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Autophagy induction in cancer is involved in cancer progression and the acquisition of resistance to anticancer agents. Therefore, autophagy is considered a potential therapeutic target in cancer therapy. In this study, we found that long-chain fatty acids (LCFAs) have inhibitory effects on Atg4B, which is essential for autophagosome formation, through screening based on the pharmacophore of 21f, a recently developed Atg4B inhibitor. Among these fatty acids, docosahexaenoic acid (DHA), a polyunsaturated fatty acid, exhibited the most potent Atg4B inhibitory activity. DHA inhibited autophagy induced by androgen receptor signaling inhibitors (ARSI) in LNCaP and 22Rv1 prostate cancer cells and significantly increased apoptotic cell death. Furthermore, we investigated the effect of DHA on resistance to ARSI by establishing darolutamide-resistant prostate cancer 22Rv1 (22Rv1/Dar) cells, which had developed resistance to darolutamide, a novel ARSI. At baseline, 22Rv1/Dar cells showed a higher autophagy level than parental 22Rv1 cells. DHA significantly suppressed Dar-induced autophagy and sensitized 22Rv1/Dar cells by inducing apoptotic cell death through mitochondrial dysfunction. These results suggest that DHA supplementation may improve prostate cancer therapy with ARSI., Competing Interests: Declaration of competing interest The authors disclose any conflicts of interest related to the work., (Copyright © 2024 Elsevier Inc. All rights reserved.)

In this study, we monitored the effect of Al 3+ ions on mesenchymal cells (KUSA-A1) and human fibroblasts (NHDF) by means of in vitro experiments by culturing the cells with addition of small concentrations of aluminum ions (i.e., 0.1, 1, 10, and 100 ppm). Bone formation test was then conducted using KUSA-A1. Small concentrations of aluminum ions delayed but did not completely inhibit cell proliferation. The amount of bone tissue decreased as the concentration of Al 3+ increased and crystallinity changes were also detected by Raman spectroscopic experiments. Moreover, Al 3+ ions greatly affected both structure and chemistry of bone tissues with mineral nodules becoming larger and atomic substitution of Ca with Al in bone tissue being more preponderant with increasing Al 3+ concentration. Such effects in turn impaired the balance between mineral and collagen in the formed bone tissue., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: The purpose of this study was to evaluate the effect of preoperative motor and cognitive activities of daily living (ADL) on long-term outcomes of patients with chronic limb-threatening ischemia (CLTI) after distal bypass., Methods: A retrospective review was performed for patients who underwent distal bypass for CLTI from 2013 to 2019 at multiple centers in Japan. Comparisons were made among patients with high and low motor and cognitive ADL based on the functional independence measure (FIM). The primary endpoint was limb salvage and the secondary endpoints were survival, amputation free survival (AFS), major adverse limb events (MALE), readmission, and wound healing., Results: A total of 226 distal bypasses were performed in 185 patients (169 males; median age, 76 years; diabetes mellitus, 70%; end-stage renal disease with hemodialysis, 40%). The patients were divided into high (n = 93, 50%) and low (n = 92, 50%) FIM-motor cases, and high (n = 157, 85%) and low (n = 28, 15%) FIM-cognitive cases. FIM-motor (high vs. low) and FIM-cognitive (high vs. low) were not significantly associated with limb salvage, freedom from MALE, freedom from readmission, and wound healing. The 1- and 3-year survival rates were significantly lower in low FIM-motor cases (93% vs. 70% at 1 year, 73% vs. 46% at 3 years, P = 0.0011); and in low FIM-cognitive cases (87% vs. 50% at 1 year, 63% vs. 45% at 3 years, P < 0.001). The 1- and 3-year AFS rates were significantly lower in low FIM-motor cases (92% vs. 67% at 1 year, 69% vs. 44% at 3 years, P < 0.001); and in low FIM-cognitive cases (85% vs. 49% at 1 year, 59% vs. 44% at 3 years, P < 0.001). In multivariate analysis, independent risk factors for survival were hemodialysis (HR = 2.17; 95% confidence interval (CI), 1.23-3.83; P = .0078), low FIM-cognitive (HR = 3.45; 95% CI, 1.78-6.71; P < 0.001), and ejection fraction (HR = 0.98; 95% CI, 0.95-0.99; P = 0.019)., Conclusions: FIM-motor and FIM-cognitive were predictive factors for long-term survival and AFS of CLTI patients after distal bypass, but had no influence on limb salvage, MALE, readmission, and wound healing. These results suggest that the motor and cognitive status of ADL should be assessed using FIM before distal bypass for patients with CLTI., (Copyright © 2021. Published by Elsevier Inc.)

