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症例は64歳男性,発熱,右季肋部痛を主訴とし画像診断にて肝S8領域に占拠性病変を認め,腫瘍摘出術を施行した.腫瘍は径8×7×6cmで病理検査にて肝原発悪性リンパ腫diffuselarge cell typeの診断を得た.術後VEPA療法を行い,5年経過の現在,再発の徴候はない.肝原発性悪性リンパ腫は極めて稀な疾患であり,当症例を含め18例の切除報告をみるにすぎない.男女比は14:4,平均年齢は42.2歳であった.術式は肝部分切除3例,区域切除1例,葉切除6例,3葉切除8例であり,化学療法は14例に行われた.Lymphoma Study Group分類では1例を除きdiffuse typeであった.術後の合併症で1例,再発で1例死亡し,予後は良好であった.本疾患の進展形式はリンパ節転移,周辺臓器への直接浸潤が主であり,肝硬変の合併率も低く,大量肝切除が可能である.他の節外性悪性リンパ腫と同様,逮隔リンパ節転移陽性となる前に手術を行い術後化学療法を併用すれば,予後は充分に期待できると考えられた.

Objective: To evaluate the effect of ultrasound irradiation when used alongside standard care in the treatment of pressure ulcers; outcome measures were reduction in wound size and exudate weight. Method: Five patients (two male and three female, age range: 76–92 years) with seven ulcers participated in this study. They had National Pressure Ulcer Advisory Panel (NPUAP) stage III or IV pressure ulcers. We conducted an ABABA study (A: standard treatment with dressings that promote a moist wound healing environment; B: ultrasound irradiation administered to the pressure ulcer through the same dressing used in period A; each period lasted 2–4 weeks). Six ulcers each were randomised to either the treatment group or control group. One ulcer was not randomised, but was the first to receive ultrasound in the BABA sequence, with a view to determining if the pilot was feasible. The control group received sham ultrasound in period B. Pulsed ultrasound (20% duty cycle, 0.5W/cm2 on the wound surface, 1MHz or 3MHz, for 10 minutes) was applied five times weekly. Results: In the treatment group, two ulcers markedly decreased in size after 3–4 weeks of US treatment, one ulcer decreased in size soon after initiation of treatment and one ulcer showed no clear reduction in size. The volume of exudate was greater in period B than A in two ulcers that reduced markedly in size after 3–4 weeks of US treatment. None of the ulcers in the control group decreased markedly in size. Conclusion: This pilot study suggests that US used alongside standard treatment might promote the healing of pressure ulcers. However, larger studies are required to determine the effi cacy and mechanism of US treatment for PUs. Declaration of interest: None.

A familiarization study was conducted on the "Combined Repeat Dose and Reproductive/Developmental Toxicity Screening Test (ReproTox)" proposed by the OECD. Cyclophosphamide (CP) at doses of 6.7, 4.5, 3, 2, and 0 mg/kg body wt was given daily by gavage to groups of 12 male and 12 female Sprague-Dawley rats. As a result, anemia and leukopenia were evident in treated males. The absolute and relative thymus and spleen weights were decreased in treated rats. Histopathologically, atrophy of the thymus, spleen, and bone marrow was observed. With respect to the reproductive/developmental toxicity, dose-dependent increases in postimplantation loss of fetuses and postnatal death were found in dams given CP. The body weight of pups treated with CP was significantly lowered in a dose-related manner. Thus the results demonstrated most of the known toxicological properties of CP, except the adverse effects on spermatogenesis and fertility. Therefore ReproTox can be considered as a useful screening test for assessing repeat dose and reproductive/developmental toxicity of existing chemicals of high production volume.

高齢者消化器手術の治療成績を周術期管理の面から検討した.教室で22年間に切除術を施行した食道癌, 胃癌, 肝胆膵悪性腫瘍, 大腸癌および胆石症の3,481症例を用い, 前期, 後期のそれぞれ11年間の術前術後合併症, リスク年齢の検討を行い, 全体としては65歳を基準に, 疾患別では食道癌根治術と膵頭十二指腸切除術は60歳, 胃癌胃全摘術は70歳を基準に, 高齢群と若年群に分け, 手術侵襲と血中指標との関連, 術後の血清蛋白, 免疫指標の変動を比較し, 長期予後の比較検討を行った.術前有合併症例は高齢群で多かった.高齢群の術後合併症発生頻度は前期の55%から後期の49%へ, 術死率は10%から3%へ減少し, 長期生存率の改善を認めた.その機序として, 術中出血量の減少や麻酔法の改善による手術侵襲の抑制, 積極的な代謝栄養管理により, 侵襲ホルモン分泌や代謝変動がよくコントロールされていることより, それが予後の改善をもたらしたと考えられた.

