance of integrity of barrier function during EVLP translates into significant attenuation of reperfusion injury and improved graft performance after transplantation. Integrity of functional metabolic pathways during normothermic perfusion was confirmed by effective gene transfer and GFP protein synthesis by lung alveolar cells. In conclusion, EVLP prevents ongoing injury associated with prolonged ischemia and accelerates lung recovery.

A new SOBP-formation method by superposing specially shaped Bragg curves formed by a mini-ridge filter for spot scanning in proton beam therapy.

PURPOSE: We propose a new spread-out Bragg peak (SOBP) formation method for low-energy regions of spot-scanning proton therapy in order to reduce the required number of energy layers while maintaining high dose uniformity, while maintaining the distal falloff as sharp as possible. METHODS: We use only one specially shaped mini-ridge filter (MRF) to create new trapezoidal Bragg curves (TBCs) from very sharp pristine Bragg curves (PBCs) of low-energy proton beams. The TBC has three pre-designed dose regions of proximal, flat-top, and distal components. These components are designed to have nearly equal depth lengths and good linearity. Then, the required SOBP is formed by superposing the TBCs with the correct spacing and beam intensity weights. We then compare the performance of the TBC-based SOBPs with those formed by PBCs. RESULTS: The dose uniformities of the SOBP formed by the proposed method are kept within the design tolerance, and are equivalent to those of conventional SOBPs. The sharpness of the distal falloff is reasonably kept by the deepest TBC. The required number of energy layers is significantly reduced compared with that of conventional PBC-based SOBP. CONCLUSIONS: The proposed method enables shortening of the irradiation time of spot-scanning proton beam therapy in low-energy regions with a reduced number of energy layers. It can be realized by using only one specially shaped MRF, which can be easily installed at any facility.

Effects of a combined exercise plus diet program on cardiorespiratory fitness of breast cancer patients.

BACKGROUND: Decreases in cardiorespiratory fitness among breast cancer patients have often been reported in previous studies, affecting patients' health and survival. Peak oxygen uptake ([Formula: see text]) is the gold standard for assessing cardiorespiratory fitness and is inversely correlated with cardiovascular disease among women with breast cancer. Some previous studies have reported that aerobic exercise and proper diet positively influence [Formula: see text]. However, almost all studies have been conducted in the Western countries, and few studies are investigating on Asian women who have lower BMI compared with Western ones. PURPOSE: Investigating the effects of a combined exercise and diet program among Japanese cancer patients undergoing therapy on [Formula: see text]. METHODS: Thirty-two Japanese women with breast cancer undergoing endocrine therapy (age; 50 ± 6 years, body weight; 59 ± 10 kg) were voluntarily assigned to either intervention group (n = 21) or control group (n = 11). The intervention group completed a 12-week combined exercise plus diet program, consisting of weekly aerobic exercise and maintaining a nutritionally well-balanced 1200 kcal/day diet. The control group was instructed to continue with their usual activities. Anthropometric indices and [Formula: see text] were measured at baseline and after the 12-week program. RESULTS: All 21 women completed the 12-week program. The [Formula: see text] significantly increased from 26.7 to 30.4 mL/kg/min (1.57-1.62 L/min) in the intervention group, while it remained unchanged (26.9-26.9 mL/kg/min) in the control group. Mean reduction of body mass index was - 2.1 in the intervention group (P < .001) and + 0.1 in the control group. CONCLUSIONS: Our combined exercise plus diet program may contribute to improvement in cardiorespiratory fitness and body weight compared with control group.

Correction to: Effects of a combined exercise plus diet program on cardiorespiratory fitness of breast cancer patients.

In the original publication of this article, Table 1 was published incorrectly. The correct Table 1 is given in the following page.

Effects of menstrual cycle on gene transfection through mouse vagina for DNA vaccine.

