Acute and chronic toxicity of Nigella sativa fixed oil

We investigated the toxicity of the fixed oil of Nigella sativa L seeds in mice and rats through determination of LD50 values and examination of possible biochemical, hematological and histopathological changes. The acute toxicity of Nigella sativa fixed oil was investigated in mice. LD50 values, obtained by single doses, orally and intraperitoneally administered in mice, were 28.8 ml/kg body wt. p.o. [26.2-31.6] and 2.06 ml/kg body wt. i.p. [1.86-2.26], respectively. Chronic toxicity was studied in rats treated daily with an oral dose of 2 ml/kg body wt. for 12 weeks. Changes in key hepatic enzymes levels, including aspartate-aminotransferase, alanine-aminotranferase, and gamma-glutamyltransferase and histopathological modifications (heart, liver, kidneys and pancreas) were not observed in rats treated with Nigella sativa after 12 weeks of treatment. The serum cholesterol, triglyceride and glucose levels and the count of leukocytes and platelets decreased significantly, compared to control values, while hematocrit and hemoglobin levels increased significantly. A slowing of body weight gain was also observed in Nigella sativa treated rats, as compared to control animals. The low toxicity of Nigella sativa fixed oil, evidenced by high LD50 values, key hepatic enzyme stability and organ integrity, suggests a wide margin of safety for therapeutic doses of Nigella sativa fixed oil, but the changes in hemoglobin metabolism and the fall in leukocyte and platelet count must be taken into consideration.     

Lethal synergism of phencyclidine with a precursor and contaminant, 1-piperidinocyclohexanecarbonitrile

The acute median lethal dose (LD50) was determined in rats (p.o.) and mice (i.p.) for phencyclidine (PCP) or 1-piperidinocyclohexanecarbonitrile (PCC) individually and in combination. The LD50 (mg/kg) of PCP was 2.3 or 2.5 times higher than the LD50 of PCC in mice or rats, respectively. For mice, combinations (8:1 or 4:1) of PCP + PCC showed additive synergism, while in rats a 2:1 combination caused sub-additive synergism. Dogs receiving an intravenous infusion of PCP + PCC (4:1) died at a one-third lower dosage of PCP than when PCP was given alone. Prior treatment with NaNO₂ significantly elevated the i.p. LD50's in mice for PCC alone or PCP + PCC (3:1) as it did for KCN. These data support the inference that toxicity from PCC arises primarily from the cyanide ion which it releases. The possibility of a toxic synergism in the course of human exposure to PCC-contaminated “street PCP” is raised.     

Chemical composition and acaricidal properties of Deverra scoparia essential oil (Araliales: Apiaceae) and blends of its major constituents against Tetranychus urticae (Acari: Tetranychidae)

The essential oil of Deverra scoparia Coss. & Durieu was investigated for its acaricidal activity against the worldwide pest twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae). The essential oil was analyzed by fast gas chromatography (GC) and GC-mass spectrometry. The activities of its individual and blended constituents were determined. Our study showed that female mortality increased with increasing D. scoparia oil concentrations, with LD50 and LD90 values at 1.79 and 3.2 mg liter(-1), respectively. A reduction in fecundity had already been observed for concentrations of 0.064, 0.08, and 0.26 mg liter(-1) D. scoparia essential oil. Ten major components, comprising 98.52% of the total weight, were identified; a-pinene was the most abundant constituent (31.95%) followed by sabinene (17.24%) and delta3-carene (16.85%). The 10 major constituents of D. scoparia oil were individually tested against T. urticae females. The most potent toxicity was found with alpha-pinene, delta3-carene, and terpinen-4-ol. The presence of all constituents together in the artificial mixture caused a significant decrease in the number of eggs laid by females, at 0.26 mg liter(-1) (11 eggs), compared with the control (50 eggs). The toxicity of blends of selected constituents indicated that the presence of all constituents was necessary to reproduce the toxicity level of the natural oil.     