Delayed granulocyte apoptosis is implicated in persistent inflammation. Although it is known that males develop sepsis more easily than females, the mechanism for this is not fully understood. Serum IL-18 levels correspond to severity of systemic inflammation. The purpose of this study was to elucidate gender differences and the effects of IL-18 on granulocyte dynamics and apoptosis. Male and female wild-type (WT) and IL-18 knockout (KO) mice were injected intraperitoneally with LPS. Bone marrow cells, peripheral blood, and peritoneal cells were then collected at different times up to 24 h after injection. Apoptosis was assessed by annexin V staining using three-color flow cytometry with differentiated granulocyte-specific Gr-1, B-lymphocyte-specific B220, and macrophage-specific F4/80 antibodies. Male WT mice were more susceptible to endotoxin than female WT mice (survival rate, 41% in male WT and 84% in female WT). Male WT mice showed stronger responses than females in myeloid differentiation, release of myeloid cells from the bone marrow into the periphery, and migration into the inflammatory peritoneal cavity. Male mice also showed greater inhibition of granulocyte apoptosis in the peritoneal cavity, which might also contribute to the higher numbers of those cells present. A comparison between WT and KO mice revealed that the gender difference in these myeloid cells/granulocytes' behaviors was independent of endogenous IL-18. Nevertheless, levels of IL-18 in the blood of WT mice were significantly higher in males than in females after intraperitoneal LPS, and the survival rate was significantly higher in KO mice compared with WT mice only in males. This indicates that endogenous IL-18 production decreases survival only in males. Thus, excessive myeloid/granulocyte immunity independent of IL-18 and other IL-18-dependent life-threatening factors in males should be taken into account as therapeutic targets during systemic inflammatory states.

The effect of methionine-deprived nutrition on cell growth and cell kinetics was investigated in cell cultures and in tumor-bearing rats using the total parenteral nutrition (TPN) technique. A simultaneous flow cytometric measurement of the cellular DNA content and the amount of 5-bromodeoxyuridine incorporated into cellular DNA was performed for analysis of cell kinetics. The methionine-free medium demonstrated a cytocidal effect on the growth of SLC cells after 6 hours of culturing. It decreased viability from 80% in the control medium to 23%, and it decreased the S phase and increased the G0/G1 phase of the cell cycles. The methionine-deprived medium showed a concentration-dependent inhibition in cellular growth. Methionine-deprived TPN was seen to inhibit AH109A and SLC tumor growth compared with conventional TPN and decreased the S phase and increased the G0/G1 phase of cell cycles. These results confirm that methionine deprivation blocks cells from processing into the G1 phase and recycling, and that it is effective in inhibiting tumor growth in cultures and in vivo.

Utility of a new method of bilirubin fractionation was evaluated in monitoring the effectiveness of plasma exchange (PE) performed in 10 postoperative cases with liver failure. Fractionation of serum bilirubin by high-performance liquid chromatography demonstrated a higher delta bilirubin (B delta) peak against lower conjugated bilirubin peaks [monoconjugated bilirubin (MCB) and diconjugated bilirubin (DCB)] in the three recovered cases. The calculated ratio of MCB/B delta and the ratio of B delta/(MCB + DCB + B delta) in the recovered cases showed statistically significant different against seven unrecovered cases (p less than 0.01). These results suggest that the recovered cases had a different quality hyperbilirubinemia and a different disease entity before PE as well as a different response to PE, and this novel method for serum bilirubin subfraction is considered a useful marker in selecting a patient responsive to PE.

To establish effective dietary self-management behavior for type II male and female diabetics, by focusing on how dietary intake and dietary self-management affect control indices of type II diabetics.The study group comprised 170 type II diabetics, 93 men (mean age, 63.1+/-8.8 years old) and 77 women (mean age 63.4 +/- 10.2 years old). The final analyses of data collected from subjects comprised three control indices of type II diabetics, several factors related to control indices, and several items on a dietary self-management behavior questionnaire. Multiple regression analyses used control indices as dependent variables, and related factors as independent variables. Multiple regression analyses were also used for the relationships between dietary intakes as dependent variables and dietary self-management factors as independent variables.Males showed a significant correlation between total energy intake per standard body weight per day and body mass index, and a significantly positive correlation between body mass index and waist circumference. Carbohydrate intake was significantly related to HbA1c. Other results showed characteristic relationships between various questionnaire items and "total energy intake, lipids intake, and carbohydrate intake", respectively. Females showed significant correlation between carbohydrate intake and both body mass index and waist circumference, as well as characteristic correlations between various questionnaire items and both reductions and increases in carbohydrate intake.Our study discloses significant differences in dietary self-management behavior between males and females with type II diabetes. We anticipate that educational support using findings of our study can make a significant contribution to improvement in control indices of type II diabetics.

To report on the establishment of the Japanese version website of the Weekly Epidemiological Record (WER) by the faculty and to evaluate its accessibility and the educational outcome: all articles from the WER since 2000 have been translated into Japanese by graduate students with teachers' guidance, verified by the committee members, and delivered to the website. The server log files and retrieval keywords were analyzed using Analog 6.0. An on-line questionnaire survey of visitors to the website was performed. Opinion sheets reported by the students for translation were evaluated as the educational outcome. Over 6 years, there were 820,571 requests to the website and, the number of requests increased with disease outbreaks. According to domain analysis, most requests were during daytime on weekdays, and the website was utilized by users in educational institutions and the Japanese government and by overseas visitors. Among respondents to the questionnaire, 47% were laypersons and 69% found the website easy to understand. SARS and HIV/AIDS were the terms most frequently used for retrieval. The students recognized the importance of the World Health Organization (WHO) and had broadened their perspective on international health. The website is useful for Japanese. The translating process was effective for international health education.