Human immunodeficiency virus (HIV) infections mainly occur through the vaginal and rectal mucosal membranes. In the present study, to develop a DNA vaginal vaccine against viral and bacterial infections, the effects of the menstrual cycle on DNA transfection through the vaginal mucosa in female mice and transfection enhancement by electroporation, a chelating agent, cell-penetrating peptides (CPP) and nuclear localizing signals (NLS) were investigated. The transfection efficiencies of a marker plasmid DNA (pDNA), pCMV-Luc, on the vaginal mucosal membrane in mice at the stages of metestrus and diestrus were significantly higher than those at the stages of proestrus and estrus. The gene expression was markedly enhanced by electroporation and by pretreatment with the chelating agent. The highest level of expression was obtained by 2h pretreatment with 5% citric acid solution combined with electroporation with 15 pulses at 250 V/cm for 5 milliseconds (ms). Furthermore, a synergistic promoting effect on pDNA transfection was obtained by co-administration of CPP, the Tat peptide analog, and NLS, the NF-kappaB analog. These results indicate that effective DNA vaccination administered through the vaginal tract is possible by selecting the menstrual stage and overcoming the mucosal barrier using a combination of methods that promotes uptake.

Rapid Bactericidal Action of Propolis against Porphyromonas gingivalis.

Propolis, a resinous substance produced by bees, is used as a folk medicine for treatment of periodontal diseases. However, its mode of the action and the compounds responsible for its activities remain obscure. In the present study, we comprehensively investigated the antibacterial activities of ethanol-extracted propolis (EEP) and EEP-derived compounds toward Porphyromonas gingivalis, a keystone pathogen for periodontal diseases. Broth microdilution and agar dilution assays were used to determine the minimum inhibitory concentrations of EEP against a range of oral bacterial species, of which P. gingivalis showed a higher level of sensitivity than oral commensals such as streptococci. Its antibacterial activity toward P. gingivalis was maintained even after extensive heat treatment, demonstrating a high level of thermostability. EEP also induced death of P. gingivalis cells by increasing membrane permeability within 30 min. Spatiotemporal analysis based on high-speed atomic force microscopy revealed that EEP immediately triggered development of aberrant membrane blebs, followed by bleb fusion events on the bacterial surface. Furthermore, we isolated artepillin C, baccharin, and ursolic acid from EEP as antibacterial compounds against P. gingivalis. Of those, artepillin C and baccharin showed bacteriostatic activities with membrane blebbing, while ursolic acid showed bactericidal activity with membrane rupture. In particular, ursolic acid demonstrated a greater ability to affect bacterial membrane potential with increased membrane permeability, probably because of its highly lipophilic nature as compared with other compounds. Taken together, these findings provide mechanistic insight into the antibacterial activities of EEP and its exquisite membrane-targeting antibacterial compounds and imply the applicability of narrow-spectrum therapeutics with EEP for treatment of periodontitis. In addition, the advanced technology utilized in the present study to visualize the nanometer-scale dynamics of microorganisms will contribute to expanding our understanding of the activities of antimicrobials and the mechanism of drug resistance in bacteria.

Relationship between copper and lipids and atherogenic indices soon after birth in Japanese preterm infants of 32-35 weeks.

Several studies have reported association of altered levels of lipids and some trace elements with risk factors for cardiovascular disease development in adulthood. Accordingly, the present study aimed to determine the relationship among the serum levels of copper (Cu), zinc (Zn), lipids, lipoproteins and apolipoproteins in preterm infants through an assessment of atherogenic indices shortly after birth. Blood samples were collected within 20 min of birth from 45 preterm infants with gestational ages ranging from 32 to 35 weeks. Serum Cu, Zn, total cholesterol (TC), low-density lipoprotein cholesterol (LDLc), high-density lipoprotein cholesterol (HDLc), apolipoprotein-A1 (apoA1) and apolipoprotein-B (apoB) levels were measured, and the TC/HDLc, LDLc/HDLc and apoB/apoA1 ratios were calculated. Upon determining the correlation between the levels of Cu, Zn and these indices of lipid metabolism, triglyceride (TG) and Cu were found to correlate negatively with birth weight (BW) and the standard deviation (s.d.) score for body weight. Furthermore, Cu levels correlated positively with the TG level and TC/HDLc, LDLc/HDLc and apoB/apoA1 ratios and negatively with the HDLc level and HDLc/apoA1 ratios. However, a stepwise multiple regression analysis indicated that the s.d. score for BW and TG level were significant independent determinants of the Cu level. In contrast, Zn did not correlate with any of these indices. In conclusion, intrauterine growth restriction and the TG level at birth influence Cu levels in preterm infants, whereas atherogenic indices do not affect this parameter.

