Effects of Recombinant Murine Interferon-{gamma} on Pregnant Mice and Their Fetuses

Effects of Recombinant Murine Interferon- on Pregnant Mice andTheir Fetuses. KATO, I., KIMURA, S., FURUHASHI, T., NAKAYOSHI,H., TAKAYAMA, S., AND UENISHI, N. (1990). Fundam. Appl. Toxicol.14, 658–665. Recombinant murine interferon- (Rec-MuIFN-)was administered intramuscularly to C3H/HeNCrj mice on Days6-15 of gestation at dosage levels of 8 x 10⁵, 4 x 10⁶, and2 x 10⁷ u/kg/day. Dams were killed for examination of fetuseson Day 18 of gestation. Pregnant females that received 2 x 10⁷u/kg/day of Rec-MuIFN- showed uterine bleeding on Days 10–15of gestation and could not maintain their pregnancy. These damsdied on Days 13–17orwerekilled/ieWrem/sonDays 10–15for examination, and therefore no fetal data were availablefor this group. In the 2 x 10⁷ u/kg/day group, the mean absoluteweights of the lung and spleen increased and the mean absoluteweight of the liver, red blood cells (RBC), hematocrit, andhemoglobin decreased significantly. Surviving dams in the 8x 10³ and 4 x 10⁶ u/kg/day groups showed significant increasesin the mean absolute weights of the lung, liver, kidneys, andspleen and a decrease in platelet count. Significant increasesin the weights of the heart and ovaries and decreases in RBC,hematocrit, and hemoglobin were observed in the 4 x 10⁶ u/kg/daygroup. Histopathological examination revealed increased extramedullaryhema-topoiesis in the spleen of the 4 x 10⁶ and 2 x 10⁷ u/kg/daygroups. Fetuses showed no external, visceral, or skeletal malformationsand variations caused by the administration of Rec-MuIFN-7 inany of the treated groups. Although a slight but statisticallysignificant decrease in fetal body weight and a delay in ossificationwere seen in fetuses in the 4 x 10⁷ u/kg/day group, these findingswere considered to be the result of maternal toxicity and notfetal toxicity. No fetal effects were observed i n the 8 x 10⁵u/kg/day group.     

d-Limonene Induced Hyaline Droplet Nephropathy in {alpha}2u-Globulin Transgenic Mice

d-Limonene is a hyaline droplet inducing agent and producesnephrotoxicity in male rats when the 1,2-epoxide metabolitebinds to 2u-globulin. Mice, which do not synthesize 2u-globulin,are resistant to hyaline droplet nephropathy. In this study,the ability of d-limonene to cause hyaline droplet nephropathyin a transgenic mouse engineered to express 2u-globulin wasevaluated. The C57BL/6-derived mice excreted 0.4 ± 0.1mg 2u-globulin/day, or approximately 16 mg 2u-globulin/kg bodywt. This represents about 30% of the amount excreted by adultmale rats (11.9 ± 1.1 mg/day or approximately 48 mg/kg).Transgenic mice excreted less mouse urinary protein (9.3 ±1.2 mg/day) than normal mice (15.1 ± 1.6 mg/day). Unlikenormal male rats, untreated transgenic mice did not show significantspontaneous hyaline droplet formation. Liver microsomes fromnaive transgenic mice oxidized d-limonene to the cis- and trans-isomersof the 1,2-epoxide, and following oral treatment with [¹⁴C]d-limonenereversible binding of d-limonene equivalents to renal cytosolicproteins was observed. Furthermore, with d-limonene treatment,hyaline droplets were observed in the transgenic mouse kidneys.These droplets, however, were much smaller in size than thoseseen in d-limonene-treated male rats. The accumulation of 2u-globulinin the kidneys of transgenic mice and normal male rats beforeand after d-limonene treatment was analyzed by Western blotting.These results indicated that 2u-globulin was present in thekidneys of the control transgenic mice, despite the lack ofspontaneous hyaline droplet formation. After d-limonene treatment,approximately a three fold increase in 2u-globulin in the transgenicmouse kidney was observed, a response similar in magnitude tothat seen in d-limonene-treated male rats. These results indicatethat expression of 2u-globulin in a species that does not normallydevelop hyaline droplet nephropathy is necessary and sufficientto render that species sensitive to this renal toxicity.     