BACKGROUND: Developmental toxicity of selenium (Se) is a nutritional, environmental and medicinal concern. Here, we investigated Se embryotoxicity by proteomic analysis of cultured rat embryos. METHODS: Rat embryos at day 9.5 or 10.5 of gestation were cultured for 48 or 24 h, respectively, in the presence of sodium selenate (100 or 150 µM) or sodium selenite (20 or 30 µM). Proteins from the embryo proper and yolk sac membrane were analyzed by two-dimensional electrophoresis for quantitative changes from those in control embryos. Proteins with quantitative changes were identified by mass spectrometric analysis. RESULTS: Growth inhibition and morphological abnormalities of cultured embryos were observed in all the Se treatment groups. By the analysis of the embryo proper, actin-binding proteins were identified as proteins with quantitative changes by selenate: increased phosphorylated-cofilin 1, increased phosphorylated-destrin, decreased drebrin E, and decreased myosin light polypeptide 3. Many proteins showed similar changes between selenate and selenite, including increased ATP-synthase, decreased acidic ribosomal phosphoprotein P0, and decreased pyrroline-5-carboxylate reductase-like. In the yolk sac membrane, antioxidant proteins were identified for protein spots with quantitative changes by selenite: increased peroxiredoxin 1 and increased glutathione S-transferase. CONCLUSION: The identified proteins with quantitative changes by selenate or selenite were considered to be candidate proteins involved in Se embryotoxicity: the actin-binding proteins for selenate embryotoxicity, proteins with the similar changes for the common Se embryotoxicity and antioxidant proteins for modification of Se embryotoxicity by redox-related treatments. These proteins may also be used as biomarkers in developmental toxicity studies. Birth Defects Res (Part B), 2008. © 2008 Wiley-Liss, Inc.

Pathogenesis of indium-caused tail malformations was investigated by in vivo and in vitro experiments. In the in vivo experiment, pregnant Wistar rats received single intravenous administration of indium trichloride at 0.4 mg/kg on day 10 of gestation, and their embryos were examined on days 11, 12 and 13. Embryos in the indium group showed caudal hypoplasia from day 11. Increased apoptosis was observed in their tailbud on day 11. Similar effects were observed in the in vitro experiment, when day 10 rat embryos were cultured in the presence of indium trichloride at 50 μM for 24 h and for further 24 h in the absence of indium. It was considered from these results that caudal hypoplasia probably due to excessive cell loss by increased apoptosis in the tailbud accounted for indium-caused tail malformations in rat fetuses, and that indium-caused embryotoxic effects were direct effects on the conceptus.

Changes in both behavior and gene expression occur in Caenorhabditis elegans following exposure to sex hormones such as estrogen and progesterone, and to bisphenol A (BPA), an estrogenic endocrine-disrupting compound. However, only one steroid hormone receptor has been identified. Of the 284 known nuclear hormone receptors (NHRs) in C. elegans, we selected nhr-14, nhr-69, and nhr-121 for analysis as potential estrogenic hormone receptors, because they share sequence similarity with the human estrogen receptor. First, the genes were cloned and expressed in Escherichia coli, and then the affinity of each protein for estrogen was determined using a surface plasmon resonance (SPR) biosensor. All three NHRs bound estrogen in a dose-dependent fashion. To evaluate the specificity of the binding, we performed a solution competition assay using an SPR biosensor. According to our results, only NHR-14 was able to interact with estrogen. Therefore, we next examined whether nhr-14 regulates estrogen signaling in vivo. To investigate whether these interactions actually control the response of C. elegans to hormones, we investigated the expression of vitellogenin, an estrogen responsive gene, in an nhr-14 mutant. Semi-quantitative RT-PCR showed that vitellogenin expression was significantly reduced in the mutant. This suggests that NHR-14 is a C. elegans estrogenic hormone receptor and that it controls gene expression in response to estrogen.

BACKGROUND: Proteomic analysis of cultured postimplantation rat embryos is expected to be useful for investigation into embryonic development. Here we analyzed protein expression in cultured postimplantation rat embryos by two-dimensional electrophoresis (2-DE) and mass-spectrometric protein identification. METHODS: Rat embryos were cultured from day 9.5 for 48 h or from day 10.5 for 24 h. Proteins of the embryo proper and yolk sac membrane were isolated by 2-DE and differentially analyzed with a 2-D analysis software. Selected protein spots in the 2-DE gels were identified by matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometric analysis and protein database search. RESULTS: About 800 and 1,000 protein spots were matched through the replicate 2-DE gels each from one embryo in the embryo proper and yolk sac membrane, respectively, and virtually the same protein spots were observed irrespective to the length of culture period. From protein spots specific to the embryo proper (126 spots) and yolk sac membrane (304 spots), proteins involved in tissue-characteristic functions, such as morphogenesis and nutritional transfer, were identified: calponin, cellular retinoic acid binding protein, cofilin, myosin, and stathmin in the embryo proper, and Ash-m, dimerization cofactor of hepatocyte nuclear factor, ERM-binding phosphoprotein, cathepsin, and legumain in the yolk sac membrane. CONCLUSION: Proteomic analysis of cultured postimplantation rat embryos will be a new approach in developmental biology and toxicology at the protein level. Birth Defects Res (Part B), 2007. © 2007 Wiley-Liss, Inc.

