METHYL MERCURY INHIBITION OF SYNAPTOSOME AND BRAIN SLICE PROTEIN SYNTHESIS: IN VIVO AND IN VITRO STUDIES

Subacute methyl mercury (MeHg) intoxication was induced in adult rats following the daily intragastric administration of 1 mg MeHg/100 g body weight. Decreased [¹⁴C]leucine incorporation into cerebral and cerebellar slice protein was found. Weight loss occurred during the latent and neurotoxic phases but pair feeding did not reveal a significant defect in amino acid incorporation into slice protein. There was no decline in synaptosome protein synthesis in vitro during the latent phase but a significant decline in cerebellar and cerebral synaptosome synthesis was found during the neurotoxic phase. MeHg in vitro inhibited cerebral slice and synaptosome protein synthesis at half maximal concentrations of 7.5 and 12.5 μM respectively. Inhibition of synthesis in synaptosomes was non-competitive with K₁ of 4 × 10^?6M. MeHg had no effect on [¹⁴C]leucine or [¹⁴C]proline uptake into synaptosomes. There was no significant inhibition of synaptosome basal ATPase or Na + K ATPase at concentrations of MeHg (12 μM) giving half maximal inhibition of protein synthesis. No preferential inhibition of the chloramphenicol (55S) or cycloheximide sensitive components of synaptosome fraction protein synthesis was found, suggesting that the inhibition is common to both mitochondrial and extramitochondrial protein synthesizing systems. Addition of nucleotides and/or atractylate failed to influence protein synthesis and did not reverse the MeHg inhibition. Mannitol, as a replacement for the predominant cation species of the incubation medium, gave 40% inhibition of protein synthesis in the control but protected against further inhibition by MeHg.     

COMPARATIVE EFFECTS OF ORGANIC AND INORGANIC MERCURY ON BRAIN SLICE RESPIRATION AND METABOLISM

—Respiration and carbohydrate metabolism were measured in guinea-pig brain slices exposed to organic and inorganic mercury. Organic mercury decreased oxygen uptake and ¹⁴CO₂ production at consistently lower concentrations than inorganic mercury. Organic mercury also caused a striking elevation of pyruvate and lactate at low doses where inorganic mercury had no effect on respiration or metabolism. A unique inhibition of tricarboxylic acid cycle function is suggested and might partially explain the distinctive neurotoxicity of organic mercury.     

吩噻嗪衍生物在体内外对肠道细菌的作用

本工作测试了７种吩噻嗪衍生物在体外对５株好氧或兼性厌氧菌和８９株厌氧菌的最小抑菌浓度,结果表明,吩噻嗪衍生物对细菌（尤其是球菌和厌氧菌）具有一定的抑制作用,但携带可拮抗艰难梭菌肠道定植屏障菌群的悉生小鼠接受２周或４周的Ｃｈｌｏｒｐｒｏｍａｚｉｎｅ（０．２ｍｇ／小鼠／天）并不会使菌群屏障遭破坏。     

EFFECTS OF GAMMA-HYDROXYBUTYRIC ACID AND OTHER HYPNOTICS ON GLUCOSE UPTAKE IN VIVO AND IN VITRO

Abstract— (1) The effects of gamma-hydroxybutyrate, imidazole-4-acetic acid and pento-barbitone on mouse brain glucose, glycogen and lactate levels have been studied. All the drugs significantly increased the brain glucose content, but did not significantly alter brain glycogen levels. The increase in brain glucose following imidazole-4-acetic acid or hypnotic doses of pentobarbitone was matched by corresponding decreases in the lactate level; this was not the case with gamma-hydroxybutyrate where the total glucose equivalents in the brain, expressed as the tissue level of (glucose) + (lactate/2), were significantly increased.
(2) All drugs except imidazole-4-acetic acid significantly decreased the rate of appearance of [¹⁴C]glucose into the bloodstream in vivo but had no effect on the uptake of glucose into rat diaphragm in vitro when present at 2·5 mM concentration.
(3) Only imidazole-4-acetic acid significantly inhibited glucose uptake into the brain in vivo but at 2·5 mM had no significant effect on glucose uptake into rat cerebral cortical slices in vitro.
(4) It is concluded that the very large increase in brain glucose level observed following the injection of hypnotic doses of gamma-hydroxybutyrate cannot be explained in terms of an increased net uptake of glucose into the brain.     