Effective cancer immunotherapy requires physical contact of T cells with cancer cells. However, tumors often constitute special microenvironments that exclude T cells and resist immunotherapy. Cholesterol sulfate (CS) is a product of sulfotransferase SULT2B1b and acts as an endogenous inhibitor of DOCK2, a Rac activator essential for migration and activation of lymphocytes. We have recently shown that cancer-derived CS prevents tumor infiltration by effector T cells. Therefore, SULT2B1b may be a therapeutic target to dampen CS-mediated immune evasion. Here, we identified 3β-hydroxy-5-cholenoic acid (3β-OH-5-Chln) as a cell-active inhibitor of SULT2B1b. 3β-OH-5-Chln inhibited the cholesterol sulfotransferase activity of SULT2B1b in vitro and suppressed CS production from cancer cells expressing SULT2B1b. In vivo administration of 3β-OH-5-Chln locally reduced CS level in murine CS-producing tumors and increased infiltration of CD8 + T cells. When combined with immune checkpoint blockade or antigen-specific T cell transfer, 3β-OH-5-Chln suppressed the growth of CS-producing tumors. These results demonstrate that pharmacological inhibition of SULT2B1b can promote antitumor immunity through suppressing CS-mediated T cell exclusion., Competing Interests: Declaration of competing interest Kyushu University, Keio University and The University of Tokyo have a patent application related to this work, in which T.U., Y.S., K.O., T. Togo, M.K., and Y.F. are listed as co-inventors., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The acylation of anthocyanins contributes to their structural diversity. Aromatic acylation is responsible for the blue color of anthocyanins and certain flowers. Aromatic acyltransferase from Gentiana triflora Pall. (Gentianaceae) (Gt5,3'AT) catalyzes the acylation of glucosyl moieties at the 5 and 3' positions of anthocyanins. Anthocyanin acyltransferase transfers an acyl group to a single position, such that Gt5,3'AT possesses a unique enzymatic activity. Structural investigation of this aromatic acyl group transfer is fundamental to understand the molecular mechanism of the acylation of double positions. In this study, structural analyses of Gt5,3'AT were conducted to identify the underlying mechanism. The crystal structure indicated that Gt5,3'AT shares structural similarities with other BAHD family enzymes, consisting of N and C terminal lobes. Structural comparison revealed that acyl group preference (aromatic or aliphatic) for the enzymes was determined by four amino acid positions, which are well conserved in aromatic and aliphatic CoA-binding acyltransferases. Although a complex structure with anthocyanins was not obtained, the binding of delphinidin 3,5,3'-triglucoside to Gt5,3'AT was investigated by evaluating the molecular dynamics. The simulation indicated that acyl transfer by Gt5,3'AT preferentially occurs at the 5-position rather than at the 3'-position, with interacting amino acids that are mainly located in the C-terminal lobe. Subsequent assays of chimeric enzymes (exchange of the N-terminal lobe and the C-terminal lobe between Gt5,3'AT and lisianthus anthocyanin 5AT) demonstrated that acyl transfer selectivity may be caused by the C-terminal lobe., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: In the revised World Health Organization 2016 classification of central nervous system tumors, "diffuse midline glioma, H3 K27M-mutant" has been added as a new diagnostic entity. However, some confusion exists concerning this diagnostic entity because H3 K27M-mutant diffuse midline glioma is diagnosed with grade IV regardless of morphologic phenotype. Furthermore, the significance of H3 K27M mutation in tumors that aren't typical "diffuse midline glioma, H3 K27M-mutant," such as those with an unusual location and nontypical histology, remains unclear., Case Description: To elucidate further such unusual tumors, we describe here a rare case of pediatric low-grade glioma located in the tectum, which was morphologically a pilocytic astrocytoma (PA) with genetically H3 K27M mutation but no microvascular proliferation, necrosis, mitoses, or other genetic alterations, insofar as we were able to observe. At the latest follow-up, 28 months after surgery, radiotherapy, and chemotherapy, the patient was found to be free from any neurologic deficits and MRI demonstrated that the tumor was stable without tumor regrowth. This case might be identified as "diffuse midline glioma, H3 K27M-mutant", grade IV, when applying only the current World Health Organization 2016 classification. In addition, we discuss the morphologically benign gliomas harboring the H3 K27M mutation based on the literature., Conclusions: We describe here a rare case and present a short literature review of circumscribed/nondiffuse gliomas, particularly in PA with H3 K27M mutation. However, the significance of H3 K27M mutation for PA remains unclear, so further studies and clinical data are needed to elucidate the biology and optimal treatment of such tumors., (Copyright © 2020 Elsevier Inc. All rights reserved.)