The effects of indium on bone and cartilage development in rat fetuses were examined. Pregnant Sprague Dawley (SD) rats were treated with indium trichloride (0.1, 0.2, or 0.3 mg/kg) by single intravenous administration on Day 10 of gestation, and their fetuses were examined on Day 21. Half of each litter was prepared for skeletal examinations using a skeletal double-staining technique to allow evaluation of cartilage as well as bone. Dose-related increased incidences of external and skeletal fetal malformations occurred at doses of 0.2 mg/kg or more. The incidences of cartilage malformations in the vertebrae, ribs, and forepaw phalanges were significantly increased at 0.3 mg/kg. Malformations of the axial bone were accompanied by cartilage malformations. It was concluded from these results that indium produced cartilage malformations, that were considered to be the underlying cause for the majority of fetal skeletal malformations observed in rats in this study.

The gut microbiota has the capacity to produce a diverse range of compounds that play a major role in regulating the activity of distal organs and the liver is strategically positioned downstream of the gut. Gut microbiota linked compounds such as short chain fatty acids, bile acids, choline metabolites, indole derivatives, vitamins, polyamines, lipids, neurotransmitters and neuroactive compounds, and hypothalamic-pituitary-adrenal axis hormones have many biological functions. This review focuses on the gut microbiota and host metabolism in liver cirrhosis. Dysbiosis in liver cirrhosis causes serious complications, such as bacteremia and hepatic encephalopathy, accompanied by small intestinal bacterial overgrowth and increased intestinal permeability. Gut dysbiosis in cirrhosis and intervention with probiotics and synbiotics in a clinical setting is reviewed and evaluated. Recent studies have revealed the relationship between gut microbiota and host metabolism in chronic metabolic liver disease, especially, non-alcoholic fatty liver disease, alcoholic liver disease, and with the gut microbiota metabolic interactions in dysbiosis related metabolic diseases such as diabetes and obesity. Recently, our understanding of the relationship between the gut and liver and how this regulates systemic metabolic changes in liver cirrhosis has increased. The serum lipid levels of phospholipids, free fatty acids, polyunsaturated fatty acids, especially, eicosapentaenoic acid, arachidonic acid, and docosahexaenoic acid have significant correlations with specific fecal flora in liver cirrhosis. Many clinical and experimental reports support the relationship between fatty acid metabolism and gut-microbiota. Various blood metabolome such as cytokines, amino acids, and vitamins are correlated with gut microbiota in probiotics-treated liver cirrhosis patients. The future evaluation of the gut-microbiota-liver metabolic network and the intervention of these relationships using probiotics, synbiotics, and prebiotics, with sufficient nutrition could aid the development of treatments and prevention for liver cirrhosis patients.

A simple method for two-dimensional electrophoresis (2-DE) of rat embryonic protein was described. Rat embryos cultured for 24 h from day 10.5 of gestation were used as protein samples. Protein samples were lysed in rehydration buffer and separated by isoelectric focusing with immobilized pH gradient for the first dimension and by sodium dodecyl sulfate–polyacrylamide gel electrophoresis for the second dimension. The use of the DeStreak Reagent as an antioxidant in the lysis buffer and electrode pads in the isoelectric focusing greatly improved the 2-DE pattern. When an embryo was used as a protein sample, about 800 protein spots were detected by silver staining in a 2-DE gel of the standard format. Eighty-one protein spots were identified by mass spectrometry for a primary 2-DE map. The same method could be applied to yolk sac membranes from the cultured embryos. The present method was considered to be suitable for a concomitant 2-DE analysis in in vitro developmental toxicity studies.

Most cosmopolitans who are concerned about world poverty assume that for citizens of affluent societies, justice beyond national borders is a matter of their positive duty to provide aid to distant people suffering from severe poverty. This assumption is challenged by some authors, notably Tomas Pogge, who maintains that these citizens are actively involved in the incidence of poverty abroad and therefore neglect their negative duty of refraining from harming others. This paper examines the extent to which it is pertinent to contend that citizens in economically advanced countries are morally liable for the impoverishment of a sizable population of the developing world. The contention in question can be interpreted in two nonexclusive ways. First, it might imply that historical injustices, including colonialism and slavery, contributed to both contemporary affluence in some parts of the world and poverty in others. Second, it could imply that the present global economic system, instituted and implemented by the governments of rich and powerful countries acting in the name of their citizens, is benefiting the citizens while harming the world’s disadvantaged. The author argues that the idea of reparation for historical injustices suffers from serious philosophical difficulties, including the non-identity problem presented by Derek Parfit, and thus fails to provide a satisfactory approach to the existing problem of poverty. This paper then examines the alleged liability of citizens in affluent countries, with a special reference to empirical observations on the policy process. The paper concludes by suggesting a twofold theory of global justice, which combines material, managerial, and moral assistances for a society lacking a competent government and proposes institutional reforms in the global order in order to achieve poverty reduction.