EFFECTS OF ANOXIA IN VITRO ON CELLULAR RESPIRATION OF BRAIN CORTEX

Abstract— –Anaerobic preincubation of slices of brain cortex from adult rats in saline at 37°C, but not at 20°C, impaired their capacity to oxidize glucose and other substrates. This effect was to a large extent prevented by including either glucose + oxaloacetate, or l -glutamine + l -aspartate +α-oxoglutarate in the medium during anaerobic preincubation, and in addition the anaerobic uptake of water by the tissue was decreased. The respiratory mechanism in slices of brain cortex from 1-day-old rats was much less affected by anoxia and sensitivity to anoxia develops with age.     

EFFECTS OF LITHIUM TREATMENT IN VITRO AND IN VIVO ON ACETYLCHOLINE METABOLISM IN RAT BRAIN

Abstract— The effects of LiCl on cholinergic function in rat brain in vitro and in vivo have been investigated. The high affinity transport of choline and the synthesis of acetylcholine in synaptosomes were reduced when part (25-75%) of the NaCl in the buffer was replaced with LiCl or sucrose. This appeared to be due to lack of Na⁺ rather than to Li⁺, as addition of LiCl to normal buffer had little effect. Following an injection of LiCl (10mmol/kg, i.p.) into rats the concentration of a pulsed dose of [²H₄]choline (20 μmol/kg, i.v., 1 min) and its conversion to [²H₄]acetylcholine, and the concentrations of [²H₂]acetylcholine and [²H₀]choline were measured in the striatum, cortex, hippocampus and cerebellum. The [²H₄]choline and [²H₄]acetylcholine were initially (15 min after LiCl) reduced (to ?30% in the cortex) and later (24 h after LiCl) increased (to + 50% in the striatum). There was a corresponding initial increase (to +50% in the cerebellum) and later decrease (to ?30% in the hippocampus) of the endogenous acetylcholine and choline. These results indicate an initial decrease and later increase in the utilization of acetylcholine after acute treatment with LiCl. Following 10 days of treatment with LiCl there was an increased rate of synthesis of [²H₄]acetylcholine from pulsed [²H₄]choline in the striatum, hippocampus and cortex (P < 0.05). The high affinity transport of [²H₄]choline and its conversion to [²H₄]acetylcholine was activated (131% of control; P < 0.01) in synaptosomes isolated from brains of 10-day treated rats. Investigation of synaptosomes isolated from striatum, hippocampus and cortex revealed that only striatal [²H₄]acetylcholine synthesis was significantly stimulated. Kinetic analysis demonstrated that the apparent K_T for choline was decreased by 30% in striatal synaptosomes isolated from rats treated for 10 days with LiCl. Striatal synaptosomes from 10-day treated rats compared to striatal synaptosomes from untreated rats also released acetylcholine at a stimulated rate in a medium containing 35 mM-KCl. These results indicate that LiCl treatment stimulates cholinergic activity in certain brain regions and this may play a significant role in the therapeutic effect of LiCl in neuropsychiatric disorders.     

α-METHYLTRYPTOPHAN: EFFECTS ON CEREBRAL MONOOXYGENASES IN VITRO AND IN VIVO

Following administration of x-methyltryptophan (AMTP) (50 mg/kg) there was a time dependent decrease of serotonin and a concomitant increase of α-methyl-5-hydroxy-tryptamine (AM-5-HT) in most cerebral areas. AMTP is hydroxylated to α-methyl-5-hydroxytryptophan (AM-5-HTP) by cerebral tryptophan hydroxylase in vitro and in vivo. Hydroxylation of AMTP in vitro and in vivo was markedly inhibited in p-chlorophenylalanine (p-CP) treated rats. After administration of AMTP, the conversion in vivo of tyrosine to norepinephrine was inhibited. This inhibition was not apparent in p-CP pretreated animals. p-Chloroamphetamine (p-CA) (10 mg/kg) lowered serotonin and AM-5-HT levels in some areas of the brain, but unlike p-CP, alone or in combination with AMTP it did not significantly inhibit hydroxylation of tryptophan (Trp). AMTP, as substrate of tryptophan hydroxylase, has a K_m of 1.08 × 10^-4 M (using 6-MPH₄, as cofactor) and as competitive inhibitor, a K₁ of 2.09 × 10^-4M with L-Trp as substrate. AMTP becomes an uncompetitive inhibitor when its concentration is equal to or exceeds that of L-Trp. D-AMTP is neither a substrate nor an inhibitor of tryptophan hydroxylase. DL-AM-5-HTP (K₁, 1.5 × 10^-5 M) and AM-5-HT (K¹ 4.0 × 10^-5 M) are competitive inhibitors.     