An effective method for removing selenium (Se) from water is required from the viewpoint of environmental preservation. To establish this method, a cation exchange resin that adsorbed ferric ions was applied as an adsorbent. In this study, the adsorption behavior of Se to the adsorbent was examined by both batch and column methods. The batch experiment confirmed that selenite ions (Se(IV)) are effectively adsorbed but selenate ions (Se(VI)) are hardly adsorbed. To elucidate the adsorption mechanism, the Fe in the adsorbent and the Fe in the adsorbent after the adsorption of Se(IV) were characterized by Fe K-edge X-ray absorption spectroscopy and 57 Fe Mӧssbauer spectroscopy. The analytical result of Se K-edge EXAFS spectra for the Se(IV) adsorbed on the adsorbent suggests that Se(IV) are adsorbed specifically to the adsorbent through the formation of Fe-O-Se bonds. The breakthrough curve obtained by the column experiment showed that Se(IV) in 3 tons of synthetic solution containing 0.1 ppm Se can be efficiently removed using a column in which 12.8 g (10.4 cm 3 ) of the adsorbent was packed., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The alternative oxidase (AOX) is a monotopic diiron carboxylate protein which catalyzes the four-electron reduction of dioxygen to water by ubiquinol. Although we have recently determined the crystal structure of Trypanosoma brucei AOX (TAO) in the presence and absence of ascofuranone (AF) derivatives (which are potent mixed type inhibitors) the mechanism by which ubiquinol and dioxygen binds to TAO remain inconclusive. In this article, ferulenol was identified as the first competitive inhibitor of AOX which has been used to probe the binding of ubiquinol. Surface plasmon resonance reveals that AF is a quasi-irreversible inhibitor of TAO whilst ferulenol binding is completely reversible. The structure of the TAO-ferulenol complex, determined at 2.7 Å, provided insights into ubiquinol binding and has also identified a potential dioxygen molecule bound in a side-on conformation to the diiron center for the first time., (Crown Copyright © 2019. Published by Elsevier B.V. All rights reserved.)