Objective Involvement of lipoxygenase (LOX) and cyclo-oxygenase (COX) on cellular differentiation or apoptosis induced by butyrate has been reported recently, but the effect on tight junction (TJ) permeability has not been reported. One major activity of butyrate and, to a lesser extent, propionate is to modulate gene transcription via histone acetylation by their histone deacetylase inhibitor activity. In this study, we evaluated the activation of LOX and COX in TJ permeability changes by short-chain fatty acids, butyrate, propionate, and acetate in intestinal monolayer cells and their possible mechanism by histone acetylation. Methods The effects of LOX and COX inhibitors on TJ permeability and the expression of LOX or COX mRNA induced by short-chain fatty acids were investigated in Caco-2 cells using Transwell chambers. The effects of hydroxyeicosatetraenoic acid (a product of LOX) on TJ permeability were also evaluated. The effects of short-chain fatty acids were compared with those of trichostatin A (histone deacetylase inhibitor). Results A LOX inhibitor clearly inhibited the effect of butyrate on TJ permeability, whereas COX inhibitors did not. The LOX and COX inhibitors partly inhibited the effects of propionate but not of acetate. Butyrate increased LOX mRNA expression, and hydroxyeicosatetraenoic acid and trichostatin A mimicked its effect. Conclusion These results suggest that short-chain fatty acids, especially butyrate, induce TJ permeability changes through LOX activation through histone acetylation.

The level of (1-->3)-beta-D-glucan in blood is a diagnostic index of fungal infection because it is released from the fungal cell wall. However, high levels of plasma (1-->3)-beta-D-glucan in patients administered blood components may give false positive results. High levels of (1-->3)-beta-D-glucan have been detected in blood components. We suspected that (1-->3)-beta-D-glucan from cellulose filters had been eluted into blood components by filtration in the manufacturing process. To investigate the contamination of blood components by (1-->3)-beta-D-glucan from cellulose filters, in vitro experiments were performed by using six cellulose filters and a nylon filter. Human serum albumin (HSA) solution (100 ml) was flowed through each filter after rinsing with 100 ml of distilled water, and (1-->3)-beta-D-glucan in each fraction was determined by Fungitec G test MK. The concentration of (1-->3)-beta-D-glucan eluted from cellulose filters in 100-ml distilled water fractions ranged from 6 to 207 pg/ml, and that of HSA fractions ranged from 33 to 20,784 pg/ml. These data showed that remarkably higher (1-->3)-beta-D-glucan levels were detected in HSA fractions flowed through cellulose filters in spite of advance rinsing with 100 ml of distilled water. In the case of a nylon filter, (1-->3)-beta-D-glucan was not eluted in either fraction. These results indicate that (1-->3)-beta-D-glucan contamination in blood components is caused by filtration with cellulose filters in the manufacturing process.

Background The false-positive elevation of plasma (1-->3)-beta-D-glucan level, a serodiagnostic test for deep-seated mycosis, is suspected in patients administered with blood components. Study design and methods (1-->3)-beta-D-Glucan and endotoxin levels in blood components consisting of 12 albumins, 8 immunoglobulins, and 3 blood coagulation factors were measured by fungal infection tests (Fungitec G-test, Seikagaku Co.; the Wako WB003 test, Wako Pure Chemical Industries; and the Endospec ES test, Seikagaku Co.). In vitro release of (1-->3)-beta-D-glucan from the depth-type filters made by cellulose membrane to process blood components was analyzed through an in vitro filtration process as a source of (1-->3)-beta-D-glucan in blood components. Results The amounts of (1-->3)-beta-D-glucan in blood components ranged from 0 to 7510 pg per mL in the Fungitec G-test, with wide variations among brands. The positive rates over 20 pg per mL were 75 percent in albumin solutions, 40 percent in blood coagulation factors, and 63 percent in immunoglobulin solutions. (1-->3)-beta-D-Glucan levels released from the five depth filters ranged from 5 to 2516 pg per mL. The (1-->3)-beta-D-glucan level in filtration fluid was decreased by rinsing with distilled water, but rebounded again during the albumin filtration process. Conclusion Depth filters are considered the source of (1-->3)-beta-D-glucan content in some blood components.

We have developed a simple assay method for the evaluation of estrogen receptor (ER) binding capacity of chemicals without the use of radio- or fluorescence-labeled compounds. We used the solution competition assay by the BIACORE biosensor, a surface plasmon resonance biosensor, with estradiol as a ligand, human recombinant ER(alpha) (hrER(alpha)) as a high molecular weight (hmw) interactant and test chemicals as analytes. For the ligand, aminated estradiol with a spacer molecule (E2-17PeNH) was synthesized and immobilized on a carboxymethyl dextran-coated sensor chip by the amine coupling method. The injection of the hmw interactant hrER(alpha) to the biosensor raised the sensorgram, indicating its binding to the ligand E2-17PeNH. The binding of test chemicals to hrERalpha was determined as a reduction in the hrER(alpha) binding to E2-17PeNH. The dissociation constant for the binding to hrER(alpha) was calculated for estrone (4.29 x 10(-9)M), estradiol (4.04 x 10(-10)M), estriol (8.35 x 10(-10)M), tamoxifen (2.16 x 10(-8)M), diethylstilbestrol (1.46 x 10(-10)M), bisphenol A (1.35 x 10(-6)M) and 4-nonylphenol (7.49 x 10(-6)M), by plotting the data according to an equation based on mass action law. This method can also be used as a high throughput screening method.