Inhibition of Human Plasma and Serum Butyrylcholinesterase (EC 3.1.1.8) by {alpha}-Chaconine and {alpha}-Solanine

The purpose of these experiments was to determine the reversibilityof -chaconine and -solanine inhibition of human plasma butyrylcholinesterase(BuChE). For the substrate -naphthylacetate, optimal assay conditionswere 0.50 M sodium phosphate buffer and a substrate concentrationof 3–5 ' 10^–4 M. Dibucaine (1 ' 10^–5 M) indicatedthe usual phenotype for all subjects; -chaconine and -solanineat 2.88 ' 10^–6 M inhibited BuChE about 70 and 50%, respectively.One-and 24-hr incubations at 1 ' 10^–1 M with -chaconine,-solanine, paraoxon, eserine, and ethanol yielded reversibleinhibition with dilution except for paraoxon. Twenty-four-hourdialyses of incubations showed no inhibition except for paraoxon.PAGE enzyme activity gels of 1-and 24-hr incubations also showedno inhibition except for paraoxon. -Chaconine and -solanineare reversible inhibitors of human butyrylcholinesterase. Atestimated tissue levels, -chaconine, -solanine, and solanidineinhibited BuChE 10–86%. In assays which combined -chaconine,-solanine, and solanidine, inhibition of BuChE was less thanadditive. No inhibition of albumin -naphthylacetate esterase(an arylesterase) was noted with any inhibitor. The importanceof these data to adverse toxicological effects of potato alkaloidsis discussed.     

Phytoestrogens and Their Human Metabolites Show Distinct Agonistic and Antagonistic Properties on Estrogen Receptor {alpha} (ER{alpha})and ER{beta} in Human Cells

Figure 3 and Table 3 for this article should have run as presentedbelow. The Publisher regrets the error. View larger version (35K): [in this window] [in a new window]   FIG. 3. Transactivational potencies and     

The Perturbation of Hepatic Glutathione by {alpha}2-Adrenergic Agonists

The Perturbation of Hepatic Glutathione by ₂ -Adrenergic Agonists.James, Robert C, Roberts, Stephen M. and Harbison, Raymond D.(1983). Fundam. Appl. Toxicol.. 3: 303–308. Single injectionsof epinephrine significantly lowered the hepatocellular levelsof reduced glutathione (GSH) while producing small but significantelevations in serum glutamic-pyruvic transaminase (SGPT) activity.Hormones, i.e. glucagon and the corticosteroids, were also foundto depress significantly hepatic glutathione. Based upon theagonist-antagonist studies performed, the hepatic GSH loweringeffects of epinephrine appear to be mediated solely by ₂ receptors.Adrenergic antagonists with ₂ receptor blocking properties,phenotolamine and yohimbine, prevented the epinephrine-inducedlowering of GSH while agonists with ₂ activity, clonidine andguanabenz, mimicked epinephrine's response. Antagonists witheither ₁or ß activity, i.e. prazosin, phenoxybenzamineand propranolol, did not prevent the epinephrine-induced loweringof hepatic GSH. Contrary to these findings antagonists witheither or P receptor blocking activity significantly reducedthe epinephrine-induced elevations in SGPT activity. Thus, therewas no apparent relationship between the elevation of SGPT activityand the reduction in hepatic glutathione levels. It is concludedthat the therapeutic administration of these compounds, or physiologicresponses to stress or pain, may exacerbate the hepatotoxicityof compounds detoxified by GSH or alter important glutathione-mediatedhepatocellular processes.     