曲马朵与二氢埃托啡协同镇痛并推迟大鼠急性耐受

本文旨在研究曲马朵（tramadol,TRA）和二氢埃托啡（dihydroetorphine,DHE）联合用药是否可产生协同镇痛并延缓耐受的发生。TRA（mg,腹腔注射）与DHE（ng,皮下注射）按固定比率给药（1：6．25,1：12．5,1：25,1：50,l：100,1：200）,用热辐射甩尾法评价镇痛效应,采用等高线法评估药物的协同作用。在急性耐受实验中,连续6次注射TRA（20mg／kg）、DHE（1000ng／kg）或两药的组合（TRA20mg／kg＋DHE250ng／kg）。结果显示：（1）除1mg：6．25ng和1mg：50ng两个比例外,其他所有比例用药均产生显著的协同镇痛效应;（2）TRA与DHE联合用药的疗效在连续给药中持续时间明显延长,提示二者联合使用可延缓耐受。以上结果提示：TRA与DHE在一定剂量比范围内可产生协同镇痛效应,并可推迟耐受的形成。     

狼毒活性部位复合乳油功效

为增强瑞香狼毒制剂的杀虫药效、充分开发利用以瑞香狼毒为主的植物源农药,进行了瑞香狼毒杀虫活性部位与其它药物的复配实验。采用瑞香狼毒杀虫活性部位(LD)分别与烟碱乳油(YJR)、CT-901A(Bz)的不同质量比,在室内对家蝇、酢酱草茹叶螨(Petrbia hartiEwing)进行毒力测定,利用软件进行毒力回归分析。实验结果显示,LD和YJR、LD和Bz的两两复配及三者(LD YJR Bz)的复配能够起显著增效作用,特别是20:1:1(LD:YJR:Bz)、30:1:1(LD:YJR:Bz)对家蝇的毒力效果更加明显,LC50达到了0.005 mg/L,20:1(LD:Bz)、30:1(LD:Bz)、30:1:1(LD:YJR:Bz)对酢酱草茹叶螨(Petrbia hartiEwing)的毒力效果也同样显著,LC50也达到了0.005 mg/L。并对瑞香狼毒活性部位进行初步分离检测,其有效成分为黄酮类物质。     

Insecticidal activity of selected monoterpenoids and rosemary oil to Agriotes obscurus (Coleoptera: Elateridae)

Acute toxicities of three naturally occurring monoterpenoid essential oil constituents and the essential oil of rosemary were tested against late instars of Agriotes obscurus (L.) (Coleoptera: Elateridae). Both contact and volatile toxicities of thymol, citronellal, eugenol, and rosemary oil were determined. Also, phytotoxicity of these compounds was evaluated on corn germination and seedling development. Thymol had the greatest contact toxicity (LD50 = 196.0 microg/larva), whereas citronellal and eugenol were less toxic (LD50 = 404.9 and 516.5 microg/larva, respectively). Rosemary oil did not show any significant contact toxicity, even at 1,600 microg/larva. In terms of volatile toxicity, citronellal was the most toxic to wireworm larvae (LC50 = 6.3 microg/cm3) followed by rosemary oil (LC50 = 15.9 microg/cm3), thymol (LC50 = 17.1 microg/cm3), and eugenol (LC50 = 20.9 microg/cm3). Thymol, eugenol, and citronellal significantly inhibited corn seed germination and development, whereas rosemary oil had only minimal phytotoxic effects.     

Toxicity and pathological effects of orally and intraperitoneally administered ivermectin on sea bass Dicentrarchus labrax

The toxicity and histopathology of ivermectin was studied in 3 and 35 g sea bass Dicentrarchus labrax L. following in-feed, oral intubation and injection administration at dose rates ranging from 0.5 to 3.5 mg kg(-1). Estimated LD50 values for 3 g fish were 0.335 and 0.106 mg kg(-1) following oral intubation and injection administration respectively, for fish reared at 11 degrees C; and 0.839 and 1.023 mg kg(-1) following oral intubation and injection administration, respectively for fish reared at 20 degrees C. For 35 g fish reared at 11 degrees C, the estimated LD50 was 0.523 and 0.361 mg kg(-1) following oral intubation and injection administration respectively. No signs of toxicity were observed when the compound was administered via the feed at 0.5 and 0.7 mg kg(-1). However, toxicity (> 10%) was observed at dose rates of 0.2 mg kg(-1) and higher when the compound was administered via oral intubation and at 0.5 mg kg(-1) when administered via injection. The compound was significantly more toxic to fish reared at 11 degrees C than at 20 degrees C. Further, ivermectin was more toxic to 3 g than to 35 g sea bass when administered via injection. Histopathological examination of the major organs revealed pathology was largely restricted to gills and intestinal tissue. In 3 g sea bass, lesions were also found in the kidneys.     