The purpose of this study is to evaluate the effect of C18 and C20 long chain fatty acids on tight junction permeability in a model of intestinal epithelium.Confluent Caco-2 cells on porous filters with double chamber system were used to measure fluorescein sulfonic acid (FS) permeability and transepithelial electrical resistance (TEER). Lactate dehydrogenase release and ultrastructure were evaluated. Effect of 200 microM eicosapentaenoic acid (EPA, C20:5 n-3), arachidonic acid (AA, C20: 4 n-6), alpha-linoleic acid (ALA, C18: 3 n-3), linoleic acid (LA, C18: 2 n-6), or oleic acid (OA, C18: 1 n-9) enrichment in the culture medium during 24 hours were compared. The effect of the cyclooxygenase inhibitor, indomethacin, lipoxygenase inhibitors, NDGA or AA861, and antioxidant, BHT, was evaluated as a mechanism to change tight junction permeability.Caco-2 cells formed polarized columnar epithelial cells with densely packed microvilli and well developed junctional complexes. Addition of EPA enhanced FS permeability to 3.0+/-1.6-fold and lowered TEER to 0.59+/-1.2-fold vs. control with concentration dependency without cell injury (P0.01-0.05). OA, AA or LA did not change, but ALA enhanced tight junction permeability. Indomethacin and AA861 normalized the changes mediated by EPA.EPA affects tight junction permeability in intestinal monolayer cells specifically and concentration dependently via cyclooxygenase and lipoxygenase products.

The developmental toxicity of indium was examined in both rats and mice using comparable experimental protocols. Pregnant rats received a single intravenous administration of indium trichloride (InCl3) at 0.4 mg In/kg, on day 9, 10, or 11 of pregnancy and their fetuses were examined on day 20. Pregnant mice were treated in the same manner at 0.8 or 1.6 mg In/kg on day 7, 8, or 9 of pregnancy and their fetuses were examined on day 18. In rats, indium caused fetal weight decrease and fetal gross malformations, such as brachyury, kinked tail, cleft palate, and oligodactyly, most severely by the administration on day 10. In mice, however, indium did not cause fetal gross malformations, although it caused fetal weight decrease at 0.8 mg In/kg or more and fetal death at 1.6 mg In/kg, most severely by the administration on day 8. It was concluded from these results that rats and mice were susceptible to the embryotoxicity of indium at similar developmental stages in the early organogenetic period, but mice were less susceptible to the teratogenicity of indium than rats in terms of gross malformation. Toxicokinetic factors may be involved in this different susceptibility. Teratogenesis Carcinog. Mutagen. 20:219–227, 2000. © 2000 Wiley-Liss, Inc.

Toxicokinetics of 2-mercaptobenzimidazole (MBI) and 2-mercaptomethylbenzimidazole (MMBI), rubber antioxidants with thioureylene structure, were compared after single oral administration in rats. Male Wistar rats received single oral administration of 2, 10, 50 and 250 mg/kg of MBI or MMBI. The serum and urine concentrations of MBI and MMBI were determined by HPLC. MBI and MMBI showed similar Cmax values, but the former disappeared slower in the serum than the latter and resulted in its larger AUC values. Analyses of MBI, MMBI and their desulfurated metabolites in urine suggested that these differences were due to their metabolic elimination rates. On the other hand, MBI and MMBI caused similar acute toxicities, such as loss of locomotive activity, ataxic gait, adoption of prone or side position and coma, being severer with higher serum concentrations at the moment. Similar acute toxicities between MBI and MMBI were explained by similar Cmax values at the same dose. It was suggested from these results that the slower disappearance and larger AUC values of MBI in the serum compared to MMBI might explain the strong thyroid toxicity which has been observed by repeated administration of MBI, but very weak thyroid toxicity by MMBI.

The present study was conducted to reveal effects of in ovo injection of nonsteroidal aromatase inhibitor (Fadrozole) or estradiol at day 3 of incubation on mRNA levels of P45017ahydroxylase (P450c17), P450 aromatase (P450arom) and anti- Mullerian hormone (AMH) in the chicken gonads. The mRNA levels in the gonads at days 4-8 of incubation were assessed by in situ hybridization analysis using digoxigenin labeling method. The in situ hybridization data were analyzed by relative expression of specific hybridizable signals of each mRNA corrected by the non-specific background by employing an image ana- lyzer. P450c17 mRNA expression increased rapidly at day 6 of incubation in the male but decreased thereaf- ter. In contrast to the transient expression in the male, the expression was gradually increased in the female. P450arom mRNA was not expressed in the male but was detectable in the female as early as day 6 and increased subsequently with days of incubation. AMH mRNA was expressed as early as day 5 of incubation followed by a sharp increase on day 6, which was maintained in the male thereafter. In contrast, the female showed very little expression. The injection of Fadrozole caused no effect on P450c17 mRNA expres- sion, while it suppressed P450arom mRNA expression but increased AMH mRNA expression in the female. In contrast, the injection of estradiol induced P450arom mRNA expression significantly but suppressed AMH mRNA expression in the male. These results indicate that expression of P450arom and AMH is sexually dimorphic and is reciprocally regulated during early ontogenic life in chicken gonads. Mol. Reprod. Dev. 55:20-30, 2000. r 2000 Wiley-Liss, Inc.