DEVELOPMENTAL EFFECTS ON PROTEIN SYNTHESIS RATES IN REGIONS OF THE CNS IN VIVO AND IN VITRO

Abstract— Protein synthesis rates have been determined quantitatively in several regions of the nervous system of rats of various ages. The developmental changes in these regions are generally similar with a high rate maintained from several days before birth to about 4 days of age (1.9–2.1% h⁻¹). A decline in the rate ensues thereupon which continues till approx 30 days of age, whence the curve flattens though continuing slowly downward with increasing age. In the young three regions, cerebellum, pineal and pituitary, exhibit exceptionally higher rates (40–50%) than the cerebral hemispheres, pons-medulla, mid brain or cord, which all display curves of similar magnitude and shape. While the rate in the cerebellum eventually declines with age to within 10% of the rate in cerebral hemisphere, rates in the pineal and pituitary though decreasing remain far above (100%) rates in cerebral hemisphere even in adults.
The rate in vitro for slices of cerebellum follows a pattern similar to that shown previously for cerebral hemispheres: in the very young rates are 70–80% of the in vivo value but decline much more rapidly with age and in adult represent only 10–15% of the rate in vivo.
A markedly different pattern is seen in whole (unsliced) pituitaries wherein in vitro rates parallel in vivo rates with increasing age at approx 70–80% of the in vivo rate. Pineals appear to follow a similar pattern.     

STABILITY OF SYNAPTOSOMAL GABA LEVELS AND THEIR USE IN DETERMINING THE IN VIVO EFFECTS OF DRUGS: CONVULSANT AGENTS

—The stability of the GABA content of synaptosomal-enriched fractions was evaluated by two approaches. Firstly, the addition of 10^?3m -aminooxyacetic acid to the homogenizing medium totally inhibited the GABA-degrading enzyme in the fractions but did not affect the GABA levels. This indicated that GABA was not being metabolized during the normal preparation of the synaptosomal-enriched fraction. Secondly, when synaptosomal-enriched fractions were re-fractionated by discontinuous density gradient centrifugation, the GABA contents of the fractions before and after the second fractionation were very similar provided they were expressed on a per mg protein basis. It was therefore concluded that the GABA content of the organelles was not subject to change during the fractionation procedures. On the basis of these findings and others it was suggested that the synaptosomal-enriched fraction could be used as a model to evaluate drug-induced changes in GABA levels in nerve endings. In vivo experimentation indicated that the convulsant agents hydrazine, isonicotinic acid hydrazide and aminooxyacetic acid brought about similar decreases in the GABA content of the synaptosomal-enriched fractions prepared from tissue at the onset of seizures despite the fact that no correlation was observed between seizure activity and whole brain GABA levels.     

半乳糖寡糖在体内和体外对双歧杆菌生长的影响

目的:利用固定化产β-半乳糖苷酶的嗜热脂肪芽孢杆菌连续合成的产物。方法:经柱层析分离纯化得到了纯半乳糖三寡糖和半乳糖四寡糖,用于体外双歧杆菌培养和动物试验。结果:证实了半乳糖寡糖能高效、专一地促进双歧杆菌的生长繁殖。结论:半乳糖寡糖具有改善小鼠肠道内菌群分布,降低小鼠盲肠内pH值的生理功能。     

低聚果糖体内外对肠道菌的影响

目的 :了解低聚果糖 (FOS)体内外对肠道菌双歧杆菌、类杆菌和肠杆菌科菌的增殖作用。方法 :FOS 1g/ (kg· bw· d) ,灌服 (ig)小鼠 ,连续 14 d后 ,取粪便 ,选择性培养基平板菌落计数法测各菌群。体外试验中 ,按 1%的 FOS添加到各人肠道分离菌培养液中 ,培养 2 4h后测其吸光度 A值和 p H值变化。结果 :ig FOS的小鼠肠道双歧杆菌和肠杆菌数量分别是 9.16± 0 .67和 8.3 3± 0 .70 (log10 N CFU/ g) ,高于对照组(P<0 .0 5)。 FOS体外对各肠道分离菌均有增殖作用 ,依大小分别是双歧杆菌、类杆菌和肠杆菌 ,其 A值分别增加了 0 .8～ 1.192、0 .80 2和 0 .198～ 0 .461,其 p H值分别降低了 1.5～ 1.68、1.2和 0 .2～ 0 .58。结论 :FOS体内外有相对选择性增殖双歧杆菌的作用 ,对类杆菌和肠杆菌也有调整作用     

