Individual and combined effects of methamphetamine and ketamine on conditioned place preference and NR1 receptor phosphorylation in rats

Methamphetamine (MA), a commonly abused psychostimulant, induces the drug dependence by enhancing the dopamine-mediated neurotransmission. Ketamine (KET) is a non-competitive N-methyl-D-aspartate receptor antagonist, which can be actually mixed with MA for polydrug abuse. In the present study, the individual and combined effects of KET (10 mg/kg, i.p.) and MA (1 mg/kg, i.p.) on conditioned place preference in rats were investigated. The alterations of serine 897 phosphorylations of NR1 receptors in the striatum and ventral tegmental area of after-conditioning rats were measured immunochemically. The results showed repeated administrations of MA, KET and their combination, at the doses studied, all could induce psychological dependences evaluated by conditioned place preference. KET was not able to suppress the MA-induced place preference. The modulations of NR1 phosphorylations in basal ganglia were partly responsible to place preference. Although the alterations induced by KET were not significant in most areas we studied, MA showed a significant increase in the ventral tegmental area but a marked decrease in caudate putamen and nucleus accumbens. Such alterations were much more significant when KET and MA were combined. These results have important implications for public awareness of harm with combined drug abuse. Further investigations toward the specific interaction of the two drugs are necessary.     

Involvement of calcium mobilization from caffeine-sensitive stores in mechanically induced cell cycle arrest in the dinoflagellate Crypthecodinium cohnii

Mechanical loads can profoundly alter cell growth and cell proliferation. The dinoflagellates are especially sensitive to mechanical stimulation. Many species will be arrested in cell cycle in response to turbulence or shear stress. We demonstrate here that mechanical shaking and caffeine, the ryanodine-receptor agonist, induced an elevation of cytosolic calcium in the dinoflagellate Crypthecodinium cohnii. Dantrolene, a ryanodine-receptor antagonist, dose-dependently inhibited both shaking-induced and caffeine-induced calcium release. Similar to the effect of mechanical shaking, caffeine alone dose-dependently and reversibly induced cell cycle arrest in dinoflagellates. Prolonged shaking substantially abolished the magnitude of caffeine-induced calcium release and vice-versa, suggesting that both agents released calcium from similar stores through ryanodine receptors. Fluorescence-conjugated ryanodine gave positive labeling, which could be blocked by ryanodine, in the cortice of C. cohnii cells. In addition, caffeine or shaking mobilized intracellular chlortetracycline (CTC)-positive membrane-bound calcium, which could be similarly depleted by t-BuBHQ, a SERCA pump inhibitor. Prior treatment with shaking or caffeine also inhibited the ability of the other agent in mobilizing CTC-positive calcium. CTC-positive microsomal fractions could also be induced to release calcium by caffeine and cADPR, the ryanodinee receptor modulator. t-BuBHQ, but not calcium ionophores, induced cell cycle arrest, and the calcium chelator BAPTA-AM was unable to rescue caffeine-induced cell cycle arrest. These data culminate to suggest that mobilization or depletion of caffeine-sensitive calcium stores, but not calcium elevation per se, is involved in the induction of cell cycle arrest by mechanical stimulation. The present study establishes the role of caffeine-sensitive calcium stores in the regulation of cell cycle progression.     

Cytokinin-mediated cell cycling arrest of pericycle founder cells in lateral root initiation of Arabidopsis

In Arabidopsis, lateral root formation is a post-embryonic developmental event, which is regulated by hormones and environmental signals. In this study, via analyzing the expression of cyclin genes during lateral root (LR) formation, we report that cytokinins (CTKs) inhibit the initiation of LR through blocking the pericycle founder cells cycling at the G(2) to M transition phase, while the promotion by CTK of LR elongation is due to the stimulation of the G(1) to S transition. No significant difference was detected in the inhibitory effect of CTK on LR formation between wild-type plants and mutants defective in auxin response or transport. In addition, exogenously applied auxin at different concentrations could not rescue the CTK-mediated inhibition of LR initiation. Our data suggest that CTK and auxin might control LR initiation through two separate signaling pathways in Arabidopsis. The CTK-mediated repression of LR initiation is transmitted through the two-component signal system and mediated by the receptor CRE1.     