Hepatic ischemia/reperfusion is a common problem encountered under many clinical conditions, and it occurs in a variety of surgical procedures. Possible consequences include liver failure and/or multi-organ system failure, both of which have high rates of morbidity and mortality.1,2 Efforts to minimize the extent of injury incurred during the ischemic period or to maximize reparative process following ischemia have importance for survival.3 In vivo 31P-nuclear magnetic resonance (NMR) spectroscopy has been used to follow the response of hepatic high-energy phosphates during hepatic ischemia/reperfusion.4 It measures the continuous changes of hepatic adenosine triphosphate (ATP) levels and their metabolites, and is an effective tool to evaluate the serial data in the individual following drug treatment.

Recent investigations of enterai nutritients or artificial feedings with nucleotides and nucleosides suggested that they potentiate the bio-defense activity in immunologically compromised patients in the intensive care units.1 Decreased T lymphocyte functions with dietary nuclear acid deficiency and reversal by ribonucleic acid (RNA) supplement are reported.2–4 In surgical stress, macrophages and/or polymorphonuclear leukocytes (PMNs) are considered as key factors in the pathogenesis of systemic inflammatory response leading to septic shock by producing humoral mediators under endotoxin stimulation. We reported that dietary nucleic acid deficiency inhibited lipopolysaccharide (LPS) uptake and humoral mediators secretion by peritoneal macrophages in rats.5 However, it is not known whether dietary nucleic acid influences on PMN functions. Blocking of Mac-1 (CDllb/CD18) and/or lymphocyte function-associated antigen-1 (LFA-1, CD 11 a/CD 18) which are the adhesion proteins involved in the integrin superfamily, reduce the number of PMNs accumulating in the tissue and attenuate the oxidant stress of infiltrated PMNs.6,11 Phagocytes, such as PMNs and macrophages, undergo an oxidative burst in response to phagocytic or membrane stimuli, with generation and release of a variety of reactive oxygen metabolites.

The usefulness of rabbit serum as a culture medium for postimplantation rat embryos was examined. Rat embryos at day 9.5 of gestation were cultured in a mixture of rat and rabbit sera at various ratios (v/v) for 48 h. In 100% rat serum, a usual medium, rat embryos grew well. On the contrary, rat embryos died with little growth in 100% rabbit serum. In 75% rabbit and 25% rat sera, rat embryos grew but were morphologically abnormal. In 50% rabbit and 50% rat serum, however, rat embryos grew well showing no morphological abnormalities, as in 100% rat serum. It was concluded from these results that rabbit serum could be used at a proportion up to 50% as a medium for postimplantation rat embryo culture in a mixed form with rat serum. The rat embryo culture using rabbit serum as a medium would be useful in developmental toxicity studies, especially those involving species differences and toxicokinetics.

Purine and pyrimidine metabolism is a key process after hepatic surgery. To evaluate the effect of purine and pyrimidinesupplementation on hepatic regeneration, the following clinical in vitro and in vivo experiments were performed. Changes in blood nucleotides, nucleosides and nucleobase were analyzed by high performance liquid chromatography in patients and rats after partial hepatectomy. The effect of supplementation of nucleotide-nucleoside solution (OG-VI) or their components on nucleic acids syntheses in primary monolayer cultured hepatocytes and preoperative intraperitoneal injection of OG-VI on hepatic regeneration in the partially hepatectomized rats were evaluated. Blood purine and pyrimidine levels in patients change after hepatectomy and their changes indicate increased salvage synthesis of purine and pyrimidine in the regenerating liver. Addition of appropriate amounts of inosine, GMP, uridine, or thymidine, the substrates for salvage purine and pyrimidine syntheses, to primary cultures of hepatocytes enhanced both DNA and RNA syntheses by the salvage and de novo pathways. The OG-VI mixture also enhanced the syntheses of DNA and RNA. Preoperative administration of OG-VI to partially-hepatectomized rats enhances hepatic DNA synthesis in a way similar to the in vitro study.