Leydig Cell Hyperplasia and Adenomas in Mice Treated with Finasteride, a 5{alpha}-Reductase Inhibitor: A Possible Mechanism

Finasteride is a selective inhibitor of the enzyme 5-reductasewhich is responsible for the conversion of testosterone (T)to dihydrotestosterone (DHT). Finasteride is indicated for thetreatment of benign prostatic hyperplasia in man ({small tilde}0.1mg/kg/day). The effect of long-term treatment was studied inmice given high doses (2.5, 25, and 250 mg/kg/day) of finasteridefor 83 weeks. In finasteride-treated mice, increased incidencesof testicular Leydig cell hyperplasia (52% compared to 24% incontrol group) at doses equal to or greater than 25 mg/kg/dayand Leydig cell adenomas (32% compared to 0.5% in control group)at 250 mg/kg/day were observed. There were no drug-related effectson the seminiferous tubules. Since luteinizing hor mone (LH)is a trophic hormone for Leydig cells, short-term studies (5to 14 weeks) were done to investigate the relationship betweenLeydig cell hyperplasia and serum LH levels in finasteride-treatedmice. In these studies, there was a positive correlation betweenthe drug-related increased incidence of Leydig cell hyperplasiaand a statistically significant (p 0.05) increase in serum LHlevels in finasteride-treated (250 mg/kg/day) mice. Furthermore,studies in castrated male mice showed that the suppression ofserum LH levels by T is reversible by inhibition of conversionof T to DHT with finasteride (250 mg/kg/day), supporting thehypothesis that DHT is involved in the regulation of LH releasein mice. The data presented support the conclusion that theeffects of finasteride on Leydig cells appear to be secondaryto increased serum LH levels and that they occur only at veryhigh doses when compared to the therapeutic dose (approximately0.1 mg/kg/day) in man.     

Application of Molecular Encapsulation for Toxicology Studies: Comparative Toxicity of p-Chloro-{alpha},{alpha},{alpha}-trifluorotoluene in {alpha}-Cyclodextrin Vehicle versus Corn Oil Vehicle in Male and Female Fischer 344 Rats and B6C3F1 Mice

The application of -cyclodextrin (-CD) as an alternative vehiclefor water insoluble and volatile chemicals was investigatedin toxicity studies of p-chloro-,,-trifluorotoluene (CTFT).Groups of F344 rats and B6C3F1 mice of each sex were administeredCTFT (97% pure) by gavage in either corn oil or -CD aqueousformulations daily for 14 consecutive days. The dose levelsused were 10 (mice only), 50, 400, and 1000 mg/kg for corn oilvehicle and 10, 50, and 400 mg/kg (maximum achievable dose atgavage volume of 5 ml/kg) for -CD vehicle. With both vehiclesCTFT and a_2u-globulin were found to accumulate in the male ratkidney after 14 days of exposure and a dose-related toxic nephropathywas observed at dose of 50 mg/kg or higher. The hepatocellularhypertrophy and cytoplasmic vacuolation of the adrenal cortexwhich appeared in dosed male and female rats were also foundto be independent of vehicle. Clinical pathology findings suggesteda mild anemia and cholestasis in rats. With both vehicles notissue bioaccumulation of CTFT was found in male or female mice.Vehicle-independent hepatocellular hypertrophy and cholestasiswere also observed in mice at doses of 400 and 1000 mg/kg. Inconclusion, the -CD vehicle does not affect the toxic responsesof CTFT in both sexes of both species. The results of the studiessuggest that -CD may be an appropriate alternative vehicle fortoxicity studies.     