Extracranial-to-intracranial (EC-IC) bypass surgery may be necessary in patients with moyamoya disease and other ischemic conditions. However, there is a potential risk of wound-related complications in some cases. In this study, we report our approach to the prevention of wound-related complications in EC-IC bypass. Technical considerations and pitfalls of surgery are also discussed. This study included 89 patients with ischemic-onset moyamoya disease and atherosclerotic disease who underwent 108 superficial temporal artery (STA)-to-middle cerebral artery bypass procedures. Our study emphasized 3 major features. First, 3-dimensional simulation imaging was used to confirm STA anatomy. Second, the STA was meticulously dissected on the epigaleal layer to protect the galeal layer. Third, scalp skin ischemia was taken into consideration at each step until skin closure. There was no neurologic morbidity or mortality in this series. There were 2 cases of major wound-related complications requiring plastic surgical intervention, and 4 cases of minor complications that were treated conservatively. In major complication cases, the scalp defect was treated with pedicle flap reconstruction. In EC-IC bypass surgery, interdisciplinary management involving neurosurgery, plastic surgery, and radiology should reduce wound-related complications and achieve safe surgery., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: It has not yet been determined whether hepatic arterial infusion (HAI) chemotherapy improves survival in patients with early hepatocellular carcinoma (HCC). We evaluated the effectiveness of HAI with high-concentration cisplatin (DDP-H) for the treatment of HCC by comparing outcomes between patients who received HAI with DDP-H before radical local treatment of early-stage HCC [Japan Integrated Staging (JIS) score 0/1] and patients who did not receive HAI chemotherapy., Patients and Methods: Survival was analyzed in 114 patients with early-stage HCC who underwent radical local treatment. The patients were divided into two groups: a HAI group (n = 79) who received DDP-H infusion into the whole liver via the proper hepatic artery, and a non-HAI group (n = 35) who did not receive HAI chemotherapy., Results: The cumulative survival rates at 1, 3, and 5 years were 77.4%, 69.2%, and 55.3% in the non-HAI group and 97.4%, 87.0%, and 84.4% in the HAI group, respectively. Survival time prolonged significantly in the HAI group compared with the non-HAI group (log-rank test: P = 0.023; generalized Wilcoxon test: P = 0.012) Multivariate analysis using the Cox proportional hazards model identified HAI with DDP-H as the most important factor affecting survival., Conclusions: Whole-liver HAI with DDP-H before radical local treatment can improve the prognosis of patients with early-stage HCC., (© Crown copyright 2014.)

Tissue infiltration of activated lymphocytes is a hallmark of transplant rejection and organ-specific autoimmune diseases. Migration and activation of lymphocytes depend on DOCK2, an atypical Rac activator predominantly expressed in hematopoietic cells. Although DOCK2 does not contain Dbl homology domain typically found in guanine nucleotide exchange factors, DOCK2 mediates the GTP-GDP exchange reaction for Rac through its DHR-2 domain. Here, we have identified 4-[3'-(2″-chlorophenyl)-2'-propen-1'-ylidene]-1-phenyl-3,5-pyrazolidinedione (CPYPP) as a small-molecule inhibitor of DOCK2. CPYPP bound to DOCK2 DHR-2 domain in a reversible manner and inhibited its catalytic activity in vitro. When lymphocytes were treated with CPYPP, both chemokine receptor- and antigen receptor-mediated Rac activation were blocked, resulting in marked reduction of chemotactic response and T cell activation. These results provide a rational of and a chemical scaffold for development of the DOCK2-targeting immunosuppressant., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A new screening method using fluorescent correlation spectroscopy was developed to select kinase inhibitors that competitively inhibit the binding of a fluorescently labeled substrate peptide. Using the method, among approximately 700 candidate compounds selected by virtual screening, we identified a novel Pim-1 kinase inhibitor targeting its peptide binding residues. X-ray crystal analysis of the complex structure of Pim-1 with the inhibitor indicated that the inhibitor actually binds to the ATP-binding site and also forms direct interactions with residues (Asp128 and Glu171) that bind the substrate peptide. These interactions, which cause small side-chain movements, seem to affect the binding ability of the fluorescently labeled substrate. The compound inhibited Pim-1 kinase in vitro, with an IC(50) value of 150 nM. Treatment of cultured leukemia cells with the compound reduced the amount of p21 and increased the amount of p27, due to Pim-1 inhibition, and then triggered apoptosis after cell-cycle arrest at the G(1)/S phase. This screening method may be widely applicable for the identification of various new Pim-1 kinase inhibitors targeting the residues that bind the substrate peptide., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A fluorescence correlation spectroscopy (FCS)-based competitive binding assay to screen fragment-size compounds that weakly and slowly inhibit protein-peptide interactions was established. The interactions were detected by the increased diffusion time of a fluorescently labeled peptide probe after binding to its interacting protein. We analyzed the interactions between the c-Cbl TKB domain and phosphopeptides derived from ZAP-70, APS, and EGFR with the FCS assay and obtained 6 hit fragments that bound to the c-Cbl interaction sites. The binding amounts of the fragments were measured by direct binding measurements using surface plasmon resonance, and 5 fragments were found to bind selectively. The effect of 2 of the 5 fragments on the interaction with c-Cbl and the peptide exhibited strong time dependency. Furthermore, the inhibition by the selected 5 fragments on the protein-peptide interaction was confirmed by their effect on pull-down assays of c-Cbl with the biotin-conjugated interaction peptides. These results indicate the advantage of our FCS-based assay to study the time-dependent binding of compounds to their target protein., (2010 Elsevier Inc. All rights reserved.)