Phase variation and genomic architecture changes in Azospirillum

The plant growth-promoting rhizobacterium Azospirillum lipoferum 4B generates in vitro at high frequency a stable nonswimming phase variant designated 4V(I), which is distinguishable from the wild type by the differential absorption of dyes. The frequency of variants generated by a recA mutant of A. lipoferum 4B was increased up to 10-fold. The pleiotropic modifications characteristic of the phase variant are well documented, but the molecular processes involved are unknown. Here, the objective was to assess whether genomic rearrangements take place during phase variation of strain 4B. The random amplified polymorphic DNA (RAPD) profiles of strains 4B and 4V(I) differed. RAPD fragments observed only with the wild type were cloned, and three cosmids carrying the corresponding fragments were isolated. The three cosmids hybridized with a 750-kb plasmid and pulse-field gel electrophoresis analysis revealed that this replicon was missing in the 4V(I) genome. The same rearrangements took place during phase variation of 4BrecA. Large-scale genomic rearrangements during phase variation were demonstrated for two additional strains. In Azospirillum brasilense WN1, generation of stable variants was correlated with the disappearance of a replicon of 260 kb. For Azospirillum irakense KBC1, the variant was not stable and coincided with the formation of a new replicon, whereas the revertant recovered the parental genomic architecture. This study shows large-scale genomic rearrangements in Azospirillum strains and correlates them with phase variation.     

The effect of magnesium supplementation on lactate levels of sportsmen and sedanter

This study was performed to assess how magnesium supplementation affects plasma lactate levels at rest and exhaustion in sportsmen and sedentary. Research was performed on 30 healthy subjects varying between 18-22 years of age for a four-week period. Subjects were separated into 3 groups: Group 1; sedentary taking magnesium supplementation only (10 mg/kg/day) (Mg + S), Group 2; subjects magnesium supplemented + training 90-120 min 5 days a week (Mg + Training), Group 3; training 90-120 min 5 days a week. Lactate levels of the groups were measured 4 times; at rest and exhaustion in the beginning of the research and after the end of the research. At the end of the research, exhaustion measurements both before and after supplement were found significantly higher than rest measurements in terms of lactate levels (p < 0.05). An important decrease was determined in the lactate levels of the 1st and 2nd groups when compared to their first measurements (p < 0.05). The results of this research indicate that lactate increases with exhaustion. However, magnesium supplement may positively affect performance of sportsmen by decreasing their lactate levels.     

Nitric oxide inhibits an interaction between JNK1 and c-Jun through nitrosylation

Nitric oxide (NO) has been shown to negatively regulate c-Jun N-terminal kinase (JNK) through S-nitrosylation. Here, we show that disruption of an interaction between JNK and its substrate c-Jun is an important mechanism underlying the NO-mediated inhibition of JNK signaling. Endogenous NO, which was generated by interferon-gamma treatment, suppressed anisomycin-stimulated JNK activity in microglial BV-2 cells. The interferon-gamma-induced suppression of JNK1 activation in BV-2 cells was prevented completely by treatment with N(G)-nitro-l-arginine, an inhibitor of NO synthase. A NO donor S-nitro-N-acetyl-dl-penicillamine (SNAP) inhibited JNK activity in vitro, and this inhibition was reversed by a thiol-reducing agent, dithiothreitol. Nitric oxide disrupts a physical interaction between JNK and its substrate c-Jun both in vitro and in intact cells without affecting an interaction between SEK1 and JNK. Collectively, our results suggest that the inhibition of the interaction between JNK and c-Jun may be an integral part of the mechanism underlying the negative regulation of the JNK signaling pathway by NO.     