After hepatectomy, purine and pyrimidine metabolism is a key process in the synthesis of DNA and RNA and maintaining cellular energy metabolism. The purpose of this study is to evaluate changes in blood purine and pyrimidine levels after partial hepatectomy and the effect of purine and pyrimidine nucleoside solution injection on hepatic regeneration under the hypothesis that the rat after partial hepatectomy requires substrates for salvage nucleotide synthesis and changes blood nucleoside and nucleobase levels. Blood levels of nucleotides, nucleosides, and nucleobase by high-performance liquid chromatography method and liver ATP level by enzymatic analysis, and the effect of preoperative injection of nucleoside solution (OG-VI) on hepatic regeneration ratio and hepatocytes DNA synthesis, were assessed in rats after 70% partial hepatectomy. Decreased liver adenosine triphosphate and increased plasma xanthine and hypoxanthine after partial hepatectomy indicated an increase in catabolism of purine nucleotides in regenerating liver. Plasma thymidine and cytidine levels increased, then returned to the prevalue, suggesting that the thymidine and cytidine pool was enlarged. OG-VI increased labeling indices of hepatocytes at postoperative d 1 (POD) and hepatic regeneration ratio at POD 14. Blood purine nucleobase and pyrimidine nucleoside levels change after partial hepatectomy and preoperative supply of nucleoside solution is effective for increasing hepatocytes DNA synthesis and hepatic regeneration after partial hepatectomy.

Embryonic susceptibility to selenium (Se) teratogenicity was examined in rats using postimplantation embryo culture. Rat embryos at day 9.5 of gestation were cultured by the roller bottle method for 48 hr in the presence of Se compounds. Sodium selenite, sodium selenate, seleno-DL-methionine, and seleno-DL-cystine were embryolethal at 20, 300, 1,000, and 1,000 microM Se, respectively. All of these compounds caused abnormalities such as deformed optic vesicle and swollen rhombencephalon in the viable embryos. These abnormalities were considered to correspond to in vivo malformations caused by Se in hamster fetuses or in bird embryos. These results indicate that rat embryos are susceptible to Se teratogenicity. It seems that there are differences in potency ranking of Se compounds between rat and bird embryos.

The effect of nitrobenzene on several spermatogenic endpoints was examined to determine which endpoints were affected by chemicals, how changes in spermatogenic endpoints are related to decreases in the rate of male fertility, and how long a treatment period is needed before damage can be detected. An experimental group of male Sprague-Dawley rats was given nitrobenzene (60 mg/kg, per os), a known inhibitor of male fertility, and a control group was given a diet of sesame oil (1 ml/kg, per os) alone. The rats were mated with normal proestrus females on days 7, 14, 21, 28, 42, 56 and 70 of treatment. Male rats were sacrificed on the day after mating, and sperm motility, progressive motility of sperm, sperm count and sperm morphology were examined. After 7 days of treatment, no changes were observed in the endpoints examined. However, the testicular and epididymal weights, sperm count, sperm motility and progressive motility decreased significantly by day 14. By day 21 sperm viability and the fertility index decreased, while increases were observed in the abnormal sperm rate and instances in which no motile sperm existed. However, even after 70 days of treatment, the copulation index was not affected. The most sensitive spermatogenic endpoints were determined to be sperm count and sperm motility, followed by progressive motility, viability, abnormal sperm rate and fertility index. The copulation index is not a meaningful endpoint for male fertility. Furthermore, a period of at least 14 days is necessary for evaluation of the effect of nitrobenzene on male fertility.

Purine metabolism is a key metabolic process after partial hepatectomy in the synthesis of DNA and RNA, and in the energy metabolism for hepatic regeneration1. However it is not evaluated in clinical situations due to the difficulty in measuring changes of purine levels in the liver itself. In order to get a basic understanding of purine metabolism in clinical hepatectomy cases, blood samples are used for analysis in this study. Also, the liver is the main nondietary source of purines in mammalian cells2 and changes in blood level may affect purine metabolism in various tissues.

Purine and pyrimidine are essential for cellular proliferation1, 2, 3. But, purine and pyrimidine are synthesized from glutamine in de novo synthesis pathway and cultured cancer cells do not require them in the medium. This is the reason that utilization of nucleosides by human cancer cells and their changes with 5 fluorouracil (5FU), the fluorinated analogue of uracil which is used in chemotherapy for various tumors2, 3, has not been reported. In this study, as a source of purine and pyrimidine, a solution containing a mixture of nucleosides and a nucleotide (OG-VI), developed recently as a nutritional supplement4, was used in the medium. It consists of purines, guanosine 5’-mono-phosphate (5’-GMP) and inosine, and pyrimidines, cytidine, uridine, and thymidine (TdR). It is known to increases hepatic regeneration and improve protein metabolism after partial hepatectomy in rats5, 6, and increase DNA synthesis in primary cultured hepatocytes7.

The importance of dietary nucleic acid in modulation of immune function without disease has been reported in nutritional disorder except under surgical stress. Dietary nucleic acid deficiency decreases T lymphocytes function1 and dietary ribonucleic acid (RNA) enhances immune response2. However, it is not known whether dietary nucleic acid influences on macrophage function. Recently, monocytes/macrophages are considered as key factors in the pathogenesis of systemic inflammatory response leading to septic shock by producing humoral mediators under endotoxin stimulation3, 4. Therefore, the aim of this study is to evaluate the influence of dietary nucleic acid deficiency on endotoxin shock and macrophage functions stimulated with lipopolysaccharide (LPS), which is the major component of endotoxin. Then, the effect of intraperitoneal administration of a nucleotide and nucleosides mixture solution (OG-VI)5 was evaluated.