Synergistic effects of short peptides and antibiotics against bacterial and fungal strains

There is a rising tide of concern about the antibiotic resistance issue. To reduce the possibility of antibiotic-resistant infections, a new generation of antimicrobials must be developed. Antimicrobial peptides are potential alternatives to antibiotics that can be used alone or together with conventional antibiotics to combat antimicrobial resistance. In this work, lead compounds LP-23, DP-23, SA4, and SPO from previously published studies were synthesized by solid-phase peptide synthesis and their antimicrobial evaluation was carried out against various bacterial and fungal strains. Peptide combinations with antibiotics were evaluated by using the checkerboard method and their minimal inhibitory concentration (MIC) in combination was calculated by using the fractional inhibitory concentration (FIC) index. Cytotoxicity evaluations of these peptides further confirmed their selectivity toward microbial cells. Based on the FIC values, LP-23, DP-23, and SPO demonstrated synergy in combination with gentamicin against a gentamicin-resistant clinical isolate of Escherichia coli. For Staphylococcus aureus, Escherichia coli, and Salmonella typhimurium, seven combinations exhibited synergistic effects between peptide/peptoids and the tested antibiotics. Additionally, almost all the combinations of peptides/peptoids with amphotericin B and fluconazole also showed effective synergy against Aspergillus niger and Aspergillus flavus. The synergy found between LP-23, DP-23, SA4, and SPO with the selected antibiotics may have the potential to be used as a combination therapy against various microbial infections.     

Effect of Some Variables on theIn VivoDetermination of Scorpion and Viper Venom Toxicities

An adequate assessment of scorpion and snake venom LD50 is an important step for accurate evaluation of antivenom sera potencies and the optimization of serotherapy. The LD50 variation of Tunisian scorpion (Androctonus australis garzonii: Aag and Buthus occitanus tunetanus: Bot) venoms with body weight, sex and strain (Swiss or C57BI/6) of mice used, the route of venom injection, the venom-milking procedures (manually or electrically) and the venom batches have been studied over a 7-year period (1990-1996). Aag venom is 3-4 times more toxic than Bot venom. However for both venoms, the LD50 determined in C57BI/6 mice, in small body weight animal or by intraperitoneal route were respectively significantly lower than those determined in Swiss mice, in high body weight or by subcutaneous route. Significant LD50 variations (25-50%) were also seen from one electrically prepared batch to another. A good correlation (r = 0.982) was observed between the concentrations of the crude venom toxic fraction determined by ELISA and LD50 values when assessed in vivo. The LD50 variation of Tunisian viper (Cerastes cerastes: Cc and Vipera lebetina: VI) venoms with the strain (Swiss or BALB/c), sex and body weight of mice used, the season and the year of venom milking were also investigated over a 3-year period (1990-1992). No significant LD50 variations were observed with the mouse strain, the sex or the season of venom milking. However, LD50 varies significantly with the year of the venom collection and the body weight of mice used. Furthermore, SDS-PAGE analysis shows annual variation for VI venom composition where no such variations were observed for Cc venom. These results stress the need either for the standardization of the venom LD50 evaluation or of the venom quality used for the development of an efficient antivenom.     

Insecticide susceptibility of Nezara viridula (Heteroptera: Pentatomidae) and three other stink bug species composing a soybean pest complex in Japan

The susceptibility of the stink bug species Nezara viridula (L.), Nezara antennata Scott, Piezodorus hybneri (Gmelin), and Riptortus pedestris (F.) to insecticides was tested, establishing their 50% lethal dose (LD50) values as baseline data. Third instars and adults of the four species were treated by topical application with seven insecticides: fenitrothion, fenthion, etofenprox, silafluofen, dinotefuran, clothianidin, and ethiprole. The weight of the stink bug and weight of the insecticide applied to each bug were used as explanatory variables in the probit regression analysis. The effect of the body weight on the dose-response relationship, the proportional model, was not uniform among the tested insecticide-stink bug combinations. However, the basic model fit all combinations and could estimate LD50 values successfully. Therefore, LD50 values at the medium (average) weight estimated by the basic model were selected to describe the susceptibility of the stink bugs. The LD50 value of silafluofen for N. viridula adults, and that of silafluofen and etofenprox for N. antennata adults, was at least 2,338 ng greater than the other species exposed to each insecticide. Almost all of the LD50 values for adults were over 10 times greater than those of the same species' nymphs treated with the same insecticide. Thus monitoring of occurring species and their developmental stages is important for controlling effectively the stink bug pest complex by insecticides, especially by silafluofen or etofenprox. The estimated LD50 values can be used as baseline data to compare the susceptibility of the species collected in another year or location.     