A rapid and simple method for determination of cyanamide in fertilizer, soil and plants has been developed. In this method, cyanamide is extracted with 2% acetic acid and the extract separated by centrifugation. It is then purified by passing through a membrane filter. The extract was derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate and the derivatized compound separated by ultra-high-pressure liquid chromatography. It is then detected with a UV detector at 260 nm by the same method as is used for amino acid analysis. The proposed method is fast, simple and cheap and also has good selectivity and sensitivity for the determination of cyanamide in a wide range of biotic and abiotic materials.

A dual FIB/SEM provides solutions to many challenges in atom probe specimen preparation. When combined with an in situ lift-out capability, the versatility of this tool allows almost any region of interest, in almost any geometry, to be placed at the apex of a specimen tip. Several preparation techniques have been developed in response to specific application requirements; for example, in cases where materials are not suitable for electropolishing, or where site-specific analysis is required. Two general techniques, with wide-ranging potential applications, are described in detail here. The first is a 'cut-out' technique that provides a relatively quick means of micro-tip specimen preparation from bulk material samples. The second method is a 'lift-out' technique that can be used in an in situ or ex situ mode and does not require the preparation of pre-sharpened mounting points.

Here, we describe a synthetic approach for generating artificial proteins by the assemblage of naturally occurring peptide motifs. Two motifs respectively related to apoptosis induction and protein transduction were encrypted into different reading frames of an artificial gene (microgene), which was then polymerized; random frame shifts at the junctions between the microgene units yielded combinatorial polymers of three reading frames. Among the proteins created, #284 was found to penetrate through cell membranes and exert a strong apoptotic effect on several cancer cell lines. Because a simple linkage of these motifs was not sufficient to construct a bifunctional peptide, and the successful reconstitution was dependent on how they were joined together, the combinatorial strategy is important for reconstituting functions from mixtures of motifs. This microgene-based approach represents a novel system for creating proteins with desired functions.

An 18-month-old boy developed seizures at 3 months of age. He developed normally and, his EEG and brain CT revealed no abnormal findings. The blood sugar level was normal at that time, thus he was diagnosed as having benign infantile convulsion. At 7 months of age seizures reappeared, and hypoglycaemia associated with hyperinsulinism was observed during the seizures. With conservative therapy his blood sugar level was well controlled and he had no further seizures. Hypoglycaemic seizures are sometimes misdiagnosed as epilepsy. We have to pay attention to hyperinsulinemic hypoglycaemia when we see seizures with normal EEG even in infants.

A comparison of the plant growth retardant activity of the chlamydocin analogues, compound 1, six derivatives from 1 and 2, and two synthetic analogues revealed that there are two types of retardant in chlamydocin analogues. One, for example in compound 1, requires an oxygen atom at C-8 of the 2-aminodecanoic acid moiety to show retardant activity. The other, for example in compound 8, requires no oxygen atom at C-8 but requires a specific alkyl group chain length for activity. To determine the differences in mode of action of both types of retardant, rice seedlings were separately treated with compounds 1 and 8, and after appearance of dwarfism, their endogenous ABA and GA(1) levels were determined and compared to those of the control. Treatment with 1 (10 nmol/plant) increased ABA levels 4 times higher than that of the control and decreased GA(1) levels to 20% of that of the control. Treatment with 8 (30 nmol/plant) did not affect the ABA level but decreased GA(1) content to 5% of that of the control.

V-1 positively controls catecholamine synthetic gene transcription to promote catecholamine production in PC12D cells. In this study, immunohistochemical analysis revealed that in Wistar rats, V-1 immunoreactivity was localized not only in sympathetic axons but also in the cytoplasm of cardiomyocytes, and that the immunoreactivity in atrial myocytes was more intense than that in ventricular myocytes. Western blot analysis also showed that V-1 expression level in the atrium was higher than that in the ventricle of Wistar rat hearts. When Dahl salt-sensitive (DS) rats were fed an 8% NaCl diet after the age of 6 weeks, blood pressure was raised 230mm Hg at 18 weeks. V-1 expression was shown to be increased in the atrial myocytes of these DS rats, but not in the sympathetic axons, when assayed by immunohistochemistry. These results suggest that in normotensive rats, V-1 is preferentially expressed in the cytoplasm of cardiomyocytes in the atrium rather than in the ventricle. It is also suggested that V-1 expression is increased by hypertension in DS rat atrium.