Metabolism of {alpha}-Olefin Sulfonate (AOS) in Rats

Metabolism of -Olefin Sulfonate (AOS) in Rats. Inoue, S., O'Grodnick,J.S. and Tomizawa, S. (1982). Fundam. Appl. Toxicol. 2:130-138.The metabolic fate of ¹⁴C-AOS (a mixture of ¹⁴C-sodium alkenyl(2)sulfonate and ¹⁴C-sodium 3-hydroxy alkane sulfonate) has beenstudied in rats by a single oral and intravenous injection of100 mg (50 µCi)/kg and 10 mg (5 µCi)/kg, respectively.After oral administration, ¹⁴C-AOS was rapidly absorbed fromthe gastrointestinal tract. The blood level of ¹⁴C-activityreached its peak 3 hr after dosing and then declined. Twenty-fourhr after the dose, about 0.8%/g of the ¹⁴C-AOS given was detectedin cecum content, but in other tissues the figures were under0.02% dose/g. Within 24 hr after the dose, 72% of the dose wasexcreted in the urine and 22% in the feces, while the excretionin the bile was 4.3% within 12 hr. The administered radioactivitywas rapidly eliminated from the whole body within 24 hr. Afterintravenous injection, half of the administered dose of radioactivitywas excreted within 1 hr. In the 0–6 hr interval post-dose,90% of the dose was excreted in the urine. No intact ¹⁴C-AOSwas detected in any of the urine samples after oral and intravenousdoses. The metabolite was apparently more polar than intact¹⁴C-AOS, and results from data of electrophoresis and equilibriumdialysis indicated that intact ¹⁴C-AOS can bind with proteins,while the metabolites cannot. The metabolite was found to containalcoholic, unsaturated and sulfonic acid functionalities. Itis suggested that the metabolite may be a hydroxylated or polyhydroxylatedsulfonic acid of shorter chain length than AOS. These resultssuggest that ¹⁴C-AOS is rapidly absorbed, metabolized and excreted,therefore, no accumulation of ¹⁴C-AOS occurs.     

重组人干扰素大肠杆菌高效表达系统的建立

目的:构建重组人干扰素-α2b(rhIFN-α2b)原核表达系统,以获得rhIFN-α2b在大肠杆菌中的高效表达。方法:依据大肠杆菌遗传密码子频率表,在不改变氨基酸组成的情况下,人工半合成适于在大肠杆菌中表达的rhIFN-α2b编码序列;经PCR扩增后,克隆人表达型质粒载体pDH;筛选阳性菌落,温度诱导表达,表达产物行聚丙烯酰胺凝胶电泳(SDS-PAGE)、蛋白质印迹(WesternBlot)鉴定,并以人羊膜细胞-水泡性口炎病毒(WISH-VSV)系统鉴定rhIFN-α2b抗病毒活性。结果:PCR产物经琼脂糖凝胶电泳鉴定可见与预期大小符合的512bp的DNA条带;重组质粒pDH/IFN-α2b经序列分析,证实含有与预期相符且读码框架正确的hIFN-2b编码序列;SDS-PAGE可见与IFN-α2b分子质量大小一致的蛋白质条带,WesternBlot鉴定此条带为rhIFN-2b,其比活性达(1.8～2)×108U/mg。结论:rhIFN-α2b在大肠杆菌中的高效表达系统构建成功,且能高水平地表达出具有生物活性的rhIFN-α2b。     

Possible Involvement of Prostaglandins in Increases in Rat Plasma Adrenocorticotropic Hormone and Corticosterone Levels Induced by Radiation and Interleukin-1{alpha} Alone or Combined

Exposing rats to 1–10 Gy of ionizing radiation increasedplasma adrenocorticotropic hormone (ACTH) and corticoste-rone(CORT) levels. In both irradiated and nonirradiated rats, recombinanthuman interleukin-1 (rhIL-1 1 hr before radiation/sham exposure)enhanced plasma ACTH and CORT levels. Indomethacin, a cyclooxygenaseinhibitor, attenuated plasma ACTH and CORT levels induced byradiation. Indomethacin also attenuated ACTH and CORT levelsinduced by radiation and interleukin-1 alone or combined. Theseresults suggest that prostaglandins are involved in the increasein plasma ACTH and CORT levels induced by radiation and rhIL-1alone or combined.     

海人酸诱导的颞叶癫痫小鼠慢性期行为学变化

目的:研究海人酸(Kainic acid)所致的颞叶癫痫小鼠慢性期行为学改变及意义。方法:用海人酸构建癫痫小鼠模型作为实验组,对照组用生理盐水处理,2个月后分别进行Morris水迷宫实验、条件恐惧实验、高架十字实验和旷场实验。结果:Morris水迷宫实验中,后3d实验组小鼠的定位航行潜伏期长于对照组(P0.01),实验组目标象限滞留时间短于对照组(P0.01);条件恐惧实验中,场景联系性记忆实验实验组小鼠僵直时间百分比小于对照组(P0.01),声音联系性记忆实验实验组小鼠僵直时间百分比小于对照组(P0.01);旷场实验中,实验组小鼠活动路程长于对照组(P0.01),实验组小鼠中间区域滞留时间短于对照组(P0.01),实验组小鼠大小便次数比对照组多(P0.01);高架十字实验中,实验组和对照组小鼠进入开臂的次数比无显著差别(P0.05),两组小鼠在开臂的活动时间比较也无差异(P0.05),但实验组小鼠进入两种臂的总次数较对照组小鼠多(P0.05)。结论:海人酸诱导的颞叶癫痫小鼠慢性期有认知功能和情绪方面的障碍。     