[New CT program simulating kidney displacement during retroperitoneal laparoscopic nephrectomy].

A total of 12 patients with malignant localized renal or ureteral neoplasms underwent multi-slice computed tomography. Imaging data were sent to the dedicated workstation to create volume rendering and virtual laparoscopic images of the kidney which was displaced ventrally with retroperitoneal balloon. These findings were compared with video images obtained during retroperitoneal laparoscopic nephrectomy. The kidney displacement simulator depicted all renal arteries (100% sensitivity) and 13 of 14 renal veins (93% sensitivity). Hilar anatomy, including the tumor, major vessels and their relationships were visualized as in the actual laparoscopic views. The desired portions of major vessels as well as the left adrenal and gonadal veins visualized with this system completely corresponded with the actual laparoscopic images during surgery. The kidney displacement simulator is useful to foresee desired portions of major vessels and branched small vessels such as the adrenal or gonadal veins in advance of surgery. It is thus able to guide surgeons and reduce operative risks and possible complications.

LR11, a mosaic LDL receptor family member, mediates the uptake of ApoE-rich lipoproteins in vitro.

Since the molecular identification of the low density lipoprotein receptor (LDLR), an ever increasing number of related proteins have been discovered. These receptors belonging to the LDLR family are thought to play key roles in lipoprotein metabolism in a variety of tissues, including the arterial wall. We have discovered that the expression of a 250-kDa mosaic LDLR-related protein, which we termed LR11 for the presence of 11 LDLR ligand-binding repeats, is markedly induced in smooth muscle cells in the hyperplastic intima of animal models used for the study of atherosclerosis. Here, we demonstrate that the human LR11, when overexpressed in hamster cells, binds and internalizes 39-kDa receptor-associated protein (RAP), an in vitro ligand for all receptors belonging to the LDLR family. Furthermore, LR11 binds the apolipoprotein E (apoE)-rich lipoproteins, beta-very low density lipoproteins (VLDLs), with a high affinity similar to that of other members, such as the LDLR and VLDL receptor. RAP and beta-VLDL compete with each other; however, other serum lipoproteins are not able to inhibit their binding. LR11 shows specific binding of apoE-enriched HDL prepared from human cerebrospinal fluid as well as of beta-VLDL, suggesting that the apoE content of lipoproteins is most likely important for mediating the high-affinity binding to the receptor. LR11-overexpressing cells are able to internalize and degrade the bound beta-VLDL; these cells also show increased accumulation of cholesteryl esters when incubated with beta-VLDL. Incubation for 48 hours with beta-VLDL of LR11-overexpressing cells, but not of control cells, promotes the appearance of numerous intracellular lipid droplets. Taken together, LR11, a mosaic LDLR family member whose expression in smooth muscle cells is markedly induced in atheroma, has all the properties of a receptor for the endocytosis of lipoproteins, particularly for the incorporation of apoE-rich lipoproteins.

Transitional changes in immunophenotypic subpopulations of human peripheral blood CD34+ cells expanded in vitro.