Temperature-dependent variation in toxicity of insecticides against Earias vitella (Lepidoptera: Noctuidae)

The toxicity of synthetic pyrethroids was found to be negatively correlated with temperature, whereas contrasting correlation was observed with the toxicity of organophosphorous compounds chlorpyriphos and quinalphos, which was most toxic at higher temperature. A similar phenomenon was observed in endosulfan at higher temperature and humidity combination. The insecticide molecules indoxacarb and spinosad were effective among the insecticides tested. Indoxacarb was effective at lower temperature, and spinosad was effective at all the temperature and relative humidity combinations with minor difference in LD50 values. During both the years, however, the levels of resistance were higher in second year compared with previous year.     

Coexistence of Two Distinct Diurnal Rhythms in Latency of Licking and Jumping Responses in Mice Hot Plate Tests

Clinical pain from several diseases has been reported to peak at different times along the day. This diverse reactivity to pain could be due to different contributions of the perception of (threshold) and reaction to (tolerance) pain to the pain perceived in various clinical conditions. To gain more insight into this subject we studied the mice hot plate licking and jumping responses, as indicative of pain threshold and pain tolerance, respectively. Both responses were measured every two hours for 24 h, and rhythmometrically analysed in groups of mice maintained for 21 days: (1) under different light-dark cycles (LD 12:12, LD 1:23 and LD 23:1) and tested in the absence or presence of naloxone (1 mg/kg, s.c.); (2) under LD 12:12 and tested in March and September; and, (3) under LD 12:12 and tested after being injected for 21 days with amitriptyline (10 mg/kg/day, i.p.). The main findings were: (1) the co-existence of two distinct diurnal rhythms for licking and jumping responses, whose phases differed by 6 hours under LD 12:12; (2) both rhythms were disrupted or considerably weakened under LD 23:1 and LD 1:23; (3) the endogenous opioid system seems to be involved in suppressing both diurnal rhythms under LD 23:1; (4) the mesor and amplitude, but not the acrophase, of the jumping rhythm seem to be seasonally modulated by opioid influences; and (5) chronic amitriptyline administration induced a significant phase delay of the diurnal rhythm for licking and abolished that for jumping. The data suggest that pain threshold and pain tolerance follow different diurnal rhythms, which might be important for explaining the diverse 24 hour time courses of the reactivity to painful clinical affections.     

Toxicity of propylene oxide at low pressure against life stages of four species of stored product insects

The relative toxicity of propylene oxide (PPO) at a low pressure of 100 mm Hg to four species of stored product insect at 30 degrees C over a 4-h exposure period was investigated. PPO at 100 mm Hg was toxic to all four species tested: Tribolium castaneum (Herbst), Plodia interpunctella (Hübner), Ephestia cautella (Wlk.), and Oryzaephilus surinamensis (L.). There were differences in susceptibility between the life stages of the tested insect species. Mortality tests on all life stages of the insects resulted in LD99 values ranging from 4.7 to 26.1 mg/liter. The pupal stage of E. cautella, O. surinamensis, and T. castaneum was the most tolerant stage with LD99 values of 14.4, 26.1, and 25.7 mg/liter, respectively. For P. interpunctella, the egg stage was most tolerant, with a LD99 value of 15.3 mg/liter. Generally, PPO at 100 mm Hg was more toxic to P. interpunctella and E. cautella than to O. surinamensis and T. castaneum. A 99% mortality of all life stages of the tested species was achieved at a concentrations x time product of 104.4 mg h/liter. These findings indicate that a combination of PPO with low pressure can render the fumigant a potential alternative to methyl bromide for rapid disinfestation of commodities.     