乳糖诱导重组人SOD基因在大肠杆菌中的表达

利用乳糖代替IPTG诱导rhCu/Zn-SOD在大肠杆菌(Escherichia coli）BL21（DE3）中的表达,并将菌体生长情况及产物表达规律与IPTG诱导进行比较。对诱导所需的乳糖浓度、金属离子浓度等因素进行了研究,并分析了低温下诱导对重组蛋白表达的影响。最终在摇瓶发酵条件的基础上,实现了乳糖诱导重组菌的5L全自动发酵罐培养,rhCu/Zn-SOD酶活性达到1810U/ml(发酵液）。结果表明,乳糖可以诱导rhCu/Zn-SOD在Escherichia coli中的表达,并且酶活性达到IPTG诱导时的89%。     

Pharmacological dose of {alpha}-tocopherol induces cardiotoxicity in Wistar rats determined by echocardiography and histology

The effect of pharmacological dose of α-tocopherol on heart health was determined in Wistar rats. Animals were randomly assigned to either C (control, n = 11) or E (α-tocopherol, n = 11) group. Animals received corn oil (C) or α-tocopherol dissolved in corn oil (250 mg α-tocopherol/[kg body wt/day]) (E) by gavage for a 7-week period. Rats underwent echocardiogram and were analyzed for cardiomyocyte histology and cardiac α-tocopherol absorption at the end of the study period. As compared to the C group, α-tocopherol-supplemented group showed significantly (p < 0.05) lower body weight (E, 412.8 g vs C, 480.3 g) and total cardiac weight (E, 0.94 g vs C, 1.08 g); cardiomyocyte histological impairment; smaller left ventricle (LV) (LV end-diastolic diameter (E, 7.22 mm vs C, 7.37 mm), lower LV systolic [left ventricle fractional shortening (E, 47.6% vs C, 53.6%) and ejection fraction ratio (E, 85.4 vs C, 89.9)] and diastolic [early peak velocities of diastolic transmitral flow (E, 64.6 cm/sec vs C, 75.1 cm/sec)] function. The α-tocopherol uptake in target tissue was confirmed by determination of α-tocopherol concentration medians in cardiac tissue (E, 109.91 nmol/kg vs C, 52.09 nmol/kg). The current study indicates that pharmacological dose of α-tocopherol supplementation can induce cardiotoxicity in healthy rats.     

Recombinant TCR Ligand Reverses Clinical Signs and CNS Damage of EAE Induced by Recombinant Human MOG

Increasing evidence suggests that in addition to T cell-dependent effector mechanisms, autoantibodies are also involved in the pathogenesis of MS, including demyelinating antibodies specific for myelin oligodendrocyte glycoprotein (MOG). Our previous studies have demonstrated that recombinant T cell receptor ligands (RTLs) are very effective for treating T cell-mediated experimental autoimmune encephalomyelitis (EAE). In order to expand the scope of RTL therapy in MS patients, it was of interest to study RTL treatment of EAE involving a demyelinating antibody component. Therefore, we evaluated the therapeutic effects of RTL551, specific for T cells reactive to mouse (m)MOG-35-55 peptide, on EAE induced with recombinant human (rh)MOG in C57BL/6 mice. We report that RTL551 therapy can reverse disease progression and reduce demyelination and axonal damage induced by rhMOG without suppressing the anti-MOG antibody response. This result suggests that T cell-mediated inflammation and associated blood–brain barrier dysfunction are the central contributors to EAE pathogenesis and that successful regulation of these key players restricts potential damage by demyelinating antibodies. The results of our study lend support for the use of RTL therapy for treatment of MS subjects whose disease includes inflammatory T cells as well as those with an additional antibody component.     