We determined the appropriate incubation period to expand human peripheral blood (PB) CD34+ cells for clinical application and the role of recombinant human (rh) interleukin-3 (rhIL-3) in the expansion and differentiation of these cells. The cells were purified up to 40 +/- 16% and transitional changes in immunophenotypic subpopulations in suspension culture were examined following stimulation with three different combinations of rh colony-stimulating factors (rhCSFs): 1) rhIL-3 alone, 2) rhIL-6, rhSCF, rhG-CSF, plus rhGM-CSF, and 3) the four CSFs plus rhIL-3. With all three CSF combinations, the total cells increased continuously after day 5 until day 14, and a combination of the five CSFs always gave rise to the highest number of total cells. Immunophenotypic analysis of the expanded cells showed that with all three CSF combinations CD34+ cells reached a maximal rate on day 5 and then decreased in an inverse correlation between the logarithm of CD34 positive rate and the duration of suspension culture. The maximum expansion of CD34+ cells and PB progenitor cells (PBPC) with rhIL-3 alone, the four CSFs, or the five CSFs was observed on day 5, 10, or 7, respectively. The combination of the five CSFs was identified as the most potent stimulus for expansion of PBPC and CD34+ cells, as it increased non-erythroid PBPC 89 +/- 69-fold, with a range of 24 to 204-fold on day 7. However, differences in the expansion rate of these cells on days 5, 7, and 10 were not statistically significant. The majority of purified CD34+ cells coexpressed CD38 (91 +/- 3%) but were negative for CD33 (85 +/- 4%), and one-half coexpressed CD13. With all three combinations of CSFs, the mature CD34+ cells that coexpressed CD38, CD33, or CD13 expanded in parallel with the total CD34+ cells, while an increase in relatively immature CD34+ cells, which do not express CD38, CD33, or CD13, was only statistically significant with the five CSFs. Thus, rhIL-3 played a critical role when combined with the four CSFs by increasing both mature and immature CD34+ cells.

Comparison of in vivo binding of aromatic nitro and amino compounds to rat hemoglobin.

The hemoglobin (Hb) binding of five nitroarenes, i.e. nitrobenzene (NB), 4-nitrobiphenyl (4-NBP), 1-nitropyrene (1-NP), 2-nitronaphthalene (2-NN) and 2-nitrofluorene (2-NF), and the corresponding amines, administered p.o. to male S.D. rats, was determined by HPLC, to evaluate the extent of in vivo reductive and oxidative activations of these compounds to N-hydroxylamines, which covalently bind to Hb to form acid-labile sulfinamides. Hb binding of the nitroarenes, except for NB, was significantly lower than that of the corresponding amines. Among the aromatic amines, 4-aminobiphenyl exhibited extremely high Hb binding. Hb binding of NB and 4-NBP decreased markedly after pretreatment with a mixture of antibiotics, but the binding of the others did not decrease appreciably. 1-Aminopyrene and 1-NP bound abundantly to plasma proteins, although the Hb binding was slight. Based on the Hb binding and the in vitro metabolism by liver microsomes and intestinal bacteria, the extent of in vivo reductive activation of nitroarenes is discussed.

Developmental regulation of LR11 expression in murine brain.

Receptors belonging to the low density lipoprotein receptor (LDLR) superfamily play important biological roles in addition to mediating lipoprotein metabolism. The recent discovery of a novel mosaic LDLR family member by us (Yamazaki H., Bujo, H., Kusunoki, J., Seimiya, K., Kanaki, T., Morisaki, N., Schneider, W.J., and Saito, Y. (1996) J. Biol. Chem. 271, 24761-24768) and others, which we termed LR11, offers the opportunity to gain new insights into receptor multifunctionality. The predominant expression of LR11 in brain and the presence of elements found in neural adhesion molecules suggested a function(s) in the central nervous system (CNS). In order to gain information about this complex receptor in an accessible system, we have molecularly characterized the murine LR11 and report on its detailed localization and developmental expression pattern. The primary sequence of the murine protein further establishes that LRlls are among the closest relatives within the LDLR family and that brain is the predominant site of expression. In situ hybridization showed that neuronal bodies such as Purkinje cells in the cerebellum and other neurons in the hippocampal formations and the cerebral cortex are particularly rich in LR11 transcripts. The developmental pattern of LR11 expression in brain, which peaks at 2 weeks, is in contrast to those of two other LDLR family members, the very low density lipoprotein receptor and the LDLR. During early development, murine LR11 expression levels are highly dependent on neural cell types. These findings are compatible with function(s) of LR11 in neural organization and, possibly, pathogenesis of degenerative brain diseases. In addition, detailed knowledge of LR11 biology will help to elucidate the roles of other mosaic proteins that share with LR11 elements whose function is not yet known.