槲皮素体内外抗氧化作用的比较研究

目的 :测定槲皮素的体外总抗氧化力 ,进一步观察槲皮素灌胃后大鼠外周血总抗氧化力的变化 ,并与芦丁、维生素C、维生素E相比较。方法 :总抗氧化力采用Fe3 还原法 ,槲皮素、芦丁分析采用紫外分光光度法及高效液相色谱法。结果 :相同浓度条件下槲皮素的体外总抗氧化力显著强于芦丁 ,与传统的抗氧化剂维生素C、维生素E相当。槲皮素 4 0mg/kg灌胃 1h后大鼠外周血总抗氧化力及槲皮素含量 (紫外分光光度法 )较灌胃前升高最为明显。维生素C也有显著提高外周血总抗氧化力的作用 ,芦丁与维生素E未表现出显著作用。血浆高效液相分析表明槲皮素灌胃后未出现明显的槲皮素吸收峰 ,而与其峰相邻的两个未知峰的面积增大。结论 :槲皮素的体外抗氧化作用强于芦丁 ,与传统的抗氧化剂维生素C、维生素E相当 ;槲皮素吸收后经代谢形成衍生物 ,提高血浆总抗氧化力的程度与维生素C相近     

乳酸杆菌体内外对阴道菌群及pH值生物学影响的研究

本研究利用从正常妇女阴道内分离出来的乳酸杆菌,从微生态学角度进行体内外生物拮抗试验,以探索其对阴道菌群及生境ｐＨ的生物学影响。结果表明：乳酸杆菌于体内外对阴道的葡萄球菌、肠杆菌、酵母菌有生物抑制作用;对生境的ｐＨ有生物学影响,主要表现为使生境ｐＨ下降。通过上述生物学影响调节生境菌群平衡及内环境的稳定。     

EFFECTS OF AMPHETAMINE ADMINISTRATION IN VIVO ON IN VITRO PROTEIN SYNTHESIZING SYSTEM FROM RAT BRAIN 1

Abstract— A highly active in vitro protein synthesizing system (S-28) has been prepared from rat brain. Poly (U)-dependent [³H] phenylalanine incorporation by brain S-28 system is significantly inhibited by D-amphetamine. The extent of inhibition by amphetamine is significantly higher than by other biogenic amines such as dopamine and serotonin. At the 100°g level of amphetamine, the inhibition is about 70°. Experiments with ribosomes and soluble enzymes from control and amphetamine-treated systems indicate that the observed inhibition may be due to the effect of the drug on the ribosomes. Kinetic analysis of the reaction mixture in the presence as well as absence of D-amphetamine indicate that this sympathomimetic drug inhibits polysome formation in vitro.     

EMBRYO GROWTH AND SEED SIZE IN RAPHANUS SATIVUS: MATERNAL AND PATERNAL EFFECTS IN VIVO AND IN VITRO

Theories on the evolution of the angiosperm seed disagree as to the effects of different plant tissues on embryo growth. To examine the relative contributions of maternal and paternal genes on embryo growth, we conducted controlled crosses in the greenhouse with wild radish plants (Raphanus sativus), looked for maternal, paternal, and interaction effects on embryo development, and compared the performance of embryos within fruits and in embryo culture. Maternal plant identity affected fruit set, seeds per fruit, embryo developmental stage, and mean seed weight. In embryo culture, maternal effects were found for cotyledon size and embryo weight. Paternal effects were fewer or smaller in magnitude than maternal effects. The identity of the pollen donor affected embryo developmental stage and mean seed weight. In culture, paternal effects were detected for cotyledon size and embryo weight. Our results demonstrate that both maternal and paternal elements affect embryo growth. The fact that maternal effects are greater than paternal effects on embryo development in culture may result from cytoplasmic elements or maternal nuclear genes. Embryo performance in vivo compared to that in vitro varied among maternal plants. The interaction between an embryo and its endosperm and maternal tissues may be either positive or negative, depending upon the maternal plant and the embryo's developmental stage.