Effects of Lifelong Administration of {beta}-Phenylisopropylhydrazine Hydrochloride and Thiocarbamylhydrazine in Mice

Effects of Lifelong Administration of ß-Phenylisopropyl-hydrazinehydrochloride and thiocarbamylhydrazine in mice. Toth, B. (1982).Fundam. Appl. Toxicol. 2:173-176. ß-Phenylisopropylhydrazinehydrochloride and thiocarbamylhydrazine were separately administeredas 0.0312 and 0.0156% solutions in drinking water for life torandomly bred Swiss mice. The consumption of the chemicals resultedin no detectable tumorigenic effects in the treated animals,and it is therefore concluded that these compounds are apparentlynoncarcinogenic under the present experimental conditions.     

重组人促红细胞生成素不良反应149例文献分析

摘 要 目的：分析重组人促红细胞生成素（rHuEPO）药品不良反应（ADR）的发生情况,为临床安全用药提供参考。方法: 收集国内外公开报道的rHuEPO的ADR文献资料,按患者年龄、性别、原患疾病、ADR发生时间、累及器官及临床表现、转归等进行整理并分析。结果:经检索,rHuEPO的ADR共计149例,累及器官主要有心血管系统损害（43.4%）、血液系统损害（20.8%）、皮肤及其附件损害（12.7%）。临床表现前3位分别是高血压、再生障碍性贫血、高钾血症。发生时间多集中在用药后5~12周内（43.0％）。结论:rHuEPO在临床使用过程中应关注其ADR、发生时间,注意患者用药教育及随访,避免严重ADR的发生。     

细胞周期在沥青烟致小鼠肺损伤的变化

目的观察沥青烟对小鼠肺组织细胞周期的影响，研究沥青致俯和致突变作用的机制。方法用沥青烟对小鼠进行不同剂量和不同时间的染毒，应用流式细胞仪进行小鼠肺组织细胞DNA含量的检测和细胞周期的分析。结果随着沥青烟染毒时间和剂量的增加，小鼠肺组织细胞G1期细胞数下降，S期阻滞，进入G2／M期的细胞减少，细胞增殖指数（PI）增加，异倍体指数（DI）升高。结论沥青烟可以对小鼠肺组织细胞周期产生影响，主要表现在S期DNA合成增加。细胞的增殖能力增强。     

Toxicokinetics of Nickel in Mice Studied with the {gamma}-Emitting Isotope 57Ni

The -emitting isotope ⁵⁷Ni was generated in a cyclotron to allowwhole-body counting of laboratory animals dosed with nickel.⁵⁷NiCl₂ was administered either orally by gastric intubationor by intraperitoneal injection to groups of mice in doses equivalentto the average human daily dietary nickel intake per mass unit.When given orally, the whole-body retention (WBR) was 0.02–0.36%of the administered dose at 45–75 hr. When given intraperitoneally,the WBR was 1–6% at 20–50 hr. After adjustment forthe rapid excretion of systemic nickel, the intestinal absorptioncould be estimated to be 1.7–10%. The relative WBR didnot vary with the magnitude of the dose within 0.05–5µmol Ni/kg given orally or 0.005–0.5 µmol/kggiven intraperitoneally. At 8 hr, the tissue concentration washighest in the kidneys, followed by the carcass, lungs, testicles,liver, and the spleen. After 20 hr, the highest concentrationswere still found in the kidneys followed by the lungs, the liver,and the carcass. At 20 hr after oral administration, 50–70%of ⁵⁷Ni retained in the body was within the carcass. The secondhighest nickel content was found in the kidneys, followed bythe liver and lungs. Whereas nickel in the kidneys was rapidlyexcreted, the elimination from the lungs and liver was relativelyslow, thereby, after 40 hr, resulting in a higher nickel contentin the liver than that in the kidneys. When nickel was givenintraperitoneally, practically no nickel was transported viathe portal vein to the liver after 20 hr, resulting in a lownickel content in the liver and a higher content in the kidney.These results document that the use of ⁵⁷Ni for studies on nickeltoxicokinetics is feasible and useful, and that the method isespecially well suited for comparative studies with a durationof up to 6 days.