Mobilization of peripheral blood progenitor cells following CHOP treatment combined with delayed granulocyte colony-stimulating factor administration in patients with non-Hodgkin's lymphoma.

The kinetic change in peripheral blood progenitor cells (PBPC) during 3 to 6 cycles of standard CHOP regimen supported with human recombinant granulocyte colony-stimulating factor (rG-CSF) was investigated in three patients with newly diagnosed intermediate grade, diffuse large cell type, non-Hodgkin's lymphoma (NHL) without bone marrow invasion. Patients were given rG-CSF subcutaneously (2 mu g/kg/day) initiated when total leukocytes was < 3.0 x 10(9)/1. When the leukocyte count remained at >3.0 x 10(9)/1, rG-CSF was started 10 days following the prior CHOP. Treatment with rG-CSF was discontinued after the leukocyte count reached >10.0 x 10(9)/1, and CHOP was started the next day (CHOP-G regimen). The number of PBPC was monitored by clonal assay in patients 1-3. No severe leukopenia with <0.5 x 10(9)/1 of neutrophils was seen in any patient. Colony-forming unit granulocyte-macrophage (CFU-GM) significantly increased after 2-3 days of consecutive administration of rG-CSF. The magnitudes of maximum amplification of CFU-GM in patients 1, 2, and 3, were 56-fold (during 3 cycles of CHOP-G), 216-fold (during 2 cycles), and 67-fold (during 4 cycles), respectively, and the absolute numbers of the maximum CFU-GM/ml blood were 983, 7,568, 9,865, respectively. In one patient who was given 6 cycles of CHOP-G, the peak values of mobilized CFU-GM in each cycle did not substantially decrease until 6 cycles of CHOP-G had been completed. Thus, the CHOP-G regimen described here seems to be very efficient increasing the circulating CFU-GM prior to harvesting PBPC.

Detection of hydroxyl radical in intact cells of Chlorella vulgaris.

Using ESR with 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin-trapping reagent, we measured the levels of free radical species generated from living cells of Chlorella vulgaris var. vulgails (IAM C-534). To investigate the production of free radicals in the living Chlorella vulgaris cells, the influence of DMPO toward the intact cells of the Chlorella vulgaris using the O2 evolution rate was first studied as a guide. Since the O2 evolution rate was not changed by DMPO, it was judged that DMPO has no toxicity toward the intact cells of Chlorella vulgaris. Only hydroxyl radicals (.OH) were detected as the DMPO-OH adduct in the suspension of intact cells of Chlorella vulgaris irradiated with visible light. Moreover, since production of .OH was inhibited by some hydroxyl radical scavengers such as KI and ethanol, production of .OH was proved to be due to hydroxyl radicals. It was also clear that the intensity of .OH increased with increasing irradiation intensity of visible light. Therefore, it was suggested that .OH might be one of the photoinhibition factors of the intact Chlorella vulgaris cells in severe light conditions.

Evaluation of active oxygen effect on photosynthesis of Chlorella vulgaris.

The relationship between O2 and an active oxygen scavenging system in Chlorella vulgaris var.vulgaris (IAM C-534) was investigated. When Chlorella vulgaris was exposed to 2% O2, only traces of active oxygen scavenging enzymes were found. When the Chlorella vulgaris was treated with 20% or 50% O2, it was shown that the level of enzyme activity increased as the O2 concentration increased. An increase in enzyme activity was not found in any specific enzyme but in all of the enzymes, but the level of glutathione and ascorbate remained the same in all the cases. In addition, the photosynthetic efficiency also decreased as the concentration of O2 was increased. These results suggest that an O2 enriched environment can lead to an increase in the production of active oxygen species such as O2.- and H2O2 and to a decrease in the photosynthetic efficiency in Chlorella vulgaris. The hydroxyl radical (.OH) was detected directly in the Chlorella vulgaris suspension with a spin trapping reagent. It was also clear that the increase in the .OH intensity as the visible light intensity increased was unrelated to the O2 concentration. It was suggested that the conditions for producting .OH and the other active oxygen species were different, and that two types of oxygen stress should exist in the Chlorella vulgaris.