Fumigant toxicity of plant essential oils to Thrips palmi (Thysanoptera: Thripidae) and Orius strigicollis (Heteroptera: Anthocoridae)

The fumigant toxicity of 92 plant essential oils to adult Thrips palmi Karny (Thysanoptera: Thripidae) and Orius strigicollis Poppius (Heteroptera: Anthocoridae) was examined by using a vapor phase toxicity bioassay and compared with those of dichlorvos, emamectin benzoate, spinosad, and thiamethoxam, four commonly used insecticides. Responses varied according to oil type and insect species. As judged by 24-h LC50 values, pennyroyal oil (2.63 mg/liter air) was the most toxic fumigant and was 23.6-fold more toxic than dichlorvos (62.09 mg/liter air) against adult T. palmi. Potent fumigant toxicity (LC50, 11.03-19.21 mg/liter air) was observed in armoise, basil, cedarleaf, coriander, cypress, howood, hyssop, marjoram, myrtle, niaouli, rosemary, and sage (Dalmatia) oils. Neither emamectin benzoate, spinosad, nor thiamethoxam exhibited fumigant action. Against adult O. strigicollis, dichlorvos (LC50, 6.3 x 10(-6) mg/liter air) was the most toxic fumigant, whereas the LC50 values of the 13 essential oils ranged from 17.29 to 158.22 mg/liter air. O. strigicollis was 1.4-22.1 times less susceptible than T. palmi to the essential oils. The essential oils described merit further study as potential fumigants for the control of T. palmi in greenhouses.     

Generalized concentration addition: A method for examining mixtures containing partial agonists

Environmentally relevant toxic exposures often consist of simultaneous exposure to multiple agents. Methods to predict the expected outcome of such combinations are critical both to risk assessment and to an accurate judgment of whether combinations are synergistic or antagonistic. Concentration addition (CA) has commonly been used to assess the presence of synergy or antagonism in combinations of similarly acting chemicals, and to predict effects of combinations of such agents. CA has the advantage of clear graphical interpretation: Curves of constant joint effect (isoboles) must be negatively sloped straight lines if the mixture is concentration additive. However, CA cannot be directly used to assess combinations that include partial agonists, although such agents are of considerable interest. Here, we propose a natural extension of CA to a functional form that may be applied to mixtures including full agonists and partial agonists. This extended definition, for which we suggest the term “generalized concentration addition,” encompasses linear isoboles with slopes of any sign. We apply this approach to the simple example of agents with dose-response relationships described by Hill functions with slope parameter n=1. The resulting isoboles are in all cases linear, with negative, zero and positive slopes. Using simple mechanistic models of ligand-receptor systems, we show that the same isobole pattern and joint effects are generated by modeled combinations of full and partial agonists. Special cases include combinations of two full agonists and a full agonist plus a competitive antagonist.     

Coexistence of Two Distinct Diurnal Rhythms in Latency of Licking and Jumping Responses in Mice Hot Plate Tests

Clinical pain from several diseases has been reported to peak at different times along the day. This diverse reactivity to pain could be due to different contributions of the perception of (threshold) and reaction to (tolerance) pain to the pain perceived in various clinical conditions. To gain more insight into this subject we studied the mice hot plate licking and jumping responses, as indicative of pain threshold and pain tolerance, respectively. Both responses were measured every two hours for 24 h, and rhythmometrically analysed in groups of mice maintained for 21 days: (1) under different light-dark cycles (LD 12:12, LD 1:23 and LD 23:1) and tested in the absence or presence of naloxone (1 mg/kg, s.c.); (2) under LD 12:12 and tested in March and September; and, (3) under LD 12:12 and tested after being injected for 21 days with amitriptyline (10 mg/kg/day, i.p.). The main findings were: (1) the co-existence of two distinct diurnal rhythms for licking and jumping responses, whose phases differed by 6 hours under LD 12:12; (2) both rhythms were disrupted or considerably weakened under LD 23:1 and LD 1:23; (3) the endogenous opioid system seems to be involved in suppressing both diurnal rhythms under LD 23:1; (4) the mesor and amplitude, but not the acrophase, of the jumping rhythm seem to be seasonally modulated by opioid influences; and (5) chronic amitriptyline administration induced a significant phase delay of the diurnal rhythm for licking and abolished that for jumping. The data suggest that pain threshold and pain tolerance follow different diurnal rhythms, which might be important for explaining the diverse 24 hour time courses of the reactivity to painful clinical affections.     