The present study was performed to investigate comparatively the toxic effects of inhalation exposure of 1-bromopropane, 2-bromopropane, and 1,2-dichloropropane on reproductive physiology, particularly on the estrous cycle and spontaneous ovulation in female F344 rats. The rats received inhalation exposure to different halogenated propanes, and were exposed daily for 8 h throughout almost 3 weeks to 0,50,200 and 1000 ppm of 1-bromopropane or 2-bromopropane, or to 0,50,100 and 200 ppm of 1,2-dichloropropane. Throughout the exposure period of 1-bromopropane or 2-bromopropane, the ratio of the number of estrous cycle of 6 days or longer to the number of all cycles in both 1000 ppm groups were about two-fold the ratio in each control group, however, no significant difference was found between the ratios of exposed and control groups. The ratios of such long estrous cycles in groups exposed to 100 or 200 ppm of 1,2-dichloropropane were six- to seven-fold higher than that of the control group. These ratios in exposed rats differed significantly from those of controls. The number of ovulated ova in rats exposed to 1,2-dichloropropane decreased in a dose-dependent manner, and the number of ovulated ova in the 200 ppm group was significantly different from that of control rats. Such significant changes in ovulation were not observed in rats exposed to 1-bromopropane or 2-bromopropane. The absolute and relative weights of the ovaries and uterus in rats exposed to three halogenated propanes were not significantly different from those in each control. Therefore, the present study clarified that: (1) 1,2-dichloropropane prolonged the length of the estrous cycle and inhibited spontaneous ovulation in F344 rats; and (2) the potency of 1,2-dichloropropane to disturb the female reproductive physiology appeared to be greater compared with that of 1-bromopropane and 2-bromopropane under the present conditions of inhalation exposure.

The biosynthesis of poly(3-hydroxyalkanoates) (PHAs) by Pseudomonas cichorii YN2 cultured with C6-C12 1-alkenes was studied. PHAs containing repeating units with terminal epoxide groups were obtained when C7-C12 1-alkenes were fed separately as the only carbon source, but no epoxidized unit was detected when 1-hexene was fed. The content of epoxidized units in the polymers was in the range of 4-20 mol%, which was not dependent on the C atom length of the 1-alkene used as a substrate. The polymers produced undergo a glass transition at around -40 degrees C, and number average molecular weights were in the range of 150000-200000 as determined by GPC relative to polystyrene, with M(w)/M(n) ratios of 1.9-2.5. As an intermediate, the corresponding 1,2-epoxyalkane was found in the culture medium. According to this result, the epoxidation of the 1-alkene is the initial step in the synthetic pathway of the epoxy unit in the polymer.

The effects of trichloroethylene (TCE) administration on aldehyde dehydrogenase (ALDH) and cytochrome P450 isozymes were studied in rats and compared with those of methanol. Intragastric administration of TCE to rats at 0.05 or 0.2 ml/kg for 1 week significantly inhibited ALDH activity for aliphatic aldehydes of short chains in the mitochondrial and cytosolic fractions of rat liver, respectively, but had no effect on the activity for long chain aliphatic aldehydes. ALDH activity catalyzing the metabolism of some aromatic aldehydes was even induced by TCE. Microsomal ALDH activity was not decreased by TCE treatment. A kinetic study showed that the low-Km isozyme of ALDH for propionaldehyde in mitochondrial and cytosolic fractions was inhibited by TCE treatment. Addition of TCE, trichloroethanol or trichloroacetic acid to the in vitro assay system did not affect the activity for acetaldehyde, but chloral hydrate at 0.02 mM decreased the activity by 42 and 35% in cytosol and the 700 x g supernatant, respectively. Methanol treatment, on the other hand, had no effect on any ALDH activity. Both TCE and methanol significantly induced CYP2E1 in rat liver. The combined effects of TCE on ALDH and cytochrome P450 may account for the degreasers' flush. Exposure to TCE and methanol may result in a change in the metabolism and toxicity of other chemicals.