Marked elevation in serum apolipoprotein E in a case of heterozygous cholesteryl ester transfer protein deficiency.

The subject was a 57-year-old Japanese woman with a body mass index of 21.2 kgm(-2). Her serum total cholesterol (TC), triglycerides (TG) and HDL-cholesterol levels were 7.11 mmoll(-1), 0.53 mmoll(-1) and 2.05 mmoll(-1), respectively. She had a marked increase of serum apolipoprotein (Apo) E concentration of 25 mgdl(-1) with normal concentrations of serum Apo A-I, A-II, B, C-II and C-III. Polymerase chain reaction-restriction fragments length polymorphism analysis of the cholesteryl ester transfer protein (CETP) gene from this subject revealed the heterozygous nucleotide change causing a Asp442 to Gly substitution (D442G) in the CETP protein. For comparison, 11 unrelated female subjects with this mutation (age, 57+/-5.1 years; BMI, 22+/-1.5 kgm(-2); TC, 7.23+/-1.16 mmoll(-1); TG, 1.44+/-0.80 mmoll(-1); HDL-C, 2.47+/-0.53 mmoll(-1)) were found to have a serum Apo E concentration of 7+/-1.5 mgdl(-1), about a third of the patient's concentration. The lipoprotein profile of the proband's serum analyzed by disk polyacrylamide gel electrophoresis showed a trace amount of VLDL. A vitamin A fat-loading test showed little increase in serum triglycerides and retinyl palmitate levels compared with control subjects at 2, 4 and 6 h after fat loading. Ultracentrifugation analysis of her serum revealed no detectable Apo E in the VLDL fraction but showed a large amount of Apo E in the HDL fraction, in contrast to a normal control, who had Apo E in the VLDL fraction as well as in the HDL fraction. Sequence analysis of the Apo E gene from the subject showed no nucleotide changes in exon 3 and exon 4, which code the mature Apo E protein, indicating there is no structural abnormality in the Apo E protein. Direct sequence analysis of the LDL receptor gene also did not show any nucleotide change. Based on these findings, it was hypothesized that the marked increase of Apo E in the patient's serum was caused by a decreased transfer of Apo E from HDL particles to TG-rich lipoproteins or impaired uptake of Apo E-containing HDL by LDL receptor or remnant receptor, due presumably to a dysfunction of these receptors in the patient.

Tumor targeting by arterial administration of lipids: rabbit model with VX2 carcinoma in the liver.

We previously found that a lipid contrast medium, Lipiodol, remained selectively in liver tumors for a prolonged time after injection via the hepatic artery. We now extend this technique to other lipids including linoleic acid, olive oil, tea seed oil, and medium-chain triglyceride (MCT). MCT is a semisynthetic triglyceride with a lower viscosity than that of the other lipids. Each lipid was mixed with 14C linoleic acid, and the mixture with or without anticancer agent smancs was injected via the proper hepatic artery of rabbits with VX2 carcinoma in the liver. The 14C count in the tumor tissue was always more than 100 times that of the plasma or kidney, and the radioactivity 15 min after injection of MCT was more than 2500 times greater than that of blood plasma. At 24 hr after injection, the radioactivity recovered in the tumor was 4.9-37.0% (average 19.6%) of the dose, which indicated the greatest retention in the tumor tissue; there was rapid clearance from the rest of the body primarily via the bile. Advantages of the administration of lipid-solubilized drugs are: (i) remarkable tumor-selective targeting (a decisive anticancer effect with fewer side effects); (ii) prolonged drug retention (resulting in infrequent administration); (iii) various choices of different fatty acids with various pharmacokinetic characteristics as the carriers of the anticancer agents; and (iv) arterial administration is practicable in most hospitals.