Fumigant Toxicity of Plant Essential Oils to Plutella xylostella (Lepidoptera: Yponomeutidae) and Cotesia glomerata (Hymenoptera: Braconidae)

The fumigant toxicity of 66 plant essential oils to Plutella xylostella (L.) larvae and Cotesia glomerata (L.) adults was examined using a vapor-phase toxicity bioassay and compared with that of dichlorvos. Responses varied according to oil and insect species used. Based on 24 h LD₅₀ values, pennyroyal oil [10.77 mg/filter paper (4.25 cm diameter)] was the most toxic fumigant, followed by rosemary and sage (Dalmatin) oils (15.15 mg/paper). Potent fumigant toxicity was also produced from armoise, buchu leaf, cedarleaf, coriander, eucalyptus, howood, lavender, myrtle, niaouli, peppermint, and rosewood oils (LD₅₀, 21.29–27.31 mg/paper). All essential oils were less effective than dichlorvos (LD₅₀, 0.52 mg/paper). Against adult C. glomerata, dichlorvos (LD₅₀, 0.03 mg/paper) was the most toxic fumigant, whereas the LD₅₀ values of the 14 essential oils ranged from 1.59 to 8.51 mg/paper. Based on selective toxicity ratio (STR, P. xylostella LD₅₀/C. glomerata LD₅₀), the 14 essential oils (STR, 2.5–14.5) are more selective than dichlorvos (STR, 17.3). The essential oils tested merit further study as potential fumigants for the control of P. xylostella in greenhouses because of their selective toxicity to adult C. glomerata and their much greater activity as a fumigant.     

Effect of L-carnitine against acute mitoxantrone toxicity in mice

Various experiments were performed in our laboratory to define a possible role for carnitine derivatives in mitoxantrone (MX) therapy. We report here the results of the effect of L-carnitine (LCAR) on the lethal toxicity of MX in mice. MX was administered intravenously at doses of 7, 9, 11, 13, 15 or 17 mg kg⁻¹ either alone or in combination with LCAR at a single intravenous dose of 200 mg kg⁻¹. The dependence of the probability of death on various doses was evaluated for the MX-LCAR combination compared to MX alone. From these experiments, the following lethal dose fifty (LD₅₀) values were calculated: LD₅₀ for MX alone was 15.2 mg kg⁻¹, whereas in combination with LCAR it increases to 21.8 mg kg⁻¹. The relative toxicity given as the ratio of the LD50 of both MX alone and the combination of MX-LCAR was 69.7%. The results of our experiments unequivocally show the effect of LCAR on acute toxic doses of MX.     

Diurnal variations in the toxicity and tissue levels of spermidine in mice

The LD50 of spermidine shows significant diurnal variability when given IV or IP to mice. It is most toxic during the light phase of a programmed light-dark cycle (12L:12D) and least toxic during the transition from light to dark when motor activity is highest and when whole brain, myelencephalon and liver tissue levels of spermidine and histamine are maximal. Pretreatment with either triplennamine or cyproheptadine significantly reduced the LD50 of spermidine.     

Protection of mice against mixed fission neutron-gamma (n: gamma = 1:1) irradiation by WR-2721, 16,16-dimethyl PGE2, and the combination of both agents

The survival of mice after whole-body exposure to a modified fission neutron-gamma field (n: gamma = 1:1) was used to examine radiation protection by WR-2721, 16,16-dimethyl PGE2(DiPGE2), and the combination of both agents. Administration of WR-2721 (453 mg/kg) increased the LD50/30 from 5.24 to 7.17 Gy (DMF = 1.37), whereas pretreatment with DiPGE2 (1.6 mg/kg) increased the LD50/30 to 5.77 Gy (dose modification factor (DMF) = 1.10). The combination of 453 mg/kg WR-2721 and 0.4 mg/kg DiPGE2 resulted in an LD50/30 of 7.33 Gy, yielding a DMF of 1.39. However, no significant difference in protection was obtained with the combination of the two agents compared to that seen with WR-2721 alone.