Changes of cytochrome P450 isozymes in livers of rats after exposure to four solvents at 4000 ppm for 6 h, were studied by enzyme assays and immunochemical detection using antibodies to cytochrome P450 isozymes. Toluene, benzene and trichloroethylene (TRI) exposure resulted in a significant increase in the activities of nitrosodimethylamine demethylase (152%, 134% and 118%) and 7-pentoxyresorufin O-depentylase (14-, 5- and 2.5-fold), respectively. 1,1,1-Trichloroethane (TCE) showed little effect on the activities of the enzymes. Anti-CYP2E1 and anti-CYP2B1/2 inhibitable activity of toluene side-chain oxidase was significantly enhanced in toluene-, benzene- and TRI-treated rats. Anti-CYP2C11 inhibitable activity was greatly reduced as compared with control. The change in CYP2E1 and CYP2C11 was confirmed by the increase and decrease in the activities inhibited by 4-methylpyrazole and cimetidine, respectively. Western blot analysis revealed that the increase in peak area of bands recognized by anti-CYP2E1 was consistent with toluene inhibition results. CYP2B1/2 was not detectable in control rats, but it was strongly induced by toluene, followed by benzene and TRI. Some increases in the peak areas of bands recognized by anti-CYP2A1 and CYP-4A1 were also observed in the three solvents exposed rat microsomes. Little immunoreactivity was found with anti-CYP1A1 in all microsomes, and no obvious change in peak area of bands recognized by anti-CYP3A and anti-CYP2C13 was observed. TCE exposure showed little effect on these bands. The formation of phenol and hydroquinone from benzene was enhanced to different degree by toluene, benzene and TRI. The hydroxylation of testosterone at 6 beta and 7 alpha was increased by benzene, and benzene and TRI, respectively. However, the metabolism at 16 alpha and 2 alpha was profoundly suppressed by the solvents except TCE. These results showed that the four solvents have different effects on specific cytochrome P450 isozymes and on the metabolism of both endogenous and exogenous substances.

The mechanism by which prostacyclin acts to prevent in vivo reperfusion injury is still uncertain. This study was therefore undertaken to assess the effect of a stable prostacyclin analogue (OP 41483-alpha-CD [OP]) on oxygen-derived free radicals after heart-lung transplantation. OP was administered to the heart-lung graft through the pulmonary artery for 25 minutes encompassing the reperfusion process. Free radicals were directly measured by electron spin resonance spectroscopy. The radical intensities of pulmonary venous blood were significantly lower in the OP group than in the control group, suggesting that fewer free radicals were generated in the lungs of the OP group. The cardiac and respiratory function were better in the OP group than in the control group. The lung is the primary source of oxygen free radical attack, and the beneficial action of OP on free radical generation is almost exclusively restricted to the lung and does not apply to the heart. This result suggested that OP probably is effective in inhibiting free radical generation from the endothelium.

Rats were exposed to methyl bromide (MB) for 24 h at 10-120 ppm or for 3 weeks at 1-10 ppm. Changes in free amino acid contents of rat midbrain were measured by high-performance liquid chromatography (HPLC). MB increased glutamine and aspartic acid contents dose-dependently by short-and long-term exposure. Alanine content was markedly increased by long-term exposure to 10 ppm MB. Glycine was dose-dependently increased, except at 120 ppm. The harmful effect of MB on the central nervous system is discussed in relation to the changes in amino acid metabolism.

To clarify the effects of organic solvents on the central nervous system (CNS), we exposed rats to toluene or n-hexane gas or a mixture of the two compounds at 1000-8000 ppm for 8 h, and measured acetylcholine (ACh) content, choline acetyltransferase (ChAT) and acetylcholine esterase (AChE) activities in homogenized rat hippocampus. ACh was increased at low concentrations of the solvents, but was markedly reduced at high concentrations. ChAT activity was significantly reduced at high concentrations of solvent mixtures. AChE activity was increased by solvents. We concluded that both the decrease in ChAT activity and the increase in AChE activity were the cause of the decrease in ACh content.