Biology, life table and predation of Feltiella acarisuga (Diptera: Cecidomyiidae) feeding on Tetranychus cinnabarinus eggs (Acari: Tetranychidae)

Feltiella acarisuga (Vallot) is a common predatory gall midge, which feeds on many species of spider mites. All major life history and life table parameters of F. acarisuga were determined using the carmine spider mite, Tetranychus cinnabarinus (Boisduval) eggs as prey under laboratory conditions [26.7 ± 2 °C, 75 ± 5% RH, and a photoperiod of l4:10 (L:D) h]. Developmental times of F. acarisuga were 2.6, 7.1, and 6.7 d for eggs, larvae, and pupae, respectively, with an average of 16.4 d from egg to adult emergence. Female immatures development took ≈1 d longer than male immatures did. Adult F. acarisuga lived an average of 12.8 d, and the female adults (13.3 d) lived significantly longer than male adults did (11.9 d). After an average of 1.5 d preoviposition period, each female laid an average of 27.3 eggs in its life span with an average of 2.1 eggs per day and an average of 2.8 eggs on each of its oviposition day. The raw data were analyzed using an age-stage, two-sex life table method that takes into consideration of the variable developmental rates among individuals and between sexes. The intrinsic rate of natural population increase (r), net reproductive rates (R_o), gross reproductive rate (∑m_x), generation time (T), and doubling time (DT), and the finite rate of increase (λ) of F. acarisuga were estimated using the age-stage, two-sex (male and female) life table analysis as 0.122 d⁻¹, 16.19 eggs per female, 20.81 eggs per female, 22.81 d, 1.1298 d, and 5.7 d⁻¹, respectively. Each of the first, second, and third instar larvae of F. acarisuga consumed an average of 35.5, 54.0 and 86.9 T. cinnabarinus eggs per day, respectively. Larvae of F. acarisuga could consume an average of 175.4, T. cinnabarinus eggs, and female larvae consumed 14% more spider mite eggs (187.6 eggs) than male larvae (165.1 eggs). The significance of other life table parameters related to the population and the potential of using F. acarisuga as a biological control agent are discussed.     

Oxygen-independent poly(dimethylsiloxane)-based carbon-paste glucose biosensors

Several silicone oils have been assessed and compared as an internal source of oxygen in connection to their use as binders for carbon-paste glucose biosensors. All four poly(dimethylsiloxane) (PDMS) oils tested a dramatic increase in the oxygen capacity of carbon-paste enzyme electrodes to allow convenient biosensing under severe oxygen-deficit conditions. The resulting oxygen independence is better than that exerted by perfluorocarbon binders or that displayed by mediator-based bioelectrodes. The resistance to oxygen effects is indicated from the identical response (observed in the presence and absence of oxygen) up to 2 x 10(-2) M glucose and the slight (12%) sensitivity loss at 4 x 10(-2) M. The influence of the viscosity of the PDMS binder upon the internal oxygen supply is examined. The PDMS carbon-paste enzyme electrode displays a stable glucose response over prolonged (15 h) operation in an oxygen-free solution. On-line continuous testing indicates favorable dynamic properties with no carry-over effects over the physiological and pathophysiological range (3-12 mM glucose).     

mtCOI PCR-RFLP技术鉴别中国入侵型和土著型烟粉虱种群的有效性分析

烟粉虱Bemisia tabaci (Gennadius)是世界范围内最重要的入侵生物之一,准确地鉴别烟粉虱入侵生物型和土著生物型具有十分重要的现实意义。mtCOI PCR-RFLP技术具有快速、高效的特点,是当前应用最广泛的烟粉虱生物型鉴定技术,但是,不同限制性内切酶为基础的mtCOI PCR-RFLP技术在鉴定我国烟粉虱种群中的有效性仍不明了。本文比较了已报导的5种mtCOI PCR-RFLP技术在鉴别中国烟粉虱种群入侵生物型和土著生物型(B型、Q型,ZHJ1型、ZHJ2型、ZHJ3型)的有效性。结果表明,内切酶AluI不能区分B型和ZHJ2型烟粉虱;内切酶TaqI不能准确区分ZHJ3型和Q型烟粉虱个体;而内切酶VspI不仅不能准确区分ZHJ1型和Q型烟粉虱个体,也不能准确区分B型和ZHJ2型烟粉虱;内切酶MseI和Tru9I则不能有效鉴别上述5种烟粉虱生物型,因此不适宜推广使用。